Impact of Viral Infection on Absorption and Scattering Properties of Marine Bacteria and Phytoplankton

DTIC Science & Technology

2001-09-30

Opt. Eng. 2963: 260-265. 5 Bratbak, G., J. K. Egge, and M. Heldal. 1993. Viral mortality of the marine alga Emiliania huxleyi (Haptophyceae...and termination of algal blooms. Mar. Ecol. Prog. Ser. 93: 39-48. Bratbak, G., W. Wilson, and M. Heldal. 1996. Viral control of Emiliania huxleyi...relation to Emiliania huxleyi blooms: a mechanism of DMSP release? Mar. Ecol. Prog. Ser. 128: 133-142. Brussaard, C. P. D., R. S. Kempers, A. J

Elastin hydrolysate derived from fish enhances proliferation of human skin fibroblasts and elastin synthesis in human skin fibroblasts and improves the skin conditions.

PubMed

Shiratsuchi, Eri; Nakaba, Misako; Yamada, Michio

2016-03-30

Recent studies have shown that certain peptides significantly improve skin conditions, such as skin elasticity and the moisture content of the skin of healthy woman. This study aimed to investigate the effects of elastin hydrolysate on human skin. Proliferation and elastin synthesis were evaluated in human skin fibroblasts exposed to elastin hydrolysate and proryl-glycine (Pro-Gly), which is present in human blood after elastin hydrolysate ingestion. We also performed an ingestion test with elastin hydrolysate in humans and evaluated skin condition. Elastin hydrolysate and Pro-Gly enhanced the proliferation of fibroblasts and elastin synthesis. Maximal proliferation response was observed at 25 ng mL(-1) Pro-Gly. Ingestion of elastin hydrolysate improved skin condition, such as elasticity, number of wrinkles, and blood flow. Elasticity improved by 4% in the elastin hydrolysate group compared with 2% in the placebo group. Therefore, elastin hydrolysate activates human skin fibroblasts and has beneficial effects on skin conditions. © 2015 Society of Chemical Industry.

MiR-29b affects the secretion of PROG and promotes the proliferation of bovine corpus luteum cells

PubMed Central

Zhang, Li-Qun; Sun, Xu-Lei; Luo, Dan; Fu, Yao; Gao, Yan; Zhang, Jia-Bao

2018-01-01

The regulatory role of miRNAs has been explored in ovarian cells, and their effects on gonadal development, apoptosis, ovulation, steroid production and corpus luteum (CL) development have been revealed. In this study, we analyzed the expression of miR-29b at different stages of bovine CL development and predicted the target genes of miR-29b. We confirmed that miR-29b reduces the expression of the oxytocin receptor (OXTR), affects progesterone (PROG) secretion and regulates the function of the CL. RT-PCR showed that the expression of miR-29b was significantly higher in functional CL phases than in the regressed CL phase. Immunohistochemistry showed that OXTR was expressed in both large and small CL cells and was mainly located in the cell membrane and cytoplasm of these cells. We analyzed the expression levels of OXTR and found that transfection with a miR-29b mimic decreased OXTR expression, but transfection with the inhibitor had a limited effect on the expression of the OXTR protein. At the same time, the secretion of PROG was significantly increased in the miR-29b mimic-transfected group. We also analyzed the effect of miR-29b on the apoptosis of CL cells. Finally, we found that miR-29b could promote the proliferation of bovine CL cells. In conclusion, we found that miR-29b reduces the expression of OXTR and can promote PROG secretion and the proliferation of CL cells via OXTR. PMID:29617446

Majorana-Fermions, Their-Own Antiparticles, Following Non-Abelian Anyon/Semion Quantum-Statistics : Solid-State MEETS Particle Physics Neutrinos: Spin-Orbit-Coupled Superconductors and/or Superfluids to Neutrinos; Insulator-Heisenberg-Antiferromagnet MnF2 Majorana-Siegel-Birgenau-Keimer - Effect

NASA Astrophysics Data System (ADS)

Majorana-Fermi-Segre, E.-L.; Antonoff-Overhauser-Salam, Marvin-Albert-Abdus; Siegel, Edward Carl-Ludwig

2013-03-01

Majorana-fermions, being their own antiparticles, following non-Abelian anyon/semion quantum-statistics: in Zhang et.al.-...-Detwiler et.al.-...``Worlds-in-Collision'': solid-state/condensed-matter - physics spin-orbit - coupled topological-excitations in superconductors and/or superfluids -to- particle-physics neutrinos: ``When `Worlds' Collide'', analysis via Siegel[Schrodinger Centenary Symp., Imperial College, London (1987); in The Copenhagen-Interpretation Fifty-Years After the Como-Lecture, Symp. Fdns. Mod.-Phys., Joensu(1987); Symp. on Fractals, MRS Fall-Mtg., Boston(1989)-5-papers!!!] ``complex quantum-statistics in fractal-dimensions'', which explains hidden-dark-matter(HDM) IN Siegel ``Sephirot'' scenario for The Creation, uses Takagi[Prog.Theo.Phys. Suppl.88,1(86)]-Ooguri[PR D33,357(85)] - Picard-Lefschetz-Arnol'd-Vassil'ev[``Principia Read After 300 Years'', Not.AMS(1989); quantum-theory caveats comment-letters(1990); Applied Picard-Lefschetz Theory, AMS(2006)] - theorem quantum-statistics, which via Euler- formula becomes which via de Moivre- -formula further becomes which on unit-circle is only real for only, i.e, for, versus complex with imaginary-damping denominator for, i.e, for, such that Fermi-Dirac quantum-statistics for

Recent advances in the multimodel hydrologic ensemble forecasting using the HydroProg system in the Nysa Klodzka river basin (southwestern Poland)

NASA Astrophysics Data System (ADS)

Niedzielski, Tomasz; Mizinski, Bartlomiej; Swierczynska-Chlasciak, Malgorzata

2017-04-01

The HydroProg system, the real-time multimodel hydrologic ensemble system elaborated at the University of Wroclaw (Poland) in frame of the research grant no. 2011/01/D/ST10/04171 financed by National Science Centre of Poland, has been experimentally launched in 2013 in the Nysa Klodzka river basin (southwestern Poland). Since that time the system has been working operationally to provide water level predictions in real time. At present, depending on a hydrologic gauge, up to eight hydrologic models are run. They are data- and physically-based solutions, with the majority of them being the data-based ones. The paper aims to report on the performance of the implementation of the HydroProg system for the basin in question. We focus on several high flows episodes and discuss the skills of the individual models in forecasting them. In addition, we present the performance of the multimodel ensemble solution. We also introduce a new prognosis which is determined in the following way: for a given lead time we select the most skillful prediction (from the set of all individual models running at a given gauge and their multimodel ensemble) using the performance statistics computed operationally in real time as a function of lead time.

Theory and High-Energy-Density Laser Experiments Relevant to Accretion Processes in Cataclysmic Variables

NASA Astrophysics Data System (ADS)

Krauland, Christine; Drake, R.; Loupias, B.; Falize, E.; Busschaert, C.; Ravasio, A.; Yurchak, R.; Pelka, A.; Koenig, M.; Kuranz, C. C.; Plewa, T.; Huntington, C. M.; Kaczala, D. N.; Klein, S.; Sweeney, R.; Villete, B.; Young, R.; Keiter, P. A.

2012-05-01

We present results from high-energy-density (HED) laboratory experiments that explore the contribution of radiative shock waves to the evolving dynamics of the cataclysmic variable (CV) systems in which they reside. CVs can be classified under two main categories, non-magnetic and magnetic. In the process of accretion, both types involve strongly radiating shocks that provide the main source of radiation in the binary systems. This radiation can cause varying structure to develop depending on the optical properties of the material on either side of the shock. The ability of high-intensity lasers to create large energy densities in targets of millimeter-scale volume makes it feasible to create similar radiative shocks in the laboratory. We provide an overview of both CV systems and their connection to the designed and executed laboratory experiments preformed on two laser facilities. Available data and accompanying simulations will likewise be shown. Funded by the NNSA-DS and SC-OFES Joint Prog. in High-Energy-Density Lab. Plasmas, by the Nat. Laser User Facility Prog. in NNSA-DS and by the Predictive Sci. Acad. Alliances Prog. in NNSA-ASC, under grant numbers are DE-FG52-09NA29548, DE-FG52-09NA29034, and DE-FC52-08NA28616.

Reverse Radiative Shock Experiments Relevant to Accreting Stream-Disk Impact in Interacting Binaries

NASA Astrophysics Data System (ADS)

Krauland, Christine; Drake, R. P.; Kuranz, C. K.; Huntington, C. M.; Grosskopf, M. J.; Marion, D. C.; Young, R.; Plewa, T.

2011-05-01

In many Cataclysmic Binary systems, mass onto an accretion disk produces a `hot spot’ where the infalling supersonic flow obliquely strikes the rotating accretion disk. This collision region has many ambiguities as a radiation hydrodynamic system, but shock development in the infalling flow can be modeled. Depending upon conditions, it has been argued (Armitage & Livio, ApJ 493, 898) that the shocked region may be optically thin, thick, or intermediate, which has the potential to significantly alter the hot spot's structure and emissions. We report the first experimental attempt to produce colliding flows that create a radiative reverse shock at the Omega-60 laser facility. Obtaining a radiative reverse shock in the laboratory requires producing a sufficiently fast flow (> 100 km/s) within a material whose opacity is large enough to produce energetically significant emission from experimentally achievable layers. We will discuss the experimental design, the available data, and our astrophysical context. Funded by the NNSA-DS and SC-OFES Joint Prog. in High-Energy-Density Lab. Plasmas, by the Nat. Laser User Facility Prog. in NNSA-DS and by the Predictive Sci. Acad. Alliances Prog. in NNSA-ASC, under grant numbers are DE-FG52-09NA29548, DE-FG52-09NA29034, and DE-FC52-08NA28616.

Geovisualization in the HydroProg web map service

NASA Astrophysics Data System (ADS)

Spallek, Waldemar; Wieczorek, Malgorzata; Szymanowski, Mariusz; Niedzielski, Tomasz; Swierczynska, Malgorzata

2016-04-01

The HydroProg system, built at the University of Wroclaw (Poland) in frame of the research project no. 2011/01/D/ST10/04171 financed by the National Science Centre of Poland, has been designed for computing predictions of river stages in real time on a basis of multimodelling. This experimental system works on the upper Nysa Klodzka basin (SW Poland) above the gauge in the town of Bardo, with the catchment area of 1744 square kilometres. The system operates in association with the Local System for Flood Monitoring of Klodzko County (LSOP), and produces hydrograph prognoses as well as inundation predictions. For presenting the up-to-date predictions and their statistics in the online mode, the dedicated real-time web map service has been designed. Geovisualisation in the HydroProg map service concerns: interactive maps of study area, interactive spaghetti hydrograms of water level forecasts along with observed river stages, animated images of inundation. The LSOP network offers a high spatial and temporal resolution of observations, as the length of the sampling interval is equal to 15 minutes. The main environmental elements related to hydrological modelling are shown on the main map. This includes elevation data (hillshading and hypsometric tints), rivers and reservoirs as well as catchment boundaries. Furthermore, we added main towns, roads as well as political and administrative boundaries for better map understanding. The web map was designed as a multi-scale representation, with levels of detail and zooming according to scales: 1:100 000, 1:250 000 and 1:500 000. Observations of water level in LSOP are shown on interactive hydrographs for each gauge. Additionally, predictions and some of their statistical characteristics (like prediction errors and Nash-Sutcliffe efficiency) are shown for selected gauges. Finally, predictions of inundation are presented on animated maps which have been added for four experimental sites. The HydroProg system is a strictly scientific project, but the web map service has been designed for all web users. The main objective of the paper is to present the design process of the web map service, following the cartographic and graphic principles.

Sediment and erosion control laboratory facility expansion.

DOT National Transportation Integrated Search

2016-08-01

The Sediment and Erosion Control Laboratory (SEC Lab), formerly the Hydraulics, Sedimentation, and : Erosion Control Laboratory, is operated by the Texas A&M Transportation Institutes Environment and : Planning Program. Performance evaluation prog...

Child restraint device loaner programs

DOT National Transportation Integrated Search

1981-06-01

The child restraint device (CRD) loaner programs in Tennessee were evaluated. In-Lerviews were conducted with loaner program clients in Memphis, Chattanooga, and Knoxville. Administrators of programs in all three sites also were interviewed. The prog...

Statistical evaluation of blood alcohol measurements

DOT National Transportation Integrated Search

1981-10-01

The U.S. Department of Transportation, National Highway Traffic Safety Administration (NHTSA) has instituted a voluntary program to evaluate the proficiency of laboratories measuring the amount of alcohol in blood. In this report, data from that prog...

Evaluation of the Oregon DMV driver improvement program.

DOT National Transportation Integrated Search

2007-04-01

This report provides an evaluation of the Oregon Department of Transportation-Driver and Motor Vehicle : (DMV) Services Driver Improvement Program (DIP), which was substantially changed in 2002. Prior to 2002, : the DIP was organized around four prog...

Effectiveness of Oregon's teen licensing program.

DOT National Transportation Integrated Search

2008-06-01

Significant changes in Oregons teen licensing laws went into effect on March 1, 2000. The new laws expanded the provisional driving license program which had been in effect since October 1989 and established a graduated driver licensing (GDL) prog...

MODELING TREE LEVEL PROCESSES

EPA Science Inventory

An overview of three main types of simulation approach (explanatory, abstraction, and estimation) is presented, along with a discussion of their capabilities limitations, and the steps required for their validation. A process model being developed through the Forest Response Prog...

Iran Sanctions

DTIC Science & Technology

2016-05-18

Manufacturing Group (Iran, missile prog.) Aerospace Industries Organization (AIO) (Iran) September 2007 Korea Mining and Development Corp. ( N . Korea... Vitaly Sokolenko (general manager of Ferland) April 29, 2014 (for connections to deceptive oil dealings for Iran) Saeed Al Aqili (co-owner of Al

NCORP Gets Underway

Cancer.gov

NCI has awarded 53 new 5-year grants to researchers across the country to conduct multi-site cancer clinical trials and cancer care delivery research studies in their communities. The grants are being awarded under the NCI Community Oncology Research Prog

Accuracy Of LTPP Traffic Loading Estimates

DOT National Transportation Integrated Search

1998-07-01

The accuracy and reliability of traffic load estimates are key to determining a pavement's life expectancy. To better understand the variability of traffic loading rates and its effect on the accuracy of the Long Term Pavement Performance (LTPP) prog...

Effect of Sex Differences on Brain Mitochondrial Function and Its Suppression by Ovariectomy and in Aged Mice.

PubMed

Gaignard, Pauline; Savouroux, Stéphane; Liere, Philippe; Pianos, Antoine; Thérond, Patrice; Schumacher, Michael; Slama, Abdelhamid; Guennoun, Rachida

2015-08-01

Sex steroids regulate brain function in both normal and pathological states. Mitochondria are an essential target of steroids, as demonstrated by the experimental administration of 17β-estradiol or progesterone (PROG) to ovariectomized female rodents, but the influence of endogenous sex steroids remains understudied. To address this issue, mitochondrial oxidative stress, the oxidative phosphorylation system, and brain steroid levels were analyzed under 3 different experimental sets of endocrine conditions. The first set was designed to study steroid-mediated sex differences in young male and female mice, intact and after gonadectomy. The second set concerned young female mice at 3 time points of the estrous cycle in order to analyze the influence of transient variations in steroid levels. The third set involved the evaluation of the effects of a permanent decrease in gonadal steroids in aged male and female mice. Our results show that young adult females have lower oxidative stress and a higher reduced nicotinamide adenine dinucleotide (NADH)-linked respiration rate, which is related to a higher pyruvate dehydrogenase complex activity as compared with young adult males. This sex difference did not depend on phases of the estrous cycle, was suppressed by ovariectomy but not by orchidectomy, and no longer existed in aged mice. Concomitant analysis of brain steroids showed that pregnenolone and PROG brain levels were higher in females during the reproductive period than in males and decreased with aging in females. These findings suggest that the major male/female differences in brain pregnenolone and PROG levels may contribute to the sex differences observed in brain mitochondrial function.

Effectiveness of Oregon's teen licensing program : final report.

DOT National Transportation Integrated Search

2008-06-01

Significant changes in Oregons teen licensing laws went into effect on March 1, 2000. The new laws expanded the provisional driving license program which had been in effect since October 1989 and established a graduated driver licensing (GDL) prog...

Siletz River nutrients: Effects of biosolids application

EPA Science Inventory

Stream water nutrients were measured in the Siletz River, Oregon, with the goal of comparing dissolved nutrient concentrations, primarily the nitrogenous nutrients nitrate and ammonium, with previously collected data for the Yaquina and Alsea Rivers for the nutrient criteria prog...

SITE-SPECIFIC DIAGNOSTIC TOOLS

EPA Science Inventory

US EPA's Office of Water is proposing Combined Assessment and Listing Methods (CALM) to
meet reporting requirements under both Sections 305b and 303d for chemical and nonchemical
stressors in the nation's waterbodies. Current Environmental Monitoring and Assessment
Prog...

Satellite Remote Sensing for Monitoring and Assessment

EPA Science Inventory

Remote sensing technology has the potential to enhance the engagement of communities and managers in the implementation and performance of best management practices. This presentation will use examples from U.S. numeric criteria development and state water quality monitoring prog...

Optical Limiting Materials Based on Gold Nanoparticles

DTIC Science & Technology

2014-04-30

of the electromagnetic spectrum. 2. Functionalization of the surface of the gold nanoparticles with selected organic and inorganic materials, with...F. A Review of Optical Limiting Mechanisms and Devices Using Organics, Fullerenes , Semiconductors and Other Materials. Prog. Quant. Electr. 1993

Effects of polarization direction on laser-assisted free-free scattering

NASA Astrophysics Data System (ADS)

deHarak, B. A.; Kim, B. N.; Weaver, C. M.; Martin, N. L. S.; Siavashpouri, Mahsa; Nosarzewski, Benjamin

2016-06-01

This work will detail the effects of laser polarization direction (relative to the momentum transfer direction) on laser-assisted free-free scattering. Such processes play a role in the gas breakdown that occurs in electric discharges as well as providing a method for the laser heating of a plasma (Musa et al 2010 J. Phys. B: At. Mol. Opt. Phys. 43 175201, Mason 1993 Rep. Prog. Phys. 56 1275). Experimental results will be presented for electron-helium scattering in the presence of an Nd:YAG laser field (hν =1.17 eV) where the polarization direction was varied in a plane that is perpendicular to the scattering plane. To date, all of our experimental results are well described by the Kroll-Watson approximation (KWA) (Kroll and Watson 1973 Phys. Rev. A 8 804). The good agreement between our experiments and calculations using the KWA includes the case where the polarization is perpendicular to the momentum transfer direction, for which the KWA predicts vanishing cross section; other workers have found that the KWA tends to be inaccurate for cases where it predicts small cross sections (e.g. Musa et al 2010 J. Phys. B: At. Mol. Opt. Phys. 43 175201). We also present simulations of the effects that multiple scattering might have on experimental measurements. In particular, we examine conditions that are expected to be similar to those of the experiments reported by Wallbank and Holmes (Wallbank and Holmes 1993 Phys. Rev. A 48 R2515).

us9805_dni

Science.gov Websites

STYLE="text-align:Left;">

MTBE REMOVAL FROM DRINKING WATER - PHASE I

EPA Science Inventory

The 1990 Federal Clean Air Act mandated the incorporation of oxygenates into gasoline in ozone and carbon monoxide nonattainment areas. Methyl tertiary butyl ether (MTBE) is the oxygenate of choice due to economic and supply considerations. Despite federal and state prog...

Transportation improvement program : Richland, Ohio : fiscal year 1997-2000

DOT National Transportation Integrated Search

1996-06-01

As part of the Urban Transportation Planning Process, under the Federal Planning regulations (Title 23 U.S.C. and Title 49 U.S.C.), the Metropolitan Planning Organization (MPO) is required to develop and keep current a Transportation Improvement Prog...

Comparative Analysis Of River Conservation In The United States And South Africa

EPA Science Inventory

Both the United States and South Africa are recognized for their strong and innovative approaches to the conservation of river ecosystems. These national programs possess similar driving legislation and ecoregional classification schemes supported by comprehensive monitoring prog...

LASP-01: Distribution of Mouse Embryonic Stem Cells Expressing MicroRNAs | Frederick National Laboratory for Cancer Research

Cancer.gov

The Laboratory Animal Sciences Program manages the expansion, processing, and distribution of1,501 genetically engineered mouse embryonic stem cell (mESC) linesharboring conditional microRNA transgenes. The Laboratory Animal Sciences Prog

Minnesota urban partnership agreement national evaluation : surveys, interviews, and focus groups test plan.

DOT National Transportation Integrated Search

2009-11-17

This report presents the test plan for developing, conducting, and analyzing surveys, interviews, and focus groups for evaluating the Minnesota Urban Partnership Agreement (UPA) under the United States Department of Transportation (U.S. DOT) UPA Prog...

Experimental use of geogrids as an alternative to gravel placement : fourth year interim and final report, October 2006.

DOT National Transportation Integrated Search

2006-10-01

With the ongoing demand for improved infrastructure, the Maine Department of Transportation : (MaineDOT) continues to identify and evaluate new and innovative construction methods and materials. : The Departments Collector Highway Improvement Prog...

GLOBAL TRANSITION TO SUSTAINABLE DEVELOPMENT

EPA Science Inventory

Global transition to sustainable development is possible but many obstacles lie in the way and it will require acts of political will on the part of both the developed and developing nations to become a reality. In this paper, sustainable development is defined as continuous prog...

An Open Letter to the Cancer Community Regarding Community Clinical Trials

Cancer.gov

The National Cancer Institute (NCI) is in the process of combining its two community-based research networks to create a single network that builds on the strengths of the Community Clinical Oncology Program/Minority-Based Community Clinical Oncology Prog

SMALL DRINKING WATER SYSTEM PEER REVIEW PROGRAM

EPA Science Inventory

The United South and Eastern Tribes, Inc., which is made up of twenty-four (24) tribes, ranging in location, geographically, from Maine to Texas, AND three (3) states, Mississippi, Kentucky, and Georgia, participated in a program, "The Small Drinking Water System Peer Review Prog...

Results of Special Accident Study Teams/ASAP Coordination Conference

DOT National Transportation Integrated Search

1974-07-01

The second Special Accident Study Teams / ASAP Coordination Conference was held in Washington, D.C. on June 12-13, 1974 to continue coordination of activities and to report recent findings. The objectives of the conference were: (1) To report on prog...

Anticipation and consumption of food each increase the concentration of neuroactive steroids in rat brain and plasma.

PubMed

Pisu, Maria Giuseppina; Floris, Ivan; Maciocco, Elisabetta; Serra, Mariangela; Biggio, Giovanni

2006-09-01

Stressful stimuli and anxiogenic drugs increase the plasma and brain concentrations of neuroactive steroids. Moreover, in rats trained to consume their daily meal during a fixed period, the anticipation of food is associated with changes in the function of various neurotransmitter systems. We have now evaluated the effects of anticipation and consumption of food in such trained rats on the plasma and brain concentrations of 3alpha-hydroxy-5alpha-pregnan-20-one (3alpha,5alpha-TH PROG) and 3alpha,21-dihydroxy-5alpha-pregnan-20-one (3alpha,5alpha-TH DOC), two potent endogenous positive modulators of type A receptors for gamma-aminobutyric acid (GABA). The abundance of these neuroactive steroids was increased in both the cerebral cortex and plasma of the rats during both food anticipation and consumption. In contrast, the concentration of their precursor, progesterone, was increased in the brain only during food consumption, whereas it was increased in plasma only during food anticipation. Intraperitoneal administration of the selective agonist abecarnil (0.1 mg/kg) 40 min before food presentation prevented the increase in the brain levels of 3alpha,5alpha-TH PROG and 3alpha,5alpha-TH DOC during food anticipation but not that associated with consumption. The change in emotional state associated with food anticipation may thus result in an increase in the plasma and brain levels of 3alpha,5alpha-TH PROG and 3alpha,5alpha-TH DOC in a manner sensitive to the activation of GABA(A) receptor-mediated neurotransmission. A different mechanism, insensitive to activation of such transmission, may underlie the changes in the concentrations of these neuroactive steroids during food consumption.

Dopaminergic modulation of effort-related choice behavior as assessed by a progressive ratio chow feeding choice task: pharmacological studies and the role of individual differences.

PubMed

Randall, Patrick A; Pardo, Marta; Nunes, Eric J; López Cruz, Laura; Vemuri, V Kiran; Makriyannis, Alex; Baqi, Younis; Müller, Christa E; Correa, Mercè; Salamone, John D

2012-01-01

Mesolimbic dopamine (DA) is involved in behavioral activation and effort-related processes. Rats with impaired DA transmission reallocate their instrumental behavior away from food-reinforced tasks with high response requirements, and instead select less effortful food-seeking behaviors. In the present study, the effects of several drug treatments were assessed using a progressive ratio (PROG)/chow feeding concurrent choice task. With this task, rats can lever press on a PROG schedule reinforced by a preferred high-carbohydrate food pellet, or alternatively approach and consume the less-preferred but concurrently available laboratory chow. Rats pass through each ratio level 15 times, after which the ratio requirement is incremented by one additional response. The DA D(2) antagonist haloperidol (0.025-0.1 mg/kg) reduced number of lever presses and highest ratio achieved but did not reduce chow intake. In contrast, the adenosine A(2A) antagonist MSX-3 increased lever presses and highest ratio achieved, but decreased chow consumption. The cannabinoid CB1 inverse agonist and putative appetite suppressant AM251 decreased lever presses, highest ratio achieved, and chow intake; this effect was similar to that produced by pre-feeding. Furthermore, DA-related signal transduction activity (pDARPP-32(Thr34) expression) was greater in nucleus accumbens core of high responders (rats with high lever pressing output) compared to low responders. Thus, the effects of DA antagonism differed greatly from those produced by pre-feeding or reduced CB1 transmission, and it appears unlikely that haloperidol reduces PROG responding because of a general reduction in primary food motivation or the unconditioned reinforcing properties of food. Furthermore, accumbens core signal transduction activity is related to individual differences in work output.

Hormone Metabolism During Potato Tuber Dormancy

USDA-ARS?s Scientific Manuscript database

At harvest and for an indeterminate period thereafter potato tubers will not sprout and are physiologically dormant. The length of tuber dormancy is dependent on cultivar and pre- and postharvest environmental conditions. Plant hormones have been shown to be involved in all phases of dormancy prog...

IMPACT OF LEAD ACID BATTERIES AND CADMIUM STABILIZERS ON INCINERATOR EMISSIONS

EPA Science Inventory

The Waste Analysis Sampling, Testing and Evaluation (WASTE) Program is a multi-year, multi-disciplinary program designed to elicit the source and fate of environmentally significant trace materials as a solid waste progresses through management processes. s part of the WASTE Prog...

Assessing genomic selection prediction accuracy in a dynamic barley breeding

USDA-ARS?s Scientific Manuscript database

Genomic selection is a method to improve quantitative traits in crops and livestock by estimating breeding values of selection candidates using phenotype and genome-wide marker data sets. Prediction accuracy has been evaluated through simulation and cross-validation, however validation based on prog...

Efficient Computations and Representations of Visible Surfaces.

DTIC Science & Technology

1979-12-01

position as stated. The smooth contour generator may lie along a sharp ridge, for instance. Richards & Stevens -28- 6m lace contout s ?S ,.......... ceoonec...From understanding computation to understanding neural circuitry. Neurosci. Res. Prog. Bull. 13. 470-488. Metelli, F. 1970 An algebraic development of

Serum microRNA biomarker identification in a residential cohort with elevated polychlorinated biphenyl exposures.

EPA Science Inventory

Toxicant-associated steatohepatitis (TASH) is a form of liver disease associated with both industrial [1] and environmental [2] chemical exposures. Like other forms of Non-alcoholic Fatty Liver Disease (NAFLD), TASH can contribute to systemic metabolic disease states and may prog...

us9805_latilt

Science.gov Websites

;meta http-equiv=Content-Type content="text/html; charset=iso-8859-1"> <meta name=ProgId content=Word.Document> <meta name=Generator content="Microsoft Word 11"> <meta name /dublin_core"> <meta name=dc.title content="Alaska Solar Resource: Flat Plate Collector, Facing

EVALUATION OF USFILTER CORPORATION'S RETEC® MODEL SCP-6 SEPARATED CELL PURIFICATION SYSTEM FOR CHROMIC ACID ANODIZE BATH SOLUTION

EPA Science Inventory

The USEPA has created the Environmental Technology Verification (ETV) Program to facilitate the deployment of innovative or improved environmental technologies through performance verification and dissemination of information. The ETV P2 Metal Finishing Technologies (ETV-MF) Prog...

HD gas purification for polarized HDice targets production at Jefferson Lab

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

Whisnant, Charles; D'Angelo, Annalisa; Colaneri, Luca

2014-06-01

Solid, frozen-spin targets of molecular HD were rst developed for nuclear physics by a collaboration between Syracuse University and Brookhaven National Lab. They have been successfully used in measurements with photon beams, rst at the Laser-Electron-Gamma-Source [1] and most recently at Je erson Lab during the running of the E06-101 (g14) experiment [2]. Preparations are underway to utilize the targets in future electron experiments after the completion of the 12 GeV JLab upgrade [3]. HD is an attractive target since all of the material is polarizable, of low Z, and requires only modest holding elds. At the same time, themore » small contributions from the target cell can be subtracted from direct measurements. Reaching the frozen-spin state with both high polarization and a signi cant spin relaxation time requires careful control of H2 and D2 impurities. Commercially available HD contains 0.5 - 2% concentrations of H2 and D2. Low-temperature distillation is required to reduce these concentrations to the 104 level to enable useful target production. This distillation is done using a column lled with heli-pack C [4] to give good separation e ciency. Approximately 12 moles of commercial HD is condensed into the mechanically refrigerated system at the base temperature of 11K. The system is then isolated and the temperature stabilized at 18K producing liquid HD, which is boiled by a resistive heater. The circulation established by the boil-o condensing throughout the column then ltering back down produces a steady-state isotopic separation permitting the extraction of HD gas with very low H2 and D2 content. A residual gas analyzer initially monitors distillation. Once the H2 concentration falls below its useful operating range, samples are periodically collected for analysis using gas chromatography [5] and Raman scattering. Where the measurement techniques overlap, good agreement is obtained. The operation of the distillery and results of gas analysis will be discussed. References [1] Phy. Rev. Lett. 101 (2009) 172002. [2] www.jlab.org/exp_prog/proposals/06/PR-06-101.pdf [3] www.jlab.org/exp_prog/proposals/12/PR12-12-009.pdf, www.jlab.org/exp_prog/proposals/12/PR12-12-010.pdf, and www.jlab.org/exp_prog/proposals/11/PR12-11-111.pdf [4] Nucl. Inst. Meth. 664 (2012) 347, www.wilmad-labglass.com/Products/LG-6730-104/ [5] Rev. Sci. Instrum. 82, 024101 (2011).« less

Semi-automatic handling of meteorological ground measurements using WeatherProg: prospects and practical implications

NASA Astrophysics Data System (ADS)

Langella, Giuliano; Basile, Angelo; Bonfante, Antonello; De Mascellis, Roberto; Manna, Piero; Terribile, Fabio

2016-04-01

WeatherProg is a computer program for the semi-automatic handling of data measured at ground stations within a climatic network. The program performs a set of tasks ranging from gathering raw point-based sensors measurements to the production of digital climatic maps. Originally the program was developed as the baseline asynchronous engine for the weather records management within the SOILCONSWEB Project (LIFE08 ENV/IT/000408), in which daily and hourly data where used to run water balance in the soil-plant-atmosphere continuum or pest simulation models. WeatherProg can be configured to automatically perform the following main operations: 1) data retrieval; 2) data decoding and ingestion into a database (e.g. SQL based); 3) data checking to recognize missing and anomalous values (using a set of differently combined checks including logical, climatological, spatial, temporal and persistence checks); 4) infilling of data flagged as missing or anomalous (deterministic or statistical methods); 5) spatial interpolation based on alternative/comparative methods such as inverse distance weighting, iterative regression kriging, and a weighted least squares regression (based on physiography), using an approach similar to PRISM. 6) data ingestion into a geodatabase (e.g. PostgreSQL+PostGIS or rasdaman). There is an increasing demand for digital climatic maps both for research and development (there is a gap between the major of scientific modelling approaches that requires digital climate maps and the gauged measurements) and for practical applications (e.g. the need to improve the management of weather records which in turn raises the support provided to farmers). The demand is particularly burdensome considering the requirement to handle climatic data at the daily (e.g. in the soil hydrological modelling) or even at the hourly time step (e.g. risk modelling in phytopathology). The key advantage of WeatherProg is the ability to perform all the required operations and calculations in an automatic fashion, except the need of a human interaction upon specific issues (such as the decision whether a measurement is an anomaly or not according to the detected temporal and spatial variations with contiguous points). The presented computer program runs from command line and shows peculiar characteristics in the cascade modelling within different contexts belonging to agriculture, phytopathology and environment. In particular, it can be a powerful tool to set up cutting-edge regional web services based on weather information. Indeed, it can support territorial agencies in charge of meteorological and phytopathological bulletins.

Project Work Assignment | Sustainable Stormwater Funding Project

EPA Pesticide Factsheets

2010-11-30

... J'l.wroprl 8OOljJf!'l fOI 8..... n """ Nwo I.1n DCN N. e_ e_ prog~;"':x~~ment ObI~~Cl~·· unt I ICents) SI:~~~O:t COSI~~od. Max6 Mo•• Ma:t6 ,,",7 1 ...

CARRY-OVER EFFECTS OF OZONE ON ROOT GROWTH AND CARBOHYDRATE CONCENTRATIONS OF PONDEROSA PINE SEEDLINGS

EPA Science Inventory

Ozone exposure decreases belowground carbon allocation and root growth of plants;however,the extent to which these effects persist and the cumulative impact of ozone stress on plant growth are poorly understood.To evaluate the potential for plant compensation,we followed the prog...

The Physiology of Growth Hormone-Releasing Hormone (GHRH) in Breast Cancer

DTIC Science & Technology

2003-06-01

production of growth hormone-releasing factor by carcinoid and pancreatic islet tumors associated with acromegaly . Prog Clin Biol Res 1981; 74:259-271. (16...promotion of apop- cause of acromegaly . More recently, expression has been tosis. These results indicate that disruption of enaog- demonstrated in tumors

Highway concrete pavement technology development and testing : volume III, field evaluation of Strategic Highway Research Program (SHRP) C-205 test sites (high-performance concrete).

DOT National Transportation Integrated Search

2006-05-01

This research study, sponsored by the Federal Highway Administration, summarizes the field performance of eight high-early-strength (HES) : concrete patches between 1994 and 1998. The patches were constructed under the Strategic Highway Research Prog...

THE WORKSHOP ON THE SOURCE APPORTIONMENT OF PM HEALTH EFFECTS: INTER-COMPARISON OF RESULTS AND IMPLICATIONS

EPA Science Inventory

While the association between exposure to ambient fine particulate matter mass (PM2.5) and human mortality is well established, the most responsible particle types/sources are not yet certain. In May 2003, the U.S. Environmental Protection Agency's Particulate Matter Centers Prog...

Performance Characteristics of Automotive Engines in the United States : Third Series - Report No. 14 - 1978 Buick 196 CID (3.2L)

DOT National Transportation Integrated Search

1981-02-01

Experimental data were obtained in dynamometer tests of a 1978 Buick 231 CID turbocharged to determine fuel consumption and emissions (hydrocarbon, carbon monoxide, oxides of nitrogen) at steady-state engine operating modes. The objective of the prog...

Patterns of Genetic Variation among Populations of the Asian Longhorned Beetle in China and Korea

USDA-ARS?s Scientific Manuscript database

Central to the study of invasive species is identifying source populations in their native ranges. Source populations of invasive species can provide important information about species life cycles, host use and species-specific predators and parasites which could be deployed in a pest control prog...

Dirk Jordan | NREL

Science.gov Websites

quantify module degradation rates. Statistical analysis of reported degradation rates of PV modules degradation rates," Prog. in PV 24(7), 2016, DOI: 10.1002/pip.2744 Jordan D.C., Silverman T.J PV, 2017, DOI: 10.1002/pip.2866 Jordan D.C., Silverman T.J., Sekulic B., Kurtz S.R., "PV

hawaii_9805_latilt

Science.gov Websites

;meta http-equiv=Content-Type content="text/html; charset=iso-8859-1"> text-decoration:underline; text-underline:single;} a:visited, span.MsoHyperlinkFollowed {color:blue; text-decoration:underline; text-underline:single;} p {mso-margin-top-alt:auto; margin-right:0in

An Assessment of Decision-Making Processes: Evaluation of Where Land Protection Planning Can Incorporate Climate Change Information (Final Report)

EPA Science Inventory

EPA announced the availability of the final report, An Assessment of Decision-Making Processes: Evaluation of Where Land Protection Planning Can Incorporate Climate Change Information. This report is a review of decision-making processes of selected land protection prog...

Statistical basis and outputs of stable isotope mixing models: Comment on Fry (2013)

EPA Science Inventory

A recent article by Fry (2013; Mar Ecol Prog Ser 472:1−13) reviewed approaches to solving underdetermined stable isotope mixing systems, and presented a new graphical approach and set of summary statistics for the analysis of such systems. In his review, Fry (2013) mis-characteri...

Juvenile Survival in Common Loons Gavia Immer: Effects of Natal Lake Size and pH

EPA Science Inventory

Survival is a vexing parameter to measure in many young birds because of dispersal and delayed impacts of natal rearing conditions on fitness. Drawing upon marking and resighting records from an 18-year study of territorial behavior, we used Cormack-Jolly-Seber analysis with Prog...

Risk of Pore Water Hydrogen Sulfide Toxicity in Dredged Material Bioassays

DTIC Science & Technology

1995-11-01

Prog. Ser. 101, 147-155. Moore, D. W., and Dillon, T. M. (1993). “Chronic sublethal effects of San Francisco Bay sediments on Neris (Neanthes...metabolism of Arctica isfandica L. (Bivalvia),” J. Exp. Mar. Biol. Ecol. 170, 213-226. Oritz, J. A., Rueda, A., Carbonell, G., Camargo , J. A., Nieto, F

Performance Characteristics of Automotive Engines in the United States, Third Series, Report No. 8, 1978 Buick, 231 CID (3.8 Liters), 4V, Turbocharge

DOT National Transportation Integrated Search

1979-02-01

Experimental data were obtained in dynamometer tests of a 1978 Buick 231 CID turbocharged to determine fuel consumption and emissions (hydrocarbon, carbon monoxide, oxides of nitrogen) at steady-state engine operating modes. The objective of the prog...

Hampton Roads, Virginia eight-hour ozone maintenance area transportation conformity analysis : 2030 long range transportation plan and FY 09-12 transportation improvement program, draft executive summary.

DOT National Transportation Integrated Search

2010-05-01

This report presents the regional conformity analysis and recommendation for a finding of conformity for the Hampton Roads 2030 Long Range Transportation Plan (LRTP, or "Plan") and associated Fiscal Year (FY) 2009-2012 Transportation Improvement Prog...

My life in the watershed: then, now, and beyond

USDA-ARS?s Scientific Manuscript database

"My Life in the Watershed" tells the first-hand account of a young girl growing up in southwestern Oklahoma, the impact growing up in a watershed had on her life, and the vision she sees for her children and her children's children, so they will continue to benefit from the USDA Small Watershed Prog...

A Review of Water Mist Technology for Fire Suppression

DTIC Science & Technology

1994-09-30

Smith, D.P., and Ball, D.N. (1993), "New Applications of Aqueous Agents for Fire Suppression," Halen Alternatives Technical Working Conference...Flows," Prog. Energy, Combust. ScL, 14, 1988, pp. 171-194. Dabros, T., and Van de Ven, T.G.M. (1992), "Hydrodynamic Interactions between Two Spheres Near

Quantifying long-term risks to sea otters from the 1989 'Exxon Valdez' oil spill: reply to Harwell & Gentile (2013)

USGS Publications Warehouse

Ballachey, Brenda E.; Bodkin, James L.; Monson, Daniel H.

2013-01-01

Recovery of sea otter populations in Prince William Sound (PWS), Alaska, has been delayed for more than 2 decades following the 1989 ‘Exxon Valdez’ oil spill. Harwell & Gentile (2013; Mar Ecol Prog Ser 488:291–296) question our conclusions in Bodkin et al. (2012; Mar Ecol Prog Ser 447:273-287) regarding adverse effects that oil lingering in the environment may have on sea otters. They agree that exposure may continue, but disagree that it constitutes a significant risk to sea otters. In Bodkin et al. (2012), we suggested that subtle effects of chronic exposure were the most reasonable explanation for delayed recovery of the sea otter population in areas of western PWS, where shorelines were most heavily oiled. Here, we provide additional information on the ecology of sea otters that clarifies why the toxicological effects of oral ingestion of oil do not reflect all effects of chronic exposure. The full range of energetic, behavioral, and toxicological concerns must be considered to appraise how chronic exposure to residual oil may constrain recovery of sea otter populations.

Antioxidant and ACE-inhibitory activities of hemp (Cannabis sativa L.) protein hydrolysates produced by the proteases AFP, HT, Pro-G, actinidin and zingibain.

PubMed

Teh, Sue-Siang; Bekhit, Alaa El-Din A; Carne, Alan; Birch, John

2016-07-15

Hemp protein isolates (HPIs) were hydrolysed by proteases (AFP, HT, ProG, actinidin and zingibain). The enzymatic hydrolysis of HPIs was evaluated through the degree of hydrolysis and SDS-PAGE profiles. The bioactive properties of the resultant hydrolysates (HPHs) were accessed through ORAC, DPPḢ scavenging and ACE-inhibitory activities. The physical properties of the resultant HPHs were evaluated for their particle sizes, zeta potential and surface hydrophobicity. HT had the highest rate of caseinolytic activity at the lowest concentration (0.1 mg mL(-1)) compared to other proteases that required concentration of 100 mg mL(-1) to achieve their maximum rate of caseinolytic activity. This led to the highest degree of hydrolysis of HPIs by HT in the SDS-PAGE profiles. Among all proteases and substrates, HT resulted in the highest bioactivities (ORAC, DPPḢ scavenging and ACE-inhibitory activities) generated from alkali extracted HPI in the shortest time (2 h) compared to the other protease preparations. Copyright © 2016 Elsevier Ltd. All rights reserved.

Detailed genetic characteristics of an international large cohort of patients with Stargardt disease: ProgStar study report 8.

PubMed

Fujinami, Kaoru; Strauss, Rupert W; Chiang, John Pei-Wen; Audo, Isabelle S; Bernstein, Paul S; Birch, David G; Bomotti, Samantha M; Cideciyan, Artur V; Ervin, Ann-Margret; Marino, Meghan J; Sahel, José-Alain; Mohand-Said, Saddek; Sunness, Janet S; Traboulsi, Elias I; West, Sheila; Wojciechowski, Robert; Zrenner, Eberhart; Michaelides, Michel; Scholl, Hendrik P N

2018-06-20

To describe the genetic characteristics of the cohort enrolled in the international multicentre progression of Stargardt disease 1 (STGD1) studies (ProgStar) and to determine geographic differences based on the allele frequency. 345 participants with a clinical diagnosis of STGD1 and harbouring at least one disease-causing ABCA4 variant were enrolled from 9 centres in the USA and Europe. All variants were reviewed and in silico analysis was performed including allele frequency in public databases and pathogenicity predictions. Participants with multiple likely pathogenic variants were classified into four national subgroups (USA, UK, France, Germany), with subsequent comparison analysis of the allele frequency for each prevalent allele. 211 likely pathogenic variants were identified in the total cohort, including missense (63%), splice site alteration (18%), stop (9%) and others. 50 variants were novel. Exclusively missense variants were detected in 139 (50%) of 279 patients with multiple pathogenic variants. The three most prevalent variants of these patients with multiple pathogenic variants were p.G1961E (15%), p.G863A (7%) and c.5461-10 T>C (5%). Subgroup analysis revealed a statistically significant difference between the four recruiting nations in the allele frequency of nine variants. There is a large spectrum of ABCA4 sequence variants, including 50 novel variants, in a well-characterised cohort thereby further adding to the unique allelic heterogeneity in STGD1. Approximately half of the cohort harbours missense variants only, indicating a relatively mild phenotype of the ProgStar cohort. There are significant differences in allele frequencies between nations, although the three most prevalent variants are shared as frequent variants. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2018. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

Impact of thyroid function abnormalities on reproductive hormones during menstrual cycle in premenopausal HIV infected females at NAUTH, Nnewi, Nigeria

PubMed Central

Emelumadu, Obiageli Fidelia; Igwegbe, Anthony Osita; Monago, Ifeoma Nwamaka; Ilika, Amobi Linus

2017-01-01

Background This was a prospective study designed to evaluate the impact of thyroid function abnormalities on reproductive hormones during menstrual cycle in HIV infected females at Nnamdi Azikiwe University Teaching Hospital Nnewi, South-East Nigeria. Methods The study randomly recruited 35 Symptomatic HIV infected females and 35 Symptomatic HIV infected females on antiretroviral therapy (HAART) for not less than six weeks from an HIV clinic and 40 apparently heathy control females among the hospital staff of NAUTH Nnewi. They were all premenopausal females with regular menstrual cycle and aged between 15–45 years. Blood samples were collected at follicular and luteal phases of their menstrual cycle for assay of Thyroid indices (FT3, FT4 and TSH) and Reproductive indices (FSH, LH, Estrogen, Progesterone, Prolactin and Testosterone) using ELISA method. Results The result showed significantly higher FSH and LH but significantly lower progesterone (prog) and estrogen (E2) in the test females compared to control females at both phases of menstrual cycle (P<0.05). There was significantly lower FT3 but significantly higher TSH value in Symptomatic HIV females (P<0.05). FSH, LH and TSH values were significantly lowered while prog and FT3 were significantly higher in Symptomatic HIV on ART compared to Symptomatic HIV females (P<0.05). FT3, FT4, Prog and E2 were inversely correlated while FSH and LH were positively correlated with duration of HIV infection in HIV females (P<0.05 respectively). There was a direct correlation between CD4+ count and FT3 while inverse correlation was found between CD4+ count and TSH levels (P<0.05). Discussion The present study demonstrated hypothyroidism with a significant degree of primary hypogonadism in Symptomatic HIV infected females at both follicular and luteal phases of menstrual cycle which tends to normalize on treatments. PMID:28723963

78 FR 76319 - Notice of Invitation-Coal Exploration License Application MTM 106757, Montana

Federal Register 2010, 2011, 2012, 2013, 2014

2013-12-17

...] Notice of Invitation--Coal Exploration License Application MTM 106757, Montana AGENCY: Bureau of Land... Signal Peak Energy, LLC on a pro rata cost sharing basis in a program for the exploration of coal... Office coal Web site at http://www.blm.gov/mt/st/en/prog/energy/coal.html . A written notice to...

Photodynamic Molecular Beacons: An Image-Guided Therapeutic Approach to Breast Cancer Vertebral Metastases

DTIC Science & Technology

2012-03-01

uptake. Prog Clin Biol Res. 1984;; 170: 629-­36. 14. Wilson BC, Firnau G, Jeeves WP, Brown KL, Burns-­McCormick DM . Chromatographic analysis and...Photosensitizer-­conjugated human serum albumin nanoparticles for effective photodynamic therapy. Theranostics. 2011;; 1: 230-­9. 24. Ferreira CL, Yapp DT, Crisp

Modeling of Stability of Electrostatic and Magnetostatic Systems

DTIC Science & Technology

2017-06-01

unlimited. 13. SUPPLEMENTARY NOTES 14. ABSTRACT Electromagnetic systems undergo a variety of different instabilities. A broad class of those...15. SUBJECT TERMS electromagnetism , morphological instabilities, computational algorithm, gradient minimization, morphology patterns, motion by mean...Nordmark AB. Magnetic field and current are zero inside ideal conductors. Prog Electromagn Res B. 2011(27):187–212. 4. Stratton JA. Electromagnetic theory

78 FR 34403 - Notice of Availability of the Record of Decision for the Quartzsite Solar Energy Project, AZ

Federal Register 2010, 2011, 2012, 2013, 2014

2013-06-07

...: http://www.blm.gov/az/st/en/prog/energy/solar/quartzsite_solar_energy.html . FOR FURTHER INFORMATION... . Persons who use a telecommunications device for the deaf (TDD) may call the Federal Information Relay.... You will receive a reply during normal business hours. SUPPLEMENTARY INFORMATION: Quartzsite Solar...

Effects of Synthetic Versus Natural Colloid Resuscitation on Inducing Dilutional Coagulopathy and Increasing Hemorrhage in Rabbits

DTIC Science & Technology

2008-05-01

hemostasis, and plasma expanders: a quarter century enigma. Fed Proc. 1975;34:1429–1440. 23. Bergqvist D. Dextran and haemostasis. a review. Acta Chir ...eds. Blood Substitutes and Plasma Expanders. Prog Clin Biol Res. 1978;19:293–298. 57. Kovalik SG, Ledgewood AM, Lucas CE, Higgins RF. The cardiac

Towards Direct Simulations of Counterflow Flames with Consistent Numerical Differential-Algebraic Boundary Conditions

DTIC Science & Technology

2015-05-18

First, the gov - erning equations of the problem are presented. A detailed discussion on the construction of the initial profile of the flow follows...time from the DoD HPCMP Open Research Systems and JPL/ NASA is gratefully acknowledged. References [1] H. Tsuji, Prog. Energ. Combust.8(2) (1982) 93-119

Prevention of Mycobacterium avium subsp. paratuberculosis (MAP) infection in Balb/c mice by feeding probiotic Lactobacillus acidophilus NP-51

USDA-ARS?s Scientific Manuscript database

The objective of this study was to examine effects of feeding Lactobacillus acidophilus strain NP51 to mice challenged with Mycobacterium avium subspecies paratuberculosis (MAP), the causative agent of Johne’s disease. We hypothesized that feeding NP51 would increase Th-1 responses and decrease prog...

Prevention of Mycobacterium avium subsp. paratuberculosis (MAP) Infection in Balb/c Mice by Feeding Probiotic Lactobacillus acidophilus NP-51

USDA-ARS?s Scientific Manuscript database

The objective of this study was to examine effects of feeding Lactobacillus acidophilus strain NP51 to mice challenged with Mycobacterium avium subspecies paratuberculosis (MAP), the causative agent of Johne’s disease. We hypothesized that feeding NP51 would increase Th-1 responses and decrease prog...

Numerical Sensitivity of Trajectories Across Conformational Energy Hypersurfaces from Geometry Optimized Molecular Orbital Calculations: AM1, MNDO, and MINDO/3

DTIC Science & Technology

1988-01-01

relationship studies, it became corn- surfaces are being traversed, the molecule monly believed that compounds with higher h can go along different paths on...1975). AMPAC, and consanguineous programs 15. W. Thiel, Quantum Chem. Prog. Exchange Cata- should be done with the tightest available log, 11, 353

Report: State of Utah Drinking Water State Revolving Fund Financial Statements with Independent Auditor’s Report, June 30, 2002

EPA Pesticide Factsheets

Report #2003-1-00110, June 3, 2003.Audit of the net assets statement of the Utah Dept of Env Quality Drinking Water State Revolving Fund Prog as of June 30, 2002, and the statements of revenues, expenses and changes in fund net assets, and 2002 cash flows.

75 FR 43551 - Notice of Intent To Prepare an Environmental Impact Statement for the Proposed Mohave County Wind...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-07-26

... Dam. The project will consist of up to 335 wind turbine generators (WTGs). Construction may consist of... County Wind Farm Project, Mohave County, AZ AGENCY: Bureau of Land Management, Interior. ACTION: Notice....gov/az/st/en/prog/energy/wind/mohave.html . In order to be included in the Draft EIS, all comments...

78 FR 57877 - Notice of Intent To Prepare an Environmental Impact Statement for the Proposed Maricopa Solar...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-09-20

... Park Project, a photovoltaic (PV) solar power plant with a planned generating capacity of up to 300...; AZA35927] Notice of Intent To Prepare an Environmental Impact Statement for the Proposed Maricopa Solar... site at http://www.blm.gov/az/st/en/prog/energy/solar/maricopa-solar.html . In order to be included in...

INDUCTION OF 6-THIOGUANINE RESISTANCE IN SYNTHRONIZED HUMAN FIBROBLAST CELLS TREATED WITH METHYL METHANESULFONATE, N-ACETOXY-2-ACETHYLAMINOFLUORENE AND N-METHYL-N'-NITRO-N-NITROSOGUANIDINE

EPA Science Inventory

Chemical induction of 6-thioguanine resistance was studied in synchronized human fibroblast cells. Cells initially grown in a medium lacking arginine and glutamine for 24 h ceased DNA synthesis and failed to enter the S phase. After introduction of complete medium, the cells prog...

76 FR 66747 - Notice of Availability of the Northern Arizona Proposed Withdrawal Final Environmental Impact...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-10-27

... on the Internet at http://www.blm.gov/az/st/en/prog/mining/timeout.html . FOR FURTHER INFORMATION... million acres of Federal locatable minerals in northern Arizona from location and entry under the Mining Law of 1872, (30 U.S.C. 22-54) (Mining Law), subject to valid existing rights, by the Secretary of the...

A Theory of Electromagnetic Shielding with Applications to MIL-STD-285, IEEE-299, and EMP Simulation

DTIC Science & Technology

1985-02-01

in a building sized enclosure slot-like discontinuities may not all be small compar- ed to all wavelengths in the incident field, and slot resonan ...OFFICE OF RESEARCH/ NPP US AIR FORCE SPACE COMMAND ATTN STATE & LOCAL PROG SUPPORT O ATTN KKO 500 C STREET, SW ATTN KRQ WASHINGTON, DC 20472 ATTN XPOW

77 FR 2317 - Notice of Availability of Record of Decision for the Northern Arizona Proposed Withdrawal

Federal Register 2010, 2011, 2012, 2013, 2014

2012-01-17

... Plan for the Arizona Strip Field Office and Forest plans for the Kaibab National Forest would be... Internet at http://www.blm.gov/az/st/en/prog/mining/timeout/rod.html . FOR FURTHER INFORMATION CONTACT... General Mining Law 1,006,545 acres of Federal land and interests in land in the vicinity of the Grand...

USAF/SCEEE Summer Faculty Research Program (1979). Volume 2

DTIC Science & Technology

1979-12-01

Summer Faculty Research Program participants. The program designed to stimulate ’Ilk scientific and engineering interaction between university faculty...Prog., Dept. of Industrial Engineering Facility design and location theory University of Oklahoma and routing and distribution systems 202 W. Boyd...Theory & Assistant Professor of Management Adninistration, 1975 University of Akron S.ec aIty: Organization Design Akron, OH 44325 Assigned: AFBRMC

Effect of feeding flax or linseed meal on progesterone clearance rate in ovariectomized ewes

USDA-ARS?s Scientific Manuscript database

Ovariectomized ewes (n = 22; 68.76 ± 2.34 kg initial body weight; 2.9 ± 0.1 initial body condition score) were individually fed one of three diets: 1) Control (phytoestrogen-free; n = 7), 2) Flax containing diet (n = 8), or 3) linseed meal (LSM) containing diet (n =7) to investigate the rate of prog...

Site-Specific Dynamics of β-Sheet Peptides with (D) Pro-Gly Turns Probed by Laser-Excited Temperature-Jump Infrared Spectroscopy.

PubMed

Popp, Alexander; Scheerer, David; Chi, Heng; Keiderling, Timothy A; Hauser, Karin

2016-05-04

Turn residues and side-chain interactions play an important role for the folding of β-sheets. We investigated the conformational dynamics of a three-stranded β-sheet peptide ((D) P(D) P) and a two-stranded β-hairpin (WVYY-(D) P) by time-resolved temperature-jump (T-jump) infrared spectroscopy. Both peptide sequences contain (D) Pro-Gly residues that favor a tight β-turn. The three-stranded β-sheet (Ac-VFITS(D) PGKTYTEV(D) PGOKILQ-NH2 ) is stabilized by the turn sequences, whereas the β-hairpin (SWTVE(D) PGKYTYK-NH2 ) folding is assisted by both the turn sequence and hydrophobic cross-strand interactions. Relaxation times after the T-jump were monitored as a function of temperature and occur on a sub-microsecond time scale, (D) P(D) P being faster than WVYY-(D) P. The Xxx-(D) Pro tertiary amide provides a detectable IR band, allowing us to probe the dynamics site-specifically. The relative importance of the turn versus the intrastrand stability in β-sheet formation is discussed. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Comparative analysis of operational forecasts versus actual weather conditions in airline flight planning, volume 4

NASA Technical Reports Server (NTRS)

Keitz, J. F.

1982-01-01

The impact of more timely and accurate weather data on airline flight planning with the emphasis on fuel savings is studied. This volume of the report discusses the results of Task 4 of the four major tasks included in the study. Task 4 uses flight plan segment wind and temperature differences as indicators of dates and geographic areas for which significant forecast errors may have occurred. An in-depth analysis is then conducted for the days identified. The analysis show that significant errors occur in the operational forecast on 15 of the 33 arbitrarily selected days included in the study. Wind speeds in an area of maximum winds are underestimated by at least 20 to 25 kts. on 14 of these days. The analysis also show that there is a tendency to repeat the same forecast errors from prog to prog. Also, some perceived forecast errors from the flight plan comparisons could not be verified by visual inspection of the corresponding National Meteorological Center forecast and analyses charts, and it is likely that they are the result of weather data interpolation techniques or some other data processing procedure in the airlines' flight planning systems.

[Effect of progesterone on the expression of GLUT in the brain following hypoxic-ischemia in newborn rats].

PubMed

Li, Dong-Liang; Han, Hua

2008-08-01

To investigate the expression of GLUT1 and GLUT3 in the hippocampus after cerebral hypoxic-ischemia (HI) in newborn rats and the effect of progesterone (PROG) on them. Forty newborn SD rats were randomly divided into four groups: normal group, sham-operated group, hypoxic-ischemic group and progesterone group. Model of hypoxic-ischemia encephalopathy (HIE) was established in the 7-day-old newborn SD rats. Immunohistochemical method was applied to detect the expression of GLUT1 and GLUT3 in hippocampus. GLUT1 and GLUT3 were slightly seen in normal and sham operation group, there was no obviously difference between the two groups (P > 0.05). The expression of GLUT1 and GLUT3 in hypoxic-ischemia group were all higher than that in sham operated group (P < 0.05). Not only the expression of GLUT in progesterone group were significantly higher than that in sham operated group (P < 0.01), but also than that in hypoxic-ischemia group (P < 0.05). PROG could increase the tolerance of neuron to hypoxic-ischemia with maintaining the energy supply in the brain by up-regulating GLUT expression.

Short-Term Evaluation of Intraoral Soft Splints

DTIC Science & Technology

1994-06-01

typical orofacial pain problems (McGlynn and Cassisi, 1985; Fricton, 1991b). The objective outcome measure should both assess the degree of muscle pain ...Second Edition. Chicago, Year Book Medical Publishers, pp 218-21. Bell, W.E. (1989) Orofacial Pains Classification, Diagnosis, Management. Fourth...Fricton, J.R. (1990) Musculoskeletal measures of orofacial pain . Anesth Prog 37:136-43. Fricton, J.R. (1991a) Recent advances in teaporomandibular

Floating Ocean Platform

DTIC Science & Technology

2003-08-15

floating structures create novel habitats for subtidal epibiota?, MARINE ECOLOGY -PROGRESS SERIES, 43-52 Mar. Ecol.- Prog. Ser., 2002 Vegueria, SFJ Godoy... ECOLOGICAL APPLICATIONS, 350-366 Ecol. Appl., 2000 Niedzwecki, JM van de Lindt, JW Gage, JH Teigen, PS, Design estimates of surface wave interaction with...The ecological effects beyond the offshore platform, Coastal Zone: Proceedings of the Symposium on Coastal and Ocean Management, v 2, n pt2, 1989, p

A Comparison of Prostate Cancer Incidence Between U.S. Air Force Enlisted Aircrew

DTIC Science & Technology

2011-06-30

COL DAVID B. RHODES, RAM Prog Dir COL ROBERT E. CARROLL , Chair FE...PROGRAM ELEMENT NUMBER 6. AUTHOR( S ) Joseph A. Lopez 5d. PROJECT NUMBER 5e. TASK NUMBER 5f. WORK UNIT NUMBER 7. PERFORMING...ORGANIZATION NAME( S ) AND ADDRESS(ES) USAF School of Aerospace Medicine Aerospace Medicine Education/FEEG 2510 Fifth St. Wright-Patterson AFB, OH 45433-7913

Chalk-Ex: Transport of Optically Active Particles from the Surface Mixed Layer

DTIC Science & Technology

2001-09-30

Atlantic Ocean. Mar. Ecol. Prog. Ser. 97: 271-285. Harris, R. P. 1994. Zooplankton grazing on the coccolithophore Emiliania huxleyi and its role...Balch, and K. A. Kilpatrick. 1998. Scattering and attenuation properties of Emiliania huxleyi cells and their detached coccoliths. Limnol. Oceanogr...coccolithophore Emiliania huxleyi under steady-state light-limited growth. Marine Ecology Progress Series. 142: 87-97. Bidigare, R. R. , M

Inherited Retinal Degenerative Clinical Trial Network. Addendum

DTIC Science & Technology

2013-10-01

visual impairment usually ending in blindness. In the United States, the total number of individuals affected by retinitis pigmentosa (RP) and other...linica l trial in the NEER network for autosomal dominant retinitis pigmentosa , and the ProgSTAR studies for Stargardt disease) . As new interventions b... retinitis pigmentosa continues at six sites- the CTEC site at University of Utah and five additional recruitment sites- the Retina Foundation of the

Self-Controlled Synthesis of Hyperbranched Poly(etherketone)s from A2 + B3 Approach in Poly(phosphoric acid)

DTIC Science & Technology

2009-01-01

aromatic keto -band arisen from carboxylic acids, which could be part of terminal groups of HPEKs, ranged from 1708 to 1719 cm1. The carbonyl bands from...1999, 143 , 1–34; (d) Inoue, K. Prog Polym Sci 2000, 25, 453–571; (e) Voit, B. J Polym Sci Part A: Polym Chem 2000, 36, 2505–2525; (f) Hult, A

The Use of Modafinil in Operational Settings: Individual Difference Implications

DTIC Science & Technology

2000-03-01

them "skylark". On another treatment of narcolepsia and idiopathic hypersomnia . hand, some people are eveningness type, they have the The wakening...showed that modafinil acts as idiopathic hypersomnia and narcolepsy with modafinil. an agonist of ct1-adrenergic post-synaptic receptors (4). Prog...of During sustained operations (SUSOPS) it is impossible narcolepsia and idiopatic hypersomnia . It is synthetized for the soldier to sleep, sometimes

Serum lipid levels and steroidal hormones in women runners with irregular menses.

PubMed

Thompson, D L; Snead, D B; Seip, R L; Weltman, J Y; Rogol, A D; Weltman, A

1997-02-01

This study compared the lipid profile of women runners with menstrual cycle irregularities with their normally menstruating counterparts. Relationships among selected steroid hormones and serum lipid levels in 10 eumenorrheic (EU) and 8 oligo-/amenorrheic (O/A) women runners and 6 eumenorrheic controls (CON) were examined. Serum 17 beta-estradiol (E2), progesterone (Prog), and dehydroepiandrosterone-sulfate (DHEAS) concentrations were determined in daily blood samples for 21 days, and integrated concentrations were calculated. Fasting blood samples were analyzed for total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), HDL2, HDL3, triglycerides (Trig), and apolipoproteins A-1, A-II, and B. The O/A group had significantly lower E2 and Prog than EU or CON groups. Women in the CON group had lower HDL-C and HDL3 than the runners. With all women grouped together, E2 was not significantly correlated with any measured blood lipid parameters. On the other hand, DHEAS was significantly correlated with HDL-C, HDL2, and apolipoprotein A-I. These data demonstrate that women runners, regardless of menstrual cycle status, exhibit higher HDL-C concentrations than CON and supports previous research reporting a positive association between DHEAS and HDL-C.

Production of xylitol by a Coniochaeta ligniaria strain tolerant of inhibitors and defective in growth on xylose.

PubMed

Nichols, Nancy N; Saha, Badal C

2016-05-01

In conversion of biomass to fuels or chemicals, inhibitory compounds arising from physical-chemical pretreatment of the feedstock can interfere with fermentation of the sugars to product. Fungal strain Coniochaeta ligniaria NRRL30616 metabolizes the furan aldehydes furfural and 5-hydroxymethylfurfural, as well as a number of aromatic and aliphatic acids and aldehydes. Use of NRRL30616 to condition biomass sugars by metabolizing the inhibitors improves their fermentability. Wild-type C. ligniaria has the ability to grow on xylose as sole source of carbon and energy, with no accumulation of xylitol. Mutants of C. ligniaria unable to grow on xylose were constructed. Xylose reductase and xylitol dehydrogenase activities were reduced by approximately two thirds in mutant C8100. The mutant retained ability to metabolize inhibitors in biomass hydrolysates. Although C. ligniaria C8100 did not grow on xylose, the strain converted a portion of xylose to xylitol, producing 0.59 g xylitol/g xylose in rich medium and 0.48 g xylitol/g xylose in corn stover dilute acid hydrolysate. 2016 American Institute of Chemical Engineers Biotechnol. Prog., 2016 © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:606-612, 2016. © 2016 American Institute of Chemical Engineers.

VizieR Online Data Catalog: The YSO population of LDN 1340 in infrared (Kun+, 2016)

NASA Astrophysics Data System (ADS)

Kun, M.; Wolf-Chase, G.; Moor, A.; Apai, D.; Balog, Z.; O'Linger-Luscusk, J.; Moriarty-Schieven, G. H.

2016-07-01

L1340 was observed by the Spitzer Space Telescope using Spitzer's Infrared Array Camera (IRAC) on 2009 March 16 and the Multiband Imaging Photometer for Spitzer (MIPS) on 2008 November 26 (Prog. ID: 50691, PI: G. Fazio). The IRAC observations covered ~1deg2 in all four bands. Moreover, a small part of the cloud, centered on RNO 7, was observed in the four IRAC bands on 2006 September 24 (Prog. ID: 30734, PI: D. Figer). We selected candidate YSOs from the Spitzer Enhanced Imaging Products (SEIP) Source List, containing 19745 point sources in the target field. High angular resolution near-infrared images of two small regions of L1340 were obtained on 2002 October 24 in the JHK bands, using the near-infrared camera Omega-Cass, mounted on the 3.5m telescope at the Calar Alto Observatory, Spain. The results for IRAS 02224+7227 have been shown in Kun et al. (2014, J/ApJ/795/L26). Here we present the results for RNO 7. To classify the evolutionary status of the color-selected candidate YSOs and obtain as complete a picture of the SFR and its YSO population as possible, we supplemented the Spitzer data with photometric data available in public databases. See section 2.3 for further details. (13 data files).

Associations between Bisphenol A Exposure and Reproductive Hormones among Female Workers

PubMed Central

Miao, Maohua; Yuan, Wei; Yang, Fen; Liang, Hong; Zhou, Zhijun; Li, Runsheng; Gao, Ersheng; Li, De-Kun

2015-01-01

The associations between Bisphenol-A (BPA) exposure and reproductive hormone levels among women are unclear. A cross-sectional study was conducted among female workers from BPA-exposed and unexposed factories in China. Women’s blood samples were collected for assay of follicle-stimulating hormone (FSH), luteinizing hormone (LH), 17β-Estradiol (E2), prolactin (PRL), and progesterone (PROG). Their urine samples were collected for BPA measurement. In the exposed group, time weighted average exposure to BPA for an 8-h shift (TWA8), a measure incorporating historic exposure level, was generated based on personal air sampling. Multiple linear regression analyses were used to examine linear associations between urine BPA concentration and reproductive hormones after controlling for potential confounders. A total of 106 exposed and 250 unexposed female workers were included in this study. A significant positive association between increased urine BPA concentration and higher PRL and PROG levels were observed. Similar associations were observed after the analysis was carried out separately among the exposed and unexposed workers. In addition, a positive association between urine BPA and E2 was observed among exposed workers with borderline significance, while a statistically significant inverse association between urine BPA and FSH was observed among unexposed group. The results suggest that BPA exposure may lead to alterations in female reproductive hormone levels. PMID:26506366

Associations between Bisphenol A Exposure and Reproductive Hormones among Female Workers.

PubMed

Miao, Maohua; Yuan, Wei; Yang, Fen; Liang, Hong; Zhou, Zhijun; Li, Runsheng; Gao, Ersheng; Li, De-Kun

2015-10-22

The associations between Bisphenol-A (BPA) exposure and reproductive hormone levels among women are unclear. A cross-sectional study was conducted among female workers from BPA-exposed and unexposed factories in China. Women's blood samples were collected for assay of follicle-stimulating hormone (FSH), luteinizing hormone (LH), 17β-Estradiol (E2), prolactin (PRL), and progesterone (PROG). Their urine samples were collected for BPA measurement. In the exposed group, time weighted average exposure to BPA for an 8-h shift (TWA8), a measure incorporating historic exposure level, was generated based on personal air sampling. Multiple linear regression analyses were used to examine linear associations between urine BPA concentration and reproductive hormones after controlling for potential confounders. A total of 106 exposed and 250 unexposed female workers were included in this study. A significant positive association between increased urine BPA concentration and higher PRL and PROG levels were observed. Similar associations were observed after the analysis was carried out separately among the exposed and unexposed workers. In addition, a positive association between urine BPA and E2 was observed among exposed workers with borderline significance, while a statistically significant inverse association between urine BPA and FSH was observed among unexposed group. The results suggest that BPA exposure may lead to alterations in female reproductive hormone levels.

Ab initio Quantum Chemical and Experimental Reaction Kinetics Studies in the Combustion of Bipropellants

DTIC Science & Technology

2017-03-24

NUMBER (Include area code) 24 March 2017 Briefing Charts 01 March 2017 - 31 March 2017 Ab initio Quantum Chemical and Experimental Reaction Kinetics...Laboratory AFRL/RQRS 1 Ara Road Edwards AFB, CA 93524 *Email: ghanshyam.vaghjiani@us.af.mil Ab initio Quantum Chemical and Experimental Reaction ...Clearance 17161 Zador et al., Prog. Energ. Combust. Sci., 37 371 (2011) Why Quantum Chemical Reaction Kinetics Studies? DISTRIBUTION A: Approved for

Acoustic Directivity Patterns for Army Weapons

DTIC Science & Technology

1979-01-01

work was performed by the Environmental Division (EN), u.S. Army Construction Engineering Research Laboratory (CERL). Dr. R. K. Jain is Chief of EN...V) P.0. Schomer,,, L. M./Little I rTPRFORMING ORGANI ZATION NAME AND ADDRESS 0.PROG3RAM ELEMENT. PPOJECT, TAWF U.S. ARMY AREA & WORK UNIT NUMBERS...34Environmental Quality for Construction and Operation of Military Facilities" Task 03, "Pollution Control Technology" and Work Unit 001, "Prediction of

Optical Limiting in Photonic Crystal Fibers

DTIC Science & Technology

2004-12-01

Optical Limiting in Photonic Crystal Fibers Mark Bloemer, Michael Scalora , Wayne Davenport, and Evgeni Poliakov(NRC Postdoc) RDECOM, Aviation...Shcherbakov, E. Wintner, M. Scalora , and A. M. Zheltikov, Appl. Opt., in press. 7 21. C. M. de Sterke and J. E. Sipe, Prog. Opt. 33, 203 (1994... Scalora , J..P. Dowling, C.M. Bowden, and M.J. Bloemer, Phys. Rev. Lett. 73, 1368 (1994). 26. M.D. Tocci, M.J. Bloemer, M. Scalora , J.P. Dowling

Principles of Work Sample Testing. 4. Generalizability

DTIC Science & Technology

1979-04-01

ARI TECHNICAL REPORT TR-79-A11 Principles of Work Sample Testing: IV. Generallzability , by ,lobert M. Guion Gail H. Ironson BOWLING GREEN STATE...UNIVERSITY Bowling Green , Ohio 43403 April 1979 or -Contract DAHC 19-77-C-0007 d CD, LUa.J Prepared for -_J ;=U.S. ARMY RESEARCH INSTITUTE for the...ORGANIZATION NAME AND ADDRESS 10. PROG RAM ELEMENT. PROJECT, TASK A REA & WORK UNIT NUMBERS Bowling Green State University __ Bowling Green , Ohio 43403

Department of Defense In-House RDT and E Activities

DTIC Science & Technology

1978-10-30

SERVICF DFV CML CML- BIO OFFENSE SMOKE/OBSCURANT TEST PROGRAMS CONDUCTS R & 0 & LAB INVESTIGATIONS NECESSARY TO SUPPORT MISSIONCONOUCTS JT OP CML & CNL... BIO nEFENSE TESTS/STUEPFS FOR CTNCS, SERVICESCONDUCTS PROG TO SUP ARMY POLL ABATEMENT HAZARD EVAL E DFMIL OPNSCONDUCTS ECOLOGICAL EPIDEMIOLOGICAL C...CDRoCOL. H. F. PENNY TECHODIR.DR. DALE H. STELING PROGRAM DATA BY FISCAL YEAR (MILLION S) PROGRAM 1978 1979 (ACT UAL) (ACT + EST)TOTAL RDT&E 23&722

Integrated Strike Avionics Study. Volume 1

DTIC Science & Technology

1980-10-01

MMW Systems Targeting Studies Perf. Meas. o C02 Laser Radar Ses. St. Army Obstacle Detect Prog. Concept Demo Mobile System 20 ’ - I...Fabrication and Test o FLIR Field of View & Classification Study (FLIR FACS) Definition m Development & Test 4. Aplicability of Current Programs to...FY80 81 8283 84 85 o LANTIRN 1 n Imaoinn Sensor Autoprocessor • o Forward Looking Active Class a 4. Aplicability of Current Program Required The need

Transitioning DARPA Technology

DTIC Science & Technology

2001-05-01

logo suggests, the Institute’s work reflects the summation of technology’s effects on business and government. With a reputation for fierce objectivity... effective for "customerpull" strategies. b. Products moved along the DIS path 30 percent of the time. This path was particularlysuccessful for small...must often be "waited out." But DARPA ha s few effective mechanisms for continuing to "market" its products after the prog ram is over- particularly

Isolation, Expression Analysis, and Functional Characterization of the First Antidiuretic Hormone Receptor in Insects

DTIC Science & Technology

2010-06-01

fruitfly Drosophila melanogaster and the honey bee Apis mellifera . Prog Neurobiol 80:1–19. 28. Larkin MA, et al. (2007) Clustal W and Clustal X version...receptors predicted or annotated in the Acyrthosiphon pisum, Pediculus humanus corporis, andApis mellifera genomes. The CAPA-related peptides in insects...prolixus CAPA Receptor Gene. CAPA receptor protein sequences identified or predicted in D. melanogaster (AAS65092) (13, 14), A. mellifera (NP_001091702

Rapid Response Concentration-Controlled Desorption of Activated Carbon to Dampen Concentration Fluctuations

DTIC Science & Technology

2007-01-01

Behavior of trickle - bed air biofilter for toluene removal: Effect of non-use periods. Environ. Prog. 2005, 24, 155-161. (3) Martin, F. J.; Loehr, R. C...dampen the fluctuation in acetone concentration at high concentrations. The effect of inlet concentration and empty bed contact time (EBCT) on dampening...oxidizer. The MSA-SST system is a fixed- bed system that rapidly controls the power that heats the adsorbent/adsorbate, resulting in controlled

Design Evolution of a Fighter Training Scheduling Decision Support System.

DTIC Science & Technology

1987-03-01

SYSTEM THESIS Paul E. Trapp Jeffrey W. Grechanik Captain, USAF Captain, USAF AFIT/GST/ENS/87M-8 MAY 191987 " Approved for public release; distribution...E. Trapp, B.S., M.A. Jeffrey W. Grechanik, B.S. Captain, USAF Captain, USAF March 87 Approved for public release; distribution unlimited This work...DNIF, TDY, and other disruptions. Therefore, cycli- cal scheduling will not be used (3:1-18). ProgAMming. Arthur and Ravindran proposed a goal

Enzootic Plague Reduces Black-Footed Ferret (Mustela nigripes) Survival in Montana

DTIC Science & Technology

2010-01-01

al. Design and testing for a non-tagged F1-V fusion protein as vaccine antigen against bubonic and pneumonic plague . Biotechnol Prog 2005; 21:1490–1510...Enzootic Plague Reduces Black-Footed Ferret (Mustela nigripes) Survival in Montana Marc R. Matchett,1 Dean E. Biggins,2 Valerie Carlson,3,* Bradford...and prey. Epizootic plague kills both prairie dogs and ferrets and is a major factor limiting recovery of the highly endangered ferret. In addition to

The Role of Limited Proteolysis of Thyrotropin-Releasing Hormone in Thermoregulation.

DTIC Science & Technology

1982-01-01

exogenously. The limited proteolysis of TRH by pyroglutamate aminopeptidase from CNS results into formation of a new cyclic dipeptide, cyclo (His-Pro...amino acids (L-histidine and L-proline), and two analogues of cyclo (His-Pro), cyclo (Pro-Gly) and cyclo . (Ala-Gly). Cyclo (His-Pro) cross-reacted only...cyclo (His-Pro). Figure 3 shows the chromato- graphic profile obtained when a neutralized perchloric acid extract of rat brain was passed through DEAE

Electrochemical and Photochemical Treatment of Aqueous Waste Streams

DTIC Science & Technology

1996-01-01

TREATMENT OF AQUEOUS WASTE STREAMS Joseph C. Farmer, Richard W. Pekala, Francis T. Wang, David V. Fix, Alan M. Volpe, Daniel D. Dietrich, William H...STREAMS Joseph C. Farmer, Richard W. Pekala, Francis T. Wang, David V. Fix, Alan M. Volpe, Daniel D. Dietrich, William H. Siegel and James F. Carley...1992). Wilbourne , C. M. Wong, , W. S. Gillam, S. Johnson, R. H. Horowitz, "Electrosorb Process for Desalting Water," Res. Dev. Prog. Rept. No. 516, 16. J

Clinical Investigation Program Report Control Symbol MED 300.

DTIC Science & Technology

1982-10-01

Preeclampsia as an Aid to Further Management. (C) (PR) 41 1961 Use of C-Reactive Protein in Prediction of ARD Prog- nosiS, (C) (PR) 42 1981 The Assessment...37 Status: Completed * Title: Routine Use of Serum Uric Acid Levels at 36 Weeks Gestation as Screening Test for Preeclampsia as an Aid to Further...nvestigators: .amily Pr-&ctiU. CPT Ellis M. Knight, MC Key Words: Serum Uric Acid Preeclampsia Accumulative MZDCASI Eat Accumulative Periodic Mar 82 Cost: ]A

Reactivity of Metal Nitrates.

DTIC Science & Technology

1982-07-20

02NOCuOH Any mechanism suggested for the nitration of aromatic systems by titanium(IV) nitrate must take into account the observed similarity, in...occurs. -26- References 1. For recent reviews see (a) R. B. Moodie and K. Schofield, Accounts Chem. Res., 1976, 9, 287; (b) G. A. Olah and S. J. Kuhn...Ithaca, N.Y., 1969, Chapter VI; L. M. Stock, Prog. Phys. Org. Chem., 1976, 12, 21; J. G. Hoggett , R. B. Moodie, J. R. Penton, and K. Schofield

The United States Air Force and the Culture of Innovation, 1945-1965

DTIC Science & Technology

2002-01-01

US Dept of Transportation; typically they hover between 85 and 95 percent. 16. Kent C. Redmond and Thomas M. Smith, Project Whirlwind: A Case Histo...Washing- ton, D.C.: AF Hist and Museums Prog, 1994). 14. Thomas A . Sturm, The USAF SAB: Its First Twenty Years 1944–1964 (Washington, D.C.: USAF...allegations at Ramo-Wooldridge and the Air Force’s approach. Schriever answered them in a letter to Lt. Gen. Thomas Power, the commander of ARDC, in

Role of Human Polyomavirus Bkv in Prostate Cancer

DTIC Science & Technology

2007-12-01

D. L. Walker. 1976 . New human papovaviruses. Prog Med Virol. 22:1-35. 59. Palapattu, G. S., S. Sutcliffe, P. J. Bastian, E. A. Platz, A. M. De Marzo ...J. Imperiale. 2004. Detection and expression of human BK virus sequences in neoplastic prostate tissues. Oncogene 23:7031-7046. 4. De Marzo , A. M...virus sequences in neoplastic prostate tissues. Oncogene 23:7031-7046. 41 18. De Marzo , A. M., T. L. DeWeese, E. A. Platz, A. K. Meeker, M. Nakayama

Correlation between average melting temperature and glass transition temperature in metallic glasses

NASA Astrophysics Data System (ADS)

Lu, Zhibin; Li, Jiangong

2009-02-01

The correlation between average melting temperature (⟨Tm⟩) and glass transition temperature (Tg) in metallic glasses (MGs) is analyzed. A linear relationship, Tg=0.385⟨Tm⟩, is observed. This correlation agrees with Egami's suggestion [Rep. Prog. Phys. 47, 1601 (1984)]. The prediction of Tg from ⟨Tm⟩ through the relationship Tg=0.385⟨Tm⟩ has been tested using experimental data obtained on a large number of MGs. This relationship can be used to predict and design MGs with a desired Tg.

Spontaneous Transitional Cell Carcinoma in the Urinary Bladder of a Strain 13 Guinea Pig.

DTIC Science & Technology

1985-05-01

less than I of all canine neoplasms. In the feline , the extremely low incidence of bladder tumors observed may well be due to a difference in...factor of age. Prog Exp Tumor Res 1967;9:261-85. ..................... , 12. Ediger R D, Rabstein M M. Spontaneous leukemia in a Hartley strain guinea pig...Intracisternal virus -like particles in two guinea pig mammary adenocarcinomas. Lab Anim Sci 1976;26:607-9. 17.. Yoshida A, Iqbal Z M, Epstein. S S

Development of the Wake Behind a Circular Cylinder Impulsively Started into Rotatory and Rectilinear Motion: Intermediate Rotation Rates

DTIC Science & Technology

1991-01-01

cylindre fixe ou en rotation. Effet Magnus . J. Mec. 14, 109-134. Taneda, S. 1977 Visual study of unsteady separated flows around bodies. Prog. Aero...enhancement schemes employing the Magnus effect (Swanson 1961). Rotating all or part of a body may also have applications in active or feedback control of...and yt into the governing equations in the generalized coordinate system. In this study, the body-fitted grid is simply one of cylindrical polar

Adapting Biofilter Processes to Treat Spray Painting Exhausts: Concentration and Leveling of Vapor Delivery Rates, and Enhancement of Destruction by Exhaust Recirculation

DTIC Science & Technology

2001-12-20

1992. "Consider biofiltration for decontaminating gases ," Chem. Eng. Prog. 88(4):34–40. Brunner, W., D. Staub, and T. Leisinger. 1980...Biotreatment processes, such as biofiltration , are environmentally friendly, and produce only non-hazardous by-products such as water, inorganic salts, and...biological air treatment system is biofiltration . Biofiltration is a process that utilizes microorganisms immobilized in the form of a biofilm layer on

Biostable Agonists that Match or Exceed Activity of Native Insect Kinins on Recombinant Arthropod GPCRs

DTIC Science & Technology

2009-01-01

with a modified FastMoc 0.25 procedure using an Fmoc-strategy starting from Rink amide resin (Novabiochem, San Diego, CA, 0.5 mM/g). The Fmoc protecting...In vitro release of amylase by culekinins in two insects: Opsinia arenosella (Lepidoptera) and Rhynchophorus ferrugineus (Coleoptera). Trends Life...Drosophila melanogaster and the honey bee Apis mellifera. Prog. Neurobiol. 80, 1–19. Holman, G.M., Nachman, R.J., Wright, M.S., 1990. Insect

CG-MS/MS Analyses of Biological Samples in Support of Developmental Toxic Effects on Whole-Body Exposure of Rats to GB

DTIC Science & Technology

2015-03-01

Sensitivity to Organophosphorous Anticholinesterase Compounds. Prog. Neurobiol. 1987, 28, 97–129. Shih, T-M.; Penetar, D.M.; McDonough, J.H. Jr.; Romano...J.A.; King, J.M. Age- related Differences in Soman Toxicity and in Blood and Brain Regional Cholinesterase Activity . Brain Res. Bull. 1990, 24...Organophosphates. Toxicol. Appl. Pharmacol. 2004, 198, 132–151. Sterri, S.H.; Berge, G.; Fonnum, F. Esterase Activities and Soman Toxicity in

JCL (Job Control Language) Procedures to Run the Hull Code on the Cyber 205 Computer Installed on CSIRONET.

DTIC Science & Technology

1986-11-01

START THE RUN>>> USERNUIDNUPW. CHARGEGROUPNPID. SETJOB, DC= NO . COMMENT . GET CR ATTACH THE INPUT DATA TO GO TO VSOS. GET, INDATA=DATFILE/NA. IFE...NtPW. CHARGEGROUPNPID. SETTL, 200. SETJOB. DC= NO . COMMENT . RUN SAIL ON NOS TO GENERATE THE MAIN PROGRAM. PURGE, SAl LOUT/NA. PURGE, PROG-PROBLEMID...NOSPASS. CHARGEDFCDFCPR.F. SETJOB. DC= NO . COMMENT . GET OR ATTACH THE INPUT DATA To Go To VSOS. GET. INDATA=MYDATA/NA. IFE. .NOT.FILE(INDATA.AS) .DOATT

Salmonella typhimurium gyrA mutations associated with fluoroquinolone resistance.

PubMed Central

Reyna, F; Huesca, M; González, V; Fuchs, L Y

1995-01-01

Spontaneous quinolone-resistant mutants obtained from Salmonella typhimurium Su694 were screened for mutations by direct DNA sequencing of an amplified PCR gyrA fragment. Substitutions Ser-83-->Phe (Ser83Phe), Ser83Tyr, Asp87Tyr, and Asp87Asn and double mutation Ala67Pro-Gly81Ser, which resulted in decreased sensitivities to ciprofloxacin, enoxacin, pefloxacin, norfloxacin, ofloxacin, and nalidixic acid, were found. The levels of resistance to quinolones for each mutant were determined. PMID:7492118

African Non-Human Primates Host Diverse Enteroviruses.

PubMed

Mombo, Illich Manfred; Lukashev, Alexander N; Bleicker, Tobias; Brünink, Sebastian; Berthet, Nicolas; Maganga, Gael D; Durand, Patrick; Arnathau, Céline; Boundenga, Larson; Ngoubangoye, Barthélémy; Boué, Vanina; Liégeois, Florian; Ollomo, Benjamin; Prugnolle, Franck; Drexler, Jan Felix; Drosten, Christian; Renaud, François; Rougeron, Virginie; Leroy, Eric

2017-01-01

Enteroviruses (EVs) belong to the family Picornaviridae and are responsible for mild to severe diseases in mammals including humans and non-human primates (NHP). Simian EVs were first discovered in the 1950s in the Old World Monkeys and recently in wild chimpanzee, gorilla and mandrill in Cameroon. In the present study, we screened by PCR EVs in 600 fecal samples of wild apes and monkeys that were collected at four sites in Gabon. A total of 32 samples were positive for EVs (25 from mandrills, 7 from chimpanzees, none from gorillas). The phylogenetic analysis of VP1 and VP2 genes showed that EVs identified in chimpanzees were members of two human EV species, EV-A and EV-B, and those identified in mandrills were members of the human species EV-B and the simian species EV-J. The identification of two novel enterovirus types, EV-B112 in a chimpanzee and EV-B113 in a mandrill, suggests these NHPs could be potential sources of new EV types. The identification of EV-B107 and EV90 that were previously found in humans indicates cross-species transfers. Also the identification of chimpanzee-derived EV110 in a mandrill demonstrated a wide host range of this EV. Further research of EVs in NHPs would help understanding emergence of new types or variants, and evaluating the real risk of cross-species transmission for humans as well for NHPs populations.

An enhanced Enterovirus surveillance system allows identification and characterization of rare and emerging respiratory enteroviruses in Denmark, 2015-16.

PubMed

Barnadas, Céline; Midgley, Sofie E; Skov, Marianne N; Jensen, Lotte; Poulsen, Mille W; Fischer, Thea Kølsen

2017-08-01

The potential for outbreaks due to Enteroviruses (EV) with respiratory tropism, such as EV-D68, and the detection of new and rare EV species C is a concern. These EVs are typically not detected in stool specimens and may therefore be missed by standard EV surveillance systems. Following the North American outbreak of EV-D68 in 2014, Denmark piloted an enhanced EV surveillance system that included the screening of respiratory samples. We aim to report clinical manifestations and phylogenetic descriptions from the rare and emerging EVs identified thereby demonstrating the usefulness of this system. Positive EV samples received through the enhanced non-polio EV pilot surveillance system were characterized by sequencing fragments of VP1, VP2 and VP4 capsid proteins and clinical observations were compiled. Between January 2015 and October 2016, six cases of rare genotypes EV-C104, C105 and C109 and nine cases of EV-D68 were identified. Patients presented with mild to moderately severe respiratory illness; no paralysis occurred. Distinct EV-C104, EV-C109 and EV-D68 sequences argue against a common source of introduction of these genotypes in the Danish population. The enhanced EV surveillance system enabled detection and characterization of rare EVs in Denmark. In order to improve our knowledge of and our preparedness against emerging EVs, public health laboratories should consider expanding their EV surveillance system to include respiratory specimens. Copyright © 2017 Elsevier B.V. All rights reserved.

The Natural History of the Progression of Atrophy Secondary to Stargardt Disease (ProgStar) Studies: Design and Baseline Characteristics: ProgStar Report No. 1.

PubMed

Strauss, Rupert W; Ho, Alex; Muñoz, Beatriz; Cideciyan, Artur V; Sahel, José-Alain; Sunness, Janet S; Birch, David G; Bernstein, Paul S; Michaelides, Michel; Traboulsi, Elias I; Zrenner, Eberhart; Sadda, SriniVas; Ervin, Ann-Margret; West, Sheila; Scholl, Hendrik P N

2016-04-01

To describe the design and baseline characteristics of patients enrolled into 2 natural history studies of Stargardt disease (STGD1). Multicenter retrospective and prospective cohort studies. Three hundred sixty-five unique patients aged 6 years and older at baseline harboring disease-causing variants in the ABCA4 gene and with specified ocular lesions were enrolled from 9 centers in the United States and Europe. In the retrospective study, patients contributed medical record data from at least 2 and up to 4 visits for at least 1 examination modality: fundus autofluorescence (FAF), spectral-domain (SD) optical coherence tomography (SD OCT), and/or microperimetry (MP). The total observational period was at least 2 years and up to 5 years between single visits. Demographic and visual acuity (VA) data also were obtained. In the prospective study, eligible patients were examined at baseline using a standard protocol, with 6-month follow-up visits planned for a 2-year period for serial Early Treatment Diabetic Retinopathy Study (ETDRS) best-corrected VA, SD OCT, FAF, and MP. Design and rationale of a multicenter study to determine the progression of STGD1 in 2 large retrospective and prospective international cohorts. Detailed baseline characteristics of both cohorts are presented, including demographics, and structural and functional retinal metrics. Into the retrospective study, 251 patients (458 eyes) were enrolled; mean follow-up ± standard deviation was 3.9±1.6 years. At baseline, 36% had no or mild VA loss, and 47% of the study eyes had areas of definitely decreased autofluorescence (DDAF) with an average lesion area of 2.5±2.9 mm(2) (range, 0.02-16.03 mm(2)). Two hundred fifty-nine patients (489 eyes) were enrolled in the prospective study. At baseline, 20% had no or mild VA loss, and 64% had areas of DDAF with an average lesion area of 4.0±4.4 mm(2) (range, 0.03-24.24 mm(2)). The mean retinal sensitivity with MP was 10.8±5.0 dB. The ProgStar cohorts have baseline characteristics that encompass a wide range of disease severity and are expected to provide valuable data on progression based on serial quantitative measurements derived from multiple methods, which will be critical to the design of planned clinical trials. Copyright © 2016 American Academy of Ophthalmology. Published by Elsevier Inc. All rights reserved.

Characterisation of adipocyte-derived extracellular vesicle subtypes identifies distinct protein and lipid signatures for large and small extracellular vesicles

PubMed Central

Durcin, Maëva; Fleury, Audrey; Taillebois, Emiliane; Hilairet, Grégory; Krupova, Zuzana; Henry, Céline; Truchet, Sandrine; Trötzmüller, Martin; Köfeler, Harald; Mabilleau, Guillaume; Hue, Olivier; Andriantsitohaina, Ramaroson; Martin, Patrice; Le Lay, Soazig

2017-01-01

ABSTRACT Extracellular vesicles (EVs) are biological vectors that can modulate the metabolism of target cells by conveying signalling proteins and genomic material. The level of EVs in plasma is significantly increased in cardiometabolic diseases associated with obesity, suggesting their possible participation in the development of metabolic dysfunction. With regard to the poor definition of adipocyte-derived EVs, the purpose of this study was to characterise both qualitatively and quantitatively EVs subpopulations secreted by fat cells. Adipocyte-derived EVs were isolated by differential centrifugation of conditioned media collected from 3T3-L1 adipocytes cultured for 24 h in serum-free conditions. Based on morphological and biochemical properties, as well as quantification of secreted EVs, we distinguished two subpopulations of adipocyte-derived EVs, namely small extracellular vesicles (sEVs) and large extracellular vesicles (lEVs). Proteomic analyses revealed that lEVs and sEVs exhibit specific protein signatures, allowing us not only to define novel markers of each population, but also to predict their biological functions. Despite similar phospholipid patterns, the comparative lipidomic analysis performed on these EV subclasses revealed a specific cholesterol enrichment of the sEV population, whereas lEVs were characterised by high amounts of externalised phosphatidylserine. Enhanced secretion of lEVs and sEVs is achievable following exposure to different biological stimuli related to the chronic low-grade inflammation state associated with obesity. Finally, we demonstrate the ability of primary murine adipocytes to secrete sEVs and lEVs, which display physical and biological characteristics similar to those described for 3T3-L1. Our study provides additional information and elements to define EV subtypes based on the characterisation of adipocyte-derived EV populations. It also underscores the need to distinguish EV subpopulations, through a combination of multiple approaches and markers, since their specific composition may cause distinct metabolic responses in recipient cells and tissues. PMID:28473884

High frequency of Polio-like Enterovirus C strains with differential clustering of CVA-13 and EV-C99 subgenotypes in a cohort of Malawian children.

PubMed

Brouwer, Lieke; van der Sanden, Sabine M G; Calis, Job C J; Bruning, Andrea H L; Wang, Steven; Wildenbeest, Joanne G; Rebers, Sjoerd P H; Phiri, Kamija S; Westerhuis, Brenda M; van Hensbroek, Michaël Boele; Pajkrt, Dasja; Wolthers, Katja C

2018-05-28

Enteroviruses (EVs) are among the most commonly detected viruses infecting humans worldwide. Although the prevalence of EVs is widely studied, the status of EV prevalence in sub-Saharan Africa remains largely unknown. The objective of our present study was therefore to increase our knowledge on EV circulation in sub-Saharan Africa. We obtained 749 fecal samples from a cross-sectional study conducted on Malawian children aged 6 to 60 months. We tested the samples for the presence of EVs using real time PCR, and typed the positive samples based on partial viral protein 1 (VP1) sequences. A large proportion of the samples was EV positive (89.9%). 12.9% of the typed samples belonged to EV species A (EV-A), 48.6% to species B (EV-B) and 38.5% to species C (EV-C). More than half of the EV-C strains (53%) belonged to subgroup C containing, among others, Poliovirus (PV) 1-3. The serotype most frequently isolated in our study was CVA-13, followed by EV-C99. The strains of CVA-13 showed a vast genetic diversity, possibly representing a new cluster, 'F'. The majority of the EV-C99 strains grouped together as cluster B. In conclusion, this study showed a vast circulation of EVs among Malawian children, with an EV prevalence of 89.9%. Identification of prevalences for species EV-C comparable to our study (38.5%) have only previously been reported in sub-Saharan Africa, and EV-C is rarely found outside of this region. The data found in this study are an important contribution to our current knowledge of EV epidemiology within sub-Saharan Africa.

Clinical Characterization of Children Presenting to the Hospital with Enterovirus D68 Infection During the 2014 Outbreak in St. Louis

PubMed Central

Orvedahl, Anthony; Padhye, Amruta; Barton, Kevin; O’Bryan, Kevin; Baty, Jack; Gruchala, Nancy; Niesen, Angela; Margoni, Angeliki; Srinivasan, Mythili

2016-01-01

Background The largest known outbreak of enterovirus D68 (EV-D68) infections occurred during 2014. The goal of our study is to characterize the illness severity and clinical presentation of children infected with enterovirus-D68 (EV-D68) in comparison to non-EV-D68-Human Rhino/Enteroviruses (HR/EV). Method Our study is a retrospective analysis of severity level, charges and length of stay of children who presented to St. Louis Children’s Hospital from 8/31/2014–10/31/2014 and tested positive for EV-D68 in comparison to non-EV-D68-HR/EV infected patients. Chart review was performed for all EV-D68 infected patients and age and severity matched non-EV-D68-HR/EV infected patients. Result There was a striking increase in hospital census in August of 2014 in our hospital with simultaneous increase in the number of patients with EV-D68 infection. There was no significant difference in severity of illness, length of stay or total charges between EV-D68 and non-EV-D68-HR/EV infected children. EV-D68 infection was characterized by presenting complaints of difficulty breathing (80%) and wheezing (67%), and by findings of tachypnea (65%), wheezing (71%) and retractions (65%) on examination. The most common interventions were albuterol (79%) and corticosteroid (68%) treatments and the most common discharge diagnosis was asthma exacerbation (55%). Conclusion EV-D68 caused a significant outbreak in 2014 with increased hospital admissions and associated increased charges. There was no significant difference in severity of illness caused by EV-D68 and non-EV-D68-HR/EV infections suggesting that the impact from EV-D68 was due to increased number of infected children presenting to the hospital and not necessarily due to increased severity of illness. PMID:26771663

Fluorescence triggering: A general strategy for enumerating and phenotyping extracellular vesicles by flow cytometry.

PubMed

Arraud, Nicolas; Gounou, Céline; Turpin, Delphine; Brisson, Alain R

2016-02-01

Plasma contains cell-derived extracellular vesicles (EVs) which participate in various physiopathological processes and have potential biomedical applications. Despite intense research activity, knowledge on EVs is limited mainly due to the difficulty of isolating and characterizing sub-micrometer particles like EVs. We have recently reported that a simple flow cytometry (FCM) approach based on triggering the detection on a fluorescence signal enabled the detection of 50× more Annexin-A5 binding EVs (Anx5+ EVs) in plasma than the conventional FCM approach based on light scattering triggering. Here, we present the application of the fluorescence triggering approach to the enumeration and phenotyping of EVs from platelet free plasma (PFP), focusing on CD41+ and CD235a+ EVs, as well as their sub-populations which bind or do not bind Anx5. Higher EV concentrations were detected by fluorescence triggering as compared to light scattering triggering, namely 40× for Anx5+ EVs, 75× for CD41+ EVs, and 15× for CD235a+ EVs. We found that about 30% of Anx5+ EVs were of platelet origin while only 3% of them were of erythrocyte origin. In addition, a majority of EVs from platelet and erythrocyte origin do not expose PS, in contrast to the classical theory of EV formation. Furthermore, the same PFP samples were analyzed fresh and after freeze-thawing, showing that freeze-thawing processes induce an increase, of about 35%, in the amount of Anx5+ EVs, while the other EV phenotypes remain unchanged. The method of EV detection and phenotyping by fluorescence triggering is simple, sensitive and reliable. We foresee that its application to EV studies will improve our understanding on the formation mechanisms and functions of EVs in health and disease and help the development of EV-based biomarkers. © 2015 International Society for Advancement of Cytometry.

Isolation and Characterization of Extracellular Vesicles from Adult Schistosoma japonicum.

PubMed

Liu, Juntao; Zhu, Lihui; Wang, Lihui; Chen, Yongjun; Giri, Bikash Ranjan; Li, Jianjun; Cheng, Guofeng

2018-05-22

Extracellular vesicles (EVs) are membranous vesicles released by a variety of cells into the extracellular microenvironment. EVs represent a population of heterogeneous vesicles, whose size range between 40 and 1,000 nm. Accumulated evidence indicated that EVs play important regulatory roles in pathogen-host interactions. A deep understanding of schistosome EVs should provide insights into the mechanisms underlying schistosome-host interactions, enabling development of novel strategies against schistosomiasis. Here, we aim to further study EVs functions in schistosomes by presenting a protocol for the isolation and characterization of EVs from adult Schistosoma japonicum (S. japonicum). EVs were isolated from in vitro culture medium using centrifugation combined with a commercial exosome isolation kit. The isolated S. japonicum EVs (SjEVs) typically possess a diameter of 100 - 400 nm, and are characterized by transmission electronic microscopy and western blotting. The usage of PKH67 dye-labeled SjEVs has demonstrated that SjEVs are internalized by the recipient cells. Overall, our protocol provides an alternative method for isolating EVs from adult schistosomes; the isolated SjEVs may be suitable for functional analysis.

A new bioluminescent reporter system to study the biodistribution of systematically injected tumor-derived bioluminescent extracellular vesicles in mice

PubMed Central

Gangadaran, Prakash; Li, Xiu Juan; Lee, Ho Won; Oh, Ji Min; Kalimuthu, Senthilkumar; Rajendran, Ramya Lakshmi; Son, Seung Hyun; Baek, Se Hwan; Singh, Thoudam Debraj; Zhu, Liya; Jeong, Shin Young; Lee, Sang-Woo; Lee, Jaetae; Ahn, Byeong-Cheol

2017-01-01

In vivo biodistribution and fate of extracellular vesicles (EVs) are still largely unknown and require reliable in vivo tracking techniques. In this study, in vivo bioluminescence imaging (BLI) using Renilla luciferase (Rluc) was developed and applied to monitoring of EVs derived from thyroid cancer (CAL-62 cells) and breast cancer (MDA-MB-231) in nude mice after intravenous administration and was compared with a dye-based labeling method for EV derived from CAL-62 cells. The EVs were successfully labeled with Rluc and visualized by BLI in mice. In vivo distribution of the EVs, as measured by BLI, was consistent with the results of ex vivo organ analysis. EV-CAL-62/Rluc showed strong signals at lung followed by liver, spleen & kidney (P < 0.05). EV-MDA-MB-231/Rluc showed strong signals at liver followed by lung, spleen & kidney (P < 0.05). EV-CAL-62/Rluc and EV-MDA-MB-231/Rluc stayed in animal till day 9 and 3, respectively; showed a differential distribution. Spontaneous EV-CAL-62/Rluc shown distributed mostly to lung followed by liver, spleen & kidney. The new BLI system used to show spontaneous distribution of EV-CAL-62/Rluc in subcutaneous CAL-62/Rluc bearing mice. Dye (DiR)-labeled EV-CAL-62/Rluc showed a different distribution in vivo & ex vivo compared to EV-CAL-62/Rluc. Fluorescent signals were predominately detected in the liver (P < 0.05) and spleen (P < 0.05) regions. The bioluminescent EVs developed in this study may be used for monitoring of EVs in vivo. This novel reporter-imaging approach to visualization of EVs in real time is expected to pave the way for monitoring of EVs in EV-based treatments. PMID:29299117

Immunoaffinity based methods are superior to kits for purification of prostate derived extracellular vesicles from plasma samples.

PubMed

Brett, Sabine I; Lucien, Fabrice; Guo, Charles; Williams, Karla C; Kim, Yohan; Durfee, Paul N; Brinker, C J; Chin, Joseph I; Yang, Jun; Leong, Hon S

2017-05-01

The ability to isolate extracellular vesicles (EVs) such as exosomes or microparticles is an important method that is currently not standardized. While commercially available kits offer purification of EVs from biofluids, such purified EV samples will also contain non-EV entities such as soluble protein and nucleic acids that could confound subsequent experimentation. Ideally, only EVs would be isolated and no soluble protein would be present in the final EV preparation. We compared commercially available EV isolation kits with immunoaffinity purification techniques and evaluated our final EV preparations using atomic force microscopy (AFM) and nanoscale flow cytometry (NFC). AFM is the only modality capable of detecting distinguishing soluble protein from EVs which is important for downstream proteomics approaches. NFC is the only technique capable of quantitating the proportion of target EVs to non-target EVs in the final EV preparation. To determine enrichment of prostate derived EVs relative to non-target MPs, anti-PSMA (Prostate Specific Membrane Antigen) antibodies were used in NFC. Antibody-based immunoaffinity purification generated the highest quality of prostate derived EV preparations due to the lack of protein and RNA present in the samples. All kits produced poor purity EV preparations that failed to deplete the sample of plasma protein. While attractive due to their ease of use, EV purification kits do not provide substantial improvements in isolation of EVs from biofluids such as plasma. Immunoaffinity approaches are more efficient and economical and will also eliminate a significant portion of plasma proteins which is necessary for downstream approaches. © 2017 Wiley Periodicals, Inc.

Rapid Analysis of Energetic and Geo-Materials Using Laser Induced Breakdown Spectroscopy

DTIC Science & Technology

2013-04-01

et al ., Anal Bioanal Chem ( 2006 ) 385, 316. 5. Mohamed, W. T. Y., Prog Phys (2007) 2, 42. 6. Elhassan, A., et al ., Spectrochim Acta B (2008) 63...Anal (2005) 5, 21. 20. Anzano, J. M., et al ., Anal Chim Acta ( 2006 ) 575, 230. 21. Rusak, D. A., et al ., TrAC Trend Anal Chem (1998) 17, 453. 22. Martin...Spectrosc Reviews (2004) 39, 27. 25. Winefordner, J. D., et al ., J Anal Atom Spectrom (2004) 19, 1061. 26. Cremers , D. A., and Radziemski, L. J.,

Evaluation of Quantitative Anti-F1 IgG and Anti-V IgG ELISAs for use as an in Vitro-Based Potency Assay of Plague Vaccine in Mice

DTIC Science & Technology

2008-04-01

Andrews GP, Welkos SL, Friedlander AM, et al. Protection of mice from fatal bubonic and pneu- monic plague by passive immunization with monoclonal...SL, Andrews GP, Adamovicz J, et al. Protection against experimental bubonic and pneumonic plague by a recombinant capsular F1-V antigen fusion...fusion protein as vaccine antigen against bubonic and pneumonic plague . Biotechnol Prog 2005; 21:1490e510.[21] Simpson WJ, Thomas RE, Schwan TG

A TBA approach to thermal transport in the XXZ Heisenberg model

NASA Astrophysics Data System (ADS)

Zotos, X.

2017-10-01

We show that the thermal Drude weight and magnetothermal coefficient of the 1D easy-plane Heisenberg model can be evaluated by an extension of the Bethe ansatz thermodynamics formulation by Takahashi and Suzuki (1972 Prog. Theor. Phys. 48 2187). They have earlier been obtained by the quantum transfer matrix method (Klümper 1999 Z. Phys. B 91 507). Furthermore, this approach can be applied to the study of the far-out of equilibrium energy current generated at the interface between two semi-infinite chains held at different temperatures.

A Glow Discharge Ion Source with Fourier Transform Ion Cyclotron Resonance Mass Spectrometric Detection

DTIC Science & Technology

1991-05-10

Hall, D . Mikrochim. Acta 1987, 1, 275. 26. Harrison, W.W.; Bentz , B.L. Prog. Analyt. Spectrosc. 1988, L19 53. 27. Harrison, W.W.; Barshick, C.M...Innovation, and Applications. ACS Symp. Series; Buchanan, M.V., Ed.; American Chemical Society: Washington, 1987; 359, p 1. 3. Wilkins, C.L.; Chowdhury, A.K...J.L. In Gas Phase Ion CheMistry; Bowers, M.T., Ed.; Academic: New York, 1984; Vol. 3, p 41. 6. Dunbar, R.C. In Gas Phase Ion Chemistry; Bowvers, M.T

A Novel Recombinant Enterovirus Type EV-A89 with Low Epidemic Strength in Xinjiang, China

PubMed Central

Fan, Qin; Zhang, Yong; Hu, Lan; Sun, Qiang; Cui, Hui; Yan, Dongmei; Sikandaner, Huerxidan; Tang, Haishu; Wang, Dongyan; Zhu, Zhen; Zhu, Shuangli; Xu, Wenbo

2015-01-01

Enterovirus A89 (EV-A89) is a novel member of the EV-A species. To date, only one full-length genome sequence (the prototype strain) has been published. Here, we report the molecular identification and genomic characterization of a Chinese EV-A89 strain, KSYPH-TRMH22F/XJ/CHN/2011, isolated in 2011 from a contact of an acute flaccid paralysis (AFP) patient during AFP case surveillance in Xinjiang China. This was the first report of EV-A89 in China. The VP1 coding sequence of this strain demonstrated 93.2% nucleotide and 99.3% amino acid identity with the EV-A89 prototype strain. In the P2 and P3 regions, the Chinese EV-A89 strain demonstrated markedly higher identity than the prototype strains of EV-A76, EV-A90, and EV-A91, indicating that one or more recombination events between EV-A89 and these EV-A types might have occurred. Long-term evolution of these EV types originated from the same ancestor provides the spatial and temporal circumstances for recombination to occur. An antibody sero-prevalence survey against EV-A89 in two Xinjiang prefectures demonstrated low positive rates and low titres of EV-A89 neutralization antibody, suggesting limited range of transmission and exposure to the population. This study provides a solid foundation for further studies on the biological and pathogenic properties of EV-A89. PMID:26685900

Enterovirus infections in hospitals of Ile de France region over 2013.

PubMed

Molet, Lucie; Saloum, Kenda; Marque-Juillet, Stéphanie; Garbarg-Chenon, Antoine; Henquell, Cécile; Schuffenecker, Isabelle; Peigue-Lafeuille, Hélène; Rozenberg, Flore; Mirand, Audrey

2016-01-01

The monitoring and genotyping of Enterovirus (EV) infections can help to associate particular or severe clinical manifestations with specific EV types and to identify the aetiology of infectious outbreaks. To describe the epidemiological features of EV infections diagnosed during the year 2013 in the Greater Paris area (Ile de France). During 2013, 2497 samples taken from 470 patients in 33 hospitals of Ile-de France were tested for EV genome by RT-PCR. EV genotyping was performed by the National Reference Centre (NRC) laboratories. EV infections were retrospectively reviewed by retrieving clinical and genotyping data from the NRC database. Of the 2497 samples, 490 (19.6%) was positive for EV genome detection. These EV infections represented 88.7% and 24.1%, respectively, of all reported regional and national infections. Twenty-seven different genotypes were identified. Echovirus 30 (E-30) accounted for 54.1% of all characterized strains and caused a large outbreak. Four severe neonatal infections were reported, of which two were caused by EV-A71. Respiratory infections involving EV-D68 were observed in two adults. One fatal case of Coxsackievirus A2-associated myocarditis was reported. Monitoring EV infections in combination with EV genotyping via the French EV network characterized the epidemiology of EV infections in the Ile de France region in 2013 and documented severe EV infections associated with EV-A71 or CV-A2. Copyright © 2015 Elsevier B.V. All rights reserved.

A Novel Recombinant Enterovirus Type EV-A89 with Low Epidemic Strength in Xinjiang, China.

PubMed

Fan, Qin; Zhang, Yong; Hu, Lan; Sun, Qiang; Cui, Hui; Yan, Dongmei; Sikandaner, Huerxidan; Tang, Haishu; Wang, Dongyan; Zhu, Zhen; Zhu, Shuangli; Xu, Wenbo

2015-12-21

Enterovirus A89 (EV-A89) is a novel member of the EV-A species. To date, only one full-length genome sequence (the prototype strain) has been published. Here, we report the molecular identification and genomic characterization of a Chinese EV-A89 strain, KSYPH-TRMH22F/XJ/CHN/2011, isolated in 2011 from a contact of an acute flaccid paralysis (AFP) patient during AFP case surveillance in Xinjiang China. This was the first report of EV-A89 in China. The VP1 coding sequence of this strain demonstrated 93.2% nucleotide and 99.3% amino acid identity with the EV-A89 prototype strain. In the P2 and P3 regions, the Chinese EV-A89 strain demonstrated markedly higher identity than the prototype strains of EV-A76, EV-A90, and EV-A91, indicating that one or more recombination events between EV-A89 and these EV-A types might have occurred. Long-term evolution of these EV types originated from the same ancestor provides the spatial and temporal circumstances for recombination to occur. An antibody sero-prevalence survey against EV-A89 in two Xinjiang prefectures demonstrated low positive rates and low titres of EV-A89 neutralization antibody, suggesting limited range of transmission and exposure to the population. This study provides a solid foundation for further studies on the biological and pathogenic properties of EV-A89.

Enterovirus D68 and other enterovirus serotypes identified in South African patients with severe acute respiratory illness, 2009-2011.

PubMed

Hellferscee, Orienka; Treurnicht, Florette K; Tempia, Stefano; Variava, Ebrahim; Dawood, Halima; Kahn, Kathleen; Cohen, Adam L; Pretorius, Marthi; Cohen, Cheryl; Madhi, Shabir A; Venter, Marietjie

2017-05-01

Human enteroviruses (EV) have been associated with severe acute respiratory illness (SARI) in South Africa. We aimed to describe the molecular epidemiology of EV serotypes among patients hospitalized with SARI during 2009-2011. Study samples from patients were tested for the presence of enterovirus using a polymerase chain reaction assay. 8.2% (842/10 260) of SARI cases tested positive for enterovirus; 16% (7/45) were species EV-A, 44% (20/45) EV-B, 18% (8/45) EV-C and 22% (10/45) EV-D. Seventeen different EV serotypes were identified within EV-A to EV-D, of which EV-D68 (22%; 10/45) and Echovirus 3 (11%; 5/45) were the most prevalent. EV-D68 should be monitored in South Africa to assess the emergence of highly pathogenic strains. © 2017 The Authors. Influenza and Other Respiratory Viruses Published by John Wiley & Sons Ltd.

Cellular tropism of human enterovirus D species serotypes EV-94, EV-70, and EV-68 in vitro: implications for pathogenesis.

PubMed

Smura, Teemu; Ylipaasto, Petri; Klemola, Päivi; Kaijalainen, Svetlana; Kyllönen, Lauri; Sordi, Valeria; Piemonti, Lorenzo; Roivainen, Merja

2010-11-01

Enterovirus 94 (EV-94) is an enterovirus serotype described recently which, together with EV-68 and EV-70, forms human enterovirus D species. This study investigates the seroprevalences of these three serotypes and their abilities to infect, replicate, and damage cell types considered to be essential for enterovirus-induced diseases. The cell types studied included human leukocyte cell lines, primary endothelial cells, and pancreatic islets. High prevalence of neutralizing antibodies against EV-68 and EV-94 was found in the Finnish population. The virus strains studied had wide leukocyte tropism. EV-94 and EV-68 were able to produce infectious progeny in leukocyte cell lines with monocytic, granulocytic, T-cell, or B-cell characteristics. EV-94 and EV-70 were capable of infecting primary human umbilical vein endothelial cells, whereas EV-68 had only marginal progeny production and did not induce cytopathic effects in these cells. Intriguingly, EV-94 was able to damage pancreatic islet β-cells, to infect, replicate, and cause necrosis in human pancreatic islets, and to induce proinflammatory and chemoattractive cytokine expression in endothelial cells. These results suggest that HEV-D viruses may be more prevalent than has been thought previously, and they provide in vitro evidence that EV-94 may be a potent pathogen and should be considered a potentially diabetogenic enterovirus type. © 2010 Wiley-Liss, Inc.

Shift of Enterovirus species among children in Cameroon--identification of a new enterovirus, EV-A119.

PubMed

Ayukekbong, James; Kabayiza, Jean-Claude; Lindh, Magnus; Nkuo-Akenji, Theresia; Tah, Ferdinand; Bergström, Tomas; Norder, Helene

2013-09-01

Infections caused by human enteroviruses (EVs) are often asymptomatic or mild, although they may cause more severe illnesses as meningitis and acute flaccid paralysis. EVs have globally posed a threat to children, and outbreaks of aseptic meningitis and hand, foot and mouth disease are frequently reported. To identify EV strains circulating among healthy children in a small community in Limbe, Cameroon two years apart. Species and EV types were obtained by partial 5'UTR-VP4 and VP1 sequencing of RNA from stool samples collected in October 2009 and September 2011 from 150 children in Cameroon. In all, 74 children (49%) were infected with 28 different types of EV. There were 29 (54%) infected children in 2009, and 45 (47%) in 2011. There was a significant difference between detected species of EV, with 15 (47%) children infected with EV-A in 2009, and 22 (71%) with EV-B in 2011 (p=0.0001). In 2009, one child was infected by a divergent EV, which was most similar to EV-A90. Based on the complete VP1 sequence, it was shown to be a new EV designated EV-A119. The current study shows a high heterogeneity of circulating EV types among children in Limbe, Cameroon, and a previously not described shift in predominating EV species. Copyright © 2013 Elsevier B.V. All rights reserved.

Large-scale screening and characterization of enteroviruses and kobuviruses infecting pigs in Vietnam

PubMed Central

Van Dung, Nguyen; Anh, Pham Hong; Van Cuong, Nguyen; Hoa, Ngo Thi; Carrique-Mas, Juan; Hien, Vo Be; Sharp, C.; Rabaa, M.; Berto, A.; Campbell, James; Baker, Stephen; Farrar, Jeremy; Woolhouse, Mark E.; Bryant, Juliet E.; Simmonds, Peter

2016-01-01

A recent survey of pigs in Dong Thap province, Vietnam identified a high frequency of enterovirus species G (EV-G) infection (144/198; 72.7 %). Amongst these was a plethora of EV-G types (EV-G1, EV-G6 and four new types EV-G8–EV-G11). To better characterize the genetic diversity of EV-G and investigate the possible existence of further circulating types, we performed a larger-scale study on 484 pig and 45 farm-bred boar faecal samples collected in 2012 and 2014, respectively. All samples from the previous and current studies were also screened for kobuviruses. The overall EV infection frequency remained extremely high (395/484; 81.6 %), but with comparable detection rates and viral loads between healthy and diarrhoeic pigs; this contrasted with less frequent detection of EV-G in boars (4/45; 8.9 %). EV was most frequently detected in pigs ≤ 14 weeks old (∼95 %) and declined in older pigs. Infections with EV-G1 and EV-G6 were most frequent, whilst less commonly detected types included EV-G3, EV-G4 and EV-G8–EV-G11, and five new types (EV-G12–EV-G16). In contrast, kobuvirus infection frequency was significantly higher in diarrhoeic pigs (40.9 versus 27.6 %; P = 0.01). Kobuviruses also showed contrasting epizootiologies and age associations; a higher prevalence was found in boars (42 %) compared with domestic pigs (29 %), with the highest infection frequency amongst pigs >52 weeks old. Although genetically diverse, all kobuviruses identified belonged to the species Aichivirus C. In summary, this study confirms infection with EV-G was endemic in Vietnamese domestic pigs and exhibits high genetic diversity and extensive inter-type recombination. PMID:26653281

Complete coding regions of the prototypes enterovirus B93 and C95: phylogenetic analyses of the P1 and P3 regions of EV-B and EV-C strains.

PubMed

Junttila, N; Lévêque, N; Magnius, L O; Kabue, J P; Muyembe-Tamfum, J J; Maslin, J; Lina, B; Norder, H

2015-03-01

Complete coding regions were sequenced for two new enterovirus genomes: EV-B93 previously identified by VP1 sequencing, derived from a child with acute flaccid paralysis in the Democratic Republic of Congo; and EV-C95 from a French soldier with acute gastroenteritis in Djibouti. The EV-B93 P1 had more than 30% nucleotide divergence from other EV-B types, with highest similarity to E-15 and EV-B80. The P1 nucleotide sequence of EV-C95 was most similar, 71%, to CV-A21. Complete coding regions for the new enteroviruses were compared with those of 135 EV-B and 176 EV-C strains representing all types available in GenBank. When strains from the same outbreak or strains isolated during the same year in the same geographical region were excluded, 27 of the 58 EV-B, and 16 of the 23 EV-C types were represented by more than one sequence. However, for EV-B the P3 sequences formed three clades mainly according to origin or time of isolation, irrespective of type, while for EV-C the P3 sequences segregated mainly according to disease manifestation, with most strains causing paralysis, including polioviruses, forming one clade, and strains causing respiratory illness forming another. There was no intermixing of types between these two clades, apart from two EV-C96 strains. The EV-B P3 sequences had lower inter-clade and higher intra-clade variability as compared to the EV-C sequences, which may explain why inter-clade recombinations are more frequent in EV-B. Further analysis of more isolates may shed light on the role of recombinations in the evolution of EV-B in geographical context. © 2014 Wiley Periodicals, Inc.

Severe paediatric conditions linked with EV-A71 and EV-D68, France, May to October 2016

PubMed Central

Antona, Denise; Kossorotoff, Manoëlle; Schuffenecker, Isabelle; Mirand, Audrey; Leruez-Ville, Marianne; Bassi, Clément; Aubart, Mélodie; Moulin, Florence; Lévy-Bruhl, Daniel; Henquell, Cécile; Lina, Bruno; Desguerre, Isabelle

2016-01-01

We report 59 cases of severe paediatric conditions linked with enterovirus (EV)-A71 and EV-D68 in France between May and October 2016. Fifty-two children had severe neurological symptoms. EV sequence-based typing for 42 cases revealed EV-A71 in 21 (18 subgenotype C1, detected for the first time in France) and EV-D68 in eight. Clinicians should be encouraged to obtain stool and respiratory specimens from patients presenting with severe neurological disorders for EV detection and characterisation. PMID:27918268

Surveillance and laboratory detection for non-polio enteroviruses in the European Union/European Economic Area, 2016

PubMed Central

Harvala, Heli; Jasir, Aftab; Penttinen, Pasi; Pastore Celentano, Lucia; Greco, Donato; Broberg, Eeva

2017-01-01

Enteroviruses (EVs) cause severe outbreaks of respiratory and neurological disease as illustrated by EV-D68 and EV-A71 outbreaks, respectively. We have mapped European laboratory capacity for identification and characterisation of non-polio EVs to improve preparedness to respond to (re)-emerging EVs linked to severe disease. An online questionnaire on non-polio EV surveillance and laboratory detection was submitted to all 30 European Union (EU)/European Economic Area (EEA) countries. Twenty-nine countries responded; 26 conducted laboratory-based non-polio EV surveillance, and 24 included neurological infections in their surveillance. Eleven countries have established specific surveillance for EV-D68 via sentinel influenza surveillance (n = 7), typing EV-positive respiratory samples (n = 10) and/or acute flaccid paralysis surveillance (n = 5). Of 26 countries performing non-polio EV characterisation/typing, 10 further characterised culture-positive EV isolates, whereas the remainder typed PCR-positive but culture-negative samples. Although 19 countries have introduced sequence-based EV typing, seven still rely entirely on virus isolation. Based on 2015 data, six countries typed over 300 specimens mostly by sequencing, whereas 11 countries characterised under 50 EV-positive samples. EV surveillance activity varied between EU/EEA countries, and did not always specifically target patients with neurological and/or respiratory infections. Introduction of sequence-based typing methods is needed throughout the EU/EEA to enhance laboratory capacity for the detection of EVs. PMID:29162204

Surveillance and laboratory detection for non-polio enteroviruses in the European Union/European Economic Area, 2016.

PubMed

Harvala, Heli; Jasir, Aftab; Penttinen, Pasi; Pastore Celentano, Lucia; Greco, Donato; Broberg, Eeva

2017-11-01

Enteroviruses (EVs) cause severe outbreaks of respiratory and neurological disease as illustrated by EV-D68 and EV-A71 outbreaks, respectively. We have mapped European laboratory capacity for identification and characterisation of non-polio EVs to improve preparedness to respond to (re)-emerging EVs linked to severe disease. An online questionnaire on non-polio EV surveillance and laboratory detection was submitted to all 30 European Union (EU)/European Economic Area (EEA) countries. Twenty-nine countries responded; 26 conducted laboratory-based non-polio EV surveillance, and 24 included neurological infections in their surveillance. Eleven countries have established specific surveillance for EV-D68 via sentinel influenza surveillance (n = 7), typing EV-positive respiratory samples (n = 10) and/or acute flaccid paralysis surveillance (n = 5). Of 26 countries performing non-polio EV characterisation/typing, 10 further characterised culture-positive EV isolates, whereas the remainder typed PCR-positive but culture-negative samples. Although 19 countries have introduced sequence-based EV typing, seven still rely entirely on virus isolation. Based on 2015 data, six countries typed over 300 specimens mostly by sequencing, whereas 11 countries characterised under 50 EV-positive samples. EV surveillance activity varied between EU/EEA countries, and did not always specifically target patients with neurological and/or respiratory infections. Introduction of sequence-based typing methods is needed throughout the EU/EEA to enhance laboratory capacity for the detection of EVs.

Update on Controls for Isolation and Quantification Methodology of Extracellular Vesicles Derived from Adipose Tissue Mesenchymal Stem Cells

PubMed Central

Franquesa, Marcella; Hoogduijn, Martin J.; Ripoll, Elia; Luk, Franka; Salih, Mahdi; Betjes, Michiel G. H.; Torras, Juan; Baan, Carla C.; Grinyó, Josep M.; Merino, Ana Maria

2014-01-01

The research field on extracellular vesicles (EV) has rapidly expanded in recent years due to the therapeutic potential of EV. Adipose tissue human mesenchymal stem cells (ASC) may be a suitable source for therapeutic EV. A major limitation in the field is the lack of standardization of the challenging techniques to isolate and characterize EV. The aim of our study was to incorporate new controls for the detection and quantification of EV derived from ASC and to analyze the applicability and limitations of the available techniques. ASC were cultured in medium supplemented with 5% of vesicles-free fetal bovine serum. The EV were isolated from conditioned medium by differential centrifugation with size filtration (0.2 μm). As a control, non-conditioned culture medium was used (control medium). To detect EV, electron microscopy, conventional flow cytometry, and western blot were used. The quantification of the EV was by total protein quantification, ExoELISA immunoassay, and Nanosight. Cytokines and growth factors in the EV samples were measured by multiplex bead array kit. The EV were detected by electron microscope. Total protein measurement was not useful to quantify EV as the control medium showed similar protein contents as the EV samples. The ExoELISA kits had technical troubles and it was not possible to quantify the concentration of exosomes in the samples. The use of Nanosight enabled quantification and size determination of the EV. It is, however, not possible to distinguish protein aggregates from EV with this method. The technologies for quantification and characterization of the EV need to be improved. In addition, we detected protein contaminants in the EV samples, which make it difficult to determine the real effect of EV in experimental models. It will be crucial in the future to optimize design novel methods for purification and characterization of EV. PMID:25374572

High levels of circulating extracellular vesicles with altered expression and function during pregnancy.

PubMed

Nardi, Fabiola da Silva; Michelon, Tatiana Ferreira; Neumann, Jorge; Manvailer, Luis Felipe Santos; Wagner, Bettina; Horn, Peter A; Bicalho, Maria da Graça; Rebmann, Vera

2016-07-01

Extracellular vesicles (EVs) are widely considered important modulators of cell-cell communication and may interact with target cells locally and on a systemic level. Several studies had shown that circulating EVs' levels are increased during pregnancy. However, EVs characteristics, composition and biological functions in pregnancy still need to be clarified. This study aims to determine if circulating EVs during pregnancy are modified regarding levels, markers and cytokine profile as well as their reactivity towards peripheral blood cells. 26 pregnant women (PW) being in the second gestational trimester and 59 non-pregnant women (NPW) were investigated. EVs enrichment was performed by ExoQuick™ or ultracentrifugation; nanoparticle tracking analysis, SDS-PAGE followed by Western Blotting and densitometry, and IFN-γ, IL-10 and TGF-β1 ELISA for EVs characterization; imaging flow cytometry to analyze EVs' uptake by peripheral blood cells and flow cytometry were performed to analyze EVs function regarding induction of caspase-3 activity. Circulating EVs' levels were increased during pregnancy [26.9×10(6)EVs/ml (range: 6.4-46.3); p=0.003] vs NPW [18.9×10(6)EVs/ml (range: 2.5-61.3)]. Importantly, the immunosuppressive TGF-β1 and IL-10 cytokine cargo were increased in EVs of PW even after normalization to 1 million EVs [TGF-β1: 0.25pg/10(6)EVs (range: 0.0-2.0); p<0.0001] and [IL-10: 0.21pg/10(6)EVs (range: 0.0-16.8); p=0.006] vs NPW. Although EVs derived from non-pregnant and pregnant women were taken up by NK cells, the latter exclusively enhanced the caspase-3 activity in CD56(dim) NK cells (8.2±0.9; p=0.02). The qualitative and quantitative pregnancy-related alterations of circulating EVs provide first hints for an immune modulating role of circulating EVs during pregnancy. Copyright © 2016 Elsevier GmbH. All rights reserved.

Extracellular vesicles from mice with alcoholic liver disease carry a distinct protein cargo and induce macrophage activation through heat shock protein 90.

PubMed

Saha, Banishree; Momen-Heravi, Fatemeh; Furi, Istvan; Kodys, Karen; Catalano, Donna; Gangopadhyay, Anwesha; Haraszti, Reka; Satishchandran, Abhishek; Iracheta-Vellve, Arvin; Adejumo, Adeyinka; Shaffer, Scott A; Szabo, Gyongyi

2018-05-01

A salient feature of alcoholic liver disease (ALD) is Kupffer cell (KC) activation and recruitment of inflammatory monocytes and macrophages (MØs). These key cellular events of ALD pathogenesis may be mediated by extracellular vesicles (EVs). EVs transfer biomaterials, including proteins and microRNAs, and have recently emerged as important effectors of intercellular communication. We hypothesized that circulating EVs from mice with ALD have a protein cargo characteristic of the disease and mediate biological effects by activating immune cells. The total number of circulating EVs was increased in mice with ALD compared to pair-fed controls. Mass spectrometric analysis of circulating EVs revealed a distinct signature for proteins involved in inflammatory responses, cellular development, and cellular movement between ALD EVs and control EVs. We also identified uniquely important proteins in ALD EVs that were not present in control EVs. When ALD EVs were injected intravenously into alcohol-naive mice, we found evidence of uptake of ALD EVs in recipient livers in hepatocytes and MØs. Hepatocytes isolated from mice after transfer of ALD EVs, but not control EVs, showed increased monocyte chemoattractant protein 1 mRNA and protein expression, suggesting a biological effect of ALD EVs. Compared to control EV recipient mice, ALD EV recipient mice had increased numbers of F4/80 hi cluster of differentiation 11b (CD11b) lo KCs and increased percentages of tumor necrosis factor alpha-positive/interleukin 12/23-positive (inflammatory/M1) KCs and infiltrating monocytes (F4/80 int CD11b hi ), while the percentage of CD206 + CD163 + (anti-inflammatory/M2) KCs was decreased. In vitro, ALD EVs increased tumor necrosis factor alpha and interleukin-1β production in MØs and reduced CD163 and CD206 expression. We identified heat shock protein 90 in ALD EVs as the mediator of ALD-EV-induced MØ activation. Our study indicates a specific protein signature of ALD EVs and demonstrates a functional role of circulating EVs containing heat shock protein 90 in mediating KC/MØ activation in the liver. (Hepatology 2018;67:1986-2000). © 2017 by the American Association for the Study of Liver Diseases.

Severe paediatric conditions linked with EV-A71 and EV-D68, France, May to October 2016.

PubMed

Antona, Denise; Kossorotoff, Manoëlle; Schuffenecker, Isabelle; Mirand, Audrey; Leruez-Ville, Marianne; Bassi, Clément; Aubart, Mélodie; Moulin, Florence; Lévy-Bruhl, Daniel; Henquell, Cécile; Lina, Bruno; Desguerre, Isabelle

2016-11-17

We report 59 cases of severe paediatric conditions linked with enterovirus (EV)-A71 and EV-D68 in France between May and October 2016. Fifty-two children had severe neurological symptoms. EV sequence-based typing for 42 cases revealed EV-A71 in 21 (18 subgenotype C1, detected for the first time in France) and EV-D68 in eight. Clinicians should be encouraged to obtain stool and respiratory specimens from patients presenting with severe neurological disorders for EV detection and characterisation. This article is copyright of The Authors, 2016.

An Update on in Vivo Imaging of Extracellular Vesicles as Drug Delivery Vehicles

PubMed Central

Gangadaran, Prakash; Hong, Chae Moon; Ahn, Byeong-Cheol

2018-01-01

Extracellular vesicles (EVs) are currently being considered as promising drug delivery vehicles. EVs are naturally occurring vesicles that exhibit many characteristics favorable to serve as drug delivery vehicles. In addition, EVs have inherent properties for treatment of cancers and other diseases. For research and clinical translation of use of EVs as drug delivery vehicles, in vivo tracking of EVs is essential. The latest molecular imaging techniques enable the tracking of EVs in living animals. However, each molecular imaging technique has its certain advantages and limitations for the in vivo imaging of EVs; therefore, understanding the molecular imaging techniques is essential to select the most appropriate imaging technology to achieve the desired imaging goal. In this review, we summarize the characteristics of EVs as drug delivery vehicles and the molecular imaging techniques used in visualizing and monitoring EVs in in vivo environments. Furthermore, we provide a perceptual vision of EVs as drug delivery vehicles and in vivo monitoring of EVs using molecular imaging technologies. PMID:29541030

PEGylated and targeted extracellular vesicles display enhanced cell specificity and circulation time.

PubMed

Kooijmans, S A A; Fliervoet, L A L; van der Meel, R; Fens, M H A M; Heijnen, H F G; van Bergen En Henegouwen, P M P; Vader, P; Schiffelers, R M

2016-02-28

Extracellular vesicles (EVs) are increasingly being recognized as candidate drug delivery systems due to their ability to functionally transfer biological cargo between cells. However, the therapeutic applicability of EVs may be limited due to a lack of cell-targeting specificity and rapid clearance of exogenous EVs from the circulation. In order to improve EV characteristics for drug delivery to tumor cells, we have developed a novel method for decorating EVs with targeting ligands conjugated to polyethylene glycol (PEG). Nanobodies specific for the epidermal growth factor receptor (EGFR) were conjugated to phospholipid (DMPE)-PEG derivatives to prepare nanobody-PEG-micelles. When micelles were mixed with EVs derived from Neuro2A cells or platelets, a temperature-dependent transfer of nanobody-PEG-lipids to the EV membranes was observed, indicative of a 'post-insertion' mechanism. This process did not affect EV morphology, size distribution, or protein composition. After introduction of PEG-conjugated control nanobodies to EVs, cellular binding was compromised due to the shielding properties of PEG. However, specific binding to EGFR-overexpressing tumor cells was dramatically increased when EGFR-specific nanobodies were employed. Moreover, whereas unmodified EVs were rapidly cleared from the circulation within 10min after intravenous injection in mice, EVs modified with nanobody-PEG-lipids were still detectable in plasma for longer than 60min post-injection. In conclusion, we propose post-insertion as a novel technique to confer targeting capacity to isolated EVs, circumventing the requirement to modify EV-secreting cells. Importantly, insertion of ligand-conjugated PEG-derivatized phospholipids in EV membranes equips EVs with improved cell specificity and prolonged circulation times, potentially increasing EV accumulation in targeted tissues and improving cargo delivery. Copyright © 2015. Published by Elsevier B.V.

Extracellular vesicles for liquid biopsy in prostate cancer: where are we and where are we headed?

PubMed

Minciacchi, V R; Zijlstra, A; Rubin, M A; Di Vizio, D

2017-09-01

Extracellular vesicles (EVs) are a heterogeneous class of lipid bound particles shed by any cell in the body in physiological and pathological conditions. EVs play critical functions in intercellular communication. EVs can actively travel in intercellular matrices and eventually reach the circulation. They can also be released directly in biological fluids where they appear to be stable. Because the molecular content of EVs reflects the composition of the cell of origin, they have recently emerged as a promising source of biomarkers in a number of diseases. EV analysis is particularly attractive in cancer patients that frequently present with increased numbers of circulating EVs. We sought to review the current literature on the molecular profile of prostate cancer-derived EVs in model systems and patient biological fluids in an attempt to draw some practical and universal conclusions on the use of EVs as a tool for liquid biopsy in clinical specimens. We discuss advantages and limitations of EV-based liquid biopsy approaches summarizing salient studies on protein, DNA and RNA. Several candidate biomarkers have been identified so far but these results are difficult to apply to the clinic. However, the field is rapidly moving toward the implementation of novel tools to isolate cancer-specific EVs that are free of benign EVs and extra-vesicular contaminants. This can be achieved by identifying markers that are exquisitely present in tumor cell-derived EVs. An important contribution might also derive from a better understanding of EV types that may play specific functions in tumor progression and that may be a source of cancer-specific markers. EV analysis holds strong promises for the development of non-invasive biomarkers in patients with prostate cancer. Implementation of modern methods for EV isolation and characterization will enable to interrogate circulating EVs in vivo.

Prediction of esophageal variceal bleeding in B-viral liver cirrhosis using the P2/MS noninvasive index based on complete blood counts.

PubMed

Kim, Beom Kyung; Ahn, Sang Hoon; Han, Kwang-Hyub; Park, Jun Yong; Han, Min Seok; Jo, Jung Hyun; Kim, Ja Kyung; Lee, Kwan Sik; Chon, Chae Yoon; Kim, Do Young

2012-01-01

Periodic endoscopy for esophageal varices (EVs) and prophylactic treatment of high-risk EVs, i.e., medium/large EVs, small EVs with the red-color sign or decompensation, are recommended in cirrhotic patients. We assessed the cumulative risks for future EV bleeding using the following simple P2/MS index: (platelet count)2/[monocyte fraction (%) × segmented neutrophil fraction (%)]. We enrolled 475 consecutive B-viral cirrhosis patients for 4 years, none of whom experienced EV bleeding. All underwent laboratory work-ups, endoscopy and ultrasonography. Those with EV bleeding took a nonselective β-blocker as prophylaxis. The major endpoint was the first occurrence of EV bleeding, analyzed using the Kaplan-Meier and Cox regression methods. Among patients with EV bleeding (n = 131), 25 experienced their first EV bleeding during follow-up. To differentiate the risk for EV bleeding, we divided them into two subgroups according to their P2/MS value (subgroup 1: P2/MS ≥9 and subgroup 2: P2/MS <9). The risk was significantly higher in subgroup 2 (p = 0.029). From multivariate analysis, a lower P2/MS (p = 0.040) remained a significant predictor for EV bleeding along with large varix size (p = 0.015), red-color sign (p = 0.041) and Child-Pugh classification B/C (p = 0.001). In subgroup 1, the risk for EV bleeding was similar to that of patients with low-risk EVs (p = 0.164). The P2/MS is a reliable predictor for the risk of EV bleeding among patients with EV bleeding. According to risk stratification, different prophylactic treatments should be considered for the subgroup with a P2/MS <9. Copyright © 2012 S. Karger AG, Basel.

Uncooled EuSbTe3 photodetector highly sensitive from ultraviolet to terahertz frequencies

NASA Astrophysics Data System (ADS)

Niu, Ying Y.; Wu, Dong; Su, Yu Q.; Zhu, Hai; Wang, Biao; Wang, Ying X.; Zhao, Zi R.; Zheng, Ping; Niu, Jia S.; Zhou, Hui B.; Wei, Jian; Wang, Nan L.

2018-01-01

Light probe from Uv to THz is critical in photoelectronics and has great applications ranging from imaging, communication to medicine (Woodward et al 2002 Phys. Med. Biol. 47 3853-63 Pospischil et al 2013 Nat. Photon. 7 892-6 Martyniuk and Rogalski 2003 Prog. Quantum Electron. 27 59-210). However, the room temperature ultrabroadband photodetection across visible down to far-infrared is still challenging. The challenging arises mainly from the lack of suitable photoactive materials. Because that conventional semiconductors, such as silicon, have their photosensitive properties cut off by the bandgap and are transparent to spectrum at long-wavelength infrared side (Ciupa and Rogalski 1997 Opto-Electron. Rev. 5 257-66 Tonouchi 2007 Nat. Photon. 1 97-105 Sizov and Rogalski 2010 Prog. Quantum Electron. 34 278-347 Kinch 2000 J. Electron. Mater. 29 809-17). Comparatively, the dielectrics with very narrow band-gap but maintain the semiconductor-like electrical conduction would have priorities for ultrabroadband photodetection. Here we report on EuSbTe3 is highly sensitive from ultraviolet directly to terahertz (THz) at room temperature. High photoresponsivities 1-8 A W-1 reached in our prototype EuSbTe3 detectors with low noise equivalent power (NEP) recorded, for instances ~150 pW · Hz-1/2 (at λ  =  532 nm) and ~0.6 nW · Hz-1/2 (at λ  =  118.8 µm) respectively. Our results demonstrate a promising system with direct photosensitivity extending well into THz regime at room temperature, shed new light on exploring more sophisticated multi-band photoelectronics.

Longitudinal Changes of Fixation Location and Stability within 12 Months in Stargardt Disease: ProgStar Report No. 12.

PubMed

Schönbach, Etienne M; Strauss, Rupert W; Kong, Xiangrong; Muñoz, Beatriz; Ibrahim, Mohamed A; Sunness, Janet S; Birch, David G; Hahn, Gesa-Astrid; Nasser, Fadi; Zrenner, Eberhart; Sadda, SriniVas R; West, Sheila K; Scholl, Hendrik P N

2018-06-08

To investigate the natural history of Stargardt disease (STGD1) using fixation location and fixation stability. Multicenter, international, prospective cohort study. Fixation testing was performed using the Nidek MP-1 microperimeter as part of the prospective, multicenter, natural history study on the Progression of Stargardt disease (ProgStar). A total of 238 patients with ABCA4-related STGD1 were enrolled at baseline (bilateral enrollment in 86.6 %) and underwent repeat testing at month 6 and 12. Outcome measures included the distance of the preferred retinal locus from the fovea (PRL) and the bivariate contour ellipse area (BCEA). After 12 months of follow-up, the change in the eccentricity of the PRL from the anatomical fovea was -0.0014 deg (95 % CI, - 0.27deg - 0.27 deg; p = 0.99). The deterioration in the stability of fixation as expressed by a larger BCEA encompassing 1 SD of all fixation points was 1.21 deg 2 (95 % CI, -1.23 deg 2 , 3.65 deg 2 ; p = 0.33). Eyes with increases and decreases in PRL eccentricity and/or BCEA values were observed. Our observations point to the complexity of fixation parameters. The association of increasingly eccentric and unstable fixation with longer disease duration that is typically found in cross-sectional studies may be countered within individual patients by poorly understood processes like neuronal adaptation. Nevertheless, fixation parameters may serve as useful secondary outcome parameters in selected cases and for counseling patients to explain changes to their visual functionality. Copyright © 2018 Elsevier Inc. All rights reserved.

Comfort and exertion while using filtering facepiece respirators with exhalation valve and an active venting system among male military personnel.

PubMed

Seng, Melvin; Wee, Liang En; Zhao, Xiahong; Cook, Alex R; Chia, Sin Eng; Lee, Vernon J

2017-07-06

This study aimed to determine if disposable filtering facepiece respirators (FFRs), with exhalation valve (EV) and a novel active venting system (AVS), provided greater perceived comfort and exertion when compared to standard N95 FFRs without these features among male military personnel performing prolonged essential outdoor duties. We used a randomised open-label controlled crossover study design to compare three FFR options: (a) standard FFR; (b) FFR with EV; and (c) FFR with EV+AVS. Male military personnel aged between 18 and 20 years completed a questionnaire at the beginning (baseline), after two hours of standardised non-strenuous outdoor duty and after 12 hours of duty divided into two-hour work-rest cycles. Participants rated the degree of discomfort, exertion and symptoms using a five-point Likert scale. The association between outcomes and the types of FFR was assessed using a multivariate ordered probit mixed-effects model. For a majority of the symptoms, study participants rated FFR with EV and FFR with EV+AVS with significantly better scores than standard FFR. Both FFR with EV and FFR with EV+AVS had significantly less discomfort (FFR with EV+AVS: 91.1%; FFR with EV: 57.6%) and exertion (FFR with EV+AVS: 83.5%; FFR with EV: 34.4%) than standard FFR. FFR with EV+AVS also had significantly better scores for exertion (53.4%) and comfort (39.4%) when compared to FFR with EV. Usage of FFR with EV+AVS resulted in significantly reduced symptoms, discomfort and exertion when compared to FFR with EV and standard FFR.

A platform for actively loading cargo RNA to elucidate limiting steps in EV-mediated delivery.

PubMed

Hung, Michelle E; Leonard, Joshua N

2016-01-01

Extracellular vesicles (EVs) mediate intercellular communication through transfer of RNA and protein between cells. Thus, understanding how cargo molecules are loaded and delivered by EVs is of central importance for elucidating the biological roles of EVs and developing EV-based therapeutics. While some motifs modulating the loading of biomolecular cargo into EVs have been elucidated, the general rules governing cargo loading and delivery remain poorly understood. To investigate how general biophysical properties impact loading and delivery of RNA by EVs, we developed a platform for actively loading engineered cargo RNAs into EVs. In our system, the MS2 bacteriophage coat protein was fused to EV-associated proteins, and the cognate MS2 stem loop was engineered into cargo RNAs. Using this Targeted and Modular EV Loading (TAMEL) approach, we identified a configuration that substantially enhanced cargo RNA loading (up to 6-fold) into EVs. When applied to vesicles expressing the vesicular stomatitis virus glycoprotein (VSVG) - gesicles - we observed a 40-fold enrichment in cargo RNA loading. While active loading of mRNA-length (>1.5 kb) cargo molecules was possible, active loading was much more efficient for smaller (~0.5 kb) RNA molecules. We next leveraged the TAMEL platform to elucidate the limiting steps in EV-mediated delivery of mRNA and protein to prostate cancer cells, as a model system. Overall, most cargo was rapidly degraded in recipient cells, despite high EV-loading efficiencies and substantial EV uptake by recipient cells. While gesicles were efficiently internalized via a VSVG-mediated mechanism, most cargo molecules were rapidly degraded. Thus, in this model system, inefficient endosomal fusion or escape likely represents a limiting barrier to EV-mediated transfer. Altogether, the TAMEL platform enabled a comparative analysis elucidating a key opportunity for enhancing EV-mediated delivery to prostate cancer cells, and this technology should be of general utility for investigations and applications of EV-mediated transfer in other systems.

Handling and storage of human body fluids for analysis of extracellular vesicles

PubMed Central

Yuana, Yuana; Böing, Anita N.; Grootemaat, Anita E.; van der Pol, Edwin; Hau, Chi M.; Cizmar, Petr; Buhr, Egbert; Sturk, Auguste; Nieuwland, Rienk

2015-01-01

Because procedures of handling and storage of body fluids affect numbers and composition of extracellular vesicles (EVs), standardization is important to ensure reliable and comparable measurements of EVs in a clinical environment. We aimed to develop standard protocols for handling and storage of human body fluids for EV analysis. Conditions such as centrifugation, single freeze–thaw cycle, effect of time delay between blood collection and plasma preparation and storage were investigated. Plasma is the most commonly studied body fluid in EV research. We mainly focused on EVs originating from platelets and erythrocytes and investigated the behaviour of these 2 types of EVs independently as well as in plasma samples of healthy subjects. EVs in urine and saliva were also studied for comparison. All samples were analysed simultaneously before and after freeze–thawing by resistive pulse sensing, nanoparticle tracking analysis, conventional flow cytometry (FCM) and transmission (scanning) electron microscopy. Our main finding is that the effect of centrifugation markedly depends on the cellular origin of EVs. Whereas erythrocyte EVs remain present as single EVs after centrifugation, platelet EVs form aggregates, which affect their measured concentration in plasma. Single erythrocyte and platelet EVs are present mainly in the range of 100–200 nm, far below the lower limit of what can be measured by conventional FCM. Furthermore, the effects of single freeze–thaw cycle, time delay between blood collection and plasma preparation up to 1 hour and storage up to 1 year are insignificant (p>0.05) on the measured concentration and diameter of EVs from erythrocyte and platelet concentrates and EVs in plasma, urine and saliva. In conclusion, in standard protocols for EV studies, centrifugation to isolate EVs from collected body fluids should be avoided. Freezing and storage of collected body fluids, albeit their insignificant effects, should be performed identically for comparative EV studies and to create reliable biorepositories. PMID:26563735

Handling and storage of human body fluids for analysis of extracellular vesicles.

PubMed

Yuana, Yuana; Böing, Anita N; Grootemaat, Anita E; van der Pol, Edwin; Hau, Chi M; Cizmar, Petr; Buhr, Egbert; Sturk, Auguste; Nieuwland, Rienk

2015-01-01

Because procedures of handling and storage of body fluids affect numbers and composition of extracellular vesicles (EVs), standardization is important to ensure reliable and comparable measurements of EVs in a clinical environment. We aimed to develop standard protocols for handling and storage of human body fluids for EV analysis. Conditions such as centrifugation, single freeze-thaw cycle, effect of time delay between blood collection and plasma preparation and storage were investigated. Plasma is the most commonly studied body fluid in EV research. We mainly focused on EVs originating from platelets and erythrocytes and investigated the behaviour of these 2 types of EVs independently as well as in plasma samples of healthy subjects. EVs in urine and saliva were also studied for comparison. All samples were analysed simultaneously before and after freeze-thawing by resistive pulse sensing, nanoparticle tracking analysis, conventional flow cytometry (FCM) and transmission (scanning) electron microscopy. Our main finding is that the effect of centrifugation markedly depends on the cellular origin of EVs. Whereas erythrocyte EVs remain present as single EVs after centrifugation, platelet EVs form aggregates, which affect their measured concentration in plasma. Single erythrocyte and platelet EVs are present mainly in the range of 100-200 nm, far below the lower limit of what can be measured by conventional FCM. Furthermore, the effects of single freeze-thaw cycle, time delay between blood collection and plasma preparation up to 1 hour and storage up to 1 year are insignificant (p>0.05) on the measured concentration and diameter of EVs from erythrocyte and platelet concentrates and EVs in plasma, urine and saliva. In conclusion, in standard protocols for EV studies, centrifugation to isolate EVs from collected body fluids should be avoided. Freezing and storage of collected body fluids, albeit their insignificant effects, should be performed identically for comparative EV studies and to create reliable biorepositories.

Serological detection and analysis of anti-VP1 responses against various enteroviruses (EV) (EV-A, EV-B and EV-C) in Chinese individuals.

PubMed

Gao, Caixia; Ding, Yingying; Zhou, Peng; Feng, Jiaojiao; Qian, Baohua; Lin, Ziyu; Wang, Lili; Wang, Jinhong; Zhao, Chunyan; Li, Xiangyu; Cao, Mingmei; Peng, Heng; Rui, Bing; Pan, Wei

2016-02-26

The overall serological prevalence of EV infections based on ELISA remains unknown. In the present study, the antibody responses against VP1 of the EV-A species (enterovirus 71 (EV71), Coxsackievirus A16 (CA16), Coxsackievirus A5 (CA5) and Coxsackievirus A6 (CA6)), of the EV-B species (Coxsackievirus B3 (CB3)), and of the EV-C species (Poliovirus 1 (PV1)) were detected and analyzed by a NEIBM (novel evolved immunoglobulin-binding molecule)-based ELISA in Shanghai blood donors. The serological prevalence of anti-CB3 VP1 antibodies was demonstrated to show the highest level, with anti-PV1 VP1 antibodies at the second highest level, and anti-CA5, CA6, CA16 and EV71 VP1 antibodies at a comparatively low level. All reactions were significantly correlated at different levels, which were approximately proportional to their sequence similarities. Antibody responses against EV71 VP1 showed obvious differences with responses against other EV-A viruses. Obvious differences in antibody responses between August 2013 and May 2014 were revealed. These findings are the first to describe the detailed information of the serological prevalence of human antibody responses against the VP1 of EV-A, B and C viruses, and could be helpful for understanding of the ubiquity of EV infections and for identifying an effective approach for seroepidemiological surveillance based on ELISA.

[Step Fisher discriminant analysis on severe clinical features of hand foot and mouth disease between enterovirus (EV) 71 and other EV].

PubMed

Ruan, Feng; Tan, Ai-jun; Zhang, Xue-bao; Chen, Xue-qin; Xiao, Song-jian; Ye, Zhong-wen; Wang, Song

2011-07-01

To compare the clinical features of severe hand foot and mouth disease between enterovirus (EV) 71 and other EV to find specific diagnosis index of EV71 severe hand foot and mouth disease. Case definition were adopted from national guideline of hand foot and mouth disease diagnose (Version 2010). Clinical data of severe hand foot and mouth disease came from case history and contents of questionnaire would include the ones between the time of onset and diagnoses being made. EV and EV71, Cox A16 nucleic acid tested were by RT-PCR in stool samples. Clinical features of severe hand foot and mouth disease between EV71 and other EV were compare. There appeared statistical differences between neurologic symptoms such as tremor, myoclonic jerk, listlessness, convulsion and white blood cell counts in CSF (P < 0.05). Results from the step Fisher discriminant analysis showed only tremor and white blood cell had an increase in CSF, with statistically significant differences. The discriminant equation of EV71 was Y = 3.059X(1) + 3.83X(5) - 2.742 and the equation of other EV was Y = 1.634X(1) + 1.623X(5) - 1.693. The specificity of EV71 was 91% and the specificity of other EV was 40%. The increase of clinical features of tremor and white blood cell in CSF could be used as diagnosis index of severe EV71.

Deep-level transient spectroscopy of Pd-H complexes in silicon

NASA Astrophysics Data System (ADS)

Sachse, J.-U.; Weber, J.; Lemke, H.

2000-01-01

The interaction of atomic hydrogen with substitutional palladium impurities is studied in n- and p-type Si by deep-level transient spectroscopy. After wet-chemical etching, we determine seven different electrically active and at least one passive palladium hydrogen complex. The levels belong to Pd complexes with different number of hydrogen atoms. The PdH1 complex exhibits one level E(200) at EC-0.43 eV. PdH2 has two levels E(60) at EC-0.10 eV and H(280) at EV+0.55 eV. Four levels are assigned to the PdH3 complex E(160) at EC-0.29 eV, H(140) at EV+0.23 eV, H(55) at EV+0.08 eV, and H(45) at EV+0.07 eV. An electrically passive complex is associated with a PdH4 complex. There is great similarity with the correspondent complexes in Pt-doped Si. Annealing above 650 K destroys all hydrogen related complexes and restores the original substitutional Pd concentration.

Metabolomic Profiling of Extracellular Vesicles and Alternative Normalization Methods Reveal Enriched Metabolites and Strategies to Study Prostate Cancer-Related Changes

PubMed Central

Puhka, Maija; Takatalo, Maarit; Nordberg, Maria-Elisa; Valkonen, Sami; Nandania, Jatin; Aatonen, Maria; Yliperttula, Marjo; Laitinen, Saara; Velagapudi, Vidya; Mirtti, Tuomas; Kallioniemi, Olli; Rannikko, Antti; Siljander, Pia R-M; af Hällström, Taija Maria

2017-01-01

Body fluids are a rich source of extracellular vesicles (EVs), which carry cargo derived from the secreting cells. So far, biomarkers for pathological conditions have been mainly searched from their protein, (mi)RNA, DNA and lipid cargo. Here, we explored the small molecule metabolites from urinary and platelet EVs relative to their matched source samples. As a proof-of-concept study of intra-EV metabolites, we compared alternative normalization methods to profile urinary EVs from prostate cancer patients before and after prostatectomy and from healthy controls. Methods: We employed targeted ultra-performance liquid chromatography-tandem mass spectrometry to profile over 100 metabolites in the isolated EVs, original urine samples and platelets. We determined the enrichment of the metabolites in the EVs and analyzed their subcellular origin, pathways and relevant enzymes or transporters through data base searches. EV- and urine-derived factors and ratios between metabolites were tested for normalization of the metabolomics data. Results: Approximately 1 x 1010 EVs were sufficient for detection of metabolite profiles from EVs. The profiles of the urinary and platelet EVs overlapped with each other and with those of the source materials, but they also contained unique metabolites. The EVs enriched a selection of cytosolic metabolites including members from the nucleotide and spermidine pathways, which linked to a number of EV-resident enzymes or transporters. Analysis of the urinary EVs from the patients indicated that the levels of glucuronate, D-ribose 5-phosphate and isobutyryl-L-carnitine were 2-26-fold lower in all pre-prostatectomy samples compared to the healthy control and post-prostatectomy samples (p < 0.05). These changes were only detected from EVs by normalization to EV-derived factors or with metabolite ratios, and not from the original urine samples. Conclusions: Our results suggest that metabolite analysis of EVs from different samples is feasible using a high-throughput platform and relatively small amount of sample material. With the knowledge about the specific enrichment of metabolites and normalization methods, EV metabolomics could be used to gain novel biomarker data not revealed by the analysis of the original EV source materials. PMID:29109780

Abundance of enterovirus C in RD-L20B cell culture-negative stool samples from acute flaccid paralysis cases in Nigeria is geographically defined.

PubMed

Donbraye, Emmanuel; Olasunkanmi, Oluwatayo Israel; Opabode, Babatunde Ayoola; Ishola, Temitayo Rachael; Faleye, Temitope Oluwasegun Cephas; Adewumi, Olubusuyi Moses; Adeniji, Johnson Adekunle

2018-06-01

We recently showed that enteroviruses (EVs) andenterovirus species C (EV-C) in particular were abundant in faecal samples from children who had been diagnosed with acute flaccid paralysis (AFP) in Nigeria but declared to be EV-free by the RD-L20B cell culture-based algorithm. In this study, we investigated whether this observed preponderance of EVs (and EV-Cs) in such samples varies by geographical region. One hundred and eight samples (i.e. 54 paired stool suspensions from 54 AFP cases) that had previously been confirmed to be negative for EVs by the WHO-recommended RD-L20B cell culture-based algorithm were analysed. The 108 samples were made into 54 pools (27 each from North-West and South-South Nigeria). All were subjected to RNA extraction, cDNA synthesis and the WHO-recommended semi-nested PCR assay and its modifications. All of the amplicons were sequenced, and the enteroviruses identified, using the enterovirus genotyping tool and phylogenetic analysis. EVs were detected in 16 (29.63 %) of the 54 samples that were screened and successfully identified in 14 (25.93 %). Of these, 10 were from North-West and 4 were from South-South Nigeria. One (7.14 %), 2 (14.29 %) and 11 (78.57 %) of the strains detected were EV-A, EV-B and EV-C, respectively. The 10 strains from North-West Nigeria included 7 EV types, namely CV-A10, E29, CV-A13, CV-A17, CV-A19, CV-A24 and EV-C99. The four EV types recovered from South-South Nigeria were E31, CV-A1, EV-C99 and EV-C116. The results of this study showed that the presence of EVs and consequently EV-Cs in AFP samples declared to be EV-free by the RD-L20B cell culture-based algorithm varies by geographical region in Nigeria.

MSC secretes at least 3 EV types each with a unique permutation of membrane lipid, protein and RNA.

PubMed

Lai, Ruenn Chai; Tan, Soon Sim; Yeo, Ronne Wee Yeh; Choo, Andre Boon Hwa; Reiner, Agnes T; Su, Yan; Shen, Yang; Fu, Zhiyan; Alexander, Lezhava; Sze, Siu Kwan; Lim, Sai Kiang

2016-01-01

Mesenchymal stem cell (MSC), a widely used adult stem cell candidate for regenerative medicine, has been shown to exert some of its therapeutic effects through the secretion of extracellular vesicles (EVs). These homogenously sized EVs of 100-150 ηm exhibited many exosome-like biophysical and biochemical properties and carry both proteins and RNAs. Recently, exosome-associated proteins in this MSC EV preparation were found to segregate primarily to those EVs that bind cholera toxin B chain (CTB), a GM1 ganglioside-specific ligand, and pulse-chase experiments demonstrated that these EVs have endosomal origin and carried many of the exosome-associated markers. Here, we report that only a fraction of the MSC EV proteome was found in CTB-bound EVs. Using Annexin V (AV) and Shiga toxin B subunit (ST) with affinities for phosphatidylserine and globotriaosylceramide, respectively, AV- and a ST-binding EV were identified. CTB-, AV- and ST-binding EVs all carried actin. However, the AV-binding EVs carried low or undetectable levels of the exosome-associated proteins. Only the ST-binding EVs carried RNA and EDA-containing fibronectin. Proteins in AV-binding EVs were also different from those released by apoptotic MSCs. CTB- and AV-binding activities were localized to the plasma membrane and cytoplasm of MSCs, while ST-binding activity was localized to the nucleus. Together, this study demonstrates that cells secrete many types of EVs. Specifically, MSCs secrete at least 3 types. They can be differentially isolated based on their affinities for membrane lipid-binding ligands. As the subcellular sites of the binding activities of these ligands and cargo load are different for each EV type, they are likely to have a different biogenesis pathway and possibly different functions.

Extracellular Vesicle Heterogeneity: Subpopulations, Isolation Techniques, and Diverse Functions in Cancer Progression.

PubMed

Willms, Eduard; Cabañas, Carlos; Mäger, Imre; Wood, Matthew J A; Vader, Pieter

2018-01-01

Cells release membrane enclosed nano-sized vesicles termed extracellular vesicles (EVs) that function as mediators of intercellular communication by transferring biological information between cells. Tumor-derived EVs have emerged as important mediators in cancer development and progression, mainly through transfer of their bioactive content which can include oncoproteins, oncogenes, chemokine receptors, as well as soluble factors, transcripts of proteins and miRNAs involved in angiogenesis or inflammation. This transfer has been shown to influence the metastatic behavior of primary tumors. Moreover, tumor-derived EVs have been shown to influence distant cellular niches, establishing favorable microenvironments that support growth of disseminated cancer cells upon their arrival at these pre-metastatic niches. It is generally accepted that cells release a number of major EV populations with distinct biophysical properties and biological functions. Exosomes, microvesicles, and apoptotic bodies are EV populations most widely studied and characterized. They are discriminated based primarily on their intracellular origin. However, increasing evidence suggests that even within these EV populations various subpopulations may exist. This heterogeneity introduces an extra level of complexity in the study of EV biology and function. For example, EV subpopulations could have unique roles in the intricate biological processes underlying cancer biology. Here, we discuss current knowledge regarding the role of subpopulations of EVs in cancer development and progression and highlight the relevance of EV heterogeneity. The position of tetraspanins and integrins therein will be highlighted. Since addressing EV heterogeneity has become essential for the EV field, current and novel techniques for isolating EV subpopulations will also be discussed. Further dissection of EV heterogeneity will advance our understanding of the critical roles of EVs in health and disease.

Increased Risk of Tics in Children Infected with Enterovirus: A Nationwide Population-Based Study.

PubMed

Lin, Jiun-Nong; Lin, Cheng-Li; Yen, Hung-Rong; Yang, Chi-Hui; Lai, Chung-Hsu; Lin, Hsi-Hsun; Kao, Chia-Hung

2017-05-01

Both tics and enterovirus (EV) infections are common in children. The association between EV infections and tics has been seldom evaluated. The aim of this study was to evaluate the risk of diagnosed tics after EV infections in children. A nationwide retrospective cohort study was conducted to determine the risk of tics after EV infections by analyzing data from the National Health Insurance Research Database in Taiwan. Children aged < 18 years with EV infection during 2000 to 2007 were enrolled. For comparison, non-EV-infected children were randomly selected and matched with EV-infected children at a 1:1 ratio according to sex, age, urbanization level, parental occupation, and the year of EV infection. All patients were followed up until the diagnosis of tics, death, loss to follow-up, withdrawal from the insurance system, or December 31, 2008. A total of 282,321 EV-infected and 282,317 non-EV-infected children were included in this study. The mean age was 2.39 years in both cohorts. The overall incidences of tics were 9.12 and 6.21 per 10,000 person-years in the EV-infected and non-EV-infected cohorts, respectively. Children with EV infection were significantly associated with an increased risk of tics compared with those without EV infection (adjusted hazard ratio, 1.38; 95% confidence interval, 1.27-1.5). Multivariable analyses showed that boys, children living in urbanized areas, children whose parents had white-collar jobs, and children with allergic rhinitis or bronchial asthma exhibited a significantly increased risk of tics. This study revealed an increased risk of tics after EV infection in children.

A novel multiplex bead-based platform highlights the diversity of extracellular vesicles

PubMed Central

Koliha, Nina; Wiencek, Yvonne; Heider, Ute; Jüngst, Christian; Kladt, Nikolay; Krauthäuser, Susanne; Johnston, Ian C. D.; Bosio, Andreas; Schauss, Astrid; Wild, Stefan

2016-01-01

The surface protein composition of extracellular vesicles (EVs) is related to the originating cell and may play a role in vesicle function. Knowledge of the protein content of individual EVs is still limited because of the technical challenges to analyse small vesicles. Here, we introduce a novel multiplex bead-based platform to investigate up to 39 different surface markers in one sample. The combination of capture antibody beads with fluorescently labelled detection antibodies allows the analysis of EVs that carry surface markers recognized by both antibodies. This new method enables an easy screening of surface markers on populations of EVs. By combining different capture and detection antibodies, additional information on relative expression levels and potential vesicle subpopulations is gained. We also established a protocol to visualize individual EVs by stimulated emission depletion (STED) microscopy. Thereby, markers on single EVs can be detected by fluorophore-conjugated antibodies. We used the multiplex platform and STED microscopy to show for the first time that NK cell–derived EVs and platelet-derived EVs are devoid of CD9 or CD81, respectively, and that EVs isolated from activated B cells comprise different EV subpopulations. We speculate that, according to our STED data, tetraspanins might not be homogenously distributed but may mostly appear as clusters on EV subpopulations. Finally, we demonstrate that EV mixtures can be separated by magnetic beads and analysed subsequently with the multiplex platform. Both the multiplex bead-based platform and STED microscopy revealed subpopulations of EVs that have been indistinguishable by most analysis tools used so far. We expect that an in-depth view on EV heterogeneity will contribute to our understanding of different EVs and functions. PMID:26901056

Enterovirus D68 detection in respiratory specimens: Association with severe disease.

PubMed

Engelmann, Ilka; Fatoux, Marie; Lazrek, Mouna; Alidjinou, Enagnon K; Mirand, Audrey; Henquell, Cécile; Dewilde, Anny; Hober, Didier

2017-07-01

Molecular techniques increased the number of documented respiratory infections. In a substantial number of cases the causative agent remains undetected. Since August 2014, an increase in Enterovirus(EV)-D68 infections was reported. We aimed to investigate epidemiology and clinical relevance of EV-D68. From June to December 2014 and from September to December 2015, 803 and 847 respiratory specimens, respectively, were tested for respiratory viruses with a multiplex RT-PCR. This multiplex RT-PCR does not detect EV-D68. Therefore, 457 (2014) and 343 (2015) specimens with negative results were submitted to an EV-specific-RT-PCR. EV-positive specimens were tested with an EV-D68-specific-RT-PCR and genotyped. Eleven specimens of 2014 tested positive in the EV-specific-RT-PCR and of these seven were positive in the EV-D68-specific-RT-PCR. Typing confirmed these as EV-D68. Median age of EV-D68-positive patients was 3 years (1 month-91 years). Common symptoms included fever (n = 6, 86%), respiratory distress (n = 5, 71%), and cough (n = 4, 57%). All EV-D68-positive patients were admitted to hospital, 4 (57%) were admitted to intensive care units and 6 (86%) received oxygen. One patient suffered from acute flaccid paralysis. Seven specimens of 2015 were positive in the EV-specific-RT-PCR but negative in the EV-D68-specific-RT-PCR. In conclusion, use of an EV-specific-RT-PCR allowed us to detect EV-D68 circulation in autumn 2014 that was not detected by the multiplex RT-PCR and was associated with severe disease. © 2017 Wiley Periodicals, Inc.

Prospective assessment of clinical symptoms associated with enterovirus and parechovirus genotypes in a multicenter study in Dutch children.

PubMed

de Crom, S C M; Rossen, J W A; de Moor, R A; Veldkamp, E J M; van Furth, A M; Obihara, C C

2016-04-01

Human non-polio enterovirus (EV) and human parechovirus (HPeV) are important pathogens of viral infection and aseptic meningitis in children. The aim of this study is to prospectively compare the incidence, clinical signs, blood and cerebrospinal fluid in EV and HPeV infected children. To compare the clinical symptoms and laboratory data of children with different EV and HPeV genotypes. This study is part of a multicenter prospective cohort study. Children were included in 3 different hospitals in The Netherlands from 2008 to 2011. Of 285 included patients, 140 (49%) had EV and 44 (15%) HPeV infection. Of children with EV infection 9 (6%) had EV-A, 109 (78%) EV-B, 12 (9%) had a non-type able EV and in 10 (7%) no genotyping was performed. Of children with HPeV infection, 24 (55%) had HPeV-3, 6 (14%) HPeV-1, 2 (5%) HPeV-4 and 1 (2%) HPeV-6. Meningitis was more frequent in EV than in HPeV infected children (54% vs. 36%, p=0.046), and in EV-B than EV-A infected children (60 vs. 33%). In contrast gastroenteritis was more frequent in HPeV than EV infected children (30% vs. 15%, p=0.030), and significantly more in HPeV-1 than HPeV-3 infected children (p<0.001). EV infection is more often associated with meningitis and HPeV infection more often with a gastro-enteritis. EV genotype B infection is more often associated with meningitis than EV genotype A infection. HPeV-1 infection was more often associated with gastroenteritis than HPeV-3 infection. Copyright © 2016 Elsevier B.V. All rights reserved.

Can hi-jacking hypoxia inhibit extracellular vesicles in cancer?

PubMed

Lowry, Michelle C; O'Driscoll, Lorraine

2018-06-01

Increasing evidence indicates that extracellular vesicles (EVs) are key players in undesirable cell-cell communication in cancer. However, the release of EVs is not unique to cancer cells; normal cells release EVs to perform physiological roles. Thus, selective inhibition of EV release from cancer cells is desirable. Hypoxia contributes to tumour development and aggressiveness. EV quantities and thus undesirable communications are substantially increased in hypoxia. Targeting hypoxia could selectively inhibit EV release from tumour cells without disturbing physiologically relevant EVs. The unfavourable association between hypoxia and EV release is evident in multiple tumour types; therefore, targeting hypoxia could have a broad therapeutic benefit. Copyright © 2018 Elsevier Ltd. All rights reserved.

Low-energy electron scattering from atomic hydrogen. II. Elastic and inelastic scattering

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

James, K.E. Jr.; Childers, J.G.; Khakoo, M.A.

2004-02-01

We present measurements of differential cross sections for elastic electron scattering from atomic hydrogen at 20 eV and 40 eV incident electron energies and ratios of differential cross sections for electron-impact excitation of atomic hydrogen to the n=2, 3, and 4 levels at incident electron energies of 14.6 eV, 15.6 eV, 17.6 eV, 20 eV, 25 eV, and 40 eV with scattering angles ranging from 10 deg. to 130 deg. We compare our results to available experimental measurements and recent convergent close-coupling calculations. Our results resolve significant discrepancies that existed between theory and past experiments.

Coxsackievirus B4 as a Causative Agent of Diabetes Mellitus Type 1: Is There a Role of Inefficiently Treated Drinking Water and Sewage in Virus Spreading?

PubMed

El-Senousy, Waled M; Abdel-Moneim, Adel; Abdel-Latif, Mahmoud; El-Hefnawy, Mohamed H; Khalil, Rehab G

2018-03-01

This study proposed to detect the enterovirus (EV) infection in children with type 1 diabetes mellitus (T1D) and to assess the role of insufficiently treated water and sewage as sources of viral spreading. Three hundred and eighty-two serum specimens of children with T1D, one hundred serum specimens of children who did not suffer from T1D as control, and forty-eight water and sewage samples were screened for EV RNA using nested RT-PCR. The number of genome copies and infectious units of EVs in raw and treated sewage and water samples were investigated using real-time (RT)-PCR and plaque assay, respectively. T1D markers [Fasting blood glucose (FBG), HbA1c, and C-peptide], in addition to anti-Coxsackie A & B viruses (CVs A & B) IgG, were measured in control, T1D-negative EV (T1D-EV - ), and T1D-positive EV (T1D-EV + ) children specimens. The prevalence of EV genome was significantly higher in diabetic children (26.2%, 100 out of 382) than the control children (0%, 0 out of 100). FBG and HbA1c in T1D-EV - and T1D-EV + children specimens were significantly higher than those in the control group, while c-peptide in T1D-EV - and T1D-EV + children specimens was significantly lower than that in the control (n = 100; p < 0.001). Positivity of anti-CVs A & B IgG was 70.7, 6.7, and 22.9% in T1D-EV + , T1D-EV - , and control children specimens, respectively. The prevalence of EV genome in drinking water and treated sewage samples was 25 and 33.3%, respectively. The prevalence of EV infectious units in drinking water and treated sewage samples was 8.5 and 25%, respectively. Quantification assays were performed to assess the capabilities of both wastewater treatment plants (WWTPs) and water treatment plants (WTPs) to remove EV. The reduction of EV genome in Zenin WWTP ranged from 2 to 4 log 10 , while the reduction of EV infectious units ranged from 1 to 4 log 10 . The reduction of EV genome in El-Giza WTP ranged from 1 to 3 log 10 , while the reduction of EV infectious units ranged from 1 to 2 log 10 . This capability of reduction did not prevent the appearance of infectious EV in treated sewage and drinking water. Plaque purification was performed for isolation of separate EV isolates from treated and untreated water and sewage samples. Characterization of the EV amplicons by RT-PCR followed by sequencing of these isolates revealed high homology (97%) with human coxsackievirus B4 (CV B4) in 60% of the isolates, while the rest of the isolates belonged to poliovirus type 1 and type 2 vaccine strains. On the other hand, characterization of the EV amplicons by RT-PCR followed by sequencing for T1D-EV + children specimens indicated that all samples contained CV B4 with the same sequence characterized in the environmental samples. CV B4-contaminated drinking water or treated sewage may play a role as a causative agent of T1D in children.

Ion-exchange chromatography purification of extracellular vesicles.

PubMed

Kosanović, Maja; Milutinović, Bojana; Goč, Sanja; Mitić, Ninoslav; Janković, Miroslava

2017-08-01

Despite numerous studies, isolating pure preparations of extracellular vesicles (EVs) has proven challenging. Here, we compared ion-exchange chromatography (IEC) to the widely used sucrose density gradient (SDG) centrifugation method for the purification of EVs. EVs in bulk were isolated from pooled normal human amniotic fluid (AF) by differential centrifugation followed by IEC or sucrose density gradient separation. The purity of the isolated EVs was evaluated by electrophoresis and lectin blotting/immuno blotting to monitor the distribution of total proteins, different EVs markers, and selected N-glycans. Our data showed efficient separation of negatively charged EVs from other differently charged molecules, while comparative profiling of EVs using SDG centrifugation confirmed anion-exchange chromatography is advantageous for EV purification. Finally, although this IEC-based method was validated using AF, the approach should be readily applicable to isolation of EVs from other sources as well.

Achieving the Promise of Therapeutic Extracellular Vesicles: The Devil is in Details of Therapeutic Loading.

PubMed

Sutaria, Dhruvitkumar S; Badawi, Mohamed; Phelps, Mitch A; Schmittgen, Thomas D

2017-05-01

Extracellular vesicles (EVs) represent a class of cell secreted organelles which naturally contain biomolecular cargo such as miRNA, mRNA and proteins. EVs mediate intercellular communication, enabling the transfer of functional nucleic acids from the cell of origin to the recipient cells. In addition, EVs make an attractive delivery vehicle for therapeutics owing to their increased stability in circulation, biocompatibility, low immunogenicity and toxicity profiles. EVs can also be engineered to display targeting moieties on their surfaces which enables targeting to desired tissues, organs or cells. While much has been learned on the role of EVs as cell communicators, the field of therapeutic EV application is currently under development. Critical to the future success of EV delivery system is the description of methods by which therapeutics can be successfully and efficiently loaded within the EVs. Two methods of loading of EVs with therapeutic cargo exist, endogenous and exogenous loading. We have therefore focused this review on describing the various published approaches for loading EVs with therapeutics.

Circulation of two Enterovirus C105 (EV-C105) lineages in Europe and Africa.

PubMed

Piralla, A; Daleno, C; Girello, A; Esposito, S; Baldanti, F

2015-06-01

The coding sequences of five human enterovirus (HEV)-C genotype 105 strains recovered in Italy, Romania and Burundi from patients with upper and lower respiratory tract infections were analysed and phylogenetically compared with other circulating HEV-C strains. The EV-C105 was closely related to EV-C109 and EV-C118 strains. The European strains were similar to other circulating EV-C105 strains, while the two African EV-C105 clustered in separate bootstrap-supported (>0.90) branches of the P2 and P3 region trees. Minor inconsistencies in the clustering pattern of EV-C105 in the capsid region (P1) and non-capsid region (P3) suggest that recombination may have occurred in EV-C105 group B viruses. In conclusion, phylogenetic analysis revealed the circulation of two distinct EV-C105 lineages in Europe and Africa. A different pattern of evolution could be hypothesized for the two EV-C105 lineages. © 2015 The Authors.

Proton irradiation effects on deep level states in Mg-doped p-type GaN grown by ammonia-based molecular beam epitaxy

NASA Astrophysics Data System (ADS)

Zhang, Z.; Arehart, A. R.; Kyle, E. C. H.; Chen, J.; Zhang, E. X.; Fleetwood, D. M.; Schrimpf, R. D.; Speck, J. S.; Ringel, S. A.

2015-01-01

The impact of proton irradiation on the deep level states throughout the Mg-doped p-type GaN bandgap is investigated using deep level transient and optical spectroscopies. Exposure to 1.8 MeV protons of 1 × 1013 cm-2 and 3 × 1013 cm-2 fluences not only introduces a trap with an EV + 1.02 eV activation energy but also brings monotonic increases in concentration for as-grown deep states at EV + 0.48 eV, EV + 2.42 eV, EV + 3.00 eV, and EV + 3.28 eV. The non-uniform sensitivities for individual states suggest different physical sources and/or defect generation mechanisms. Comparing with prior theoretical calculations reveals that several traps are consistent with associations to nitrogen vacancy, nitrogen interstitial, and gallium vacancy origins, and thus are likely generated through displacing nitrogen and gallium atoms from the crystal lattice in proton irradiation environment.

Recombination in the evolution of enterovirus C species sub-group that contains types CVA-21, CVA-24, EV-C95, EV-C96 and EV-C99.

PubMed

Smura, Teemu; Blomqvist, Soile; Vuorinen, Tytti; Ivanova, Olga; Samoilovich, Elena; Al-Hello, Haider; Savolainen-Kopra, Carita; Hovi, Tapani; Roivainen, Merja

2014-01-01

Genetic recombination is considered to be a very frequent phenomenon among enteroviruses (Family Picornaviridae, Genus Enterovirus). However, the recombination patterns may differ between enterovirus species and between types within species. Enterovirus C (EV-C) species contains 21 types. In the capsid coding P1 region, the types of EV-C species cluster further into three sub-groups (designated here as A-C). In this study, the recombination pattern of EV-C species sub-group B that contains types CVA-21, CVA-24, EV-C95, EV-C96 and EV-C99 was determined using partial 5'UTR and VP1 sequences of enterovirus strains isolated during poliovirus surveillance and previously published complete genome sequences. Several inter-typic recombination events were detected. Furthermore, the analyses suggested that inter-typic recombination events have occurred mainly within the distinct sub-groups of EV-C species. Only sporadic recombination events between EV-C species sub-group B and other EV-C sub-groups were detected. In addition, strict recombination barriers were inferred for CVA-21 genotype C and CVA-24 variant strains. These results suggest that the frequency of inter-typic recombinations, even within species, may depend on the phylogenetic position of the given viruses.

Transmission of HBV DNA Mediated by Ceramide-Triggered Extracellular Vesicles.

PubMed

Sanada, Takahiro; Hirata, Yuichi; Naito, Yutaka; Yamamoto, Naoki; Kikkawa, Yoshiaki; Ishida, Yuji; Yamasaki, Chihiro; Tateno, Chise; Ochiya, Takahiro; Kohara, Michinori

2017-03-01

An extracellular vesicle (EV) is a nanovesicle that shuttles proteins, nucleic acids, and lipids, thereby influencing cell behavior. A recent crop of reports have shown that EVs are involved in infectious biology, influencing host immunity and playing a role in the viral life cycle. In the present work, we investigated the EV-mediated transmission of hepatitis B virus (HBV) infection. We investigated the EV-mediated transmission of HBV infection by using a HBV infectious culture system that uses primary human hepatocytes derived from humanized chimeric mice (PXB-cells). Purified EVs were isolated by ultracentrifugation. To analyze the EVs and virions, we used stimulated emission depletion microscopy. Purified EVs from HBV-infected PXB-cells were shown to contain HBV DNA and to be capable of transmitting HBV DNA to naive PXB-cells. These HBV-DNA-transmitting EVs were shown to be generated through a ceramide-triggered EV production pathway. Furthermore, we showed that these HBV-DNA-transmitting EVs were resistant to antibody neutralization; stimulated emission depletion microscopy showed that EVs lacked hepatitis B surface antigen, the target of neutralizing antibodies. These findings suggest that EVs harbor a DNA cargo capable of transmitting viral DNA into hepatocytes during HBV infection, representing an additional antibody-neutralization-resistant route of HBV infection.

Detailed analysis of the plasma extracellular vesicle proteome after separation from lipoproteins.

PubMed

Karimi, Nasibeh; Cvjetkovic, Aleksander; Jang, Su Chul; Crescitelli, Rossella; Hosseinpour Feizi, Mohammad Ali; Nieuwland, Rienk; Lötvall, Jan; Lässer, Cecilia

2018-02-13

The isolation of extracellular vesicles (EVs) from blood is of great importance to understand the biological role of circulating EVs and to develop EVs as biomarkers of disease. Due to the concurrent presence of lipoprotein particles, however, blood is one of the most difficult body fluids to isolate EVs from. The aim of this study was to develop a robust method to isolate and characterise EVs from blood with minimal contamination by plasma proteins and lipoprotein particles. Plasma and serum were collected from healthy subjects, and EVs were isolated by size-exclusion chromatography (SEC), with most particles being present in fractions 8-12, while the bulk of the plasma proteins was present in fractions 11-28. Vesicle markers peaked in fractions 7-11; however, the same fractions also contained lipoprotein particles. The purity of EVs was improved by combining a density cushion with SEC to further separate lipoprotein particles from the vesicles, which reduced the contamination of lipoprotein particles by 100-fold. Using this novel isolation procedure, a total of 1187 proteins were identified in plasma EVs by mass spectrometry, of which several proteins are known as EV-associated proteins but have hitherto not been identified in the previous proteomic studies of plasma EVs. This study shows that SEC alone is unable to completely separate plasma EVs from lipoprotein particles. However, combining SEC with a density cushion significantly improved the separation of EVs from lipoproteins and allowed for a detailed analysis of the proteome of plasma EVs, thus making blood a viable source for EV biomarker discovery.

Development of duplex real-time RT-PCR based on Taqman technology for detecting simultaneously the genome of pan-enterovirus and enterovirus 71.

PubMed

Hwang, Seoyeon; Kang, Byunghak; Hong, Jiyoung; Kim, Ahyoun; Kim, Hyejin; Kim, Kisang; Cheon, Doo-Sung

2013-07-01

Human enterovirus (EV) 71 is the main etiological agent of hand, foot, and mouth disease (HFMD). It is associated with neurological complications, and caused fatalities during recent outbreaks in the Asia-Pacific region. Infections caused by EV71 could lead to many complications, ranging from brainstem encephalitis to pulmonary oedema, resulting in high mortality. In this study, a duplex real-time RT-PCR assay was developed in order to simultaneously detect pan-EV and EV71. EV71-specific primers and probes were designed based on the highly conserved VP1 region of EV71. Five EV71 strains were detected as positive, and no positive fluorescence signal was observed in the duplex real-time RT-PCR for other viral RNA, which showed 100% specificity for the selected panel, and no cross-reactions were observed in this duplex real-time RT-PCR. The EV71-specific duplex real-time RT-PCR was more sensitive than conventional RT-PCR, and detected viral titers that were 10-fold lower than those measured by the latter. Of the 381 HFMD clinical specimens, 196 (51.4%) cases were pan-EV-positive, of which 170 (86.7%) were EV71-positive when tested by pan-EV and EV71-specific duplex real-time RT-PCR. EV71-specific duplex real-time RT-PCR offers a rapid and sensitive method to detect EV71 from clinical specimens, and will allow quarantine measures to be taken more effectively during outbreaks. Copyright © 2013 Wiley Periodicals, Inc.

Association between Antioxidant Enzyme Activities and Enterovirus-Infected Type 1 Diabetic Children.

PubMed

Abdel-Moneim, Adel; El-Senousy, Waled M; Abdel-Latif, Mahmoud; Khalil, Rehab G

2018-01-01

To examine the effect of infection with Enterovirus (EV) in children with type 1 diabetes (T1D) on the activities of serum antioxidant enzymes in diabetic and nondiabetic controls. Three hundred and eighty-two diabetic and 100 nondiabetic children were tested for EV RNA using reverse transcriptase (RT)-PCR. The activities of serum superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) were also estimated in diabetic patients infected with EV (T1D-EV+), those not infected with EV (T1D-EV-), and in nondiabetic controls. The frequency of EV was higher in diabetic children (100/382; 26.2%) than in healthy controls (0/100). Levels of fasting blood glucose (FBG), glycosylated hemoglobin (HbA1c) and C-reactive protein (CRP) were significantly higher but C-peptide was significantly lower in diabetic children than in controls. CRP levels were higher in the T1D-EV+ group than in the T1D-EV- group, and higher in all diabetic children than in nondiabetic controls. The activities of the antioxidant enzymes GPx, SOD, and CAT decreased significantly in diabetic children compared to in controls. Moreover, the activities of the enzymes tested were significantly reduced in the T1D-EV+ group compared to in the T1D-EV- group. Our data indicate that EV infection correlated with a decrease in the activity of antioxidant enzymes in the T1D-EV+ group compared to in the T1D-EV- group; this may contribute to β cell damage and increased inflammation. © 2018 The Author(s) Published by S. Karger AG, Basel.

Extracellular vesicles for liquid biopsy in prostate cancer: where are we and where are we headed?

PubMed Central

Minciacchi, V R; Zijlstra, A; Rubin, M A; Di Vizio, D

2017-01-01

Background: Extracellular vesicles (EVs) are a heterogeneous class of lipid bound particles shed by any cell in the body in physiological and pathological conditions. EVs play critical functions in intercellular communication. EVs can actively travel in intercellular matrices and eventually reach the circulation. They can also be released directly in biological fluids where they appear to be stable. Because the molecular content of EVs reflects the composition of the cell of origin, they have recently emerged as a promising source of biomarkers in a number of diseases. EV analysis is particularly attractive in cancer patients that frequently present with increased numbers of circulating EVs. Methods: We sought to review the current literature on the molecular profile of prostate cancer-derived EVs in model systems and patient biological fluids in an attempt to draw some practical and universal conclusions on the use of EVs as a tool for liquid biopsy in clinical specimens. Results: We discuss advantages and limitations of EV-based liquid biopsy approaches summarizing salient studies on protein, DNA and RNA. Several candidate biomarkers have been identified so far but these results are difficult to apply to the clinic. However, the field is rapidly moving toward the implementation of novel tools to isolate cancer-specific EVs that are free of benign EVs and extra-vesicular contaminants. This can be achieved by identifying markers that are exquisitely present in tumor cell-derived EVs. An important contribution might also derive from a better understanding of EV types that may play specific functions in tumor progression and that may be a source of cancer-specific markers. Conclusions: EV analysis holds strong promises for the development of non-invasive biomarkers in patients with prostate cancer. Implementation of modern methods for EV isolation and characterization will enable to interrogate circulating EVs in vivo. PMID:28374743

Long-term Durability of Infrainguinal Endovascular and Open Revascularization for Disabling Claudication.

PubMed

Khan, Sikandar Z; Rivero, Mariel; Cherr, Gregory S; Harris, Linda M; Dryjski, Maciej L; Dosluoglu, Hasan H

2018-05-15

Infrainguinal revascularization for disabling claudication (DC) is frequently performed, but long-term results are still unknown. In this study, we compared clinical outcomes of infrainguinal endovascular (EV) and open interventions for DC after the failure of medical management. One hundred ninety-four patients with DC (Rutherford category 3) who had open (n = 53) or EV (n = 141) interventions were grouped as open-great saphenous vein (GSV) (n = 21), open-prosthetic (n = 32), EV-Trans-Atlantic Inter-Society Consensus II (TASC II) A and B (AB) (n = 48), and EV-TASC II C and D (CD) (n = 93). Patency, primary clinical success (PCS; sustained improvement in symptoms without reintervention), and secondary clinical success (SCS; sustained improvement in symptoms with reintervention) rates were compared. Mean follow-up was 57 ± 33 months. Five-year primary patency was 58% in open-GSV, 40% in open-prosthetic, 72% in EV-AB, and 38% in EV-CD (P < 0.001). Five-year secondary patency was 77% in open-GSV, 50% in open-prosthetic, 96% in EV-AB, and 61% in EV-CD (P < 0.001). Freedom from major adverse limb events was 73% in open-GSV, 77% in EV-AB, 70% in EV-CD, and 67% in open-prosthetic (P = 0.279). Five-year PCS was 46% in open-GSV, 40% in open-prosthetic, 57% in EV-AB, and 44% in EV-CD (P = 0.02). Five-year SCS was 78% in open-GSV, 78% in open-prosthetic, 85% in EV-AB, and 84% in EV-CD (P = 0.732). A total of 116 reinterventions were performed, 10 in 6 limbs (27%) in open-GSV, 18 in 12 limbs (36%) in open-prosthetic, 26 in 15 limbs (24%) in EV-AB, and 62 in 39 limbs (36%) in EV-CD. Reinterventions included 71 (61%) EV and 45 (39%) open procedures. Durability of infrainguinal interventions in claudicants depends mainly on anatomic complexity of disease. Good long-term clinical success can be achieved with both open and EV interventions, albeit with high reintervention rates, especially in patients with TASC II C and D disease. A considerable subset of EV patients will eventually require surgical revascularization to maintain clinical benefit. In this study, almost 20% of patients undergoing EV for TASC II C and D disease eventually required surgical bypass. Copyright © 2018 Elsevier Inc. All rights reserved.

Is your article EV-TRACKed?

PubMed

Van Deun, Jan; Hendrix, An

2017-01-01

The EV-TRACK knowledgebase is developed to cope with the need for transparency and rigour to increase reproducibility and facilitate standardization of extracellular vesicle (EV) research. The knowledgebase includes a checklist for authors and editors intended to improve the transparency of methodological aspects of EV experiments, allows queries and meta-analysis of EV experiments and keeps track of the current state of the art. Widespread implementation by the EV research community is key to its success.

Analysis of Electric Vehicle Charging Impact on the Electric Power Grid

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

Jiang, Zeming; Tian, Hao; Beshir, Mohammed J.

2016-09-24

In order to evaluate the impact of electric vehicles (EVs) on the distribution grid and assess their potential benefits to the future smart grid, it is crucial to study the EV charging patterns and the usage charging station. Though EVs are not yet widely adopted nationwide, a valuable methodology to conduct such studies is the statistical analysis of real-world charging data. This paper presents actual EV charging behavior of 64 EVs (5 brands, 8 models) from EV users and charging stations at Los Angeles Department of Water and Power for more than one year. Twenty-four-hour EV charging load curves havemore » been generated and studied for various load periods: daily, monthly, seasonally and yearly. Finally, the effect and impact of EV load on the California distribution network are evaluated at different EV penetration rates.« less

Phylogenetic Characterizations of Highly Mutated EV-B106 Recombinants Showing Extensive Genetic Exchanges with Other EV-B in Xinjiang, China.

PubMed

Song, Yang; Zhang, Yong; Fan, Qin; Cui, Hui; Yan, Dongmei; Zhu, Shuangli; Tang, Haishu; Sun, Qiang; Wang, Dongyan; Xu, Wenbo

2017-02-23

Human enterovirus B106 (EV-B106) is a new member of the enterovirus B species. To date, only three nucleotide sequences of EV-B106 have been published, and only one full-length genome sequence (the Yunnan strain 148/YN/CHN/12) is available in the GenBank database. In this study, we conducted phylogenetic characterisation of four EV-B106 strains isolated in Xinjiang, China. Pairwise comparisons of the nucleotide sequences and the deduced amino acid sequences revealed that the four Xinjiang EV-B106 strains had only 80.5-80.8% nucleotide identity and 95.4-97.3% amino acid identity with the Yunnan EV-B106 strain, indicating high mutagenicity. Similarity plots and bootscanning analyses revealed that frequent intertypic recombination occurred in all four Xinjiang EV-B106 strains in the non-structural region. These four strains may share a donor sequence with the EV-B85 strain, which circulated in Xinjiang in 2011, indicating extensive genetic exchanges between these strains. All Xinjiang EV-B106 strains were temperature-sensitive. An antibody seroprevalence study against EV-B106 in two Xinjiang prefectures also showed low titres of neutralizing antibodies, suggesting limited exposure and transmission in the population. This study contributes the whole genome sequences of EV-B106 to the GenBank database and provides valuable information regarding the molecular epidemiology of EV-B106 in China.

Phylogenetic Characterizations of Highly Mutated EV-B106 Recombinants Showing Extensive Genetic Exchanges with Other EV-B in Xinjiang, China

PubMed Central

Song, Yang; Zhang, Yong; Fan, Qin; Cui, Hui; Yan, Dongmei; Zhu, Shuangli; Tang, Haishu; Sun, Qiang; Wang, Dongyan; Xu, Wenbo

2017-01-01

Human enterovirus B106 (EV-B106) is a new member of the enterovirus B species. To date, only three nucleotide sequences of EV-B106 have been published, and only one full-length genome sequence (the Yunnan strain 148/YN/CHN/12) is available in the GenBank database. In this study, we conducted phylogenetic characterisation of four EV-B106 strains isolated in Xinjiang, China. Pairwise comparisons of the nucleotide sequences and the deduced amino acid sequences revealed that the four Xinjiang EV-B106 strains had only 80.5–80.8% nucleotide identity and 95.4–97.3% amino acid identity with the Yunnan EV-B106 strain, indicating high mutagenicity. Similarity plots and bootscanning analyses revealed that frequent intertypic recombination occurred in all four Xinjiang EV-B106 strains in the non-structural region. These four strains may share a donor sequence with the EV-B85 strain, which circulated in Xinjiang in 2011, indicating extensive genetic exchanges between these strains. All Xinjiang EV-B106 strains were temperature-sensitive. An antibody seroprevalence study against EV-B106 in two Xinjiang prefectures also showed low titres of neutralizing antibodies, suggesting limited exposure and transmission in the population. This study contributes the whole genome sequences of EV-B106 to the GenBank database and provides valuable information regarding the molecular epidemiology of EV-B106 in China. PMID:28230168

A novel inactivated enterovirus 71 vaccine can elicit cross-protective immunity against coxsackievirus A16 in mice.

PubMed

Yang, Lisheng; Liu, Yajing; Li, Shuxuan; Zhao, Huan; Lin, Qiaona; Yu, Hai; Huang, Xiumin; Zheng, Qingbing; Cheng, Tong; Xia, Ningshao

2016-11-21

Hand, foot, and mouth disease (HFMD) is a highly contagious disease that mainly affects infants and children. Enterovirus 71 (EV71) and coxsackievirus A16 (CA16) are the major pathogens of HFMD. Two EV71 vaccines were recently licensed in China and the administration of the EV71 vaccines is believed to significantly reduce the number of HFMD-related severe or fatal cases. However, a monovalent EV71 vaccine cannot cross-protect against CA16 infection, this may result in that it cannot effectively control the overall HFMD epidemic. In this study, a chimeric EV71, whose VP1/210-225 epitope was replaced by that of CA16, was constructed using a reverse genetics technique to produce a candidate EV71/CA16 bivalent vaccine strain. The chimeric EV71 was infectious and showed similar growth characteristics as its parental strain. The replacement of the VP1/210-225 epitope did not significantly affect the antigenicity and immunogenicity of EV71. More importantly, the chimeric EV71 could induce protective immunity against both EV71 and CA16, and protect neonatal mice against either EV71 or CA16 lethal infections, the chimeric EV71 constructed in this study was shown to be a feasible and promising candidate bivalent vaccine against both EV71 and CA16. The construction of a chimeric enterovirus also provides an alternative platform for broad-spectrum HFMD vaccines development. Copyright © 2016 Elsevier Ltd. All rights reserved.

Making change easy: A peer-to-peer guide on transitioning to new hand hygiene products.

PubMed

Amirov, Chingiz M; Candon, Heather L; Jacob, Latha

2017-01-01

This report summarizes our experiences planning and implementing the transition to a new commercial line of hand hygiene products and their dispensing systems in a large academic health care facility in Toronto, Canada. Our lessons learned are organized into a practical guide made available in 2 different formats: this article and an illustrated peer-to-peer guide (http://www.baycrest.org/wp-content/uploads/HCE-PROG-HH_HighQuality.pdf). Copyright © 2017 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.

Hemorrhagic Fever with Renal Syndrome (Korean Hemorrhagic Fever)

DTIC Science & Technology

1988-06-30

camp 5/19(26%) n~t. n.t. 23 - 28 US Army camp 1/31(3%) 2/31(7%) 4/34 (13%) OK Army cimp 4/30(13%) 6/30(20%) 2/30(7%) 9 - 14 Chinhae 12/3,1 (35%) 2/34... data showed that 15% of the laboratory rats assayed from the Microbiology Department, School of Medicine, University of Buenos Aires, and 70% of the...Lee, M. C. and Cho, K. S. Management of Korean haemorrhagic fever. Med. Prog. 2: 15-21, 1980. 28. Lee, H. W., Bark, D. H., Baek, L. H., Choi, K. S

Integrated Artillery Recoil Mechanism and Automated Handling Design for 155mm Self-Propelled Howitzer

DTIC Science & Technology

1980-04-01

3.8" iv, ?J Oo -r- p,Ik-- 1. 346.57T ,.~ ~ ~~ q 1-- - 44t_ L m50 PCF-RM-1284 A-3 PACIFIC CAR AND FOUNDRY COMPANY lit) ENGINEERING DEPARTMENT .1 NAME...c . / KBEARSTR _ 14138 11/19/79 MONDAY I06 THIS PROG. CALCS. BEARING STRESS IN HOLES DUE TO BOLT BEARING & IOMtENT. ENTER HOLT (SHAFT) & HOLE...Ref 2. Alternativ ly, the calculated Survival after 10 million cycles of ASI-SA| 4340 steel with tensile strengths of "S, 1320, stress may be adjusted

A new efficient mixture screening design for optimization of media.

PubMed

Rispoli, Fred; Shah, Vishal

2009-01-01

Screening ingredients for the optimization of media is an important first step to reduce the many potential ingredients down to the vital few components. In this study, we propose a new method of screening for mixture experiments called the centroid screening design. Comparison of the proposed design with Plackett-Burman, fractional factorial, simplex lattice design, and modified mixture design shows that the centroid screening design is the most efficient of all the designs in terms of the small number of experimental runs needed and for detecting high-order interaction among ingredients. (c) 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009.

Updated Review of Planetary Atmospheric Electricity

NASA Astrophysics Data System (ADS)

Yair, Y.; Fischer, G.; Simões, F.; Renno, N.; Zarka, P.

2008-06-01

This paper reviews the progress achieved in planetary atmospheric electricity, with focus on lightning observations by present operational spacecraft, aiming to fill the hiatus from the latest review published by Desch et al. (Rep. Prog. Phys. 65:955 997, 2002). The information is organized according to solid surface bodies (Earth, Venus, Mars and Titan) and gaseous planets (Jupiter, Saturn, Uranus and Neptune), and each section presents the latest results from space-based and ground-based observations as well as laboratory experiments. Finally, we review planned future space missions to Earth and other planets that will address some of the existing gaps in our knowledge.

Updated Review of Planetary Atmospheric Electricity

NASA Astrophysics Data System (ADS)

Yair, Y.; Fischer, G.; Simões, F.; Renno, N.; Zarka, P.

This paper reviews the progress achieved in planetary atmospheric electricity, with focus on lightning observations by present operational spacecraft, aiming to fill the hiatus from the latest review published by Desch et al. (Rep. Prog. Phys. 65:955-997, 2002). The information is organized according to solid surface bodies (Earth, Venus, Mars and Titan) and gaseous planets (Jupiter, Saturn, Uranus and Neptune), and each section presents the latest results from space-based and ground-based observations as well as laboratory experiments. Finally, we review planned future space missions to Earth and other planets that will address some of the existing gaps in our knowledge.

A photometric study of the hot exoplanet WASP-19b

NASA Astrophysics Data System (ADS)

Lendl, M.; Gillon, M.; Queloz, D.; Alonso, R.; Fumel, A.; Jehin, E.; Naef, D.

2013-04-01

Context. The sample of hot Jupiters that have been studied in great detail is still growing. In particular, when the planet transits its host star, it is possible to measure the planetary radius and the planet mass (with radial velocity data). For the study of planetary atmospheres, it is essential to obtain transit and occultation measurements at multiple wavelengths. Aims: We aim to characterize the transiting hot Jupiter WASP-19b by deriving accurate and precise planetary parameters from a dedicated observing campaign of transits and occultations. Methods: We have obtained a total of 14 transit lightcurves in the r'-Gunn, I-Cousins, z'-Gunn, and I + z' filters and 10 occultation lightcurves in z'-Gunn using EulerCam on the Euler-Swiss telescope and TRAPPIST. We also obtained one lightcurve through the narrow-band NB1190 filter of HAWK-I on the VLT measuring an occultation at 1.19 μm. We performed a global MCMC analysis of all new data, together with some archive data in order to refine the planetary parameters and to measure the occultation depths in z'-band and at 1.19 μm. Results: We measure a planetary radius of Rp = 1.376 ± 0.046 RJ, a planetary mass of Mp = 1.165 ± 0.068 MJ, and find a very low eccentricity of e = 0.0077-0.0032+0.0068, compatible with a circular orbit. We have detected the z'-band occultation at 3σ significance and measure it to be δFocc,z' = 352 ± 116 ppm, more than a factor of 2 smaller than previously published. The occultation at 1.19 μm is only marginally constrained at δFocc,NB1190 = 1711-726+745 ppm. Conclusions: We show that the detection of occultations in the visible range is within reach, even for 1 m class telescopes if a considerable number of individual events are observed. Our results suggest an oxygen-dominated atmosphere of WASP-19b, making the planet an interesting test case for oxygen-rich planets without temperature inversion. Based on photometric observations made with HAWK-I on the ESO VLT/UT4 (Prog. ID 084.C-0532), EulerCam on the Euler-Swiss telescope and the Belgian TRAPPIST telescope, as well as archive data from the Faulkes South Telescope, CORALIE on the Euler-Swiss telescope, HARPS on the ESO 3.6 m telescope (Prog. ID 084-C-0185), and HAWK-I (Prog. ID 083.C-0377(A)).The photometric time series data in this work are only available at the CDS via anonymous ftp to cdsarc.u-strasbg.fr (130.79.128.5) or via http://cdsarc.u-strasbg.fr/viz-bin/qcat?J/A+A/552/A2

Protective effect of enterovirus‑71 (EV71) virus‑like particle vaccine against lethal EV71 infection in a neonatal mouse model.

PubMed

Cao, Lei; Mao, Fengfeng; Pang, Zheng; Yi, Yao; Qiu, Feng; Tian, Ruiguang; Meng, Qingling; Jia, Zhiyuan; Bi, Shengli

2015-08-01

Enterovirus-71 (EV71) is a viral pathogen that causes severe cases of hand, foot and mouth disease (HFMD) among young children, with significant mortality. Effective vaccines against HFMD are urgently required. Several EV71 virus-like particle (VLP) vaccine candidates were found to be protective in the neonatal mouse EV71 challenge model. However, to what extent the VLP vaccine protects susceptible organs against EV71 infection in vivo has remained elusive. In the present study, the comprehensive immunogenicity of a potential EV71 vaccine candidate based on VLPs was evaluated in a neonatal mouse model. Despite lower levels of neutralizing antibodies to EV71 in the sera of VLP-immunized mice compared with those in mice vaccinated with inactivated EV71, the VLP-based vaccine was shown to be able to induce immunoglobulin (Ig)G and IgA memory-associated cellular immune responses to EV71. Of note, the EV71 VLP vaccine candidate was capable of inhibiting viral proliferation in cardiac muscle, skeletal muscle, lung and intestine of immunized mice and provided effective protection against the pathological damage caused by viral attack. In particular, the VLP vaccine was able to inhibit the transportation of EV71 from the central nervous system to the muscle tissue and greatly protected muscle tissue from infection, along with recovery from the viral infection. This led to nearly 100% immunoprotective efficacy, enabling neonatal mice delivered by VLP-immunized female adult mice to survive and grow with good health. The present study provided valuable additional knowledge of the specific protective efficacy of the EV71 VLP vaccine in vivo, which also indicated that it is a promising potential candidate for being developed into an EV71 vaccine.

Microvesicle- and exosome-mediated drug delivery enhances the cytotoxicity of Paclitaxel in autologous prostate cancer cells.

PubMed

Saari, Heikki; Lázaro-Ibáñez, Elisa; Viitala, Tapani; Vuorimaa-Laukkanen, Elina; Siljander, Pia; Yliperttula, Marjo

2015-12-28

Extracellular vesicles (EVs) are naturally occurring membrane particles that mediate intercellular communication by delivering molecular information between cells. In this study, we investigated the effectiveness of two different populations of EVs (microvesicle- and exosome-enriched) as carriers of Paclitaxel to autologous prostate cancer cells. EVs were isolated from LNCaP- and PC-3 prostate cancer cell cultures using differential centrifugation and characterized by electron microscopy, nanoparticle tracking analysis, and Western blot. The uptake of microvesicles and exosomes by the autologous prostate cancer cells was assessed by flow cytometry and confocal microscopy. The EVs were loaded with Paclitaxel and the effectiveness of EV-mediated drug delivery was assessed with viability assays. The distribution of EVs and EV-delivered Paclitaxel in cells was inspected by confocal microscopy. Our main finding was that the loading of Paclitaxel to autologous prostate cancer cell-derived EVs increased its cytotoxic effect. This capacity was independent of the EV population and the cell line tested. Although the EVs without the drug increased cancer cell viability, the net effect of enhanced cytotoxicity remained. Both EV populations delivered Paclitaxel to the recipient cells through endocytosis, leading to the release of the drug from within the cells. The removal of EV surface proteins did not affect exosomes, while the drug delivery mediated by microvesicles was partially inhibited. Cancer cell-derived EVs can be used as effective carriers of Paclitaxel to their parental cells, bringing the drug into the cells through an endocytic pathway and increasing its cytotoxicity. However, due to the increased cell viability, the use of cancer cell-derived EVs must be further investigated before any clinical applications can be designed. Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.

EV-A71 vaccine licensure: a first step for multivalent enterovirus vaccine to control HFMD and other severe diseases

PubMed Central

Mao, Qunying; Wang, Yiping; Bian, Lianlian; Xu, Miao; Liang, Zhenglun

2016-01-01

Enteroviruses (EVs) are the most common viral agents in humans. Although most infections are mild or asymptomatic, there is a wide spectrum of clinical manifestations that may be caused by EV infections with varying degrees of severity. Among these viruses, EV-A71 and coxsackievirus (CV) CV-A16 from group A EVs attract the most attention because they are responsible for hand, foot and mouth disease (HFMD). Other EV-A viruses such as CV-A6 and CV-A10 were also reported to cause HFMD outbreaks in several countries or regions. Group B EVs such as CV-B3, CV-B5 and echovirus 30 were reported to be the main pathogens responsible for myocarditis and encephalitis epidemics and were also detected in HFMD patients. Vaccines are the best tools to control infectious diseases. In December 2015, China's Food and Drug Administration approved two inactivated EV-A71 vaccines for preventing severe HFMD.The CV-A16 vaccine and the EV-A71-CV-A16 bivalent vaccine showed substantial efficacy against HFMD in pre-clinical animal models. Previously, research on EV-B group vaccines was mainly focused on CV-B3 vaccine development. Because the HFMD pathogen spectrum has changed, and the threat from EV-B virus-associated severe diseases has gradually increased, it is necessary to develop multivalent HFMD vaccines. This study summarizes the clinical symptoms of diseases caused by EVs, such as HFMD, myocarditis and encephalitis, and the related EV vaccine development progress. In conclusion, developing multivalent EV vaccines should be strongly recommended to prevent HFMD, myocarditis, encephalitis and other severe diseases. PMID:27436364

Time-resolved radiation chemistry: Dynamics of electron attachment to uracil following UV excitation of iodide-uracil complexes

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

King, Sarah B.; Yandell, Margaret A.; Stephansen, Anne B.

Electron attachment to uracil was investigated by applying time-resolved photoelectron imaging to iodide-uracil (I{sup –}U) complexes. In these studies, an ultraviolet pump pulse initiated charge transfer from the iodide to the uracil, and the resulting dynamics of the uracil temporary negative ion were probed. Five different excitation energies were used, 4.00 eV, 4.07 eV, 4.14 eV, 4.21 eV, and 4.66 eV. At the four lowest excitation energies, which lie near the vertical detachment energy of the I{sup –}U complex (4.11 eV), signatures of both the dipole bound (DB) as well as the valence bound (VB) anion of uracil were observed.more » In contrast, only the VB anion was observed at 4.66 eV, in agreement with previous experiments in this higher energy range. The early-time dynamics of both states were highly excitation energy dependent. The rise time of the DB anion signal was ∼250 fs at 4.00 eV and 4.07 eV, ∼120 fs at 4.14 eV and cross-correlation limited at 4.21 eV. The VB anion rise time also changed with excitation energy, ranging from 200 to 300 fs for excitation energies 4.00–4.21 eV, to a cross-correlation limited time at 4.66 eV. The results suggest that the DB state acts as a “doorway” state to the VB anion at 4.00–4.21 eV, while direct attachment to the VB anion occurs at 4.66 eV.« less

Enterovirus 71 infection in children with hand, foot, and mouth disease in Shanghai, China: epidemiology, clinical feature and diagnosis.

PubMed

Wang, Ying; Zou, Gang; Xia, Aimei; Wang, Xiangshi; Cai, Jiehao; Gao, Qianqian; Yuan, Shilin; He, Guimei; Zhang, Shuyi; Zeng, Mei; Altmeyer, Ralf

2015-06-03

In 2012 a large outbreak of hand, foot, and mouth disease (HFMD) widely spread over China, causing more than 2 million cases and 567 deaths. Our purpose was to characterize the major pathogens responsible for the 2012 HFMD outbreak and analyze the genetic characterization of the enterovirus 71 (EV71) strains in Shanghai; also, to analyze the dynamic patterns of neutralizing antibody (NAb) against EV71 and evaluate the diagnostic value of several methods for clinical detection of EV71. Clinical samples including stool, serum and CSF were collected from 396 enrolled HFMD inpatients during the peak seasons in 2012. We analyzed the molecular epidemiology, clinical feature, and diagnostic tests of EV71 infection. EV71 was responsible for 60.35 % of HFMD inpatients and 88.46 % of severe cases. The circulating EV71 strains belonged to subgenogroup C4a. The nucleotide sequences of VP1 between severe cases and uncomplicated cases shared 99.2 ~ 100 % of homology. Among 218 cases with EV71 infection, 211 (96.79 %) serum samples showed NAb positive against EV71 and NAb titer reached higher level 3 days after disease onset. Of 92 cases with EV71-associated meningitis or encephalitis, 5 (5.43 %) of 92 had EV71 RNA detected in CSF samples. The blood anti-EV71 IgM assay showed a sensitivity of 93.30 % and a specificity of 50 %. EV71 C4a remained the predominant subgenotype circulating in Shanghai. The severity of the EV71 infection is not associated with the virulence determinants in VP1. RT-PCR together with IgM detection can enhance the early diagnosis of severe EV71-associated HFMD.

EV-A71 vaccine licensure: a first step for multivalent enterovirus vaccine to control HFMD and other severe diseases.

PubMed

Mao, Qunying; Wang, Yiping; Bian, Lianlian; Xu, Miao; Liang, Zhenglun

2016-07-20

Enteroviruses (EVs) are the most common viral agents in humans. Although most infections are mild or asymptomatic, there is a wide spectrum of clinical manifestations that may be caused by EV infections with varying degrees of severity. Among these viruses, EV-A71 and coxsackievirus (CV) CV-A16 from group A EVs attract the most attention because they are responsible for hand, foot and mouth disease (HFMD). Other EV-A viruses such as CV-A6 and CV-A10 were also reported to cause HFMD outbreaks in several countries or regions. Group B EVs such as CV-B3, CV-B5 and echovirus 30 were reported to be the main pathogens responsible for myocarditis and encephalitis epidemics and were also detected in HFMD patients. Vaccines are the best tools to control infectious diseases. In December 2015, China's Food and Drug Administration approved two inactivated EV-A71 vaccines for preventing severe HFMD.The CV-A16 vaccine and the EV-A71-CV-A16 bivalent vaccine showed substantial efficacy against HFMD in pre-clinical animal models. Previously, research on EV-B group vaccines was mainly focused on CV-B3 vaccine development. Because the HFMD pathogen spectrum has changed, and the threat from EV-B virus-associated severe diseases has gradually increased, it is necessary to develop multivalent HFMD vaccines. This study summarizes the clinical symptoms of diseases caused by EVs, such as HFMD, myocarditis and encephalitis, and the related EV vaccine development progress. In conclusion, developing multivalent EV vaccines should be strongly recommended to prevent HFMD, myocarditis, encephalitis and other severe diseases.

Is your article EV-TRACKed?

PubMed Central

Van Deun, Jan; Hendrix, An

2017-01-01

ABSTRACT The EV-TRACK knowledgebase is developed to cope with the need for transparency and rigour to increase reproducibility and facilitate standardization of extracellular vesicle (EV) research. The knowledgebase includes a checklist for authors and editors intended to improve the transparency of methodological aspects of EV experiments, allows queries and meta-analysis of EV experiments and keeps track of the current state of the art. Widespread implementation by the EV research community is key to its success. PMID:29184624

Recombination in the Evolution of Enterovirus C Species Sub-Group that Contains Types CVA-21, CVA-24, EV-C95, EV-C96 and EV-C99

PubMed Central

Smura, Teemu; Blomqvist, Soile; Vuorinen, Tytti; Ivanova, Olga; Samoilovich, Elena; Al-Hello, Haider; Savolainen-Kopra, Carita; Hovi, Tapani; Roivainen, Merja

2014-01-01

Genetic recombination is considered to be a very frequent phenomenon among enteroviruses (Family Picornaviridae, Genus Enterovirus). However, the recombination patterns may differ between enterovirus species and between types within species. Enterovirus C (EV-C) species contains 21 types. In the capsid coding P1 region, the types of EV-C species cluster further into three sub-groups (designated here as A–C). In this study, the recombination pattern of EV-C species sub-group B that contains types CVA-21, CVA-24, EV-C95, EV-C96 and EV-C99 was determined using partial 5′UTR and VP1 sequences of enterovirus strains isolated during poliovirus surveillance and previously published complete genome sequences. Several inter-typic recombination events were detected. Furthermore, the analyses suggested that inter-typic recombination events have occurred mainly within the distinct sub-groups of EV-C species. Only sporadic recombination events between EV-C species sub-group B and other EV-C sub-groups were detected. In addition, strict recombination barriers were inferred for CVA-21 genotype C and CVA-24 variant strains. These results suggest that the frequency of inter-typic recombinations, even within species, may depend on the phylogenetic position of the given viruses. PMID:24722726

Indication of Horizontal DNA Gene Transfer by Extracellular Vesicles

PubMed Central

Speiseder, Thomas; Badbaran, Anita; Reimer, Rudolph; Indenbirken, Daniela; Grundhoff, Adam; Brunswig-Spickenheier, Bärbel; Alawi, Malik; Lange, Claudia

2016-01-01

The biological relevance of extracellular vesicles (EV) in intercellular communication has been well established. Thus far, proteins and RNA were described as main cargo. Here, we show that EV released from human bone marrow derived mesenchymal stromal cells (BM-hMSC) also carry high-molecular DNA in addition. Extensive EV characterization revealed this DNA mainly associated with the outer EV membrane and to a smaller degree also inside the EV. Our EV purification protocol secured that DNA is not derived from apoptotic or necrotic cells. To analyze the relevance of EV-associated DNA we lentivirally transduced Arabidopsis thaliana-DNA (A.t.-DNA) as indicator into BM-hMSC and generated EV. Using quantitative polymerase chain reaction (qPCR) techniques we detected high copy numbers of A.t.-DNA in EV. In recipient hMSC incubated with tagged EV for two weeks we identified A.t.-DNA transferred to recipient cells. Investigation of recipient cell DNA using quantitative PCR and verification of PCR-products by sequencing suggested stable integration of A.t.-DNA. In conclusion, for the first time our proof-of-principle experiments point to horizontal DNA transfer into recipient cells via EV. Based on our results we assume that eukaryotic cells are able to exchange genetic information in form of DNA extending the known cargo of EV by genomic DNA. This mechanism might be of relevance in cancer but also during cell evolution and development. PMID:27684368

Phylogenetic evidence for multiple intertypic recombinations in enterovirus B81 strains isolated in Tibet, China

PubMed Central

Hu, Lan; Zhang, Yong; Hong, Mei; Zhu, Shuangli; Yan, Dongmei; Wang, Dongyan; Li, Xiaolei; Zhu, Zhen; Tsewang; Xu, Wenbo

2014-01-01

Enterovirus B81 (EV-B81) is a newly identified serotype within the species enterovirus B (EV-B). To date, only eight nucleotide sequences of EV-B81 have been published and only one full-length genome sequence (the prototype strain) has been made available in the GenBank database. Here, we report the full-length genome sequences of two EV-B81 strains isolated in the Tibet Autonomous Region of China during acute flaccid paralysis surveillance activities, and we also conducted an antibody seroprevalence study in two prefectures of Tibet. The sequence comparison and phylogenetic dendrogram analysis revealed high variability among the global EV-B81 strains and frequent intertypic recombination in the non-structural protein region of EV-B serotypes, suggesting high genetic diversity of EV-B81. However, low positive rates and low titers of neutralizing antibodies against EV-B81 were detected. Nearly 68% of children under the age of five had no neutralizing antibodies against EV-B81. Hence, the extent of transmission and the exposure of the population to this EV type are very limited. Although little is known about the biological and pathogenic properties of EV-B81 because of few research in this field owing to the limited number of isolates, our study provides basic information for further studies of EV-B81. PMID:25112835

Indication of Horizontal DNA Gene Transfer by Extracellular Vesicles.

PubMed

Fischer, Stefanie; Cornils, Kerstin; Speiseder, Thomas; Badbaran, Anita; Reimer, Rudolph; Indenbirken, Daniela; Grundhoff, Adam; Brunswig-Spickenheier, Bärbel; Alawi, Malik; Lange, Claudia

The biological relevance of extracellular vesicles (EV) in intercellular communication has been well established. Thus far, proteins and RNA were described as main cargo. Here, we show that EV released from human bone marrow derived mesenchymal stromal cells (BM-hMSC) also carry high-molecular DNA in addition. Extensive EV characterization revealed this DNA mainly associated with the outer EV membrane and to a smaller degree also inside the EV. Our EV purification protocol secured that DNA is not derived from apoptotic or necrotic cells. To analyze the relevance of EV-associated DNA we lentivirally transduced Arabidopsis thaliana-DNA (A.t.-DNA) as indicator into BM-hMSC and generated EV. Using quantitative polymerase chain reaction (qPCR) techniques we detected high copy numbers of A.t.-DNA in EV. In recipient hMSC incubated with tagged EV for two weeks we identified A.t.-DNA transferred to recipient cells. Investigation of recipient cell DNA using quantitative PCR and verification of PCR-products by sequencing suggested stable integration of A.t.-DNA. In conclusion, for the first time our proof-of-principle experiments point to horizontal DNA transfer into recipient cells via EV. Based on our results we assume that eukaryotic cells are able to exchange genetic information in form of DNA extending the known cargo of EV by genomic DNA. This mechanism might be of relevance in cancer but also during cell evolution and development.

Circulation of multiple serotypes of highly divergent enterovirus C in the Xinjiang Uighur Autonomous Region of China

PubMed Central

Zhang, Yong; Sun, Qiang; Cui, Hui; Yan, Dongmei; Fan, Qin; Song, Yang; Zhu, Shuangli; Li, Xiaolei; Huang, Guohong; Ji, Tianjiao; Hu, Lan; Wang, Dongyan; Yang, Qian; Xu, Wenbo

2016-01-01

Poliomyelitis associated with circulating vaccine-derived polioviruses (cVDPVs) is a serious public health issue in the post-eradication era, and the occurrence of recombinant cVDPVs emphasizes the need to elucidate enterovirus C (EV-C) epidemiology. Stool samples were collected from 826 healthy children in Southern Xinjiang in 2011 to investigate EV-C circulation and epidemiology. Thirty-six EV-Cs were isolated and assigned to eight EV-C serotypes by molecular serotyping, suggesting the circulation of diverse EV-Cs in Xinjiang. Phylogenetic analysis showed that the Xinjiang EV-C strains had larger variation compared to the prototype and other modern strains. Additionally, the results showed unique characteristics of Xinjiang EV-Cs, such as the cytopathicity of CV-A1 strains to RD cells; the high divergence in CV-A11, CV-A13, CV-A17, and CV-A20 strains; the divergence of Xinjiang CV-A24 from AHC-related CV-A24 variant stains distributed worldwide; and the circulation of two novel EV-C serotypes (EV-C96 and EV-C99). Evaluations of this dense and diverse EV-C ecosystem will help elucidate the processes shaping enteroviral biodiversity. This study will improve our understanding of the evolution of enteroviruses and the recombination potential between polioviruses and other EV-Cs. PMID:27642136

The effect of storage temperature on the biological activity of extracellular vesicles for the complement system.

PubMed

Park, Sang June; Jeon, Hyungtaek; Yoo, Seung-Min; Lee, Myung-Shin

2018-05-10

Extracellular vesicles (EVs) are mediators of intercellular communication by transporting cargo containing proteins, lipids, mRNA, and miRNA. There is increasing evidence that EVs have various roles in regulating migration, invasion, stemness, survival, and immune functions. Previously, we have found that EVs from Kaposi's sarcoma-associated herpesvirus (KSHV)-infected human endothelial cells have the potential to activate the complement system. Although many studies have shown that the physical properties of EVs can be changed by their storage condition, there have been few studies for the stability of biological activity of EVs in various storage conditions. In this study, we investigated various conditions to identify the best conditions to store EVs with functional stability for 25 d. Furthermore, the correlation between the function and other characteristics of EVs, including the expression of EV markers, size distribution, and particle number, were also analyzed. Our results demonstrated that storage temperature is an important factor to maintain the activity of EVs and would be useful information for basic research and clinical application using EVs.

ACHIEVING THE PROMISE OF THERAPEUTIC EXTRACELLULAR VESICLES: THE DEVIL IS IN DETAILS OF THERAPEUTIC LOADING

PubMed Central

Sutaria, Dhruvitkumar S.; Badawi, Mohamed; Phelps, Mitch A.; Schmittgen, Thomas D.

2017-01-01

Extracellular vesicles (EVs) represent a class of cell secreted organelles which naturally contain biomolecular cargo such as miRNA, mRNA and proteins. EVs mediate intercellular communication, enabling the transfer of functional nucleic acids from the cell of origin to the recipient cells. In addition, EVs make an attractive delivery vehicle for therapeutics owing to their increased stability in circulation, biocompatibility, low immunogenicity and toxicity profiles. EVs can also be engineered to display targeting moieties on their surfaces which enables targeting to desired tissues, organs or cells. While much has been learned on the role of EVs as cell communicators, the field of therapeutic EV application is currently under development. Critical to the future success of EV delivery system is the description of methods by which therapeutics can be successfully and efficiently loaded within the EVs. Two methods of loading of EVs with therapeutic cargo exist, endogenous and exogenous loading. We have therefore focused this review on describing the various published approaches for loading EVs with therapeutics. PMID:28315083

PREVALENCE OF HUMAN ENTEROVIRUS AMONG PATIENTS WITH HAND, FOOT, AND MOUTH DISEASE AND HERPANGINA IN THAILAND, 2013.

PubMed

Mauleekoonphairoj, John; Puenpa, Jiratchaya; Korkong, Sumeth; Vongpunsawad, Sompong; Poovorawan, Yong

2015-11-01

Human enterovirus (EV) infection causes hand, foot, and mouth disease (HFMD) and herpangina (HA). We studied the prevalence of enterovirus (EV) among patients with HFMD and HA in Thailand during 2013. We conducted a study in archived specimens of patients sent for screening for enterovirus. A total of 203 clinical specimens from 184 individuals with painful blister in the oropharynx and on the palms, soles, knees, elbows or buttock were examined by semi-nested polymerase chain reaction (PCR) for the 5'UTR and VP1 genes of EV. Eighty-six samples were positive: EV71 was detected in 14 (30%), CV-A8 in 12 (26%) and CV-A16 in 10 (21%). Classification of EV species detected revealed that 46 specimens were EV-A, 14 specimens were EV-B, 1 specimen was EV-D, and 16 specimens were positive for unclassified enterovirus. The majority of individuals with EV infection were aged 2-6 years. Multiple EV-A serotypes were detected among HFMD and HA patients in our study.

Heparin affinity purification of extracellular vesicles

PubMed Central

Balaj, Leonora; Atai, Nadia A.; Chen, Weilin; Mu, Dakai; Tannous, Bakhos A.; Breakefield, Xandra O.; Skog, Johan; Maguire, Casey A.

2015-01-01

Extracellular vesicles (EVs) are lipid membrane vesicles released by cells. They carry active biomolecules including DNA, RNA, and protein which can be transferred to recipient cells. Isolation and purification of EVs from culture cell media and biofluids is still a major challenge. The most widely used isolation method is ultracentrifugation (UC) which requires expensive equipment and only partially purifies EVs. Previously we have shown that heparin blocks EV uptake in cells, supporting a direct EV-heparin interaction. Here we show that EVs can be purified from cell culture media and human plasma using ultrafiltration (UF) followed by heparin-affinity beads. UF/heparin-purified EVs from cell culture displayed the EV marker Alix, contained a diverse RNA profile, had lower levels of protein contamination, and were functional at binding to and uptake into cells. RNA yield was similar for EVs isolated by UC. We were able to detect mRNAs in plasma samples with comparable levels to UC samples. In conclusion, we have discovered a simple, scalable, and effective method to purify EVs taking advantage of their heparin affinity. PMID:25988257

Methods to isolate extracellular vesicles for diagnosis

NASA Astrophysics Data System (ADS)

Kang, Hyejin; Kim, Jiyoon; Park, Jaesung

2017-12-01

Extracellular vesicles (EVs) are small membrane-bound bodies that are released into extracellular space by diverse cells, and are found in body fluids like blood, urine and saliva. EVs contain RNA, DNA and proteins, which can be biomarkers for diagnosis. EVs can be obtained by minimally-invasive biopsy, so they are useful in disease diagnosis. High yield and purity contribute to precise diagnosis of disease, but damaged EVs and impurities can cause confu sed results. However, EV isolation methods have different yields and purities. Furthermore, the isolation method that is most suitable to maximize EV recovery efficiency depends on the experimental conditions. This review focuses on merits and demerits of several types of EV isolation methods, and provides examples of how to diagnose disease by exploiting information obtained by analysis of EVs.

Comprehensive Proteomic Analysis of Human Milk-derived Extracellular Vesicles Unveils a Novel Functional Proteome Distinct from Other Milk Components*

PubMed Central

van Herwijnen, Martijn J.C.; Zonneveld, Marijke I.; Goerdayal, Soenita; Nolte – 't Hoen, Esther N.M.; Garssen, Johan; Stahl, Bernd; Maarten Altelaar, A.F.; Redegeld, Frank A.; Wauben, Marca H.M.

2016-01-01

Breast milk contains several macromolecular components with distinctive functions, whereby milk fat globules and casein micelles mainly provide nutrition to the newborn, and whey contains molecules that can stimulate the newborn's developing immune system and gastrointestinal tract. Although extracellular vesicles (EV) have been identified in breast milk, their physiological function and composition has not been addressed in detail. EV are submicron sized vehicles released by cells for intercellular communication via selectively incorporated lipids, nucleic acids, and proteins. Because of the difficulty in separating EV from other milk components, an in-depth analysis of the proteome of human milk-derived EV is lacking. In this study, an extensive LC-MS/MS proteomic analysis was performed of EV that had been purified from breast milk of seven individual donors using a recently established, optimized density-gradient-based EV isolation protocol. A total of 1963 proteins were identified in milk-derived EV, including EV-associated proteins like CD9, Annexin A5, and Flotillin-1, with a remarkable overlap between the different donors. Interestingly, 198 of the identified proteins are not present in the human EV database Vesiclepedia, indicating that milk-derived EV harbor proteins not yet identified in EV of different origin. Similarly, the proteome of milk-derived EV was compared with that of other milk components. For this, data from 38 published milk proteomic studies were combined in order to construct the total milk proteome, which consists of 2698 unique proteins. Remarkably, 633 proteins identified in milk-derived EV have not yet been identified in human milk to date. Interestingly, these novel proteins include proteins involved in regulation of cell growth and controlling inflammatory signaling pathways, suggesting that milk-derived EVs could support the newborn's developing gastrointestinal tract and immune system. Overall, this study provides an expansion of the whole milk proteome and illustrates that milk-derived EV are macromolecular components with a unique functional proteome. PMID:27601599

Apigenin Inhibits Enterovirus-71 Infection by Disrupting Viral RNA Association with trans-Acting Factors

PubMed Central

Zhang, Wei; Qiao, Haishi; Lv, Yuanzi; Wang, Jingjing; Chen, Xiaoqing; Hou, Yayi; Tan, Renxiang; Li, Erguang

2014-01-01

Flavonoids are widely distributed natural products with broad biological activities. Apigenin is a dietary flavonoid that has recently been demonstrated to interact with heterogeneous nuclear ribonucleoproteins (hnRNPs) and interferes with their RNA editing activity. We investigated whether apigenin possessed antiviral activity against enterovirus-71 (EV71) infection since EV71 infection requires of hnRNP proteins. We found that apigenin selectively blocks EV71 infection by disrupting viral RNA association with hnRNP A1 and A2 proteins. The estimated EC50 value for apigenin to block EV71 infection was determined at 10.3 µM, while the CC50 was estimated at 79.0 µM. The anti-EV71 activity was selective since no activity was detected against several DNA and RNA viruses. Although flavonoids in general share similar structural features, apigenin and kaempferol were among tested compounds with significant activity against EV71 infection. hnRNP proteins function as trans-acting factors regulating EV71 translation. We found that apigenin treatment did not affect EV71-induced nucleocytoplasmic redistribution of hnRNP A1 and A2 proteins. Instead, it prevented EV71 RNA association with hnRNP A1 and A2 proteins. Accordingly, suppression of hnRNP A1 and A2 expression markedly reduced EV71 infection. As a positive sense, single strand RNA virus, EV71 has a type I internal ribosome entry site (IRES) that cooperates with host factors and regulates EV71 translation. The effect of apigenin on EV71 infection was further demonstrated using a bicistronic vector that has the expression of a GFP protein under the control of EV71 5′-UTR. We found that apigenin treatment selectively suppressed the expression of GFP, but not a control gene. In addition to identification of apigenin as an antiviral agent against EV71 infection, this study also exemplifies the significance in antiviral agent discovery by targeting host factors essential for viral replication. PMID:25330384

Innovative Approaches to Remote Sensing in NASA's Earth System Science Pathfinder (ESSP) Program

NASA Technical Reports Server (NTRS)

Peri, Frank; Volz, Stephen

2013-01-01

NASA's Earth Venture class (EV) of mission are competitively selected, Principal Investigator (PI) led, relatively low cost and narrowly focused in scientific scope. Investigations address a full spectrum of earth science objectives, including studies of the atmosphere, oceans, land surface, polar ice regions, and solid Earth. EV has three program elements: EV-Suborbital (EVS) are suborbital/airborne investigations; EV-Mission (EVM) element comprises small complete spaceborne missions; and EV-Instrument (EVI) element develops spaceborne instruments for flight as missions-of-opportunity (MoO). To ensure the success of EV, the management approach of each element is tailored according to the specific needs of the element.

Surface Glycosylation Profiles of Urine Extracellular Vesicles

PubMed Central

Gerlach, Jared Q.; Krüger, Anja; Gallogly, Susan; Hanley, Shirley A.; Hogan, Marie C.; Ward, Christopher J.

2013-01-01

Urinary extracellular vesicles (uEVs) are released by cells throughout the nephron and contain biomolecules from their cells of origin. Although uEV-associated proteins and RNA have been studied in detail, little information exists regarding uEV glycosylation characteristics. Surface glycosylation profiling by flow cytometry and lectin microarray was applied to uEVs enriched from urine of healthy adults by ultracentrifugation and centrifugal filtration. The carbohydrate specificity of lectin microarray profiles was confirmed by competitive sugar inhibition and carbohydrate-specific enzyme hydrolysis. Glycosylation profiles of uEVs and purified Tamm Horsfall protein were compared. In both flow cytometry and lectin microarray assays, uEVs demonstrated surface binding, at low to moderate intensities, of a broad range of lectins whether prepared by ultracentrifugation or centrifugal filtration. In general, ultracentrifugation-prepared uEVs demonstrated higher lectin binding intensities than centrifugal filtration-prepared uEVs consistent with lesser amounts of co-purified non-vesicular proteins. The surface glycosylation profiles of uEVs showed little inter-individual variation and were distinct from those of Tamm Horsfall protein, which bound a limited number of lectins. In a pilot study, lectin microarray was used to compare uEVs from individuals with autosomal dominant polycystic kidney disease to those of age-matched controls. The lectin microarray profiles of polycystic kidney disease and healthy uEVs showed differences in binding intensity of 6/43 lectins. Our results reveal a complex surface glycosylation profile of uEVs that is accessible to lectin-based analysis following multiple uEV enrichment techniques, is distinct from co-purified Tamm Horsfall protein and may demonstrate disease-specific modifications. PMID:24069349

Obstacles and opportunities in the functional analysis of extracellular vesicle RNA – an ISEV position paper

PubMed Central

Mateescu, Bogdan; Kowal, Emma J. K.; van Balkom, Bas W. M.; Bartel, Sabine; Bhattacharyya, Suvendra N.; Buzás, Edit I.; Buck, Amy H.; de Candia, Paola; Chow, Franklin W. N.; Das, Saumya; Driedonks, Tom A. P.; Fernández-Messina, Lola; Haderk, Franziska; Hill, Andrew F.; Jones, Jennifer C.; Van Keuren-Jensen, Kendall R.; Lai, Charles P.; Lässer, Cecilia; Liegro, Italia di; Lunavat, Taral R.; Lorenowicz, Magdalena J.; Maas, Sybren L. N.; Mäger, Imre; Mittelbrunn, Maria; Momma, Stefan; Mukherjee, Kamalika; Nawaz, Muhammed; Pegtel, D. Michiel; Pfaffl, Michael W.; Schiffelers, Raymond M.; Tahara, Hidetoshi; Théry, Clotilde; Tosar, Juan Pablo; Wauben, Marca H. M.; Witwer, Kenneth W.; Nolte-‘t Hoen, Esther N. M.

2017-01-01

ABSTRACT The release of RNA-containing extracellular vesicles (EV) into the extracellular milieu has been demonstrated in a multitude of different in vitro cell systems and in a variety of body fluids. RNA-containing EV are in the limelight for their capacity to communicate genetically encoded messages to other cells, their suitability as candidate biomarkers for diseases, and their use as therapeutic agents. Although EV-RNA has attracted enormous interest from basic researchers, clinicians, and industry, we currently have limited knowledge on which mechanisms drive and regulate RNA incorporation into EV and on how RNA-encoded messages affect signalling processes in EV-targeted cells. Moreover, EV-RNA research faces various technical challenges, such as standardisation of EV isolation methods, optimisation of methodologies to isolate and characterise minute quantities of RNA found in EV, and development of approaches to demonstrate functional transfer of EV-RNA in vivo. These topics were discussed at the 2015 EV-RNA workshop of the International Society for Extracellular Vesicles. This position paper was written by the participants of the workshop not only to give an overview of the current state of knowledge in the field, but also to clarify that our incomplete knowledge – of the nature of EV(-RNA)s and of how to effectively and reliably study them – currently prohibits the implementation of gold standards in EV-RNA research. In addition, this paper creates awareness of possibilities and limitations of currently used strategies to investigate EV-RNA and calls for caution in interpretation of the obtained data. PMID:28326170

Identification of specific antigenic epitope at N-terminal segment of enterovirus 71 (EV-71) VP1 protein and characterization of its use in recombinant form for early diagnosis of EV-71 infection.

PubMed

Zhang, Jianhua; Jiang, Bingfu; Xu, Mingjie; Dai, Xing; Purdy, Michael A; Meng, Jihong

2014-08-30

Human enterovirus 71 (EV-71) is the main etiologic agent of hand, foot and mouth disease (HFMD). We sought to identify EV-71 specific antigens and develop serologic assays for acute-phase EV-71 infection. A series of truncated proteins within the N-terminal 100 amino acids (aa) of EV-71 VP1 was expressed in Escherichia coli. Western blot (WB) analysis showed that positions around 11-21 aa contain EV-71-specific antigenic sites, whereas positions 1-5 and 51-100 contain epitopes shared with human coxsackievirus A16 (CV-A16) and human echovirus 6 (E-6). The N-terminal truncated protein of VP1, VP₁₆₋₄₃, exhibited good stability and was recognized by anti-EV-71 specific rabbit sera. Alignment analysis showed that VP₁₆₋₄₃ is highly conserved among EV-71 strains from different genotypes but was heterologous among other enteroviruses. When the GST-VP₁₆₋₄₃ fusion protein was incorporated as antibody-capture agent in a WB assay and an ELISA for detecting anti-EV-71 IgM in human sera, sensitivities of 91.7% and 77.8% were achieved, respectively, with 100% specificity for both. The characterized EV-71 VP1 protein truncated to positions 6-43 aa has potential as an antigen for detection of anti-EV-71 IgM for early diagnosis of EV-71 infection in a WB format. Copyright © 2014 Elsevier B.V. All rights reserved.

Localized excitons in fluoroperovskite LiBaF3 crystals

NASA Astrophysics Data System (ADS)

Springis, Maris; Trukhin, Anatoly N.; Tale, Ivar

2003-08-01

Two radiating processes in LiBaF3 crystals, fast valence-core transitions (5.4 - 6.5 eV) and slow, so called self-trapped exciton luminescence (about 4.3 eV), are important for practical application. Here we present a study of 4.3 eV luminescence under X-ray excitation and photoexcitation as well as under photostimulation after X-irradiation of undoped and Ag-doped LiBaF3 crystals at various temperatures. It is shown that 4.3 eV luminescence appears under X-ray excitation at least from 85 K to 400 K in both undoped and doped crystals. In all samples studied the excitation spectra of 4.3 eV luminescence contain both the main exciton like band at the edge of fundamental absorption at about 10 eV and weaker band in 7.8 - 8.6 eV region. Luminescence spectrum in the 3.8 - 4.8 eV region under 7.8 - 8.6 eV excitation differs slightly from that under 10 eV excitation. Several luminescence bands in 3.8 - 4.8 eV region arises in the temperature range 85 - 230 K under photostimulation in absorption band of F-type center at 2.9 eV created previously under X-irradiation. We propose the luminescence of LiBaF3 crystals in the 3.8 - 4.8 eV region may be caused by localized excitons formed not only under excitation near the fundamental absorption but also in result of electron recombination with localized holes thermally destroyed above 230 K.

Obstacles and opportunities in the functional analysis of extracellular vesicle RNA - an ISEV position paper.

PubMed

Mateescu, Bogdan; Kowal, Emma J K; van Balkom, Bas W M; Bartel, Sabine; Bhattacharyya, Suvendra N; Buzás, Edit I; Buck, Amy H; de Candia, Paola; Chow, Franklin W N; Das, Saumya; Driedonks, Tom A P; Fernández-Messina, Lola; Haderk, Franziska; Hill, Andrew F; Jones, Jennifer C; Van Keuren-Jensen, Kendall R; Lai, Charles P; Lässer, Cecilia; Liegro, Italia di; Lunavat, Taral R; Lorenowicz, Magdalena J; Maas, Sybren L N; Mäger, Imre; Mittelbrunn, Maria; Momma, Stefan; Mukherjee, Kamalika; Nawaz, Muhammed; Pegtel, D Michiel; Pfaffl, Michael W; Schiffelers, Raymond M; Tahara, Hidetoshi; Théry, Clotilde; Tosar, Juan Pablo; Wauben, Marca H M; Witwer, Kenneth W; Nolte-'t Hoen, Esther N M

2017-01-01

The release of RNA-containing extracellular vesicles (EV) into the extracellular milieu has been demonstrated in a multitude of different in vitro cell systems and in a variety of body fluids. RNA-containing EV are in the limelight for their capacity to communicate genetically encoded messages to other cells, their suitability as candidate biomarkers for diseases, and their use as therapeutic agents. Although EV-RNA has attracted enormous interest from basic researchers, clinicians, and industry, we currently have limited knowledge on which mechanisms drive and regulate RNA incorporation into EV and on how RNA-encoded messages affect signalling processes in EV-targeted cells. Moreover, EV-RNA research faces various technical challenges, such as standardisation of EV isolation methods, optimisation of methodologies to isolate and characterise minute quantities of RNA found in EV, and development of approaches to demonstrate functional transfer of EV-RNA in vivo . These topics were discussed at the 2015 EV-RNA workshop of the International Society for Extracellular Vesicles. This position paper was written by the participants of the workshop not only to give an overview of the current state of knowledge in the field, but also to clarify that our incomplete knowledge - of the nature of EV(-RNA)s and of how to effectively and reliably study them - currently prohibits the implementation of gold standards in EV-RNA research. In addition, this paper creates awareness of possibilities and limitations of currently used strategies to investigate EV-RNA and calls for caution in interpretation of the obtained data.

A reverse genetics system for enterovirus D68 using human RNA polymerase I.

PubMed

Pan, Minglei; Gao, Shuai; Zhou, Zhenwei; Zhang, Keke; Liu, Sihua; Wang, Zhiyun; Wang, Tao

2018-05-17

Human enterovirus D68 (EV-D68) is a highly contagious virus, which causes respiratory tract infections. However, no effective vaccines are currently available for controlling EV-D68 infection. Here, we developed a reverse genetics system to recover EV-D68 minireplicons and infectious EV-D68 from transfected plasmids using the RNA polymerase I (Pol I) promoter. The EV-D68 minireplicons contained the luciferase reporter gene, which flanked by the non-coding regions of the EV-D68 RNA. The luciferase signals could be detected in cells after transfection and Pol I promoter-mediated luciferase signal was significantly stronger than that mediated by the T7 promoter. Furthermore, recombinant viruses were generated by transfecting plasmids that contained the genomic RNA segments of EV-D68, under the control of Pol I promoter into 293T cells or RD cells. On plaque morphology and growth kinetics, the rescued virus and parental virus were indistinguishable. In addition, we showed that the G394C mutation disrupts the viral 5'-UTR structure and suppresses the viral cap-independent translation. This reverse genetics system for EV-D68 recovery can greatly facilitate research into EV-D68 biology. Moreover, this system could accelerate the development of EV-D68 vaccines and anti-EV-D68 drugs.

TLR3 Ligand Poly(I:C) Exerts Distinct Actions in Synovial Fibroblasts When Delivered by Extracellular Vesicles

PubMed Central

Frank-Bertoncelj, Mojca; Pisetsky, David S.; Kolling, Christoph; Michel, Beat A.; Gay, Renate E.; Jüngel, Astrid; Gay, Steffen

2018-01-01

Extracellular vesicles (EV) can modulate the responses of cells to toll-like receptor (TLR) ligation; conversely, TLR ligands such as double-stranded RNA (dsRNA) can enhance the release of EV and influence of the composition and functions of EV cargos. Inflamed synovial joints in rheumatoid arthritis (RA) are rich in EV and extracellular RNA; besides, RNA released from necrotic synovial fluid cells can activate the TLR3 signaling in synovial fibroblasts (SFs) from patients with RA. Since EV occur prominently in synovial joints in RA and may contribute to the pathogenesis, we questioned whether EV can interact with dsRNA, a TLR3 ligand, and modify its actions in arthritis. We have used as model the effects on RA SFs, of EV released from monocyte U937 cells and peripheral blood mononuclear cells upon stimulation with Poly(I:C), a synthetic analog of dsRNA. We show that EV released from unstimulated cells and Poly(I:C)-stimulated U937 cells [Poly(I:C) EV] differ in size but bind similar amounts of Annexin V and express comparable levels of MAC-1, the receptor for dsRNA, on the vesicular membranes. Specifically, Poly(I:C) EV contain or associate with Poly(I:C) and at least partially protect Poly(I:C) from RNAse III degradation. Poly(I:C) EV shuttle Poly(I:C) to SFs and reproduce the proinflammatory and antiviral gene responses of SFs to direct stimulation with Poly(I:C). Poly(I:C) EV, however, halt the death receptor-induced apoptosis in SFs, thereby inverting the proapoptotic nature of Poly(I:C). These prosurvival effects sharply contrast with the high toxicity of cationic liposome-delivered Poly(I:C) and may reflect the route of Poly(I:C) delivery via EV or the fine-tuning of Poly(I:C) actions by molecular cargo in EV. The demonstration that EV may safeguard extracellular dsRNA and allow dsRNA to exert antiapoptotic effects on SFs highlights the potential of EV to amplify the pathogenicity of dsRNA in arthritis beyond inflammation (by concurrently enhancing the expansion of the invasive synovial stroma). PMID:29434584

Antiviral effects of two Ganoderma lucidum triterpenoids against enterovirus 71 infection

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

Zhang, Wenjing; Tao, Junyan; Yang, Xiaoping

Highlights: • Triterpenoids GLTA and GLTB display anti-EV71 activities without cytotoxicity. • The compounds prevent EV71 infection by blocking adsorption of the virus to the cells. • GLTA and GLTB bind to EV71 capsid at the hydrophobic pocket to block EV71 uncoating. • The two compounds significantly inhibit the replication of EV71 viral RNA. • GLTA and GLTB may be used as potential therapeutic agents to treat EV71 infection. - Abstract: Enterovirus 71 (EV71) is a major causative agent for hand, foot and mouth disease (HFMD), and fatal neurological and systemic complications in children. However, there is currently no clinicalmore » approved antiviral drug available for the prevention and treatment of the viral infection. Here, we evaluated the antiviral activities of two Ganoderma lucidum triterpenoids (GLTs), Lanosta-7,9(11),24-trien-3-one,15;26-dihydroxy (GLTA) and Ganoderic acid Y (GLTB), against EV71 infection. The results showed that the two natural compounds display significant anti-EV71 activities without cytotoxicity in human rhabdomyosarcoma (RD) cells as evaluated by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cell proliferation assay. The mechanisms by which the two compounds affect EV71 infection were further elucidated by three action modes using Ribavirin, a common antiviral drug, as a positive control. The results suggested that GLTA and GLTB prevent EV71 infection through interacting with the viral particle to block the adsorption of virus to the cells. In addition, the interactions between EV71 virion and the compounds were predicated by computer molecular docking, which illustrated that GLTA and GLTB may bind to the viral capsid protein at a hydrophobic pocket (F site), and thus may block uncoating of EV71. Moreover, we demonstrated that GLTA and GLTB significantly inhibit the replication of the viral RNA (vRNA) of EV71 replication through blocking EV71 uncoating. Thus, GLTA and GLTB may represent two potential therapeutic agents to control and treat EV71 infection.« less

Alternative Fuels Data Center

Science.gov Websites

Colorado Electric Vehicle (EV) Plan The Colorado Energy Office, Regional Air Quality Council Colorado EV Plan (Plan). The Plan calls for Colorado to be a leader in the EV market and accelerate the requirements to include EV fast-charging, and ensuring economic and tourism benefits of EV charging. The Plan

Visualization and tracking of tumour extracellular vesicle delivery and RNA translation using multiplexed reporters

PubMed Central

Lai, Charles P.; Kim, Edward Y.; Badr, Christian E.; Weissleder, Ralph; Mempel, Thorsten R.; Tannous, Bakhos A.; Breakefield, Xandra O.

2015-01-01

Accurate spatiotemporal assessment of extracellular vesicle (EV) delivery and cargo RNA translation requires specific and robust live-cell imaging technologies. Here we engineer optical reporters to label multiple EV populations for visualization and tracking of tumour EV release, uptake and exchange between cell populations both in culture and in vivo. Enhanced green fluorescence protein (EGFP) and tandem dimer Tomato (tdTomato) were fused at NH2-termini with a palmitoylation signal (PalmGFP, PalmtdTomato) for EV membrane labelling. To monitor EV-RNA cargo, transcripts encoding PalmtdTomato were tagged with MS2 RNA binding sequences and detected by co-expression of bacteriophage MS2 coat protein fused with EGFP. By multiplexing fluorescent and bioluminescent EV membrane reporters, we reveal the rapid dynamics of both EV uptake and translation of EV-delivered cargo mRNAs in cancer cells that occurred within 1-hour post-horizontal transfer between cells. These studies confirm that EV-mediated communication is dynamic and multidirectional between cells with delivery of functional mRNA. PMID:25967391

Optimal Coordinated EV Charging with Reactive Power Support in Constrained Distribution Grids

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

Paudyal, Sumit; Ceylan, Oğuzhan; Bhattarai, Bishnu P.

Electric vehicle (EV) charging/discharging can take place in any P-Q quadrants, which means EVs could support reactive power to the grid while charging the battery. In controlled charging schemes, distribution system operator (DSO) coordinates with the charging of EV fleets to ensure grid’s operating constraints are not violated. In fact, this refers to DSO setting upper bounds on power limits for EV charging. In this work, we demonstrate that if EVs inject reactive power into the grid while charging, DSO could issue higher upper bounds on the active power limits for the EVs for the same set of grid constraints.more » We demonstrate the concept in an 33-node test feeder with 1,500 EVs. Case studies show that in constrained distribution grids in coordinated charging, average costs of EV charging could be reduced if the charging takes place in the fourth P-Q quadrant compared to charging with unity power factor.« less

Extracellular Vesicles in Cardiovascular Theranostics

PubMed Central

Bei, Yihua; Das, Saumya; Rodosthenous, Rodosthenis S.; Holvoet, Paul; Vanhaverbeke, Maarten; Monteiro, Marta Chagas; Monteiro, Valter Vinicius Silva; Radosinska, Jana; Bartekova, Monika; Jansen, Felix; Li, Qian; Rajasingh, Johnson; Xiao, Junjie

2017-01-01

Extracellular vesicles (EVs) are small bilayer lipid membrane vesicles that can be released by most cell types and detected in most body fluids. EVs exert key functions for intercellular communication via transferring their bioactive cargos to recipient cells or activating signaling pathways in target cells. Increasing evidence has shown the important regulatory effects of EVs in cardiovascular diseases (CVDs). EVs secreted by cardiomyocytes, endothelial cells, fibroblasts, and stem cells play essential roles in pathophysiological processes such as cardiac hypertrophy, cardiomyocyte survival and apoptosis, cardiac fibrosis, and angiogenesis in relation to CVDs. In this review, we will first outline the current knowledge about the physical characteristics, biological contents, and isolation methods of EVs. We will then focus on the functional roles of cardiovascular EVs and their pathophysiological effects in CVDs, as well as summarize the potential of EVs as therapeutic agents and biomarkers for CVDs. Finally, we will discuss the specific application of EVs as a novel drug delivery system and the utility of EVs in the field of regenerative medicine. PMID:29158817

Flow analysis of individual blood extracellular vesicles in acute coronary syndrome.

PubMed

Vagida, Murad; Arakelyan, Anush; Lebedeva, Anna; Grivel, Jean-Charles; Shpektor, Alexander; Vasilieva, Elena; Margolis, Leonid

2017-03-01

A diverse population of small extracellular vesicles (EVs) that are released by various cells has been characterized predominantly in bulk, a procedure whereby the individual characteristics of EVs are lost. Here, we used a new nanotechnology-based flow cytometric analysis to characterize the antigenic composition of individual EVs in patients with acute coronary syndrome (ACS). Plasma EVs were captured with 15-nm magnetic nanoparticles coupled to antibodies against CD31 (predominantly an endothelial marker), CD41a (a marker for platelets), and CD63 or MHC class I (common EV markers). The total amounts of EVs were higher in the ACS patients than in the controls, predominantly due to the contribution of patients with acute myocardial infarction. For all captured fractions, the differences in the EV amounts were restricted to CD41a + EVs. The increase in the numbers of EVs in the ACS patients, predominantly of platelet origin, probably reflects platelet activation and may indicate disease progression.

Extracellular Vesicles Released from Mycobacterium tuberculosis-Infected Neutrophils Promote Macrophage Autophagy and Decrease Intracellular Mycobacterial Survival

PubMed Central

Alvarez-Jiménez, Violeta D.; Leyva-Paredes, Kahiry; García-Martínez, Mariano; Vázquez-Flores, Luis; García-Paredes, Víctor Gabriel; Campillo-Navarro, Marcia; Romo-Cruz, Israel; Rosales-García, Víctor Hugo; Castañeda-Casimiro, Jessica; González-Pozos, Sirenia; Hernández, José Manuel; Wong-Baeza, Carlos; García-Pérez, Blanca Estela; Ortiz-Navarrete, Vianney; Estrada-Parra, Sergio; Serafín-López, Jeanet; Wong-Baeza, Isabel; Chacón-Salinas, Rommel; Estrada-García, Iris

2018-01-01

Tuberculosis is an infectious disease caused by Mycobacterium tuberculosis (Mtb). In the lungs, macrophages and neutrophils are the first immune cells that have contact with the infecting mycobacteria. Neutrophils are phagocytic cells that kill microorganisms through several mechanisms, which include the lytic enzymes and antimicrobial peptides that are found in their lysosomes, and the production of reactive oxygen species. Neutrophils also release extracellular vesicles (EVs) (100–1,000 nm in diameter) to the extracellular milieu; these EVs consist of a lipid bilayer surrounding a hydrophilic core and participate in intercellular communication. We previously demonstrated that human neutrophils infected in vitro with Mtb H37Rv release EVs (EV-TB), but the effect of these EVs on other cells relevant for the control of Mtb infection, such as macrophages, has not been completely analyzed. In this study, we characterized the EVs produced by non-stimulated human neutrophils (EV-NS), and the EVs produced by neutrophils stimulated with an activator (PMA), a peptide derived from bacterial proteins (fMLF) or Mtb, and observed that the four EVs differed in their size. Ligands for toll-like receptor (TLR) 2/6 were detected in EV-TB, and these EVs favored a modest increase in the expression of the co-stimulatory molecules CD80, a higher expression of CD86, and the production of higher amounts of TNF-α and IL-6, and of lower amounts of TGF-β, in autologous human macrophages, compared with the other EVs. EV-TB reduced the amount of intracellular Mtb in macrophages, and increased superoxide anion production in these cells. TLR2/6 ligation and superoxide anion production are known inducers of autophagy; accordingly, we found that EV-TB induced higher expression of the autophagy-related marker LC3-II in macrophages, and the co-localization of LC3-II with Mtb inside infected macrophages. The intracellular mycobacterial load increased when autophagy was inhibited with wortmannin in these cells. In conclusion, our results demonstrate that neutrophils produce different EVs in response to diverse activators, and that EV-TB activate macrophages and promote the clearance of intracellular Mtb through early superoxide anion production and autophagy induction, which is a novel role for neutrophil-derived EVs in the immune response to Mtb. PMID:29520273

Extracellular Vesicles Released from Mycobacterium tuberculosis-Infected Neutrophils Promote Macrophage Autophagy and Decrease Intracellular Mycobacterial Survival.

PubMed

Alvarez-Jiménez, Violeta D; Leyva-Paredes, Kahiry; García-Martínez, Mariano; Vázquez-Flores, Luis; García-Paredes, Víctor Gabriel; Campillo-Navarro, Marcia; Romo-Cruz, Israel; Rosales-García, Víctor Hugo; Castañeda-Casimiro, Jessica; González-Pozos, Sirenia; Hernández, José Manuel; Wong-Baeza, Carlos; García-Pérez, Blanca Estela; Ortiz-Navarrete, Vianney; Estrada-Parra, Sergio; Serafín-López, Jeanet; Wong-Baeza, Isabel; Chacón-Salinas, Rommel; Estrada-García, Iris

2018-01-01

Tuberculosis is an infectious disease caused by Mycobacterium tuberculosis (Mtb). In the lungs, macrophages and neutrophils are the first immune cells that have contact with the infecting mycobacteria. Neutrophils are phagocytic cells that kill microorganisms through several mechanisms, which include the lytic enzymes and antimicrobial peptides that are found in their lysosomes, and the production of reactive oxygen species. Neutrophils also release extracellular vesicles (EVs) (100-1,000 nm in diameter) to the extracellular milieu; these EVs consist of a lipid bilayer surrounding a hydrophilic core and participate in intercellular communication. We previously demonstrated that human neutrophils infected in vitro with Mtb H37Rv release EVs (EV-TB), but the effect of these EVs on other cells relevant for the control of Mtb infection, such as macrophages, has not been completely analyzed. In this study, we characterized the EVs produced by non-stimulated human neutrophils (EV-NS), and the EVs produced by neutrophils stimulated with an activator (PMA), a peptide derived from bacterial proteins (fMLF) or Mtb, and observed that the four EVs differed in their size. Ligands for toll-like receptor (TLR) 2/6 were detected in EV-TB, and these EVs favored a modest increase in the expression of the co-stimulatory molecules CD80, a higher expression of CD86, and the production of higher amounts of TNF-α and IL-6, and of lower amounts of TGF-β, in autologous human macrophages, compared with the other EVs. EV-TB reduced the amount of intracellular Mtb in macrophages, and increased superoxide anion production in these cells. TLR2/6 ligation and superoxide anion production are known inducers of autophagy; accordingly, we found that EV-TB induced higher expression of the autophagy-related marker LC3-II in macrophages, and the co-localization of LC3-II with Mtb inside infected macrophages. The intracellular mycobacterial load increased when autophagy was inhibited with wortmannin in these cells. In conclusion, our results demonstrate that neutrophils produce different EVs in response to diverse activators, and that EV-TB activate macrophages and promote the clearance of intracellular Mtb through early superoxide anion production and autophagy induction, which is a novel role for neutrophil-derived EVs in the immune response to Mtb.

Nanoplasmonic Quantification of Tumor-derived Extracellular Vesicles in Plasma Microsamples for Diagnosis and Treatment Monitoring

PubMed Central

Liang, Kai; Liu, Fei; Fan, Jia; Sun, Dali; Liu, Chang; Lyon, Christopher J.; Bernard, David W.; Li, Yan; Yokoi, Kenji; Katz, Matthew H.; Koay, Eugene J.; Zhao, Zhen; Hu, Ye

2017-01-01

Tumour-derived extracellular vesicles (EVs) are of increasing interest as a resource of diagnostic biomarkers. However, most EV assays require large samples, are time-consuming, low-throughput and costly, and thus impractical for clinical use. Here, we describe a rapid, ultrasensitive and inexpensive nanoplasmon-enhanced scattering (nPES) assay that directly quantifies tumor-derived EVs from as little as 1 μL of plasma. The assay uses the binding of antibody-conjugated gold nanospheres and nanorods to EVs captured by EV-specific antibodies on a sensor chip to produce a local plasmon effect that enhances tumour-derived EV detection sensitivity and specificity. We identified a pancreatic cancer EV biomarker, ephrin type-A receptor 2 (EphA2), and demonstrate that an nPES assay for EphA2-EVs distinguishes pancreatic cancer patients from pancreatitis patients and healthy subjects. EphA2-EVs were also informative in staging tumour progression and in detecting early responses to neoadjuvant therapy, with better performance than a conventional enzyme-linked immunosorbent assay. The nPES assay can be easily refined for clinical use, and readily adapted for diagnosis and monitoring of other conditions with disease-specific EV biomarkers. PMID:28791195

Isolation of Extracellular Vesicles: General Methodologies and Latest Trends

PubMed Central

Konoshenko, Maria Yu.; Laktionov, Pavel P.

2018-01-01

Background Extracellular vesicles (EVs) play an essential role in the communication between cells and transport of diagnostically significant molecules. A wide diversity of approaches utilizing different biochemical properties of EVs and a lack of accepted protocols make data interpretation very challenging. Scope of Review This review consolidates the data on the classical and state-of-the-art methods for isolation of EVs, including exosomes, highlighting the advantages and disadvantages of each method. Various characteristics of individual methods, including isolation efficiency, EV yield, properties of isolated EVs, and labor consumption are compared. Major Conclusions A mixed population of vesicles is obtained in most studies of EVs for all used isolation methods. The properties of an analyzed sample should be taken into account when planning an experiment aimed at studying and using these vesicles. The problem of adequate EVs isolation methods still remains; it might not be possible to develop a universal EV isolation method but the available protocols can be used towards solving particular types of problems. General Significance With the wide use of EVs for diagnosis and therapy of various diseases the evaluation of existing methods for EV isolation is one of the key problems in modern biology and medicine. PMID:29662902

Seroprevalence of enterovirus 71 and no evidence of crossprotection of enterovirus 71 antibody against the other enteroviruses in kindergarten children in Taipei city.

PubMed

Huang, Wen-Chan; Huang, Li-Min; Kao, Chuan-Liang; Lu, Chun-Yi; Shao, Pei-Lan; Cheng, Ai-Ling; Fan, Tsui-Yien; Chi, Hsin; Chang, Luan-Yin

2012-04-01

Enterovirus 71 (EV71) infection may cause severe neurological and cardiopulmonary complications, especially in preschool children. This study is to investigate the seroprevalence and seroconversion of EV71, and the crossprotection of EV71 antibody against other enteroviruses among kindergarteners. Overall 228 children in a public kindergarten were enrolled during two academic years, 2006 and 2007, in Taipei, Taiwan and we measured their EV71 neutralizing antibody. When the participants had herpangina; hand, foot and mouth disease (HFMD); febrile illness or respiratory symptoms, throat swabs were sampled and processed for viral culture and enterovirus real-time reverse transcriptase polymerase chain reaction (RT-PCR). Questionnaires, completed by the participants' guardians, surveyed the history of allergy and annual incidence of symptoms related to enterovirus infection. Seropositive rates of EV71 were 20% (32/163) in 2006 and 6% (4/65) in 2007. The rate of EV71 seropositivity increased with age (p < 0.01) in 2006 but it did not differ between genders (p = 0.14). No seroconversion was observed from 2006 to 2007. Herpangina occurred in 64% of children with EV71 seropositivity and 48% of those without EV71 antibodies (p = 0.12). Non-71 enterovirus infection, confirmed by viral study, occurred in 53% (19/36) of the EV71-seropositive children and in 53% (102/192) of EV71-seronegative children (p = 0.89). No participants had EV71 infection during the study period. EV71 did not frequently circulate in Taipei City from September 2006 to June 2008. Presence of EV71 neutralizing antibody was not associated with lower incidence of enterovirus infection caused by non-71 serotypes. Copyright © 2011. Published by Elsevier B.V.

Efficient ultrafiltration-based protocol to deplete extracellular vesicles from fetal bovine serum

PubMed Central

Kornilov, Roman; Puhka, Maija; Mannerström, Bettina; Hiidenmaa, Hanna; Peltoniemi, Hilkka; Siljander, Pia; Seppänen-Kaijansinkko, Riitta; Kaur, Sippy

2018-01-01

ABSTRACT Fetal bovine serum (FBS) is the most commonly used supplement in studies involving cell-culture experiments. However, FBS contains large numbers of bovine extracellular vesicles (EVs), which hamper the analyses of secreted EVs from the cell type of preference and, thus, also the downstream analyses. Therefore, a prior elimination of EVs from FBS is crucial. However, the current methods of EV depletion by ultracentrifugation are cumbersome and the commercial alternatives expensive. In this study, our aim was to develop a protocol to completely deplete EVs from FBS, which may have wide applicability in cell-culture applications. We investigated different EV-depleted FBS prepared by our novel ultrafiltration-based protocol, by conventionally used overnight ultracentrifugation, or commercially available depleted FBS, and compared them with regular FBS. All sera were characterized by nanoparticle tracking analysis, electron microscopy, Western blotting and RNA quantification. Next, adipose-tissue mesenchymal stem cells (AT-MSCs) and cancer cells were grown in the media supplemented with the three different EV-depleted FBS and compared with cells grown in regular FBS media to assess the effects on cell proliferation, stress, differentiation and EV production. The novel ultrafiltration-based protocol depleted EVs from FBS clearly more efficiently than ultracentrifugation and commercial methods. Cell proliferation, stress, differentiation and EV production of AT-MSCs and cancer cell lines were similarly maintained in all three EV-depleted FBS media up to 96 h. In summary, our ultrafiltration protocol efficiently depletes EVs, is easy to use and maintains cell growth and metabolism. Since the method is also cost-effective and easy to standardize, it could be used in a wide range of cell-culture applications helping to increase comparability of EV research results between laboratories. PMID:29410778

Enterovirus 71 Uses Cell Surface Heparan Sulfate Glycosaminoglycan as an Attachment Receptor

PubMed Central

Tan, Chee Wah; Poh, Chit Laa; Sam, I-Ching

2013-01-01

Enterovirus 71 (EV-71) infections are usually associated with mild hand, foot, and mouth disease in young children but have been reported to cause severe neurological complications with high mortality rates. To date, four EV-71 receptors have been identified, but inhibition of these receptors by antagonists did not completely abolish EV-71 infection, implying that there is an as yet undiscovered receptor(s). Since EV-71 has a wide range of tissue tropisms, we hypothesize that EV-71 infections may be facilitated by using receptors that are widely expressed in all cell types, such as heparan sulfate. In this study, heparin, polysulfated dextran sulfate, and suramin were found to significantly prevent EV-71 infection. Heparin inhibited infection by all the EV-71 strains tested, including those with a single-passage history. Neutralization of the cell surface anionic charge by polycationic poly-d-lysine and blockage of heparan sulfate by an anti-heparan sulfate peptide also inhibited EV-71 infection. Interference with heparan sulfate biosynthesis either by sodium chlorate treatment or through transient knockdown of N-deacetylase/N-sulfotransferase-1 and exostosin-1 expression reduced EV-71 infection in RD cells. Enzymatic removal of cell surface heparan sulfate by heparinase I/II/III inhibited EV-71 infection. Furthermore, the level of EV-71 attachment to CHO cell lines that are variably deficient in cell surface glycosaminoglycans was significantly lower than that to wild-type CHO cells. Direct binding of EV-71 particles to heparin-Sepharose columns under physiological salt conditions was demonstrated. We conclude that EV-71 infection requires initial binding to heparan sulfate as an attachment receptor. PMID:23097443

Electric vehicle energy management system

NASA Astrophysics Data System (ADS)

Alaoui, Chakib

This thesis investigates and analyzes novel strategies for the optimum energy management of electric vehicles (EVs). These are aimed to maximize the useful life of the EV batteries and make the EV more practical in order to increase its acceptability to market. The first strategy concerns the right choice of the batteries for the EV according to the user's driving habits, which may vary. Tests conducted at the University of Massachusetts Lowell battery lab show that the batteries perform differently from one manufacturer to the other. The second strategy was to investigate the fast chargeability of different batteries, which leads to reduce the time needed to recharge the EV battery pack. Tests were conducted again to prove that only few battery types could be fast charged. Test data were used to design a fast battery charger that could be installed in an EV charging station. The third strategy was the design, fabrication and application of an Electric Vehicle Diagnostic and Rejuvenation System (EVDRS). This system is based on Mosfet Controlled Thyristors (MCTs). It is capable of quickly identifying any failing battery(s) within the EV pack and rejuvenating the whole battery pack without dismantling them and unloading them. A novel algorithm to rejuvenate Electric Vehicle Sealed Lead Acid Batteries is described. This rejuvenation extends the useful life of the batteries and makes the EV more competitive. The fourth strategy was to design a thermal management system for EV, which is crucial to the safe operation, and the achievement of normal/optimal performance of, electric vehicle (EV) batteries. A novel approach for EV thermal management, based on Pettier-Effect heat pumps, was designed, fabricated and tested in EV. It shows the application of this type of technology for thermal management of EVs.

3C Protease of Enterovirus D68 Inhibits Cellular Defense Mediated by Interferon Regulatory Factor 7

PubMed Central

Xiang, Zichun; Liu, Lulu; Lei, Xiaobo; Zhou, Zhuo

2015-01-01

ABSTRACT Human enterovirus 68 (EV-D68) is a member of the EV-D species, which belongs to the EV genus of the Picornaviridae family. Over the past several years, clusters of EV-D68 infections have occurred worldwide. A recent outbreak in the United States is the largest one associated with severe respiratory illness and neurological complication. Although clinical symptoms are recognized, the virus remains poorly understood. Here we report that EV-D68 inhibits innate antiviral immunity by downregulation of interferon regulatory factor 7 (IRF7), an immune factor with a pivotal role in viral pathogenesis. This process depends on 3Cpro, an EV-D68-encoded protease, to mediate IRF7 cleavage. When expressed in host cells, 3Cpro targets Q167 and Q189 within the constitutive activation domain, resulting in cleavage of IRF7. Accordingly, wild-type IRF7 is fully active. However, IRF7 cleavage abrogated its capacity to activate type I interferon expression and limit replication of EV-D68. Notably, IRF7 cleavage strictly requires the protease activity of 3Cpro. Together, these results suggest that a dynamic interplay between 3Cpro and IRF7 may determine the outcome of EV-D68 infection. IMPORTANCE EV-D68 is a globally emerging pathogen, but the molecular basis of EV-D68 pathogenesis is unclear. Here we report that EV-D68 inhibits innate immune responses by targeting an immune factor, IRF7. This involves the 3C protease encoded by EV-D68, which mediates the cleavage of IRF7. These observations suggest that the 3Cpro-IRF7 interaction may represent an interface that dictates EV-D68 infection. PMID:26608321

Quantum Mechanical Calculations of Monoxides of Silicon Carbide Molecules

DTIC Science & Technology

2003-03-01

Data for CO Final Energy Charge Mult Basis Set (hart) EA (eV) ZPE (hart) EA (eV) w/ ZPE 0 1 DVZ -112.6850703739 2.02121 -1 2 DVZ...Energy Charge Mult Basis Set (hart) EA (eV) ZPE (hart) EA (eV) w/ ZPE 0 1 DVZ -363.7341927429 0.617643 -1 2 DVZ -363.7114852831 0 3 DVZ...Input Geometry Output Geometry Basis Set Final Energy (hart) EA (eV) ZPE (hart) EA (eV) w/ ZPE -1 2 O-C-Si Linear O-C-Si Linear DZV -401.5363

Analysis on influencing factors of EV charging station planning based on AHP

NASA Astrophysics Data System (ADS)

Yan, F.; Ma, X. F.

2016-08-01

As a new means of transport, electric vehicle (EV) is of great significance to alleviate the energy crisis. EV charging station planning has a far-reaching significance for the development of EV industry. This paper analyzes the impact factors of EV charging station planning, and then uses the analytic hierarchy process (AHP) to carry on the further analysis to the influencing factors, finally it gets the weight of each influence factor, and provides the basis for the evaluation scheme of the planning of charging stations for EV.

Functional Characterization of Four Putative δ1-Pyrroline-5-Carboxylate Reductases from Bacillus subtilis

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

Forlani, Giuseppe; Nocek, Boguslaw; Chakravarthy, Srinivas

In most living organisms, the amino acid proline is synthesized starting from both glutamate and ornithine. In prokaryotes, in the absence of an ornithine cyclodeaminase that has been identified to date only in a small number of soil and plant bacteria, these pathways share the last step, the reduction of delta(1)-pyrroline-5-carboxylate (P5C) catalyzed by P5C reductase (EC 1.5.1.2). In several species, multiple forms of P5C reductase have been reported, possibly reflecting the dual function of proline. Aside from its common role as a building block of proteins, proline is indeed also involved in the cellular response to osmotic and oxidativemore » stress conditions. Genome analysis of Bacillus subtilis identifies the presence of four genes (ProH, ProI, ProG, and ComER) that, based on bioinformatic and phylogenic studies, were defined as respectively coding a putative P5C reductase. Here we describe the cloning, heterologous expression, functional analysis and small-angle X-ray scattering studies of the four affinity-purified proteins. Results showed that two of them, namely ProI and ComER, lost their catalytic efficiency or underwent subfunctionalization. In the case of ComER, this could be likely explained by the loss of the ability to form a dimer, which has been previously shown to be an essential structural feature of the catalytically active P5C reductase. The properties of the two active enzymes are consistent with a constitutive role for ProG, and suggest that ProH expression may be beneficial to satisfy an increased need for proline.« less

Functional Characterization of Four Putative δ1-Pyrroline-5-Carboxylate Reductases from Bacillus subtilis

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

Forlani, Giuseppe; Nocek, Boguslaw; Chakravarthy, Srinivas

In most living organisms, the amino acid proline is synthesized starting from both glutamate and ornithine. In prokaryotes, in the absence of an ornithine cyclodeaminase that has been identified to date only in a small number of soil and plant bacteria, these pathways share the last step, the reduction of δ1-pyrroline-5-carboxylate (P5C) catalyzed by P5C reductase (EC 1.5.1.2). In several species, multiple forms of P5C reductase have been reported, possibly reflecting the dual function of proline. Aside from its common role as a building block of proteins, proline is indeed also involved in the cellular response to osmotic and oxidativemore » stress conditions. Genome analysis of Bacillus subtilis identifies the presence of four genes (ProH, ProI, ProG, and ComER) that, based on bioinformatic and phylogenic studies, were defined as respectively coding a putative P5C reductase. Here we describe the cloning, heterologous expression, functional analysis and small-angle X-ray scattering studies of the four affinity-purified proteins. Results showed that two of them, namely ProI and ComER, lost their catalytic efficiency or underwent subfunctionalization. In the case of ComER, this could be likely explained by the loss of the ability to form a dimer, which has been previously shown to be an essential structural feature of the catalytically active P5C reductase. The properties of the two active enzymes are consistent with a constitutive role for ProG, and suggest that ProH expression may be beneficial to satisfy an increased need for proline.« less

Amphipathic Polyproline Peptides Stimulate Cholesterol Efflux by the ABCA1 Transporter

PubMed Central

Sviridov, D.O.; Drake, S.K.; Freeman, L.A.; Remaley, A.T.

2016-01-01

ApoA-I mimetics are short synthetic peptides that contain an amphipathic αα-helix and stimulate cholesterol efflux by the ABCA1 transporter in a detergent-like extraction mechanism. We investigated the use of amphipathic peptides with a polypro helix for stimulating cholesterol efflux by ABCA1. Polypro peptides were synthesized with modified prolines, containing either a hydrophobic phenol group (Prop) or a polar N-acetylgalactosamine (Prog) attached to the pyrrolidine ring and were designated as either PP-2, 3, 4, or 5, depending on the number of 3 amino acid repeat units (Prop - Prog - Prop). Based on molecular modeling, these peptides were predicted to be relatively rigid and to bind to a phospholipid bilayer. By CD spectroscopy, PP peptides formed a Type-II polypro helix in an aqueous solution. PP-2 was inactive in promoting cholesterol efflux, but peptides with more than 2 repeat units were active. PP-4 showed a similar Vmax as a much longer amphipathic α-αhelical peptide, containing 37 amino acids, but had a Km that was approximately 20-fold lower. PP peptides were specific in that they did not stimulate cholesterol efflux from cells not expressing ABCA1 and were also non-cytotoxic. Addition of PP-3, 4 and 5 to serum promoted the formation of smaller size HDL species (7 nM) and increased its capacity for ABCA1-dependent cholesterol efflux by approximately 20-35% (p<0.05). Because of their relatively small size and increased potency, amphipathic peptides with a polypro helix may represent an alternative structural motif for the development of apoA-I mimetic peptides. PMID:26879139

Characterization of Enteroviruses from Non-Human Primates in Cameroon Revealed Virus Types Widespread in Humans along with Candidate New Types and Species

PubMed Central

Sadeuh-Mba, Serge Alain; Bessaud, Maël; Joffret, Marie-Line; Endegue Zanga, Marie-Claire; Balanant, Jean; Mpoudi Ngole, Eitel; Njouom, Richard; Reynes, Jean-Marc; Delpeyroux, Francis; Rousset, Dominique

2014-01-01

Enteroviruses (EVs) infecting African Non-Human Primates (NHP) are still poorly documented. This study was designed to characterize the genetic diversity of EVs among captive and wild NHP in Cameroon and to compare this diversity with that found in humans. Stool specimens were collected in April 2008 in NHP housed in sanctuaries in Yaounde and neighborhoods. Moreover, stool specimens collected from wild NHP from June 2006 to October 2008 in the southern rain forest of Cameroon were considered. RNAs purified directly from stool samples were screened for EVs using a sensitive RT-nested PCR targeting the VP1 capsid coding gene whose nucleotide sequence was used for molecular typing. Captive chimpanzees (Pan troglodytes) and gorillas (Gorilla gorilla) were primarily infected by EV types already reported in humans in Cameroon and elsewhere: Coxsackievirus A13 and A24, Echovirus 15 and 29, and EV-B82. Moreover EV-A119, a novel virus type recently described in humans in central and west Africa, was also found in a captive Chimpanzee. EV-A76, which is a widespread virus in humans, was identified in wild chimpanzees, thus suggesting its adaptation and parallel circulation in human and NHP populations in Cameroon. Interestingly, some EVs harbored by wild NHP were genetically distinct from all existing types and were thus assigned as new types. One chimpanzee-derived virus was tentatively assigned as EV-J121 in the EV-J species. In addition, two EVs from wild monkeys provisionally registered as EV-122 and EV-123 were found to belong to a candidate new species. Overall, this study indicates that the genetic diversity of EVs among NHP is more important than previously known and could be the source of future new emerging human viral diseases. PMID:25079078

Exposure of Piglets to Enteroviruses Investigated by an Immunoassay Based on the EV-G1 VP4 Peptide.

PubMed

Benkahla, Mehdi A; Sane, Famara; Desailloud, Rachel; Hober, Didier

2016-01-01

The aim of this study was to investigate the exposure of piglets to enteroviruses-G (EV-G) through the presence of antibodies in their serum. Serum samples were obtained from the vena cava of 10 piglets at 9 weeks of age and again 39 days later (day 39). They were tested using an immunoassay based on the EV-G1 VP4 peptide, since VP4 is highly conserved among the four Enterovirus capsid proteins, and by using a seroneutralization assay. For each serum collected on day 39 the optical density was high compared to the value obtained in serum collected earlier (p = 0.002). However, the titers of anti-EV-G1 serum neutralizing activity were not different in paired samples (p > 0.999). The sequence alignment of the EV-G1 VP4 peptide, encompassing 50 amino acids, used in the immunoassay showed 88% homology with EV-G, suggesting that antibodies directed toward other EV-G than EV-G1 may be detected. An immunoassay based on EV-G1 VP4 can detect an increased level of EV-G antibodies in piglet serum samples. Further studies are needed to determine whether this immunoassay may be useful for diagnosis and/or epidemiological studies and to monitor EV-G infection in pigs to evaluate strategies aimed to prevent enterovirus infections. © 2016 S. Karger AG, Basel.

Extracellular Vesicles in Physiology, Pathology, and Therapy of the Immune and Central Nervous System, with Focus on Extracellular Vesicles Derived from Mesenchymal Stem Cells as Therapeutic Tools

PubMed Central

Koniusz, Sylwia; Andrzejewska, Anna; Muraca, Maurizio; Srivastava, Amit K.; Janowski, Miroslaw; Lukomska, Barbara

2016-01-01

Extracellular vesicles (EVs) are membrane-surrounded structures released by most cell types. They are characterized by a specific set of proteins, lipids and nucleic acids. EVs have been recognized as potent vehicles of intercellular communication to transmit biological signals between cells. In addition, pathophysiological roles of EVs in conditions like cancer, infectious diseases and neurodegenerative disorders are well established. In recent years focus has been shifted on therapeutic use of stem cell derived-EVs. Use of stem cell derived-EVs present distinct advantage over the whole stem cells as EVs do not replicate and after intravenous administration, they are less likely to trap inside the lungs. From the therapeutic perspective, the most promising cellular sources of EVs are mesenchymal stem cells (MSCs), which are easy to obtain and maintain. Therapeutic activity of MSCs has been shown in numerous animal models and the beneficial paracrine effect of MSCs may be mediated by EVs. The various components of MSC derived-EVs such as proteins, lipids, and RNA might play a specific therapeutic role. In this review, we characterize the role of EVs in immune and central nervous system (CNS); present evidences for defective signaling of these vesicles in neurodegeneration and therapeutic role of EVs in CNS. PMID:27199663

Extracellular Vesicles Exploit Viral Entry Routes for Cargo Delivery

PubMed Central

van Dongen, Helena M.; Masoumi, Niala

2016-01-01

SUMMARY Extracellular vesicles (EVs) have emerged as crucial mediators of intercellular communication, being involved in a wide array of key biological processes. Eukaryotic cells, and also bacteria, actively release heterogeneous subtypes of EVs into the extracellular space, where their contents reflect their (sub)cellular origin and the physiologic state of the parent cell. Within the past 20 years, presumed subtypes of EVs have been given a rather confusing diversity of names, including exosomes, microvesicles, ectosomes, microparticles, virosomes, virus-like particles, and oncosomes, and these names are variously defined by biogenesis, physical characteristics, or function. The latter category, functions, in particular the transmission of biological signals between cells in vivo and how EVs control biological processes, has garnered much interest. EVs have pathophysiological properties in cancer, neurodegenerative disorders, infectious disease, and cardiovascular disease, highlighting possibilities not only for minimally invasive diagnostic applications but also for therapeutic interventions, like macromolecular drug delivery. Yet, in order to pursue therapies involving EVs and delivering their cargo, a better grasp of EV targeting is needed. Here, we review recent progress in understanding the molecular mechanisms underpinning EV uptake by receptor-ligand interactions with recipient cells, highlighting once again the overlap of EVs and viruses. Despite their highly heterogeneous nature, EVs require common viral entry pathways, and an unanticipated specificity for cargo delivery is being revealed. We discuss the challenges ahead in delineating specific roles for EV-associated ligands and cellular receptors. PMID:26935137

SIRT1 inhibits EV71 genome replication and RNA translation by interfering with the viral polymerase and 5′UTR RNA

PubMed Central

Han, Yang; Wang, Lvyin; Cui, Jin; Song, Yu; Luo, Zhen; Chen, Junbo; Xiong, Ying; Zhang, Qi; Liu, Fang; Ho, Wenzhe; Liu, Yingle; Wu, Jianguo

2016-01-01

ABSTRACT Enterovirus 71 (EV71) possesses a single-stranded positive RNA genome that contains a single open reading frame (ORF) flanked by a 5′ untranslated region (5′UTR) and a polyadenylated 3′UTR. Here, we demonstrated that EV71 activates the production of silent mating type information regulation 2 homolog 1 (SIRT1), a histone deacetylase (HDAC). EV71 further stimulates SIRT1 sumoylation and deacetylase activity, and enhances SIRT1 translocation from the nucleus to the cytoplasm. More interestingly, activated SIRT1 subsequently binds with the EV71 3Dpol protein (a viral RNA-dependent RNA polymerase, RdRp) to repress the acetylation and RdRp activity of 3Dpol, resulting in the attenuation of viral genome replication. Moreover, SIRT1 interacts with the cloverleaf structure of the EV71 RNA 5′UTR to inhibit viral RNA transcription, and binds to the internal ribosome entry site (IRES) of the EV71 5′UTR to attenuate viral RNA translation. Thus, EV71 stimulates SIRT1 production and activity, which in turn represses EV71 genome replication by inhibiting viral polymerase, and attenuates EV71 RNA transcription and translation by interfering with viral RNA. These results uncover a new function of SIRT1 and reveal a new mechanism underlying the regulation of EV71 replication. PMID:27875274

Generation and management of waste electric vehicle batteries in China.

PubMed

Xu, ChengJian; Zhang, Wenxuan; He, Wenzhi; Li, Guangming; Huang, Juwen; Zhu, Haochen

2017-09-01

With the increasing adoption of EVs (electric vehicles), a large number of waste EV LIBs (electric vehicle lithium-ion batteries) were generated in China. Statistics showed generation of waste EV LIBs in 2016 reached approximately 10,000 tons, and the amount of them would be growing rapidly in the future. In view of the deleterious effects of waste EV LIBs on the environment and the valuable energy storage capacity or materials that can be reused in them, China has started emphasizing the management, reuse, and recycling of them. This paper presented the generation trend of waste EV LIBs and focused on interrelated management development and experience in China. Based on the situation of waste EV LIBs management in China, existing problems were analyzed and summarized. Some recommendations were made for decision-making organs to use as valuable references to improve the management of waste EV LIBs and promote the sustainable development of EVs.

Chemical shift in Lα, Lβ1, Lβ3,4, Lβ2,15, Lγ1 and Lγ2,3 emission lines of 47Ag, 48Cd and 50Sn compounds

NASA Astrophysics Data System (ADS)

Singh Kainth, Harpreet; Singh, Ranjit; Singh, Gurjot; Mehta, D.

2018-01-01

Positive and negative shifts in L shell emission lines of 47Ag, 48Cd and 50Sn elements in different chemical compounds were determined from their recorded X-ray emission spectra in high resolution wavelength dispersive X-ray fluorescence (WDXRF) spectrometer. In 47Ag compounds, the measured energy shifts in Lα X-ray emission line were in the ranges from (0.12 to 0.40) eV, Lβ1 (0.27 to 0.36) eV, Lβ3,4 (1.10 to 4.89) eV, Lγ1 (-0.09 to 1.13) eV and Lγ2,3 (-2.08 to 0.59) eV. Likewise, for 48Cd compounds, the estimated shifts in Lα X-ray emission lines were in the range (-0.27 to 0.69) eV, Lβ1 (0.50 to 2.06) eV, Lβ2,15 (0.12 to 0.79), Lβ3,4 (-0.62 to 1.79) eV, Lγ1 (0.10 to 1.35) eV and Lγ2,3 (-0.73 to 1.75) eV, while for 50Sn compounds, the measured shifts in Lα X-ray emission lines were in the range of (0.02 to 1.81) eV, Lβ1 (0.11 to 0.78) eV, Lβ2,15 (0.15 to 1.40), Lβ3,4 (0.17 to 2.01) eV, Lγ1 (0.09 to 1.08) eV and Lγ2,3 (0.17 to 1.40) eV respectively. The effective charges (qP, qS, qL and qB) were calculated by four different theoretical methods (Pauling method, Suchet method, Levine method and Batsonav method) and found to be linear dependent with the chemical shift. Further, the measured chemical shifts were correlated with bond length, relative line-width (FWHM), effective charge, electronegativity, number of ligands and Coster-Kronig (CK) transition processes.

Comprehensive Proteomic Analysis of Human Milk-derived Extracellular Vesicles Unveils a Novel Functional Proteome Distinct from Other Milk Components.

PubMed

van Herwijnen, Martijn J C; Zonneveld, Marijke I; Goerdayal, Soenita; Nolte-'t Hoen, Esther N M; Garssen, Johan; Stahl, Bernd; Maarten Altelaar, A F; Redegeld, Frank A; Wauben, Marca H M

2016-11-01

Breast milk contains several macromolecular components with distinctive functions, whereby milk fat globules and casein micelles mainly provide nutrition to the newborn, and whey contains molecules that can stimulate the newborn's developing immune system and gastrointestinal tract. Although extracellular vesicles (EV) have been identified in breast milk, their physiological function and composition has not been addressed in detail. EV are submicron sized vehicles released by cells for intercellular communication via selectively incorporated lipids, nucleic acids, and proteins. Because of the difficulty in separating EV from other milk components, an in-depth analysis of the proteome of human milk-derived EV is lacking. In this study, an extensive LC-MS/MS proteomic analysis was performed of EV that had been purified from breast milk of seven individual donors using a recently established, optimized density-gradient-based EV isolation protocol. A total of 1963 proteins were identified in milk-derived EV, including EV-associated proteins like CD9, Annexin A5, and Flotillin-1, with a remarkable overlap between the different donors. Interestingly, 198 of the identified proteins are not present in the human EV database Vesiclepedia, indicating that milk-derived EV harbor proteins not yet identified in EV of different origin. Similarly, the proteome of milk-derived EV was compared with that of other milk components. For this, data from 38 published milk proteomic studies were combined in order to construct the total milk proteome, which consists of 2698 unique proteins. Remarkably, 633 proteins identified in milk-derived EV have not yet been identified in human milk to date. Interestingly, these novel proteins include proteins involved in regulation of cell growth and controlling inflammatory signaling pathways, suggesting that milk-derived EVs could support the newborn's developing gastrointestinal tract and immune system. Overall, this study provides an expansion of the whole milk proteome and illustrates that milk-derived EV are macromolecular components with a unique functional proteome. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

Oxidation of CO on a carbon-based material composed of nickel hydroxide and hydroxyl graphene oxide, (Ni4(OH)3-hGO)--a first-principles calculation.

PubMed

Yeh, Chen-Hao; Ho, Jia-Jen

2015-03-21

Nickel or nickel hydroxide clusters and graphene oxide (GO) composites are novel nanomaterials in the application of electrochemical catalysts. In this work, we calculated the energy of Ni4 adsorbed onto saturated hydroxyl graphene oxide (hGO), which forms a Ni4(OH)3 cluster on the hydroxyl graphene oxide (Ni4(OH)3-hGO) and releases 4.47 eV (5.22 eV with DFT-D3 correction). We subsequently studied the oxidation of CO on the Ni4(OH)3-hGO system via three mechanisms - LH, ER and carbonated mechanisms. Our results show that the activation energy for oxidation of the first CO molecule according to the ER mechanism is 0.14 eV (0.12 eV with DFT-D3 correction), much smaller than that with LH (Ea = 0.65 eV, 0.61 eV with DFT-D3 correction) and with carbonated (Ea = 1.28 eV, 1.20 eV with DFT-D3 correction) mechanisms. The barrier to oxidation of the second CO molecule to CO2 with the ER mechanism increases to 0.43 eV (0.37 eV with DFT-D3 correction), but still less than that via LH (Ea = 1.09 eV, 1.07 eV with DFT-D3 correction), indicating that CO could be effectively oxidized through the ER mechanism on the Ni4(OH)3/hGO catalyst.

Variations in cerebrospinal fluid viral loads among enterovirus genotypes in patients hospitalized with laboratory-confirmed meningitis due to enterovirus.

PubMed

Volle, Romain; Bailly, Jean-Luc; Mirand, Audrey; Pereira, Bruno; Marque-Juillet, Stéphanie; Chambon, Martine; Regagnon, Christel; Brebion, Amélie; Henquell, Cécile; Peigue-Lafeuille, Hélène; Archimbaud, Christine

2014-08-15

Acute enterovirus (EV) meningitis is a major cause of hospitalization among adults and children. It is caused by multiple EV genotypes assigned to 4 species (EV-A, EV-B, EV-C, and EV-D). We determined viral loads in the cerebrospinal fluid (CSF) of 156 patients of all ages with EV meningitis during a 5-year observational prospective study. The virus strains were genotyped, and their time origin was determined with Bayesian phylogenetic methods. The CSF viral loads ranged between 3.4 and 7.5 log10 copies/mL (median, 4.9 log10 copies/mL). They were higher in neonates than in infants and children (P = .02) but were comparable in adults. Viral loads were associated with EV genotypes (P < .001). The EV strains were identified in 152 of 156 patients and assigned to 23 genotypes within the EV-A and EV-B species. The most frequent genotypes, echoviruses 6 and 30, were associated with different viral loads (P < .001). The highest viral loads were in meningitis cases caused by coxsackievirus A9, B4, and B5 genotypes. Most patients infected by a same genotype were infected by a major virus variant of recent emergence. The variations in CSF viral loads in patients at the onset of EV meningitis are related to genotypic differences in the virus strains involved. © The Author 2014. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

IL-3R-alpha blockade inhibits tumor endothelial cell-derived extracellular vesicle (EV)-mediated vessel formation by targeting the β-catenin pathway.

PubMed

Lombardo, Giusy; Gili, Maddalena; Grange, Cristina; Cavallari, Claudia; Dentelli, Patrizia; Togliatto, Gabriele; Taverna, Daniela; Camussi, Giovanni; Brizzi, Maria Felice

2018-03-01

The proangiogenic cytokine Interleukin-3 (IL-3) is released by inflammatory cells in breast and ovarian cancer tissue microenvironments and also acts as an autocrine factor for human breast and kidney tumor-derived endothelial cells (TECs). We have previously shown that IL-3-treated endothelial cells (ECs) release extracellular vesicles (EVs), which serve as a paracrine mechanism for neighboring ECs, by transferring active molecules. The impact of an anti-IL-3R-alpha blocking antibody on the proangiogenic effect of EVs released from TECs (anti-IL-3R-EVs) has therefore been investigated in this study. We have found that anti-IL-3R-EV treatment prevented neovessel formation and, more importantly, also induced the regression of in vivo TEC-derived neovessels. Two miRs that target the canonical wingless (Wnt)/β-catenin pathway, at different levels, were found to be differentially regulated when comparing the miR-cargo of naive TEC-derived EVs (EVs) and anti-IL-3R-EVs. miR-214-3p, which directly targets β-catenin, was found to be upregulated, whereas miR-24-3p, which targets adenomatous polyposis coli (APC) and glycogen synthase kinase-3β (GSK3β), was found to be downregulated. In fact, upon their transfer into the cell, low β-catenin content and high levels of the two members of the "β-catenin destruction complex" were detected. Moreover, c-myc downregulation was found in TECs treated with anti-IL-3R-EVs, pre-miR-214-3p-EVs and antago-miR-24-3p-EVs, which is consistent with network analyses of miR-214-3p and miR-24-3p gene targeting. Finally, in vivo studies have demonstrated the impaired growth of vessels in pre-miR-214-3p-EV- and antago-miR-24-3p-EV-treated animals. These effects became much more evident when combo treatment was applied. The results of the present study identify the canonical Wnt/β-catenin pathway as a relevant mechanism of TEC-derived EV proangiogenic action. Furthermore, we herein provide evidence that IL-3R blockade may yield some significant advantages, than miR targeting, in inhibiting the proangiogenic effects of naive TEC-derived EVs by changing TEC-EV-miR cargo.

Complete genome characterization of a novel enterovirus type EV-B106 isolated in China, 2012.

PubMed

Tang, Jingjing; Tao, Zexin; Ding, Zhengrong; Zhang, Yong; Zhang, Jie; Tian, Bingjun; Zhao, Zhixian; Zhang, Lifen; Xu, Wenbo

2014-03-03

Human enterovirus B106 (EV-B106) is a recently identified member of enterovirus species B. In this study, we report the complete genomic characterization of an EV-B106 strain (148/YN/CHN/12) isolated from an acute flaccid paralysis patient in Yunnan Province, China. The new strain had 79.2-81.3% nucleotide and 89.1-94.8% amino acid similarity in the VP1 region with the other two EV-B106 strains from Bolivia and Pakistan. When compared with other EV serotypes, it had the highest (73.3%) VP1 nucleotide similarity with the EV-B77 prototype strain CF496-99. However, when aligned with all EV-B106 and EV-B77 sequences available from the GenBank database, two major frame shifts were observed in the VP1 coding region, which resulted in substantial (20.5%) VP1 amino acid divergence between the two serotypes. Phylogenetic analysis and similarity plot analysis revealed multiple recombination events in the genome of this strain. This is the first report of the complete genome of EV-B106.

[The genetical evolution of the full length genes of 5 EV 71 strains from 5 Shenzhen patients with hand-food-mouth disease associated with EV71 infection].

PubMed

Liu, Wei-long; Yang, Gui-lin; Wei, Qing; Zhang, Ming-xia; Chen, Xin-chun; Liu, Ying-xia; Gao, Yang; Zhou, Bo-ping

2011-02-01

To investigate the characteristics of molecular epidemiology and molecular evolution of 5 EV 71 (enterovirus 71, EV71) strains from 5 Shenzhen patients with hand-food-mouth disease associated with EV 71 infection. 5 EV 71 strains were isolated, and sequenced to analyzed the full length gene sequences in order to compare nucleotide and amino acid homology with other EV71 strains from other regions and countries as well as previous strains across the world through bioinformatics software. 5 strains of EV 71 belonged to sub-genotype C4 by analysis of nucleotide sequences of VP1 and VP4 of EV 71. The differences of nucleotide and amino acid sequences were much small with nucleotide homology of 93% and amino acid homology of 98% among these 5 strains. A phylogenetic tree analysis indicated that 2008 Shenzhen epidemic strains were the most close to 2004 Shenzhen circulating strains, and also much close to 1998 Shenzhen epidemic strains and 2008 Fuyang Anhui strains. The dead strain was very close to 2008 Fuyang Anhui epidemic strains. It can be speculated that this epidemic strains of EV 71 probably originate from the same ancient strain in the history, may from 1998 Shenzhen strain.

Coordinated EV adoption: double-digit reductions in emissions and fuel use for $40/vehicle-year.

PubMed

Choi, Dong Gu; Kreikebaum, Frank; Thomas, Valerie M; Divan, Deepak

2013-09-17

Adoption of electric vehicles (EVs) would affect the costs and sources of electricity and the United States efficiency requirements for conventional vehicles (CVs). We model EV adoption scenarios in each of six regions of the Eastern Interconnection, containing 70% of the United States population. We develop electricity system optimization models at the multidecade, day-ahead, and hour-ahead time scales, incorporating spatial wind energy modeling, endogenous modeling of CV efficiencies, projections for EV efficiencies, and projected CV and EV costs. We find two means to reduce total consumer expenditure (TCE): (i) controlling charge timing and (ii) unlinking the fuel economy regulations for CVs from EVs. Although EVs provide minimal direct GHG reductions, controlled charging provides load flexibility, lowering the cost of renewable electricity. Without EVs, a 33% renewable electricity standard (RES) would cost $193/vehicle-year more than the reference case (10% RES). Combining a 33% RES, EVs with controlled charging and unlinking would reduce combined electric- and vehicle-sector CO2 emissions by 27% and reduce gasoline consumption by 59% for $40/vehicle-year more than the reference case. Coordinating EV adoption with adoption of controlled charging, unlinked fuel economy regulations, and renewable electricity standards would provide low-cost reductions in emissions and fuel usage.

Bond Dissociation Energies of Tungsten Molecules: WC, WSi, WS, WSe, and WCl.

PubMed

Sevy, Andrew; Huffaker, Robert F; Morse, Michael D

2017-12-14

Resonant two-photon ionization spectroscopy was used to locate predissociation thresholds in WC, WSi, WS, WSe, and WCl, allowing bond dissociation energies to be measured for these species. Because of the high degree of vibronic congestion in the observed spectra, it is thought that the molecules dissociate as soon as the lowest separated atom limit is exceeded. From the observed predissociation thresholds, dissociation energies are assigned as D 0 (WC) = 5.289(8) eV, D 0 (WSi) = 3.103(10) eV, D 0 (WS) = 4.935(3) eV, D 0 (WSe) = 4.333(6) eV, and D 0 (WCl) = 3.818(6) eV. These results are combined with other data to obtain the ionization energy IE(WC) = 8.39(9) eV and the anionic bond dissociation energies of D 0 (W-C - ) = 6.181(17) eV, D 0 (W - -C) = 7.363(19) eV, D 0 (W-Si - ) ≤ 3.44(4) eV, and D 0 (W - -Si) ≤ 4.01(4) eV. Combination of the D 0 (WX) values with atomic enthalpies of formation also provides Δ f H 0K ° values for the gaseous WX molecules. Computational results are also provided, which shed some light on the electronic structure of these molecules.

Environmental implication of electric vehicles in China.

PubMed

Huo, Hong; Zhang, Qiang; Wang, Michael Q; Streets, David G; He, Kebin

2010-07-01

Today, electric vehicles (EVs) are being proposed in China as one of the potential options to address the dramatically increasing energy demand from on-road transport. However, the mass use of EVs could involve multiple environmental issues, because EVs use electricity that is generated primarily from coal in China. We examined the fuel-cycle CO(2), SO(2), and NO(x) emissions of EVs in China in both current (2008) and future (2030) periods and compared them with those of conventional gasoline vehicles and gasoline hybrids. EVs do not promise much benefit in reducing CO(2) emissions currently, but greater CO(2) reduction could be expected in future if coal combustion technologies improve and the share of nonfossil electricity increases significantly. EVs could increase SO(2) emissions by 3-10 times and also double NO(x) emissions compared to gasoline vehicles if charged using the current electricity grid. In the future, EVs would be able to reach the NO(x) emission level of gasoline vehicles with advanced emission control devices equipped in thermal power plants but still increase SO(2). EVs do represent an effective solution to issues in China such as oil shortage, but critical policy support is urgently needed to address the environmental issues caused by the use of EVs to make EVs competitive with other vehicle alternatives.

Enterovirus D68 disease and molecular epidemiology in Australia.

PubMed

Levy, Avram; Roberts, Jason; Lang, Jurissa; Tempone, Simone; Kesson, Alison; Dofai, Alfred; Daley, Andrew J; Thorley, Bruce; Speers, David J

2015-08-01

Enterovirus D68 (EV-D68) has received considerable recent attention as a cause of widespread respiratory illness. Neurological syndromes such as acute flaccid paralysis following EV-D68 infection have also been reported in a small number of cases. To summarize the clinical and epidemiological characteristics of laboratory confirmed EV-D68 cases in Australia. We combined EV-D68 data acquired through laboratory surveillance in Western Australia with cases from national enterovirus surveillance and regional acute flaccid paralysis (AFP) surveillance. Clinical data was obtained for EV-D68 cases and capsid protein sequences were used for phylogenetic analysis. Sporadic cases of EV-D68 were recorded in Australia since 2008, with peaks in activity during 2011 and 2013. EV-D68 was primarily associated with respiratory disease, but was also detected in cerebrospinal fluid of one patient and faeces of two patients presenting with AFP. EV-D68 has been circulating in Western Australia and is likely to have also been present in the wider region for a number of years, causing primarily respiratory disease. Detection of EV-D68 in cerebrospinal fluid of one patient and in faeces of two AFP cases reinforces the association between EV-D68 and neurological disease. Copyright © 2015 Elsevier B.V. All rights reserved.

Viral chimeras decrypt the role of enterovirus capsid proteins in viral tropism, acid sensitivity and optimal growth temperature

PubMed Central

Royston, Léna; Essaidi-Laziosi, Manel; Piuz, Isabelle; Geiser, Johan; Huang, Song; Kaiser, Laurent; Garcin, Dominique

2018-01-01

Despite their genetic similarities, enteric and respiratory enteroviruses (EVs) have highly heterogeneous biophysical properties and cause a vast diversity of human pathologies. In vitro differences include acid sensitivity, optimal growth temperature and tissue tropism, which reflect a preferential in vivo replication in the respiratory or gastrointestinal tract and are thus key determinants of EV virulence. To investigate the underlying cause of these differences, we generated chimeras at the capsid-level between EV-D68 (a respiratory EV) and EV-D94 (an enteric EV). Although some chimeras were nonfunctional, EV-D94 with both the capsid and 2A protease or the capsid only of EV-D68 were both viable. Using this latter construct, we performed several functional assays, which indicated that capsid proteins determine acid sensitivity and tropism in cell lines and in respiratory, intestinal and neural tissues. Additionally, capsid genes were shown to also participate in determining the optimal growth temperature, since EV-D94 temperature adaptation relied on single mutations in VP1, while constructs with EV-D68 capsid could not adapt to higher temperatures. Finally, we demonstrate that EV-D68 maintains residual binding-capacity after acid-treatment despite a loss of infectivity. In contrast, non-structural rather than capsid proteins modulate the innate immune response in tissues. These unique biophysical insights expose another layer in the phenotypic diversity of one of world’s most prevalent pathogens and could aid target selection for vaccine or antiviral development. PMID:29630666

In vitro immunotoxicity assessment of culture-derived extracellular vesicles in human monocytes

PubMed Central

Rosas, Lucia E.; Elgamal, Ola A.; Mo, Xiaokui; Phelps, Mitch A.; Schmittgen, Thomas D.; Papenfuss, Tracey L.

2016-01-01

The potential to engineer extracellular vesicles (EV) that target specific cells and deliver a therapeutic payload has propelled a growing interest in their development as promising therapeutics. These EV are often produced from cultured cells. Very little is known about the interaction of cell culture-derived EV with cells of the immune system and their potential immunomodulatory effects. The present study evaluated potential immunotoxic effects of HEK293T-derived EV on the human monocytic cell lines THP-1 and U937. Incubation of cells with different doses of EV for 16–24 h was followed by assessment of cytotoxicity and cell function by flow cytometry. Changes in cell functionality were evaluated by the capacity of cells to phagocytize fluorescent microspheres. In addition, the internalization of labeled EV in THP-1 and U937 cells was evaluated. Exposure to EV did not affect the viability of THP-1 or U937 cells. Although lower doses of the EV increased phagocytic capacity in both cell lines, phagocytic efficiency of individual cells was not affected by EV exposure at any of the doses evaluated. This study also demonstrated that THP-1 and U937 monocytic cells are highly permissive to EV entry in a dose-response manner. These results suggest that, although HEK293T-derived EV are efficiently internalized by human monocytic cells, they do not exert a cytotoxic effect or alter phagocytic efficiency on the cell lines evaluated. PMID:27075513

Electrospray ionization mass spectrometry and partial least squares discriminant analysis applied to the quality control of olive oil.

PubMed

Alves, Junia O; Botelho, Bruno G; Sena, Marcelo M; Augusti, Rodinei

2013-10-01

Direct infusion electrospray ionization mass spectrometry in the positive ion mode [ESI(+)-MS] is used to obtain fingerprints of aqueous-methanolic extracts of two types of olive oils, extra virgin (EV) and ordinary (OR), as well as of samples of EV olive oil adulterated by the addition of OR olive oil and other edible oils: corn (CO), sunflower (SF), soybean (SO) and canola (CA). The MS data is treated by the partial least squares discriminant analysis (PLS-DA) protocol aiming at discriminating the above-mentioned classes formed by the genuine olive oils, EV (1) and OR (2), as well as the EV adulterated samples, i.e. EV/SO (3), EV/CO (4), EV/SF (5), EV/CA (6) and EV/OR (7). The PLS-DA model employed is built with 190 and 70 samples for the training and test sets, respectively. For all classes (1-7), EV and OR olive oils as well as the adulterated samples (in a proportion varying from 0.5 to 20.0% w/w) are properly classified. The developed methodology required no ions identification and demonstrated to be fast, as each measurement lasted about 3 min including the extraction step and MS analysis, and reliable, because high sensitivities (rate of true positives) and specificities (rate of true negatives) were achieved. Finally, it can be envisaged that this approach has potential to be applied in quality control of EV olive oils. Copyright © 2013 John Wiley & Sons, Ltd.

Cross-reactivity of anti-EV71 IgM and neutralizing antibody in series sera of patients infected with Enterovirus 71 and Coxsackievirus A 16.

PubMed

Lin, Yulong; Wen, Kun; Pan, Yuxian; Wang, Yadi; Che, Xiaoyan; Wang, Bin

2011-01-01

To evaluate the cross-reactivity of anti-EV71 IgM and neutralizing antibody in series sera of patients infected with EV71 and CA16. Real-time RT-PCR, virus isolation, ELISA and neutralization test were used to detect enteroviruses from clinical specimens and series sera of 79 HFMD patients. 27 EV71, 37 CA16, and 11 other enterovirus-infected patients were identified by RT-PCR. Among EV71 infected patients, anti-EV71 IgM positive ratios were 87.5% during 1-3 days after onset and 100% over 4 days after onset. In CA16 infected patients, the positive ratios were 7.4%, 26.4%, and 62.5% during 1-3 days, 4-6 days, and over 6 days after onset, respectively. Meanwhile, the results of neutralization test showed 18.9% of CA16 infected patients and 11.1% of EV71 infected patients present high cross-neutralization antibody against each other. Cross-reactivity of anti-EV71 IgM in patients infected with EV71 and CA16 becomes stronger with the progress of disease. Moreover, high cross-neutralization antibody existing in part of patients suggests that the immune reactivity to EV71 infection can be recalled by CA16, and the immune reactivity to CA16 infection can be recalled by EV71. Therefore, identifying enteroviruses by neutralization test may not be an ideal selection.

Considering the dynamic refueling behavior in locating electric vehicle charging stations

NASA Astrophysics Data System (ADS)

Liu, K.; Sun, X. H.

2014-11-01

Electric vehicles (EVs) will certainly play an important role in addressing the energy and environmental challenges at current situation. However, location problem of EV charging stations was realized as one of the key issues of EVs launching strategy. While for the case of locating EV charging stations, more influence factors and constraints need to be considered since the EVs have some special attributes. The minimum requested charging time for EVs is usually more than 30minutes, therefore the possible delay time due to waiting or looking for an available station is one of the most important influence factors. In addition, the intention to purchase and use of EVs that also affects the location of EV charging stations is distributed unevenly among regions and should be considered when modelling. Unfortunately, these kinds of time-spatial constraints were always ignored in previous models. Based on the related research of refuelling behaviours and refuelling demands, this paper developed a new concept with dual objectives of minimum waiting time and maximum service accessibility for locating EV charging stations - named as Time-Spatial Location Model (TSLM). The proposed model and the traditional flow-capturing location model are applied on an example network respectively and the results are compared. Results demonstrate that time constraint has great effects on the location of EV charging stations. The proposed model has some obvious advantages and will help energy providers to make a viable plan for the network of EV charging stations.

Inflammaging and Frailty Status Do Not Result in an Increased Extracellular Vesicle Concentration in Circulation

PubMed Central

Alberro, Ainhoa; Sáenz-Cuesta, Matías; Muñoz-Culla, Maider; Mateo-Abad, Maider; Gonzalez, Esperanza; Carrasco-Garcia, Estefania; Araúzo-Bravo, Marcos J.; Matheu, Ander; Vergara, Itziar; Otaegui, David

2016-01-01

In the last decades extracellular vesicles (EVs) have emerged as key players for intercellular communication. In the case of inflammation, several studies have reported that EV levels are increased in circulation during inflammatory episodes. Based on this, we investigated whether aging results in elevated EV number, as a basal proinflammatory status termed “inflammaging” has been described in aged individuals. Moreover, we also hypothesized that frailty and dependence conditions of the elderly could affect EV concentration in plasma. Results showed that inflammaging, frailty or dependence status do not result in EV increase, at least in the total number of EVs in circulation. These results open a new perspective for investigating the role of EVs in human aging and in the inflammaging process. PMID:27447627

Inflammaging and Frailty Status Do Not Result in an Increased Extracellular Vesicle Concentration in Circulation.

PubMed

Alberro, Ainhoa; Sáenz-Cuesta, Matías; Muñoz-Culla, Maider; Mateo-Abad, Maider; Gonzalez, Esperanza; Carrasco-Garcia, Estefania; Araúzo-Bravo, Marcos J; Matheu, Ander; Vergara, Itziar; Otaegui, David

2016-07-20

In the last decades extracellular vesicles (EVs) have emerged as key players for intercellular communication. In the case of inflammation, several studies have reported that EV levels are increased in circulation during inflammatory episodes. Based on this, we investigated whether aging results in elevated EV number, as a basal proinflammatory status termed "inflammaging" has been described in aged individuals. Moreover, we also hypothesized that frailty and dependence conditions of the elderly could affect EV concentration in plasma. Results showed that inflammaging, frailty or dependence status do not result in EV increase, at least in the total number of EVs in circulation. These results open a new perspective for investigating the role of EVs in human aging and in the inflammaging process.

Opportunities for Small Satellites in NASA's Earth System Science Pathfinder (ESSP) Program

NASA Technical Reports Server (NTRS)

Peri, Frank; Law, Richard C.; Wells, James E.

2014-01-01

NASA's Earth Venture class (EV) of missions are competitively selected, Principal Investigator (PI) led, relatively low cost and narrowly focused in scientific scope. Investigations address a full spectrum of earth science objectives, including studies of the atmosphere, oceans, land surface, polar ice regions, and solid Earth. EV has three program elements: EV-Suborbital (EVS) are suborbital/airborne investigations; EV-Mission (EVM) element comprises small complete spaceborne missions; and EV-Instrument (EVI) element develops spaceborne instruments for flight as Missions-of-Opportunity (MoO). To ensure the success of EV, frequent opportunities for selecting missions has been established in NASA's Earth Science budget. This paper will describe those opportunities and how the management approach of each element is tailored according to the specific needs of the element.

The Natural Diyne-Furan Fatty Acid EV-086 Is an Inhibitor of Fungal Delta-9 Fatty Acid Desaturation with Efficacy in a Model of Skin Dermatophytosis

PubMed Central

Diefenbacher, Melanie; Greve, Katrine B. V.; Brianza, Federico; Folly, Christophe; Heider, Harald; Lone, Museer A.; Long, Lisa; Meyer, Jean-Philippe; Roussel, Patrick; Ghannoum, Mahmoud A.; Schneiter, Roger; Sorensen, Alexandra S.

2014-01-01

Human fungal infections represent a therapeutic challenge. Although effective strategies for treatment are available, resistance is spreading, and many therapies have unacceptable side effects. A clear need for novel antifungal targets and molecules is thus emerging. Here, we present the identification and characterization of the plant-derived diyne-furan fatty acid EV-086 as a novel antifungal compound. EV-086 has potent and broad-spectrum activity in vitro against Candida, Aspergillus, and Trichophyton spp., whereas activities against bacteria and human cell lines are very low. Chemical-genetic profiling of Saccharomyces cerevisiae deletion mutants identified lipid metabolic processes and organelle organization and biogenesis as targets of EV-086. Pathway modeling suggested that EV-086 inhibits delta-9 fatty acid desaturation, an essential process in S. cerevisiae, depending on the delta-9 fatty acid desaturase OLE1. Delta-9 unsaturated fatty acids—but not saturated fatty acids—antagonized the EV-086-mediated growth inhibition, and transcription of the OLE1 gene was strongly upregulated in the presence of EV-086. EV-086 increased the ratio of saturated to unsaturated free fatty acids and phosphatidylethanolamine fatty acyl chains, respectively. Furthermore, EV-086 was rapidly taken up into the lipid fraction of the cell and incorporated into phospholipids. Together, these findings demonstrate that EV-086 is an inhibitor of delta-9 fatty acid desaturation and that the mechanism of inhibition might involve an EV-086–phospholipid. Finally, EV-086 showed efficacy in a guinea pig skin dermatophytosis model of topical Trichophyton infection, which demonstrates that delta-9 fatty acid desaturation is a valid antifungal target, at least for dermatophytoses. PMID:24189258

Clinically isolated enterovirus A71 subgenogroup C4 strain with lethal pathogenicity in 14-day-old mice and the application as an EV-A71 mouse infection model.

PubMed

Xu, Yi; Ma, Shuzhi; Zhu, Limeng; Huang, Zhiqiu; Chen, Liyun; Xu, Yuhua; Yin, Haibin; Peng, Tao; Wang, Yi

2017-01-01

The Enterovirus A71 (EV-A71) subgenogroup C4 is prevalent in China. EV-A71 causes hand, foot and mouth disease (HFMD) in children and may lead to severe neurological diseases. The development of antiviral and protective vaccines against EV-A71 is significantly hindered by the lack of suitable animal models to recapitulate human neurological symptoms. In this study, GZ-CII, a highly virulent EV-A71 subgenogroup C4 strain, was isolated from hospitalized children with HFMD. Intraperitoneal infections of GZ-CII resulted in progressive neurological disease in mice as old as 14 days. Administration of an inactivated EV-A71 vaccine or an anti-EV-A71 immune serum protected the mice against the GZ-CII infection. This demonstrated that a mouse model with EV-A71 GZ-CII could be used to evaluate potential vaccine candidates and therapeutics for subgenogroup C4. Comparing the genome sequence of GZ-CII with that of the avirulent EV-A71 subgenogroup C4 strain revealed unique mutations in GZ-CII. When mutation VP2-K149I was introduced into the nonpathogenic EV-A71 subgenogroup C4 strain, the variant similar to GZ-CII significantly increased viral replication and virulence in mice. These results indicated that the VP2-K149I mutation played an important role in enhancing the virulence of the EV-A71 subgenogroup C4 strain in mice, and that mice infected with the GZ-CII strain are a promising model for evaluating vaccines and therapeutics against the EV-A71 subgenogroup C4. Copyright © 2016 Elsevier B.V. All rights reserved.

The natural diyne-furan fatty acid EV-086 is an inhibitor of fungal delta-9 fatty acid desaturation with efficacy in a model of skin dermatophytosis.

PubMed

Knechtle, Philipp; Diefenbacher, Melanie; Greve, Katrine B V; Brianza, Federico; Folly, Christophe; Heider, Harald; Lone, Museer A; Long, Lisa; Meyer, Jean-Philippe; Roussel, Patrick; Ghannoum, Mahmoud A; Schneiter, Roger; Sorensen, Alexandra S

2014-01-01

Human fungal infections represent a therapeutic challenge. Although effective strategies for treatment are available, resistance is spreading, and many therapies have unacceptable side effects. A clear need for novel antifungal targets and molecules is thus emerging. Here, we present the identification and characterization of the plant-derived diyne-furan fatty acid EV-086 as a novel antifungal compound. EV-086 has potent and broad-spectrum activity in vitro against Candida, Aspergillus, and Trichophyton spp., whereas activities against bacteria and human cell lines are very low. Chemical-genetic profiling of Saccharomyces cerevisiae deletion mutants identified lipid metabolic processes and organelle organization and biogenesis as targets of EV-086. Pathway modeling suggested that EV-086 inhibits delta-9 fatty acid desaturation, an essential process in S. cerevisiae, depending on the delta-9 fatty acid desaturase OLE1. Delta-9 unsaturated fatty acids-but not saturated fatty acids-antagonized the EV-086-mediated growth inhibition, and transcription of the OLE1 gene was strongly upregulated in the presence of EV-086. EV-086 increased the ratio of saturated to unsaturated free fatty acids and phosphatidylethanolamine fatty acyl chains, respectively. Furthermore, EV-086 was rapidly taken up into the lipid fraction of the cell and incorporated into phospholipids. Together, these findings demonstrate that EV-086 is an inhibitor of delta-9 fatty acid desaturation and that the mechanism of inhibition might involve an EV-086-phospholipid. Finally, EV-086 showed efficacy in a guinea pig skin dermatophytosis model of topical Trichophyton infection, which demonstrates that delta-9 fatty acid desaturation is a valid antifungal target, at least for dermatophytoses.

Human Enterovirus 68 Interferes with the Host Cell Cycle to Facilitate Viral Production

PubMed Central

Wang, Zeng-yan; Zhong, Ting; Wang, Yue; Song, Feng-mei; Yu, Xiao-feng; Xing, Li-ping; Zhang, Wen-yan; Yu, Jing-hua; Hua, Shu-cheng; Yu, Xiao-fang

2017-01-01

Enterovirus D68 (EV-D68) is an emerging pathogen that recently caused a large outbreak of severe respiratory disease in the United States and other countries. Little is known about the relationship between EV-D68 virus and host cells. In this study, we assessed the effect of the host cell cycle on EV-D68 viral production, as well as the ability of EV-D68 to manipulate host cell cycle progression. The results suggest that synchronization in G0/G1 phase, but not S phase, promotes viral production, while synchronization in G2/M inhibits viral production. Both an early EV-D68 isolate and currently circulating strains of EV-D68 can manipulate the host cell cycle to arrest cells in the G0/G1 phase, thus providing favorable conditions for virus production. Cell cycle regulation by EV-D68 was associated with corresponding effects on the expression of cyclins and CDKs, which were observed at the level of the protein and/or mRNA. Furthermore, the viral non-structural protein 3D of EV-D68 prevents progression from G0/G1 to S. Interestingly, another member of the Picornaviridae family, EV-A71, differs from EV-D68 in that G0/G1 synchronization inhibits, rather than promotes, EV-A71 viral replication. However, these viruses are similar in that G2/M synchronization inhibits the production and activity of both viruses, which is suggestive of a common therapeutic target for both types of enterovirus. These results further clarify the pathogenic mechanisms of enteroviruses and provide a potential strategy for the treatment and prevention of EV-D68-related disease. PMID:28229049

Pellet-free isolation of human and bovine milk extracellular vesicles by size-exclusion chromatography.

PubMed

Blans, Kristine; Hansen, Maria S; Sørensen, Laila V; Hvam, Michael L; Howard, Kenneth A; Möller, Arne; Wiking, Lars; Larsen, Lotte B; Rasmussen, Jan T

2017-01-01

Studies have suggested that nanoscale extracellular vesicles (EV) in human and bovine milk carry immune modulatory properties which could provide beneficial health effects to infants. In order to assess the possible health effects of milk EV, it is essential to use isolates of high purity from other more abundant milk structures with well-documented bioactive properties. Furthermore, gentle isolation procedures are important for reducing the risk of generating vesicle artefacts, particularly when EV subpopulations are investigated. In this study, we present two isolation approaches accomplished in three steps based on size-exclusion chromatography (SEC) resulting in effective and reproducible EV isolation from raw milk. The approaches do not require any EV pelleting and can be applied to both human and bovine milk. We show that SEC effectively separates phospholipid membrane vesicles from the primary casein and whey protein components in two differently obtained casein reduced milk fractions, with one of the fractions obtained without the use of ultracentrifugation. Milk EV isolates were enriched in lactadherin, CD9, CD63 and CD81 compared to minimal levels of the EV-marker proteins in other relevant milk fractions such as milk fat globules. Nanoparticle tracking analysis and electron microscopy reveals the presence of heterogeneous sized vesicle structures in milk EV isolates. Lipid analysis by thin layer chromatography shows that EV isolates are devoid of triacylglycerides and presents a phospholipid profile differing from milk fat globules surrounded by epithelial cell plasma membrane. Moreover, the milk EV fractions are enriched in RNA with distinct and diverging profiles from milk fat globules. Collectively, our data supports that successful milk EV isolation can be accomplished in few steps without the use of ultracentrifugation, as the presented isolation approaches based on SEC effectively isolates EV in both human and bovine milk.

Pellet-free isolation of human and bovine milk extracellular vesicles by size-exclusion chromatography

PubMed Central

Blans, Kristine; Hansen, Maria S.; Sørensen, Laila V.; Hvam, Michael L.; Howard, Kenneth A.; Möller, Arne; Wiking, Lars; Larsen, Lotte B.; Rasmussen, Jan T.

2017-01-01

ABSTRACT Studies have suggested that nanoscale extracellular vesicles (EV) in human and bovine milk carry immune modulatory properties which could provide beneficial health effects to infants. In order to assess the possible health effects of milk EV, it is essential to use isolates of high purity from other more abundant milk structures with well-documented bioactive properties. Furthermore, gentle isolation procedures are important for reducing the risk of generating vesicle artefacts, particularly when EV subpopulations are investigated. In this study, we present two isolation approaches accomplished in three steps based on size-exclusion chromatography (SEC) resulting in effective and reproducible EV isolation from raw milk. The approaches do not require any EV pelleting and can be applied to both human and bovine milk. We show that SEC effectively separates phospholipid membrane vesicles from the primary casein and whey protein components in two differently obtained casein reduced milk fractions, with one of the fractions obtained without the use of ultracentrifugation. Milk EV isolates were enriched in lactadherin, CD9, CD63 and CD81 compared to minimal levels of the EV-marker proteins in other relevant milk fractions such as milk fat globules. Nanoparticle tracking analysis and electron microscopy reveals the presence of heterogeneous sized vesicle structures in milk EV isolates. Lipid analysis by thin layer chromatography shows that EV isolates are devoid of triacylglycerides and presents a phospholipid profile differing from milk fat globules surrounded by epithelial cell plasma membrane. Moreover, the milk EV fractions are enriched in RNA with distinct and diverging profiles from milk fat globules. Collectively, our data supports that successful milk EV isolation can be accomplished in few steps without the use of ultracentrifugation, as the presented isolation approaches based on SEC effectively isolates EV in both human and bovine milk. PMID:28386391

[Embryo volume estimated by three-dimensional ultrasonography at seven to ten weeks of pregnancy].

PubMed

Filho, João Bortoletti; Nardozza, Luciano Marcondes Machado; Araujo Júnior, Edward; Rôlo, Líliam Cristine; Nowak, Paulo Martin; Moron, Antonio Fernandes

2008-10-01

to evaluate the embryo's volume (EV) between the seventh and the tenth gestational week, through tridimensional ultrasonography. a transversal study with 63 normal pregnant women between the seventh and the tenth gestational week. The ultrasonographical exams have been performed with a volumetric abdominal transducer. Virtual Organ Computer-aided Analysis (VOCAL) has been used to calculate EV, with a rotation angle of 12 masculine and a delimitation of 15 sequential slides. The average, median, standard deviation and maximum and minimum values have been calculated for the EV in all the gestational ages. A dispersion graphic has been drawn to assess the correlation between EV and the craniogluteal length (CGL), the adjustment being done by the determination coefficient (R(2)). To determine EV's reference intervals as a function of the CGL, the following formula was used: percentile=EV+K versus SD, with K=1.96. CGL has varied from 9.0 to 39.7 mm, with an average of 23.9 mm (+/-7.9 mm), while EV has varied from 0.1 to 7.6 cm(3), with an average of 2.7 cm(3) (+/-3.2 cm(3)). EV was highly correlated to CGL, the best adjustment being obtained with quadratic regression (EV=0.2-0.055 versus CGL+0.005 versus CGL(2); R(2)=0.8). The average EV has varied from 0.1 (-0.3 to 0.5 cm(3)) to 6.7 cm(3) (3.8 to 9.7 cm(3)) within the interval of 9 to 40 mm of CGL. EV has increased 67 times in this interval, while CGL, only 4.4 times. EV is a more sensitive parameter than CGL to evaluate embryo growth between the seventh and the tenth week of gestation.

Biological reference materials for extracellular vesicle studies.

PubMed

Valkonen, S; van der Pol, E; Böing, A; Yuana, Y; Yliperttula, M; Nieuwland, R; Laitinen, S; Siljander, P R M

2017-02-15

Extracellular vesicles (EVs) mediate normal physiological homeostasis and pathological processes by facilitating intercellular communication. Research of EVs in basic science and clinical settings requires both methodological standardization and development of reference materials (RM). Here, we show insights and results of biological RM development for EV studies. We used a three-step approach to find and develop a biological RM. First, a literature search was done to find candidates for biological RMs. Second, a questionnaire was sent to EV researchers querying the preferences for RM and their use. Third, a biological RM was selected, developed, characterized, and evaluated. The responses to the survey demonstrated a clear and recognized need for RM optimized for the calibration of EV measurements. Based on the literature, naturally occurring and produced biological RM, such as virus particles and liposomes, were proposed as RM. However, none of these candidate RMs have properties completely matching those of EVs, such as size and refractive index distribution. Therefore, we evaluated the use of nanoerythrosomes (NanoE), vesicles produced from erythrocytes, as a potential biological RM. The strength of NanoE is their resemblance to EVs. Compared to the erythrocyte-derived EVs (eryEVs), NanoE have similar morphology, a similar refractive index (1.37), larger diameter (70% of the NanoE are over 200nm), and increased positive staining for CD235a and lipids (Di-8-ANEPPS) (58% and 67% in NanoE vs. 21% and 45% in eryEVs, respectively). Altogether, our results highlight the general need to develop and validate new RM with similar physical and biochemical properties as EVs to standardize EV measurements between instruments and laboratories. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

Increased liver-specific proteins in circulating extracellular vesicles as potential biomarkers for drug- and alcohol-induced liver injury

PubMed Central

Cho, Young-Eun; Im, Eun-Ju; Moon, Pyong-Gon; Mezey, Esteban; Song, Byoung-Joon; Baek, Moon-Chang

2017-01-01

Drug- and alcohol-induced liver injury are a leading cause of liver failure and transplantation. Emerging evidence suggests that extracellular vesicles (EVs) are a source of biomarkers because they contain unique proteins reflecting the identity and tissue-specific origin of the EV proteins. This study aimed to determine whether potentially hepatotoxic agents, such as acetaminophen (APAP) and binge alcohol, can increase the amounts of circulating EVs and evaluate liver-specific EV proteins as potential biomarkers for liver injury. The circulating EVs, isolated from plasma of APAP-exposed, ethanol-fed mice, or alcoholic hepatitis patients versus normal control counterparts, were characterized by proteomics and biochemical methods. Liver specific EV proteins were analyzed by immunoblots and ELISA. The amounts of total and liver-specific proteins in circulating EVs from APAP-treated mice significantly increased in a dose- and time-dependent manner. Proteomic analysis of EVs from APAP-exposed mice revealed that the amounts of liver-specific and/or hepatotoxic proteins were increased compared to those of controls. Additionally, the increased protein amounts in EVs following APAP exposure returned to basal levels when mice were treated with N-acetylcysteine or glutathione. Similar results of increased amounts and liver-specific proteins in circulating EVs were also observed in mice exposed to hepatotoxic doses of thioacetamide or d-galactosamine but not by non-hepatotoxic penicillin or myotoxic bupivacaine. Additionally, binge ethanol exposure significantly elevated liver-specific proteins in circulating EVs from mice and alcoholics with alcoholic hepatitis, compared to control counterparts. These results indicate that circulating EVs in drug- and alcohol-mediated hepatic injury contain liver-specific proteins that could serve as specific biomarkers for hepatotoxicity. PMID:28225807

Longitudinal safety evaluation of electric vehicles with the partial wireless charging lane on freeways.

PubMed

Li, Ye; Wang, Wei; Xing, Lu; Fan, Qi; Wang, Hao

2018-02-01

As an environment friendly transportation mode, the electric vehicle (EV) has drawn an increasing amount of attention from governments, vehicle manufactories and researchers recently. One of the biggest issue impeding EV's popularization associates with the charging process. The wireless charging lane (WCL) has been proposed as a convenient charging facility for EVs. Due to the high costs, the application of WCL on the entire freeways is impractical in the near future, while the partial WCL (PWCL) may be a feasible solution. This study aims to evaluate longitudinal safety of EVs with PWCL on freeways based on simulations. The simulation experiments are firstly designed, including deployment of PWCL on freeways and distribution of state of charge (SOC) of EVs. Then, a vehicle behavior model for EVs is proposed based on the intelligent driver model (IDM). Two surrogate safety measures, derived from time-to-collision (TTC), are utilized as indicators for safety evaluations. Sensitivity analysis is also conducted for related factors. Results show that the distribution of EVs' SOC significantly affect longitudinal safety when the PWCL is utilized. The low SOC in traffic consisting of EVs has the negative effect on longitudinal safety. The randomness and incompliance of EV drivers worsens the safety performance. The sensitivity analysis indicates that the larger maximum deceleration rate results in the higher longitudinal crash risks of EVs, while the length of PWCL has no monotonous effect. Different TTC thresholds also show no impact on results. A case study shows the consistent results. Based on the findings, several suggestions are discussed for EVs' safety improvement. Results of this study provide useful information for freeway safety when EVs are applied in the future. Copyright © 2017 Elsevier Ltd. All rights reserved.

Comparative analysis of viral shedding in pediatric and adult subjects with central nervous system-associated enterovirus infections from 2013 to 2015 in Switzerland.

PubMed

Cordey, S; Schibler, M; L'Huillier, A G; Wagner, N; Gonçalves, A R; Ambrosioni, J; Asner, S; Turin, L; Posfay-Barbe, K M; Kaiser, L

2017-04-01

Several enterovirus (EV) genotypes can result in aseptic meningitis, but their routes of access to the central nervous system remain to be elucidated and may differ between the pediatric and adult populations. To assess the pattern of viral shedding in pediatric and adult subjects with acute EV meningitis and to generate EV surveillance data for Switzerland. All pediatric and adult subjects admitted to the University Hospitals of Geneva with a diagnosis of EV meningitis between 2013 and 2015 were enrolled. A quantitative EV real-time reverse transcriptase (rRT)-PCR was performed on the cerebrospinal fluid (CSF), blood, stool, urine and respiratory specimens to assess viral shedding and provide a comparative analysis of pediatric and adult populations. EV genotyping was systematically performed. EV positivity rates differed significantly between pediatric and adult subjects; 62.5% of pediatric cases (no adult case) were EV-positive in stool and blood for subjects for whom these samples were all collected. Similarly, the EV viral load in blood was significantly higher in pediatric subjects. Blood C-reactive protein levels were lower and the number of leucocytes/mm 3 in the CSF were higher in non-viremic than in viremic pediatric subjects, respectively. A greater diversity of EV genotypes was observed in pediatric cases, with a predominance of echovirus 30 in children ≥3 years old and adults. In contrast to adults, EV-disseminated infections are predominant in pediatric subjects and show different patterns of EV viral shedding. This observation may be useful for clinicians and contribute to modify current practices of patient care. Copyright © 2017 The Author(s). Published by Elsevier B.V. All rights reserved.

Human pancreatic islet-derived extracellular vesicles modulate insulin expression in 3D-differentiating iPSC clusters

PubMed Central

Andersson, Eva-Marie; Heath, Nikki; Persson-kry, Anette; Collins, Richard; Hicks, Ryan; Dekker, Niek; Forslöw, Anna

2017-01-01

It has been suggested that extracellular vesicles (EVs) can mediate crosstalk between hormones and metabolites within pancreatic tissue. However, the possible effect of pancreatic EVs on stem cell differentiation into pancreatic lineages remains unknown. Herein, human islet-derived EVs (h-Islet-EVs) were isolated, characterized and subsequently added to human induced pluripotent stem cell (iPSC) clusters during pancreatic differentiation. The h-islet-EVs had a mean size of 117±7 nm and showed positive expression of CD63 and CD81 EV markers as measured by ELISA. The presence of key pancreatic transcription factor mRNA, such as NGN3, MAFA and PDX1, and pancreatic hormone proteins such as C-peptide and glucagon, were confirmed in h-Islet-EVs. iPSC clusters were differentiated in suspension and at the end stages of the differentiation protocol, the mRNA expression of the main pancreatic transcription factors and pancreatic hormones was increased. H-Islet-EVs were supplemented to the iPSC clusters in the later stages of differentiation. It was observed that h-Islet-EVs were able to up-regulate the intracellular levels of C-peptide in iPSC clusters in a concentration-dependent manner. The effect of h-Islet-EVs on the differentiation of iPSC clusters cultured in 3D-collagen hydrogels was also assessed. Although increased mRNA expression for pancreatic markers was observed when culturing the iPSC clusters in 3D-collagen hydrogels, delivery of EVs did not affect the insulin or C-peptide intracellular content. Our results provide new information on the role of h-Islet-EVs in the regulation of insulin expression in differentiating iPSC clusters, and are highly relevant for pancreatic tissue engineering applications. PMID:29117231

Comparative analysis of EV isolation procedures for miRNAs detection in serum samples.

PubMed

Andreu, Zoraida; Rivas, Eva; Sanguino-Pascual, Aitana; Lamana, Amalia; Marazuela, Mónica; González-Alvaro, Isidoro; Sánchez-Madrid, Francisco; de la Fuente, Hortensia; Yáñez-Mó, María

2016-01-01

Extracellular vesicles (EVs) are emerging as potent non-invasive biomarkers. However, current methodologies are time consuming and difficult to translate to clinical practice. To analyse EV-encapsulated circulating miRNA, we searched for a quick, easy and economic method to enrich frozen human serum samples for EV. We compared the efficiency of several protocols and commercial kits to isolate EVs. Different methods based on precipitation, columns or filter systems were tested and compared with ultracentrifugation, which is the most classical protocol to isolate EVs. EV samples were assessed for purity and quantity by nanoparticle tracking analysis and western blot or cytometry against major EV protein markers. For biomarker validation, levels of a set of miRNAs were determined in EV fractions and compared with their levels in total serum. EVs isolated with precipitation-based methods were enriched for a subgroup of miRNAs that corresponded to miRNAs described to be encapsulated into EVs (miR-126, miR-30c and miR-143), while the detection of miR-21, miR-16-5p and miR-19a was very low compared with total serum. Our results point to precipitation using polyethylene glycol (PEG) as a suitable method for an easy and cheap enrichment of serum EVs for miRNA analyses. The overall performance of PEG was very similar, or better than other commercial precipitating reagents, in both protein and miRNA yield, but in comparison to them PEG is much cheaper. Other methods presented poorer results, mostly when assessing miRNA by qPCR analyses. Using PEG precipitation in a longitudinal study with human samples, we demonstrated that miRNA could be assessed in frozen samples up to 8 years of storage. We report a method based on a cut-off value of mean of fold EV detection versus serum that provides an estimate of the degree of encapsulation of a given miRNA.

Extracellular vesicle communication pathways as regulatory targets of oncogenic transformation.

PubMed

Choi, Dongsic; Lee, Tae Hoon; Spinelli, Cristiana; Chennakrishnaiah, Shilpa; D'Asti, Esterina; Rak, Janusz

2017-07-01

Pathogenesis of human cancers bridges intracellular oncogenic driver events and their impact on intercellular communication. Among multiple mediators of this 'pathological connectivity' the role of extracellular vesicles (EVs) and their subsets (exosomes, ectosomes, oncosomes) is of particular interest for several reasons. The release of EVs from cancer cells represents a unique mechanism of regulated expulsion of bioactive molecules, a process that also mediates cell-to-cell transfer of lipids, proteins, and nucleic acids. Biological effects of these processes have been implicated in several aspects of cancer-related pathology, including tumour growth, invasion, angiogenesis, metastasis, immunity and thrombosis. Notably, the emerging evidence suggests that oncogenic mutations may impact several aspects of EV-mediated cell-cell communication including: (i) EV release rate and protein content; (ii) molecular composition of cancer EVs; (iii) the inclusion of oncogenic and mutant macromolecules in the EV cargo; (iv) EV-mediated release of genomic DNA; (v) deregulation of mechanisms responsible for EV biogenesis (vesiculome) and (vi) mechanisms of EV uptake by cancer cells. Intriguingly, EV-mediated intercellular transfer of mutant and oncogenic molecules between subpopulations of cancer cells, their indolent counterparts and stroma may exert profound biological effects that often resemble (but are not tantamount to) oncogenic transformation, including changes in cell growth, clonogenicity and angiogenic phenotype, or cause cell stress and death. However, several biological barriers likely curtail a permanent horizontal transformation of normal cells through EV-mediated mechanisms. The ongoing analysis and targeting of EV-mediated intercellular communication pathways can be viewed as a new therapeutic paradigm in cancer, while the analysis of oncogenic cargo contained in EVs released from cancer cells into biofluids is being developed for clinical use as a biomarker and companion diagnostics. Indeed, studies are underway to further explore the multiple links between molecular causality in cancer and various aspects of cellular vesiculation. Copyright © 2017 Elsevier Ltd. All rights reserved.

The eye-voice span during reading aloud

PubMed Central

Laubrock, Jochen; Kliegl, Reinhold

2015-01-01

Although eye movements during reading are modulated by cognitive processing demands, they also reflect visual sampling of the input, and possibly preparation of output for speech or the inner voice. By simultaneously recording eye movements and the voice during reading aloud, we obtained an output measure that constrains the length of time spent on cognitive processing. Here we investigate the dynamics of the eye-voice span (EVS), the distance between eye and voice. We show that the EVS is regulated immediately during fixation of a word by either increasing fixation duration or programming a regressive eye movement against the reading direction. EVS size at the beginning of a fixation was positively correlated with the likelihood of regressions and refixations. Regression probability was further increased if the EVS was still large at the end of a fixation: if adjustment of fixation duration did not sufficiently reduce the EVS during a fixation, then a regression rather than a refixation followed with high probability. We further show that the EVS can help understand cognitive influences on fixation duration during reading: in mixed model analyses, the EVS was a stronger predictor of fixation durations than either word frequency or word length. The EVS modulated the influence of several other predictors on single fixation durations (SFDs). For example, word-N frequency effects were larger with a large EVS, especially when word N-1 frequency was low. Finally, a comparison of SFDs during oral and silent reading showed that reading is governed by similar principles in both reading modes, although EVS maintenance and articulatory processing also cause some differences. In summary, the EVS is regulated by adjusting fixation duration and/or by programming a regressive eye movement when the EVS gets too large. Overall, the EVS appears to be directly related to updating of the working memory buffer during reading. PMID:26441800

Improved resolution in extracellular vesicle populations using 405 instead of 488 nm side scatter

PubMed Central

McVey, Mark J.; Spring, Christopher M.; Kuebler, Wolfgang M.

2018-01-01

ABSTRACT Improvements in identification and assessment of extracellular vesicles (EVs) have fuelled a recent surge in EV publications investigating their roles as biomarkers and mediators of disease. Meaningful scientific comparisons are, however, hampered by difficulties in accurate, reproducible enumeration and characterization of EVs in biological fluids. High-sensitivity flow cytometry (FCM) is presently the most commonly applied strategy to assess EVs, yet its utility is limited by variant ability to resolve smaller EVs. Here, we propose the use of 405 nm (violet) wavelength lasers in place of 488 nm (blue) for side scatter (SSC) detection to obtain greater resolution of EVs using high-sensitivity FCM. To test this hypothesis, we modelled EV resolution by violet versus blue SSC in silico and compared resolution of reference beads and biological EVs from plasma and bronchoalveolar lavage (BAL) fluid using either violet or blue wavelength SSC EV detection. Mie scatter modelling predicted that violet as compared to blue SSC increases resolution of small (100–500 nm) spherical particles with refractive indices (1.34–1.46) similar to EVs by approximately twofold in terms of light intensity and by nearly 20% in SSC signal quantum efficiency. Resolution of reference beads was improved by violet instead of blue SSC with two- and fivefold decreases in coefficients of variation for particles of 300–500 nm and 180–240 nm size, respectively. Resolution was similarly improved for detection of EVs from plasma or BAL fluid. Violet SSC detection for high-sensitivity FCM allows for significantly greater resolution of EVs in plasma and BAL compared to conventional blue SSC and particularly improves resolution of smaller EVs. Notably, the proposed strategy is readily implementable and inexpensive for machines already equipped with 405 nm SSC or the ability to accommodate 405/10 nm bandpass filters in their violet detector arrays. PMID:29696076

Human enterovirus D68 in clinical and sewage samples in Israel.

PubMed

Weil, Merav; Mandelboim, Michal; Mendelson, Ella; Manor, Yossi; Shulman, Lester; Ram, Daniela; Barkai, Galia; Shemer, Yonat; Wolf, Dana; Kra-Oz, Zipi; Weiss, Leah; Pando, Rakefet; Hindiyeh, Musa; Sofer, Danit

2017-01-01

Since mid-August 2014, North America experienced a wide outbreak of Enterovirus D68 (EV-D68) associated with severe respiratory illness in children. Several other countries also reported cases of EV-D68 in 2014. The aim of this study was to determine whether EV-D68 circulated in Israel in 2014, caused severe respiratory illness in children and was the causative agent of Acute Flaccid Paralysis. Archived clinical respiratory samples from a cohort of 710 hospitalized pediatric patient's (<10years old) with respiratory illness were screened for clade B specific EV-D68 by real-time PCR. The patients were seen at four medical centers covering the entire country between August and November 2014. We also evaluated 49 patient stool samples from 26 AFP cases during 2014 for presence of EV-D68. In addition, RNA from sewage samples collected throughout Israel during the same study period was also tested for EV-D68. Partial VP1 sequencing was performed on all positive samples. Of the 710 clinical samples evaluated, 7 (1%) were positive for EV-D68. Two patients were from the central part of Israel, while the rest was from the southern part. The majority of the patients did not have any underlying disease. Not only that, but, none of the 26 suspected AFP cases had EV-D68 nucleic acid in their stool samples. EV-D68 RNA was detected in 9 out of 93 sewage samples, mainly from Southern Israel. Sequence analysis of EV-D68 VP1 gene from both sewage and clinical samples indicated that the Israeli EV-D68 RNA belonged to Clade B which was genetically similar to 2014 circulating European and North American EV-D68 virus. EV-D68 circulated in Israel during the 2014 summer-fall season and caused hospitalization of a small percent of the patients with respiratory illness. Copyright © 2016 Elsevier B.V. All rights reserved.

EnVision+, a new dextran polymer-based signal enhancement technique for in situ hybridization (ISH).

PubMed

Wiedorn, K H; Goldmann, T; Henne, C; Kühl, H; Vollmer, E

2001-09-01

Seventy paraffin-embedded cervical biopsy specimens and condylomata were tested for the presence of human papillomavirus (HPV) by conventional in situ hybridization (ISH) and ISH with subsequent signal amplification. Signal amplification was performed either by a commercial biotinyl-tyramide-based detection system [GenPoint (GP)] or by the novel two-layer dextran polymer visualization system EnVision+ (EV), in which both EV-horseradish peroxidase (EV-HRP) and EV-alkaline phosphatase (EV-AP) were applied. We could demonstrate for the first time, that EV in combination with preceding ISH results in a considerable increase in signal intensity and sensitivity without loss of specificity compared to conventional ISH. Compared to GP, EV revealed a somewhat lower sensitivity, as measured by determination of the integrated optical density (IOD) of the positively stained cells. However, EV is easier to perform, requires a shorter assay time, and does not raise the background problems that may be encountered with biotinyl-tyramide-based amplification systems. (J Histochem Cytochem 49:1067-1071, 2001)

Enterovirus and parechovirus infection in children: a brief overview.

PubMed

de Crom, S C M; Rossen, J W A; van Furth, A M; Obihara, C C

2016-08-01

Enterovirus and parechovirus are a frequent cause of infection in children. This review is an overview of what is known from enterovirus and parechovirus infection in children and contains information about the epidemiology, pathogenesis, clinical presentation, diagnosis, treatment, and prognosis of enterovirus and parechovirus infection in children. EV and HPeV infections are a frequent cause of infection in childhood. The clinical presentation is diverse. RT-qPCR is the best way to detect an EV or HPeV. Cerebrospinal fluid, blood and feces have the highest sensitivity for detecting an EV or HPeV. There is no treatment for EV and HPeV infections. Two vaccines against EV 71 are just licensed in China and will be available on the private market. Little is known about the prognosis of EV and HPeV infections. •EV and HPeV are a frequent cause of infection in children. What is new: •This review gives a brief overview over EV and HPeV infection in children.

Potential Role of Extracellular Vesicles in the Pathophysiology of Drug Addiction.

PubMed

Rao, P S S; O'Connell, Kelly; Finnerty, Thomas Kyle

2018-01-23

Extracellular vesicles (EVs) are small vesicles secreted by cells and are known to carry sub-cellular components including microRNA, proteins, and lipids. Due to their ability to transport cargo between cells, EVs have been identified as important regulators of various pathophysiological conditions and can therefore influence treatment outcomes. In particular, the significance of microRNAs in EV-mediated cell-cell communication is well-documented. While the influence of EVs and the cargo delivered by EVs has been extensively reviewed in other neurological disorders, the available literature on the potential role of EVs in the pathophysiology of drug addiction has not been reviewed. Hence, in this article, the known effects of commonly abused drugs (ethanol, nicotine, opiates, cocaine, and cannabinoids) on EV secretion have been reviewed. In addition, the potential role of drugs of abuse in affecting the delivery of EV-packaged microRNAs, and the subsequent impact on neuronal health and continued drug dependence, has been discussed.

Extracellular vesicles at the cross-line between basic science and clinical needs.

PubMed

Sasso, Luana; Hosamuddin, Huma; Emanueli, Costanza

2017-01-01

MiRNAs are small noncoding RNAs vital for protein regulation and gene expression. Since their discovery in the early nineties, many of their intracellular roles have been characterized. However, it is only recently that EVs loaded with miRNAs and other molecular types have started to be appreciated for their substantial involvement in cell-to-cell communication and signaling in physiological and pathological processes. EVs cell-to-cell signaling functions are complex and largely unknown, which still hampers the direct use of endogenous engineered EVs as therapeutics. However, ad hoc engineered synthetic EVs could represent new therapeutics. The potential of EV-inspired delivery carriers has now attracted the interest of the pharmaceutical industry and has challenged drug delivery researchers with new questions. This review will focus on EVs and EV-inspired drug delivery carriers, on their potential and on the challenges involved in the use of EV-inspired drug delivery systems. © 2016 John Wiley & Sons Ltd.

Production of EV71 vaccine candidates.

PubMed

Chong, Pele; Hsieh, Shih-Yang; Liu, Chia-Chyi; Chou, Ai-Hsiang; Chang, Jui-Yuan; Wu, Suh-Chin; Liu, Shih-Jen; Chow, Yen-Hung; Su, Ih-Jen; Klein, Michel

2012-12-01

Enterovirus 71 (EV71) is now recognized as an emerging neurotropic virus in Asia and with Coxsackie virus (CV) it is the other major causative agent of hand-foot-mouth diseases (HFMD). Effective medications and/or prophylactic vaccines against HFMD are urgently needed. From a scientific (the feasibility of bioprocess, immunological responses and potency in animal challenge model) and business development (cost of goods) points of view, we in this review address and discuss the pros and cons of different EV71 vaccine candidates that have been produced and evaluated in animal models. Epitope-based synthetic peptide vaccine candidates containing residues 211-225 of VP1 formulated with Freund's adjuvant (CFA/IFA) elicited low EV71 virus neutralizing antibody responses, but were protective in the suckling mouse challenge model. Among recombinant EV71 subunits (rVP1, rVP2 and rVP3) expressed in E. coli, purified and formulated with CFA/IFA, only VP1 elicited mouse antibody responses with measurable EV71-specific virus neutralization titers. Immunization of mice with either a DNA plasmid containing VP1 gene or VP1 expressed in Salmonella typhimurium also generated neutralizing antibody responses and protected animals against a live EV71 challenge. Recombinant EV71 virus-like particles (rVLP) produced from baculovirus formulated either with CFA/IFA or alum elicited good virus neutralization titers in both mice and non-human primates, and were found to be protective in the suckling mouse EV71 challenge model. Synthetic peptides or recombinant EV71 subunit vaccines (rVP1 and rVLP) formulated in alum were found to be poorly immunogenic in rabbits. Only formalin-inactivated (FI) EV71 virions formulated in alum elicited cross-neutralizing antibodies against different EV71 genotypes in mice, rabbits and non-human primates but induced weak neutralizing responses against CAV16. From a regulatory, economic and market acceptability standpoint, FI-EV71 virion vaccines are the most promising candidates and are currently being evaluated in human clinical trials. We further describe and analyze some new bioprocesses technologies that have great potential applications in EV71 vaccine development. This review also demonstrates the opportunities and challenges that the Asian vaccine industry faces today.

Systematic Methodological Evaluation of a Multiplex Bead-Based Flow Cytometry Assay for Detection of Extracellular Vesicle Surface Signatures

PubMed Central

Wiklander, Oscar P. B.; Bostancioglu, R. Beklem; Welsh, Joshua A.; Zickler, Antje M.; Murke, Florian; Corso, Giulia; Felldin, Ulrika; Hagey, Daniel W.; Evertsson, Björn; Liang, Xiu-Ming; Gustafsson, Manuela O.; Mohammad, Dara K.; Wiek, Constanze; Hanenberg, Helmut; Bremer, Michel; Gupta, Dhanu; Björnstedt, Mikael; Giebel, Bernd; Nordin, Joel Z.; Jones, Jennifer C.; EL Andaloussi, Samir; Görgens, André

2018-01-01

Extracellular vesicles (EVs) can be harvested from cell culture supernatants and from all body fluids. EVs can be conceptually classified based on their size and biogenesis as exosomes and microvesicles. Nowadays, it is however commonly accepted in the field that there is a much higher degree of heterogeneity within these two subgroups than previously thought. For instance, the surface marker profile of EVs is likely dependent on the cell source, the cell’s activation status, and multiple other parameters. Within recent years, several new methods and assays to study EV heterogeneity in terms of surface markers have been described; most of them are being based on flow cytometry. Unfortunately, such methods generally require dedicated instrumentation, are time-consuming and demand extensive operator expertise for sample preparation, acquisition, and data analysis. In this study, we have systematically evaluated and explored the use of a multiplex bead-based flow cytometric assay which is compatible with most standard flow cytometers and facilitates a robust semi-quantitative detection of 37 different potential EV surface markers in one sample simultaneously. First, assay variability, sample stability over time, and dynamic range were assessed together with the limitations of this assay in terms of EV input quantity required for detection of differently abundant surface markers. Next, the potential effects of EV origin, sample preparation, and quality of the EV sample on the assay were evaluated. The findings indicate that this multiplex bead-based assay is generally suitable to detect, quantify, and compare EV surface signatures in various sample types, including unprocessed cell culture supernatants, cell culture-derived EVs isolated by different methods, and biological fluids. Furthermore, the use and limitations of this assay to assess heterogeneities in EV surface signatures was explored by combining different sets of detection antibodies in EV samples derived from different cell lines and subsets of rare cells. Taken together, this validated multiplex bead-based flow cytometric assay allows robust, sensitive, and reproducible detection of EV surface marker expression in various sample types in a semi-quantitative way and will be highly valuable for many researchers in the EV field in different experimental contexts.

Systematic Methodological Evaluation of a Multiplex Bead-Based Flow Cytometry Assay for Detection of Extracellular Vesicle Surface Signatures.

PubMed

Wiklander, Oscar P B; Bostancioglu, R Beklem; Welsh, Joshua A; Zickler, Antje M; Murke, Florian; Corso, Giulia; Felldin, Ulrika; Hagey, Daniel W; Evertsson, Björn; Liang, Xiu-Ming; Gustafsson, Manuela O; Mohammad, Dara K; Wiek, Constanze; Hanenberg, Helmut; Bremer, Michel; Gupta, Dhanu; Björnstedt, Mikael; Giebel, Bernd; Nordin, Joel Z; Jones, Jennifer C; El Andaloussi, Samir; Görgens, André

2018-01-01

Extracellular vesicles (EVs) can be harvested from cell culture supernatants and from all body fluids. EVs can be conceptually classified based on their size and biogenesis as exosomes and microvesicles. Nowadays, it is however commonly accepted in the field that there is a much higher degree of heterogeneity within these two subgroups than previously thought. For instance, the surface marker profile of EVs is likely dependent on the cell source, the cell's activation status, and multiple other parameters. Within recent years, several new methods and assays to study EV heterogeneity in terms of surface markers have been described; most of them are being based on flow cytometry. Unfortunately, such methods generally require dedicated instrumentation, are time-consuming and demand extensive operator expertise for sample preparation, acquisition, and data analysis. In this study, we have systematically evaluated and explored the use of a multiplex bead-based flow cytometric assay which is compatible with most standard flow cytometers and facilitates a robust semi-quantitative detection of 37 different potential EV surface markers in one sample simultaneously. First, assay variability, sample stability over time, and dynamic range were assessed together with the limitations of this assay in terms of EV input quantity required for detection of differently abundant surface markers. Next, the potential effects of EV origin, sample preparation, and quality of the EV sample on the assay were evaluated. The findings indicate that this multiplex bead-based assay is generally suitable to detect, quantify, and compare EV surface signatures in various sample types, including unprocessed cell culture supernatants, cell culture-derived EVs isolated by different methods, and biological fluids. Furthermore, the use and limitations of this assay to assess heterogeneities in EV surface signatures was explored by combining different sets of detection antibodies in EV samples derived from different cell lines and subsets of rare cells. Taken together, this validated multiplex bead-based flow cytometric assay allows robust, sensitive, and reproducible detection of EV surface marker expression in various sample types in a semi-quantitative way and will be highly valuable for many researchers in the EV field in different experimental contexts.

Flow cytometric analysis of extracellular vesicle subsets in plasma: impact of swarm by particles of non-interest.

PubMed

Libregts, S F W M; Arkesteijn, G J A; Németh, A; Nolte-'t Hoen, E N M; Wauben, M H M

2018-05-20

Essentials Extracellular vesicles (EVs) in biological fluids are promising biomarkers for disease. Fluorescence-based flow cytometric analysis is suitable to detect low abundant EV subsets. Particles of non-interest can induce false-positive light scatter and fluorescent signals. Interference of particles of non-interest can be monitored by analyzing serial dilutions. Background Extracellular vesicles (EVs) in plasma are increasingly being recognized as potential biomarkers. EV analysis for diagnostic purposes should be robust and should allow analysis of EV subsets with a wide range of abundance and in a large number of patient samples. Flow cytometry offers possibilities to meet these criteria, as it allows multiparameter analysis of individual EVs. However, analysis of plasma EVs is challenging, because of their size and heterogeneity, and the presence of other submicrometer-sized particles in plasma that could interfere with EV analysis. Objectives To explore whether fluorescence-based flow cytometric analysis of EV subsets is suitable when the EVs of interest are present in low abundance in a background of non-labeled or differently labeled EVs and particles. Methods Fluorescently labeled EVs of interest were spiked at different ratios in full plasma, purified plasma components, or (non-)fluorescent polystyrene beads, and subsequently analyzed by flow cytometry with fluorescence threshold triggering. Results We found that light scatter detection of low-abundance or rare EV subsets during fluorescence threshold triggering was severely affected by particles of non-interest, owing to coincidence and swarming. Importantly, we show that interfering particles labeled with different fluorophores induced false-positive fluorescent signals on the particles of interest. These unwanted effects could only be discerned and controlled by performing serial dilutions and analyzing light scatter and fluorescence parameters. Conclusions We demonstrate how particles of non-interest in plasma can impact on the light scatter and fluorescence detection of low-abundance EVs of interest during fluorescence-based flow cytometric analysis, and provide a means to prevent erroneous data interpretation. © 2018 The Authors. Journal of Thrombosis and Haemostasis published by Wiley Periodicals, Inc. on behalf of International Society on Thrombosis and Haemostasis.

TRASANA. Force Stratification System User Handbook.

DTIC Science & Technology

1977-09-01

n - 4 u m W CL a 0-C C COCOc:11 7 k., 0 =. t.4 c-Ac( ~ lqa . U~ ~~~~~ p. c A? W 0=0 ’. 0 a - 0 - EO U=ON~.S4’ o S .0 p. . 0 0 0 ’ ft 0 LO Cl O.CCL. . k...ADGRU, USAF Institute of Technology I Chief, ADGRU, US Naval War College 1 Chief, US Elm, USAF Air-Ground Operations School 1 USA Rep, USMC Educ Cen...Marine Corps Dev and Educ Command I USA Rep, Comb Svc Spt Prog School, Atlantic I Commandant, USA Element, School of Music 1 Liaison Offices/Officers

Design of Computer-Related Workstations in Relation to Job Functions and Productivity.

DTIC Science & Technology

1984-12-01

I nadequat e Neut ra I Adequat e Mana,’,ement 36.6% 17. 3 46.2 C Compiter Prog. 45.1 25.3 29.7 Syst ems Analyst 44.6 20.0 35.4 FPmctional Analyst...2~ der md prJgrvTm~r C S 4 0 6 I 0 I S I S g tS * Ar . ± " S . l _ - ---- ,E- , ! ---- - -.-- _EA E L 2 . ... .. __., ____ I - / 1: - i - - " I...there ;ire lrina van :ibl -e; which .0 tect fati I I ty desi gn and l ayout. In a professional * oputr-rlatd ’vi ronimInt , sat i stact 110 with

Comment on ‘Numerical estimates of the spectrum for anharmonic PT symmetric potentials’

NASA Astrophysics Data System (ADS)

Amore, Paolo; Fernández, Francisco M.

2013-04-01

We show that the authors of the commented paper (Bowen et al 2012 Phys. Scr. 85 065005) draw their conclusions from the eigenvalues of truncated Hamiltonian matrices that do not converge as the matrix dimension increases. In some of the studied examples, the authors missed the real positive eigenvalues that already converge towards the exact eigenvalues of the non-Hermitian operators and focused their attention on the complex ones that do not. We also show that the authors misread Bender's argument about the eigenvalues of the harmonic oscillator with boundary conditions in the complex-x plane (Bender 2007 Rep. Prog. Phys. 70 947).

DIRECT COUPLED PROGRESSIVE STAGE PULSE COUNTER APPARATUS

DOEpatents

Kaufman, W.M.

1962-08-14

A progressive electrical pulse counter circuit was designed for the counting of a chain of input pulses of random width and/or frequency. The circuit employs an odd and even pulse input line alternately connected to a series of directly connected bistable counting stages. Each bistable stage has two d-c operative states which stage, when in its rnrtial state, prevents the next succeeding stage from changing its condition when the latter stage is pulsed. Since only altennate stages are pulsed for each incoming pulse, only one stage will change its state for each input pulse thereby providing prog essive stage by stage counting. (AEC)

Quenched-in defects in flashlamp-annealed silicon

NASA Technical Reports Server (NTRS)

Borenstein, J. T.; Jones, J. T.; Corbett, J. W.; Oehrlein, G. S.; Kleinhenz, R. L.

1986-01-01

Deep levels introduced in boron-doped silicon by heat-pulse annealing with a tungsten-halogen flashlamp are investigated using deep-level transient spectroscopy. Two majority-carrier trapping levels in the band gap, at Ev + 0.32 eV and at Ev + 0.45 eV, are observed. These results are compared to those obtained by furnace-quenching and laser-annealing studies. Both the position in the gap and the annealing kinetics of the hole trap at Ev + 0.45 eV suggest that this center is due to an interstitial iron impurity in the lattice. The deep levels are not consistently observed in all flashlamp-annealed Si crystals utilized.

A Method for Isolation of Extracellular Vesicles and Characterization of Exosomes from Brain Extracellular Space.

PubMed

Pérez-González, Rocío; Gauthier, Sebastien A; Kumar, Asok; Saito, Mitsuo; Saito, Mariko; Levy, Efrat

2017-01-01

Extracellular vesicles (EV), including exosomes, secreted vesicles of endocytic origin, and microvesicles derived from the plasma membrane, have been widely isolated and characterized from conditioned culture media and bodily fluids. The difficulty in isolating EV from tissues, however, has hindered their study in vivo. Here, we describe a novel method designed to isolate EV and characterize exosomes from the extracellular space of brain tissues. The purification of EV is achieved by gentle dissociation of the tissue to free the brain extracellular space, followed by sequential low-speed centrifugations, filtration, and ultracentrifugations. To further purify EV from other extracellular components, they are separated on a sucrose step gradient. Characterization of the sucrose step gradient fractions by electron microscopy demonstrates that this method yields pure EV preparations free of large vesicles, subcellular organelles, or debris. The level of EV secretion and content are determined by assays for acetylcholinesterase activity and total protein estimation, and exosomal identification and protein content are analyzed by Western blot and immuno-electron microscopy. Additionally, we present here a method to delipidate EV in order to improve the resolution of downstream electrophoretic analysis of EV proteins.

Multiparametric plasma EV profiling facilitates diagnosis of pancreatic malignancy.

PubMed

Yang, Katherine S; Im, Hyungsoon; Hong, Seonki; Pergolini, Ilaria; Del Castillo, Andres Fernandez; Wang, Rui; Clardy, Susan; Huang, Chen-Han; Pille, Craig; Ferrone, Soldano; Yang, Robert; Castro, Cesar M; Lee, Hakho; Del Castillo, Carlos Fernandez; Weissleder, Ralph

2017-05-24

Pancreatic ductal adenocarcinoma (PDAC) is usually detected late in the disease process. Clinical workup through imaging and tissue biopsies is often complex and expensive due to a paucity of reliable biomarkers. We used an advanced multiplexed plasmonic assay to analyze circulating tumor-derived extracellular vesicles (tEVs) in more than 100 clinical populations. Using EV-based protein marker profiling, we identified a signature of five markers (PDAC EV signature) for PDAC detection. In our prospective cohort, the accuracy for the PDAC EV signature was 84% [95% confidence interval (CI), 69 to 93%] but only 63 to 72% for single-marker screening. One of the best markers, GPC1 alone, had a sensitivity of 82% (CI, 60 to 95%) and a specificity of 52% (CI, 30 to 74%), whereas the PDAC EV signature showed a sensitivity of 86% (CI, 65 to 97%) and a specificity of 81% (CI, 58 to 95%). The PDAC EV signature of tEVs offered higher sensitivity, specificity, and accuracy than the existing serum marker (CA 19-9) or single-tEV marker analyses. This approach should improve the diagnosis of pancreatic cancer. Copyright © 2017, American Association for the Advancement of Science.

The key role of extracellular vesicles in the metastatic process.

PubMed

Zhao, Hongyun; Achreja, Abhinav; Iessi, Elisabetta; Logozzi, Mariantonia; Mizzoni, Davide; Di Raimo, Rossella; Nagrath, Deepak; Fais, Stefano

2018-01-01

Extracellular vesicles (EVs), including exosomes, have a key role in the paracrine communication between organs and compartments. EVs shuttle virtually all types of biomolecules such as proteins, lipids, nucleic acids, metabolites and even pharmacological compounds. Their ability to transfer their biomolecular cargo into target cells enables EVs to play a key role in intercellular communication that can regulate cellular functions such as proliferation, apoptosis and migration. This has led to the emergence of EVs as a key player in tumor growth and metastasis through the formation of "tumor niches" in target organs. Recent data have also been shown that EVs may transform the microenvironment of primary tumors thus favoring the selection of cancer cells with a metastatic behavior. The release of EVs from resident non-malignant cells may contribute to the metastatic processes as well. However, cancer EVs may induce malignant transformation in resident mesenchymal stem cells, suggesting that the metastatic process is not exclusively due to circulating tumor cells. In this review, we outline and discuss evidence-based roles of EVs in actively regulating multiple steps of the metastatic process and how we can leverage EVs to impair metastasis. Copyright © 2017 Elsevier B.V. All rights reserved.

Extracellular vesicles as shuttles of tumor biomarkers and anti-tumor drugs.

PubMed

Zocco, Davide; Ferruzzi, Pietro; Cappello, Francesco; Kuo, Winston Patrick; Fais, Stefano

2014-01-01

Extracellular vesicles (EV) include vesicles released by either normal or tumor cells. EV may exceed the nanometric scale (microvesicles), or to be within the nanoscale, also called exosomes. Thus, it appears that only exosomes and larger vesicles may have the size for potential applications in nanomedicine, in either disease diagnosis or therapy. This is of particular interest for research in cancer, also because the vast majority of existing data on EV are coming from pre-clinical and clinical oncology. We know that the microenvironmental features of cancer may favor cell-to-cell paracrine communication through EV, but EV have been purified, characterized, and quantified from plasma of tumor patients as well, thus suggesting that EV may have a role in promoting and maintaining cancer dissemination and progression. These observations are prompting research efforts to evaluate the use of nanovesicles as tumor biomarkers. Moreover, EVs are emerging as natural delivery systems and in particular, exosomes may represent the ideal natural nanoshuttles for new and old anti-tumor drugs. However, much is yet to be understood about the role of EV in oncology and this article aims to discuss the future of EV in cancer on the basis of current knowledge.

Extracellular Vesicles as Shuttles of Tumor Biomarkers and Anti-Tumor Drugs

PubMed Central

Zocco, Davide; Ferruzzi, Pietro; Cappello, Francesco; Kuo, Winston Patrick; Fais, Stefano

2014-01-01

Extracellular vesicles (EV) include vesicles released by either normal or tumor cells. EV may exceed the nanometric scale (microvesicles), or to be within the nanoscale, also called exosomes. Thus, it appears that only exosomes and larger vesicles may have the size for potential applications in nanomedicine, in either disease diagnosis or therapy. This is of particular interest for research in cancer, also because the vast majority of existing data on EV are coming from pre-clinical and clinical oncology. We know that the microenvironmental features of cancer may favor cell-to-cell paracrine communication through EV, but EV have been purified, characterized, and quantified from plasma of tumor patients as well, thus suggesting that EV may have a role in promoting and maintaining cancer dissemination and progression. These observations are prompting research efforts to evaluate the use of nanovesicles as tumor biomarkers. Moreover, EVs are emerging as natural delivery systems and in particular, exosomes may represent the ideal natural nanoshuttles for new and old anti-tumor drugs. However, much is yet to be understood about the role of EV in oncology and this article aims to discuss the future of EV in cancer on the basis of current knowledge. PMID:25340037

Extracellular Vesicles in Hematological Malignancies: From Biology to Therapy

PubMed Central

Caivano, Antonella; La Rocca, Francesco; Laurenzana, Ilaria; Trino, Stefania; De Luca, Luciana; Lamorte, Daniela; Del Vecchio, Luigi; Musto, Pellegrino

2017-01-01

Extracellular vesicles (EVs) are a heterogeneous group of particles, between 15 nanometers and 10 microns in diameter, released by almost all cell types in physiological and pathological conditions, including tumors. EVs have recently emerged as particularly interesting informative vehicles, so that they could be considered a true “cell biopsy”. Indeed, EV cargo, including proteins, lipids, and nucleic acids, generally reflects the nature and status of the origin cells. In some cases, EVs are enriched of peculiar molecular cargo, thus suggesting at least a degree of specific cellular packaging. EVs are identified as important and critical players in intercellular communications in short and long distance interplays. Here, we examine the physiological role of EVs and their activity in cross-talk between bone marrow microenvironment and neoplastic cells in hematological malignancies (HMs). In these diseases, HM EVs can modify tumor and bone marrow microenvironment, making the latter “stronger” in supporting malignancy, inducing drug resistance, and suppressing the immune system. Moreover, EVs are abundant in biologic fluids and protect their molecular cargo against degradation. For these and other “natural” characteristics, EVs could be potential biomarkers in a context of HM liquid biopsy and therapeutic tools. These aspects will be also analyzed in this review. PMID:28574430

Disruption of Membranes of Extracellular Vesicles Is Necessary for ELISA Determination of Urine AQP2: Proof of Disruption and Epitopes of AQP2 Antibodies

PubMed Central

Nameta, Masaaki; Saijo, Yoko; Ohmoto, Yasukazu; Katsuragi, Kiyonori; Yamamoto, Keiko; Yamamoto, Tadashi; Ishibashi, Kenichi; Sasaki, Sei

2016-01-01

Aquaporin-2 (AQP2) is present in urine extracellular vesicles (EVs) and is a useful biomarker for water balance disorders. We previously found that pre-treatment of urine with alkali/detergent or storage at −25 °C is required for enzyme-linked immunosorbent assay (ELISA) measurement. We speculated that disruptions of EVs membranes are necessary to allow for the direct contact of antibodies with their epitopes. Human urine EVs were prepared using an ultracentrifugation method. Urine EV samples were stored at different temperatures for a week. Electron microscopy showed abundant EVs with diameters of 20–100 nm, consistent with those of exosomes, in normal urine, whereas samples from alkali/detergent pre-treated urine showed fewer EVs with large swollen shapes and frequent membrane disruptions. The abundance and structures of EVs were maintained during storage at −80 °C, but were severely damaged at −25 °C. Binding and competitive inhibition assays showed that epitopes of monoclonal antibody and polyclonal antibody were the hydrophilic Loop D and C-terminus of AQP2, respectively, both of which are present on the inner surface of EVs. Thus, urine storage at −25 °C or pre-treatment with alkali/detergent disrupt EVs membranes and allow AQP2 antibodies to bind to their epitopes located inside EVs. PMID:27681727

Functional transferred DNA within extracellular vesicles

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

Cai, Jin; Department of Neurology, Jinling Hospital, Nanjing University School of Medicine, Jiangsu Province; Wu, Gengze

Extracellular vesicles (EVs) are small membrane vesicles including exosomes and shedding vesicles that mediated a cell-to-cell communication. EVs are released from almost all cell types under both physiological and pathological conditions and incorporate nuclear and cytoplasmic molecules for intercellular delivery. Besides protein, mRNA, and microRNA of these molecules, as recent studies show, specific DNA are prominently packaged into EVs. It appears likely that some of exosomes or shedding vesicles, bearing nuclear molecules are released upon bubble-like blebs. Specific interaction of EVs with susceptible recipients performs the uptake of EVs into the target cells, discharging their cargo including nuclear and cytoplasmicmore » macromolecules into the cytosol. These findings expand the nucleic acid content of EVs to include increased levels of specific DNA. Thus, EVs contain a repertoire of genetic information available for horizontal gene transfer and potential use as blood biomarkers for cancer and atherosclerosis. In this review, the focus is on the characteristics, biological functions, and roles in diseases of DNA within EVs. - Highlights: • This review is focused on the DNA within EVs including its characteristics, biological functions, and roles in diseases. • It is clear that DNA within EVs might have important physiological and pathological roles in various diseases. • Knowledge in this area may provides us alternative methods for disease diagnosis or therapy in the future.« less

Development of a magnetic bead-based method for the collection of circulating extracellular vesicles.

PubMed

Shih, Chun-Liang; Chong, Kowit-Yu; Hsu, Shih-Che; Chien, Hsin-Jung; Ma, Ching-Ting; Chang, John Wen-Cheng; Yu, Chia-Jung; Chiou, Chiuan-Chian

2016-01-25

Cells release different types of extracellular vesicles (EVs). These EVs contain biomolecules, including proteins and nucleic acids, from their parent cells, which can be useful for diagnostic applications. The aim of this study was to develop a convenient procedure to collect circulating EVs with detectable mRNA or other biomolecules. Magnetic beads coated with annexin A5 (ANX-beads), which bound to phosphatidylserine moieties on the surfaces of most EVs, were tested for their ability to capture induced apoptotic bodies in vitro and other phosphatidylserine-presenting vesicles in body fluids. Our results show that up to 60% of induced apoptotic bodies could be captured by the ANX-beads. The vesicles captured from cultured media or plasma contained amplifiable RNA. Suitable blood samples for EV collection included EDTA-plasma and serum but not heparin-plasma. In addition, EVs in plasma were labile to freeze-and-thaw cycles. In rodents xenografted with human cancer cells, tumor-derived mRNA could be detected in EVs captured from serum samples. Active proteins could be detected in EVs captured from ascites but not from plasma. In conclusion, we have developed a magnetic bead-based procedure for the collection of EVs from body fluids and proved that captured EVs contain biomolecules from their parent cells, and therefore have great potential for disease diagnosis. Copyright © 2015 Elsevier B.V. All rights reserved.

Exercise-induced circulating extracellular vesicles protect against cardiac ischemia-reperfusion injury.

PubMed

Bei, Yihua; Xu, Tianzhao; Lv, Dongchao; Yu, Pujiao; Xu, Jiahong; Che, Lin; Das, Avash; Tigges, John; Toxavidis, Vassilios; Ghiran, Ionita; Shah, Ravi; Li, Yongqin; Zhang, Yuhui; Das, Saumya; Xiao, Junjie

2017-07-01

Extracellular vesicles (EVs) serve an important function as mediators of intercellular communication. Exercise is protective for the heart, although the signaling mechanisms that mediate this cardioprotection have not been fully elucidated. Here using nano-flow cytometry, we found a rapid increase in plasma EVs in human subjects undergoing exercise stress testing. We subsequently identified that serum EVs were increased by ~1.85-fold in mice after 3-week swimming. Intramyocardial injection of equivalent quantities of EVs from exercised mice and non-exercised controls provided similar protective effects against acute ischemia/reperfusion (I/R) injury in mice. However, injection of exercise-induced EVs in a quantity equivalent to the increase seen with exercise (1.85 swim group) significantly enhanced the protective effect. Similarly, treatment with exercise-induced increased EVs provided additional anti-apoptotic effect in H 2 O 2 -treated H9C2 cardiomyocytes mediated by the activation of ERK1/2 and HSP27 signaling. Finally, by treating H9C2 cells with insulin-like growth factor-1 to mimic exercise stimulus in vitro, we found an increased release of EVs from cardiomyocytes associated with ALIX and RAB35 activation. Collectively, our results show that exercise-induced increase in circulating EVs enhances the protective effects of endogenous EVs against cardiac I/R injury. Exercise-derived EVs might serve as a potent therapy for myocardial injury in the future.

Identification of a Novel Enterovirus Species in Rhesus Macaque in China.

PubMed

Ao, Yuan-Yun; Yu, Jie-Mei; Zhang, Cui-Yuan; Xin, Yun-Yun; Li, Li-Li; Duan, Zhao-Jun

2016-06-22

Recent studies of Enterovirus (EV) in nonhuman primates (NHPs), which could act as a source of future emerging human viral diseases, have boosted interest in the search for novel EVs. Here, a highly divergent strain of EV, tentatively named SEV-gx, was identified by viral metagenomic analysis from stool samples of rhesus macaques in China. In total, 27 of 280 (9.6%) faecal samples from rhesus macaques were positive for SEV-gx. Its complete genomic sequence is 7,367 nucleotide (nt). Genomic analyses showed that it has a standard genomic organisation for EVs, being more closely related to EV-J strains (approximately 54.0%, 43.0-44.1%, 52.3-55.2%, 61.1-62.7% and 64.0% amino acids identity in polyprotein, P1, P2 and P3 and combined 2C/3CD regions, respectively). It was also shown to have genome characteristics typical of EVs. Phylogenetic analysis of P1, 2C and 3CD aa indicated that SEV-gx can be classified as a distinct cluster in the EVs. All of this evidence demonstrates SEV-gx is a novel species (tentatively named EV-K) in the EV genus, which contributes to our understanding of the genetic diversity and evolution of EVs. Further studies are needed to investigate the potential pathogenicity of SEV-gx in NHPs and humans.

Vectorization of biomacromolecules into cells using extracellular vesicles with enhanced internalization induced by macropinocytosis.

PubMed

Nakase, Ikuhiko; Noguchi, Kosuke; Fujii, Ikuo; Futaki, Shiroh

2016-10-17

Extracellular vesicles (EVs, exosomes) are approximately 30- to 200-nm-long vesicles that have received increased attention due to their role in cell-to-cell communication. Although EVs are highly anticipated to be a next-generation intracellular delivery tool because of their pharmaceutical advantages, including non-immunogenicity, their cellular uptake efficacy is low because of the repulsion of EVs and negatively charged cell membranes and size limitations in endocytosis. Here, we demonstrate a methodology for achieving enhanced cellular EV uptake using arginine-rich cell-penetrating peptides (CPPs) to induce active macropinocytosis. The induction of macropinocytosis via a simple modification to the exosomal membrane using stearylated octaarginine, which is a representative CPP, significantly enhanced the cellular EV uptake efficacy. Consequently, effective EV-based intracellular delivery of an artificially encapsulated ribosome-inactivating protein, saporin, in EVs was attained.

Enterovirus 74 Infection in Children

PubMed Central

Peacey, Matthew; Hall, Richard J.; Wang, Jing; Todd, Angela K.; Yen, Seiha; Chan-Hyams, Jasmine; Rand, Christy J.; Stanton, Jo-Ann; Huang, Q. Sue

2013-01-01

Enterovirus 74 (EV74) is a rarely detected viral infection of children. In 2010, EV74 was identified in New Zealand in a 2 year old child with acute flaccid paralysis (AFP) through routine polio AFP surveillance. A further three cases of EV74 were identified in children within six months. These cases are the first report of EV74 in New Zealand. In this study we describe the near complete genome sequence of four EV74 isolates from New Zealand, which shows only limited sequence identity in the non-structural proteins when compared to the other two known EV74 sequences. As is typical of enteroviruses multiple recombination events were evident, particularly in the P2 region and P3 regions. This is the first complete EV74 genome sequenced from a patient with acute flaccid paralysis. PMID:24098514

Exacerbation of Acute Traumatic Brain Injury by Circulating Extracellular Vesicles.

PubMed

Hazelton, Isla; Yates, Abi; Dale, Ashley; Roodselaar, Jay; Akbar, Naveed; Ruitenberg, Marc J; Anthony, Daniel C; Couch, Yvonne

2018-02-15

Inflammatory lesions in the brain activate a systemic acute-phase response (APR), which is dependent on the release of extracellular vesicles (EVs) into the circulation. The resulting APR is responsible for regulating leukocyte mobilization and subsequent recruitment to the brain. Factors that either exacerbate or inhibit the APR will also exacerbate or inhibit central nervous system (CNS) inflammation as a consequence and have the potential to influence ongoing secondary damage. Here, we were interested to discover how the circulating EV population changes after traumatic brain injury (TBI) and how manipulation of the circulating EV pool impacts on the outcome of TBI. We found the number of circulating EVs increased rapidly post-TBI, and this was accompanied by an increase in CNS and hepatic leukocyte recruitment. In an adoptive transfer study, we then evaluated the outcomes of TBI after administering EVs derived from either in vitro macrophage or endothelial cell lines stimulated with lipopolysaccharide (LPS), or from murine plasma from an LPS challenge using the air-pouch model. By manipulating the circulating EV population, we were able to demonstrate that each population of transferred EVs increased the APR. However, the characteristics of the response were dependent on the nature of the EVs; specifically, it was significantly increased when animals were challenged with macrophage-derived EVs, suggesting that the cellular origins of EVs may determine their function. Selectively targeting EVs from macrophage/monocyte populations is likely to be of value in reducing the impact of the systemic inflammatory response on the outcome of traumatic CNS injury.

[Correlation between thermostability of the xylanase EvXyn11(TS) and its N-terminal disulfide bridge].

PubMed

Min, Rou; Li, Jianfang; Gao, Shujuan; Zhang, Huimin; Wu, Jing; Wu, Minchen

2013-04-04

To reveal the correlation between thermostability of xylanase EvXyn11(TS) and its N-terminal disulfide bridge, an EvXyn11(TS)-encoding gene (Syxyn11) was synthesized and subjected to site-directed mutagenesis. Multiple homology alignment of protein primary structures between the EvXyn11(TS) and several GH family 11 xylanases displayed that, in their N-termini, only EvXyn11(TS) contained a disulfide bridge (Cys5-Cys32), whose effect on the xylanase thermostability was predicted by molecular dynamics simulation. We constructed a gene Syxyn11(M), encoding the mutated xylanase (EvXyn11(M)) without N-terminal disulfide bridge. Then, Syxyn11 and Syxyn11(M) were expressed in Pichia pastoris GS115, and temperature and pH properties of the expressed enzymes were analyzed. The analytical results displayed that the temperature optimum of EvXyn11(M) was 70 degrees C, which was 15 degrees C lower than that of EvXyn11(TS). The half-life (t1/2(90)) of EvXyn11(TS) at 90 degrees C was 32 min, while the t1/2(70) of EvXyn11(M) at 70 degrees C was only 8.0 min. The important role of the N-terminal disulfide bridge on the thermostability of EvXyn11(TS) was first predicted by molecular dynamics simulation, and confirmed by site-directed mutagenesis. This work provided a novel strategy to improve thermostabilities of the mesophilic family 11 xylanases with high specific activities.

Procoagulant activity of extracellular vesicles as a potential biomarker for risk of thrombosis and DIC in patients with acute leukaemia.

PubMed

Gheldof, Damien; Haguet, Hélène; Dogné, Jean-Michel; Bouvy, Céline; Graux, Carlos; George, Fabienne; Sonet, Anne; Chatelain, Christian; Chatelain, Bernard; Mullier, François

2017-02-01

Haemostatic complication is common for patients with hematologic malignancies. Recent studies suggest that the procoagulant activity (PCA) of extracellular vesicles (EV) may play a major role in venous thromboembolism and disseminated intravascular coagulation (DIC) in acute leukaemia. To study the impact of EVs from leukaemic patients on thrombin generation and to assess EV-PCA as a potential biomarker for thrombotic complications in patients with acute leukaemia. Blood samples from a cohort of patients with newly diagnosed acute leukaemia were obtained before treatment (D-0), 3 and 7 days after treatment (D-3 and D-7). Extracellular vesicles were isolated and concentrated by ultracentrifugation. EV-PCA was assessed by thrombin generation assay, and EV-associated tissue factor activity was measured using a commercial bio-immunoassay (Zymuphen MP-TF®). Of the 53 patients, 6 had increased EV-PCA at D-0 and 4 had a thrombotic event. Patients without thrombotic events (n = 47) had no elevated EV-PCA. One patient had increased EVs with procoagulant activity at D-3 and developed a DIC at D-5. This patient had no increased EVs-related tissue factor activity from D-0 to D-7 (<2 pg/ml). Eight patients had increased EVs with tissue factor activity (>2 pg/ml), of these, four had a thrombosis and two had haemorrhages. Procoagulant activity of extracellular vesicles could have a predictive value in excluding the risk of thrombotic events. Our findings also suggest a possible association between thrombotic events and EV-PCA.

InterEvDock2: an expanded server for protein docking using evolutionary and biological information from homology models and multimeric inputs.

PubMed

Quignot, Chloé; Rey, Julien; Yu, Jinchao; Tufféry, Pierre; Guerois, Raphaël; Andreani, Jessica

2018-05-08

Computational protein docking is a powerful strategy to predict structures of protein-protein interactions and provides crucial insights for the functional characterization of macromolecular cross-talks. We previously developed InterEvDock, a server for ab initio protein docking based on rigid-body sampling followed by consensus scoring using physics-based and statistical potentials, including the InterEvScore function specifically developed to incorporate co-evolutionary information in docking. InterEvDock2 is a major evolution of InterEvDock which allows users to submit input sequences - not only structures - and multimeric inputs and to specify constraints for the pairwise docking process based on previous knowledge about the interaction. For this purpose, we added modules in InterEvDock2 for automatic template search and comparative modeling of the input proteins. The InterEvDock2 pipeline was benchmarked on 812 complexes for which unbound homology models of the two partners and co-evolutionary information are available in the PPI4DOCK database. InterEvDock2 identified a correct model among the top 10 consensus in 29% of these cases (compared to 15-24% for individual scoring functions) and at least one correct interface residue among 10 predicted in 91% of these cases. InterEvDock2 is thus a unique protein docking server, designed to be useful for the experimental biology community. The InterEvDock2 web interface is available at http://bioserv.rpbs.univ-paris-diderot.fr/services/InterEvDock2/.

Peptidyl aldehyde NK-1.8k suppresses enterovirus 71 and enterovirus 68 infection by targeting protease 3C.

PubMed

Wang, Yaxin; Yang, Ben; Zhai, Yangyang; Yin, Zheng; Sun, Yuna; Rao, Zihe

2015-05-01

Enterovirus (EV) is one of the major causative agents of hand, foot, and mouth disease in the Pacific-Asia region. In particular, EV71 causes severe central nervous system infections, and the fatality rates from EV71 infection are high. Moreover, an outbreak of respiratory illnesses caused by an emerging EV, EV68, recently occurred among over 1,000 young children in the United States and was also associated with neurological infections. Although enterovirus has emerged as a considerable global public health threat, no antiviral drug for clinical use is available. In the present work, we screened our compound library for agents targeting viral protease and identified a peptidyl aldehyde, NK-1.8k, that inhibits the proliferation of different EV71 strains and one EV68 strain and that had a 50% effective concentration of 90 nM. Low cytotoxicity (50% cytotoxic concentration, >200 μM) indicated a high selective index of over 2,000. We further characterized a single amino acid substitution inside protease 3C (3C(pro)), N69S, which conferred EV71 resistance to NK-1.8k, possibly by increasing the flexibility of the substrate binding pocket of 3C(pro). The combination of NK-1.8k and an EV71 RNA-dependent RNA polymerase inhibitor or entry inhibitor exhibited a strong synergistic anti-EV71 effect. Our findings suggest that NK-1.8k could potentially be developed for anti-EV therapy. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Extracellular vesicles are key intercellular mediators in the development of immune dysfunction to allergens in the airways.

PubMed

Shin, T-S; Kim, J H; Kim, Y-S; Jeon, S G; Zhu, Z; Gho, Yong Song; Kim, Yoon-Keun

2010-10-01

Previous evidence indicates that inhalation of lipopolysaccharide (LPS)-containing with allergens induced mixed Th1 and Th17 cell responses in the airways. Extracellular vesicles (EVs) are nanometer-sized spherical, lipid-bilayered structures and are recently in the public eye as an intercellular communicator in immune responses. To evaluate the role of EVs secreted by LPS inhalation in the development of airway immune dysfunction in response to allergens. Extracellular vesicles in bronchoalveolar lavage fluids of BALB/c mice were isolated and characterized 24 h after applications to the airway of 10 μg of LPS for 3 days. To evaluate the role of LPS-induced EVs on the development of airway immune dysfunction, in vivo and in vitro experiments were performed using the isolated LPS-induced EVs. The inhalation of LPS enhanced EVs release into the BAL fluid, when compared to the application of PBS. Airway sensitization with allergens and LPS-induced EVs resulted in a mixed Th1 and Th17 cell responses, although that with allergens and PBS-induced EVs induced immune tolerance. In addition, LPS-induced EVs enhanced the production of Th1- and Th17-polarizing cytokines (IL-12p70 and IL-6, respectively) by lung dendritic cells. Moreover, the immune responses induced by the LPS-induced EVs were blocked by denaturation of the EV-bearing proteins. These data suggest that EVs (especially, the protein components) secreted by LPS inhalation are a key intercellular communicator in the development of airway immune dysfunction to inhaled LPS-containing allergens.

Extracellular vesicles are key intercellular mediators in the development of immune dysfunction to allergens in the airways

PubMed Central

Shin, T-S; Kim, J H; Kim, Y-S; Jeon, S G; Zhu, Z; Gho, Y S; Kim, Y-K

2010-01-01

Background Previous evidence indicates that inhalation of lipopolysaccharide (LPS)-containing with allergens induced mixed Th1 and Th17 cell responses in the airways. Extracellular vesicles (EVs) are nanometer-sized spherical, lipid-bilayered structures and are recently in the public eye as an intercellular communicator in immune responses. Objective To evaluate the role of EVs secreted by LPS inhalation in the development of airway immune dysfunction in response to allergens. Methods Extracellular vesicles in bronchoalveolar lavage fluids of BALB/c mice were isolated and characterized 24 h after applications to the airway of 10 μg of LPS for 3 days. To evaluate the role of LPS-induced EVs on the development of airway immune dysfunction, in vivo and in vitro experiments were performed using the isolated LPS-induced EVs. Results The inhalation of LPS enhanced EVs release into the BAL fluid, when compared to the application of PBS. Airway sensitization with allergens and LPS-induced EVs resulted in a mixed Th1 and Th17 cell responses, although that with allergens and PBS-induced EVs induced immune tolerance. In addition, LPS-induced EVs enhanced the production of Th1- and Th17-polarizing cytokines (IL-12p70 and IL-6, respectively) by lung dendritic cells. Moreover, the immune responses induced by the LPS-induced EVs were blocked by denaturation of the EV-bearing proteins. Conclusion These data suggest that EVs (especially, the protein components) secreted by LPS inhalation are a key intercellular communicator in the development of airway immune dysfunction to inhaled LPS-containing allergens. PMID:20337607

Design and Testing of a Thermal Storage System for Electric Vehicle Cabin Heating

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

Wang, Mingyu; WolfeIV, Edward; Craig, Timothy

Without the waste heat available from the engine of a conventional automobile, electric vehicles (EVs) must provide heat to the cabin for climate control using energy stored in the vehicle. In current EV designs, this energy is typically provided by the traction battery. In very cold climatic conditions, the power required to heat the EV cabin can be of a similar magnitude to that required for propulsion of the vehicle. As a result, the driving range of an EV can be reduced very significantly during winter months, which limits consumer acceptance of EVs and results in increased battery costs tomore » achieve a minimum range while ensuring comfort to the EV driver. To minimize the range penalty associated with EV cabin heating, a novel climate control system that includes thermal energy storage has been designed for use in EVs and plug-in hybrid electric vehicles (PHEVs). The system uses the stored latent heat of an advanced phase change material (PCM) to provide cabin heating. The PCM is melted while the EV is connected to the electric grid for charging of the electric battery, and the stored energy is subsequently transferred to the cabin during driving. To minimize thermal losses when the EV is parked for extended periods, the PCM is encased in a high performance insulation system. The electrical PCM-Assisted Thermal Heating System (ePATHS) was designed to provide enough thermal energy to heat the EV s cabin for approximately 46 minutes, covering the entire daily commute of a typical driver in the U.S.« less

Exosome and microvesicle mediated phene transfer in mammalian cells.

PubMed

Christianson, Helena C; Svensson, Katrin J; Belting, Mattias

2014-10-01

Extracellular vesicles (EVs), e.g. exosomes and microvesicles, emerge as new signaling organelles in the exchange of information between cells at the paracrine and systemic level. It is clear that these virus like particles carry complex biological information that can elicit a pleiotropic response in recipient cells with potential relevance in physiology as well as in cancer and other pathological conditions. Numerous studies convincingly show that the molecular composition of EVs closely reflects their cell or tissue of origin. Thus, the signaling status of donor cells, more specifically their endosomal compartments, may largely determine the biological output in recipient cells, a process that we then may conceptualize as vesicle mediated phene transfer. Whereas more conventional modes of cell-cell communication mostly depend on extracellular ligand concentration and cell-surface receptor availability, the magnitude of the EV signaling response relies on the capture and uptake by target cells, allowing release of the EV content. Numerous reports point at the intriguing possibility that, among thousands of mRNAs, miRNAs, and proteins, single EV constituents effectuate the biological response, e.g. stimulation of angiogenesis and cancer cell metastasis, in recipient cells; however, we find it conceivable that strategies targeted at general mechanisms of EV function should provide more rational avenues for therapeutic intervention directed at the EV system. Such strategies include manipulation of EV formation in the endolysosomal system, EV stability in the extracellular milieu, and EV entry into target cells. Here, we provide important insights into potential mechanisms of EV transport in mammalian cells and how these may be targeted. Copyright © 2014 Elsevier Ltd. All rights reserved.

Alternative Fuels Data Center: Minneapolis Makes EV-Charging History Record

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S/EV 1992: Solar and Electric Vehicles. Volume 1: Proceedings

NASA Astrophysics Data System (ADS)

These proceedings include the morning plenary sessions and then sessions on each of the following topics: Electric Vehicle (EV) fundamentals, advanced EV's, photovoltaics, policy, and fleets. Many individual topics are considered to include: EV races, environmental benefits, demonstration programs, lightweight composite materials, advanced components, and electric batteries.

Ellis-van Creveld syndrome in adulthood: extending the clinical spectrum

PubMed Central

Pérez-Andreu, Joaquín; Ray, Victor Glenn; Arribas, José María; Sánchez, Sergio Juan

2015-01-01

Ellis-van Creveld (EvC) syndrome is a rare autosomal recessive malformation disorder. Cardiac defects are observed in about 50% of EvC cases. Surgical data is lacking on the prognosis and life expectancy of EvC patients. Herein, we report the case of a 38-year-old man with EvC syndrome who underwent two surgical corrections for cardiac anomalies. This report supplements the available information on the clinical course of EvC syndrome in older patients. PMID:26106249

Extracellular Vesicles from Hypoxic Adipocytes and Obese Subjects Reduce Insulin‐Stimulated Glucose Uptake

PubMed Central

Mleczko, Justyna; Ortega, Francisco J.; Falcon‐Perez, Juan Manuel; Wabitsch, Martin; Fernandez‐Real, Jose Manuel

2018-01-01

Scope We investigate the effects of extracellular vesicles (EVs) obtained from in vitro adipocyte cell models and from obese subjects on glucose transport and insulin responsiveness. Methods and results EVs are isolated from the culture supernatant of adipocytes cultured under normoxia, hypoxia (1% oxygen), or exposed to macrophage conditioned media (15% v/v). EVs are isolated from the plasma of lean individuals and subjects with obesity. Cultured adipocytes are incubated with EVs and activation of insulin signalling cascades and insulin‐stimulated glucose transport are measured. EVs released from hypoxic adipocytes impair insulin‐stimulated 2‐deoxyglucose uptake and reduce insulin mediated phosphorylation of AKT. Insulin‐mediated phosphorylation of extracellular regulated kinases (ERK1/2) is not affected. EVs from individuals with obesity decrease insulin stimulated 2‐deoxyglucose uptake in adipocytes (p = 0.0159). Conclusion EVs released by stressed adipocytes impair insulin action in neighboring adipocytes. PMID:29292863

The combination effects of licl and the active leflunomide metabolite, A771726, on viral-induced interleukin 6 production and EV-A71 replication.

PubMed

Hung, Hui-Chen; Shih, Shin-Ru; Chang, Teng-Yuan; Fang, Ming-Yu; Hsu, John T-A

2014-01-01

Enterovirus 71 (EV-A71) is a neurotropic virus that can cause severe complications involving the central nervous system. No effective antiviral therapeutics are available for treating EV-A71 infection and drug discovery efforts are rarely focused to target this disease. Thus, the main goal of this study was to discover existing drugs with novel indications that may effectively inhibit EV-A71 replication and the inflammatory cytokines elevation. In this study, we showed that LiCl, a GSK3β inhibitor, effectively suppressed EV-A71 replication, apoptosis and inflammatory cytokines production (Interleukin 6, Interleukin-1β) in infected cells. Furthermore, LiCl and an immunomodular agent were shown to strongly synergize with each other in suppressing EV-A71 replication. The results highlighted potential new treatment regimens in suppressing sequelae caused by EV-A71 replication.

Cyclophilin A Associates with Enterovirus-71 Virus Capsid and Plays an Essential Role in Viral Infection as an Uncoating Regulator

PubMed Central

Huang, Jiaoyan; Yan, Wenzhong; Wang, Jinglan; Su, Dan; Ni, Cheng; Li, Jian; Rao, Zihe; Liu, Lei; Lou, Zhiyong

2014-01-01

Viruses utilize host factors for their efficient proliferation. By evaluating the inhibitory effects of compounds in our library, we identified inhibitors of cyclophilin A (CypA), a known immunosuppressor with peptidyl-prolyl cis-trans isomerase activity, can significantly attenuate EV71 proliferation. We demonstrated that CypA played an essential role in EV71 entry and that the RNA interference-mediated reduction of endogenous CypA expression led to decreased EV71 multiplication. We further revealed that CypA directly interacted with and modified the conformation of H-I loop of the VP1 protein in EV71 capsid, and thus regulated the uncoating process of EV71 entry step in a pH-dependent manner. Our results aid in the understanding of how host factors influence EV71 life cycle and provide new potential targets for developing antiviral agents against EV71 infection. PMID:25275585

The Antiviral Effect of Baicalin on Enterovirus 71 In Vitro

PubMed Central

Li, Xiang; Liu, Yuanyuan; Wu, Tingting; Jin, Yue; Cheng, Jianpin; Wan, Changbiao; Qian, Weihe; Xing, Fei; Shi, Weifeng

2015-01-01

Baicalin is a flavonoid compound extracted from Scutellaria roots that has been reported to possess antibacterial, anti-inflammatory, and antiviral activities. However, the antiviral effect of baicalin on enterovirus 71 (EV71) is still unknown. In this study, we found that baicalin showed inhibitory activity on EV71 infection and was independent of direct virucidal or prophylactic effect and inhibitory viral absorption. The expressions of EV71/3D mRNA and polymerase were significantly blocked by baicalin treatment at early stages of EV71 infection. In addition, baicalin could decrease the expressions of FasL and caspase-3, as well as inhibit the apoptosis of EV71-infected human embryonal rhabdomyosarcoma (RD) cells. Altogether, these results indicate that baicalin exhibits potent antiviral effect on EV71 infection, probably through inhibiting EV71/3D polymerase expression and Fas/FasL signaling pathways. PMID:26295407

Dissociation of CH4 by electron impact: Production of metastable hydrogen and carbon fragments

NASA Technical Reports Server (NTRS)

Finn, T. G.; Carnahan, B. L.; Zipf, E. C.

1974-01-01

Metastable fragments produced by electron impact excitation of CH4 have been investigated for incident electron energies from threshold to 300 eV. Only metastable hydrogen and carbon atoms were observed. Onset energies for the production of metastable hydrogen atoms were observed at electron impact energies of 22.0 + or - .5 eV, 25.5 + or - .6 eV, 36.7 + or - .6 eV and 66 + or - 3 eV, and at 26.6 + or - .6 eV for the production of metastable carbon atoms. Most of the fragments appear to have been formed in high-lying Rydberg states. The total metastable hydrogen cross section reaches a maximum value of approximately 1 X 10 to the minus 18th power sq cm at 100 eV. At the same energy, the metastable carbon cross section is 2 x 10 to the minus 19th power sq cm.

Extracellular vesicles from parasitic helminths and their potential utility as vaccines.

PubMed

Mekonnen, Gebeyaw Getnet; Pearson, Mark; Loukas, Alex; Sotillo, Javier

2018-03-01

Helminths are multicellular parasites affecting nearly three billion people worldwide. To orchestrate a parasitic existence, helminths secrete different molecules, either in soluble form or contained within extracellular vesicles (EVs). EVs are secreted by most cell types and organisms, and have varied roles in intercellular communication, including immune modulation and pathogenesis. Areas covered: In this review, we describe the nucleic acid and proteomic composition of EVs from helminths, with a focus on the protein vaccine candidates present on the EV surface membrane, and discuss the potential utility of helminth EVs and their constituent proteins in the fight against helminth infections. Expert commentary: A significant number of proteins present in helminth-secreted EVs are known vaccine candidates. The characterization of helminth EV proteomes will shed light on host-pathogen interactions, facilitate the discovery of new diagnostic biomarkers, and provide a novel approach for the development of new control measures against helminth infections.

How cancer cells dictate their microenvironment: present roles of extracellular vesicles.

PubMed

Naito, Yutaka; Yoshioka, Yusuke; Yamamoto, Yusuke; Ochiya, Takahiro

2017-02-01

Intercellular communication plays an important role in cancer initiation and progression through secretory molecules, including growth factors and cytokines. Recent advances have revealed that small membrane vesicles, termed extracellular vesicles (EVs), served as a regulatory agent in the intercellular communication of cancer. EVs enable the transfer of functional molecules, including proteins, mRNA and microRNAs (miRNAs), into recipient cells. Cancer cells utilize EVs to dictate the unique phenotype of surrounding cells, thereby promoting cancer progression. Against such "education" by cancer cells, non-tumoral cells suppress cancer initiation and progression via EVs. Therefore, researchers consider EVs to be important cues to clarify the molecular mechanisms of cancer biology. Understanding the functions of EVs in cancer progression is an important aspect of cancer biology that has not been previously elucidated. In this review, we summarize experimental data that indicate the pivotal roles of EVs in cancer progression.

Development of a sandwich ELISA for the quantification of enterovirus 71.

PubMed

Ma, Shujun; Mao, Qunying; Liang, Zhenglun; Zhang, Cuijuan; Yang, Wenxing; Sun, Zhe; Zhang, Haijiang; Shen, Xinliang; Bi, Shengli; Sun, Le

2014-05-01

Since 2008, enterovirus 71 (EV71) has been responsible for high-mortality seasonal epidemics of hand, foot and mouth disease in China. Currently many groups in the world are in the process of developing EV71 vaccines to combat this deadly disease. We have developed three EV71-specific monoclonal antibodies, and in this study we report the establishment of a fast and cost-effective sandwich ELISA kit for measurement of virus concentration in EV71 vaccines using a pair of mouse anti-EV71 monoclonal antibodies. The system is specific for EV71 virus, with no cross-reactivity to coxsackievirus A16, H1N1, rabies, and hepatitis A. Using a reference EV71 vaccine standard, the sensitivity of the assay kit was determined to be 0.82 U/ml, with a linear range between 3.75 and 120 U/ml.

Differential Interaction of Platelet-Derived Extracellular Vesicles with Leukocyte Subsets in Human Whole Blood.

PubMed

Weiss, René; Gröger, Marion; Rauscher, Sabine; Fendl, Birgit; Eichhorn, Tanja; Fischer, Michael B; Spittler, Andreas; Weber, Viktoria

2018-04-26

Secretion and exchange of biomolecules via extracellular vesicles (EVs) are crucial mechanisms in intercellular communication, and the roles of EVs in infection, inflammation, or thrombosis have been increasingly recognized. EVs have emerged as central players in immune regulation and can enhance or suppress the immune response, depending on the state of donor and recipient cells. We investigated the interaction of blood cell-derived EVs with leukocyte subpopulations (monocytes and their subsets, granulocytes, B cells, T cells, and NK cells) directly in whole blood using a combination of flow cytometry, imaging flow cytometry, cell sorting, and high resolution confocal microscopy. Platelet-derived EVs constituted the majority of circulating EVs and were preferentially associated with granulocytes and monocytes, while they scarcely interacted with lymphocytes. Further flow cytometric differentiation of monocyte subsets provided clear indications for a preferential association of platelet-derived EVs with intermediate (CD14 ++ CD16 + ) monocytes in whole blood.

How much are Chevrolet Volts in The EV Project driven in EV Mode?

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

Smart, John

2013-08-01

This report summarizes key conclusions from analysis of data collected from Chevrolet Volts participating in The EV Project. Topics include how many miles are driven in EV mode, how far vehicles are driven between charging events, and how much energy is charged from the electric grid per charging event.

Alternative Fuels Data Center: Seattle Rideshare Fleet Adds EVs, Enjoys

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Molecular identification of enteroviruses including two new types (EV-98 and EV-107) isolated from Japanese travellers from Asian countries.

PubMed

Yamashita, Teruo; Ito, Miyabi; Tsuzuki, Hideaki; Sakae, Kenji; Minagawa, Hiroko

2010-04-01

Of 58 enterovirus strains isolated from Japanese travellers returning from Asian countries, eight were non-serotypable with existing antisera. By sequencing a part of the VP1 region, six of these strains were typed as echovirus 9, enterovirus (EV)-73, EV-79 or EV-97. The nucleotide identity of the VP1 region of isolate T92-1499 to all enterovirus prototypes was <70 %. The VP1 sequence of isolate TN94-0349 was closely related to coxsackievirus (CV)-A9 (73.3 % nucleotide identity), but the virus could not be neutralized with a serum raised against the prototype CV-A9 strain. On the basis of complete molecular comparisons, T92-1499 and TN94-0349 were identified as EV-98 and EV-107, respectively, by the ICTV Picornavirus Study Group. Serum neutralization tests of Japanese individuals revealed a seroprevalence rate of 11 % for EV-73, and even lower seroprevalence rates, 1.0-3.8 %, were found for the other new enteroviruses, suggesting that prior circulation of these viruses in Japan was unlikely.

Inherited MST1 deficiency underlies susceptibility to EV-HPV infections.

PubMed

Crequer, Amandine; Picard, Capucine; Patin, Etienne; D'Amico, Aurelia; Abhyankar, Avinash; Munzer, Martine; Debré, Marianne; Zhang, Shen-Ying; de Saint-Basile, Geneviève; Fischer, Alain; Abel, Laurent; Orth, Gérard; Casanova, Jean-Laurent; Jouanguy, Emmanuelle

2012-01-01

Epidermodysplasia verruciformis (EV) is characterized by persistent cutaneous lesions caused by a specific group of related human papillomavirus genotypes (EV-HPVs) in otherwise healthy individuals. Autosomal recessive (AR) EVER1 and EVER2 deficiencies account for two thirds of known cases of EV. AR RHOH deficiency has recently been described in two siblings with EV-HPV infections as well as other infectious and tumoral manifestations. We report here the whole-exome based discovery of AR MST1 deficiency in a 19-year-old patient with a T-cell deficiency associated with EV-HPV, bacterial and fungal infections. MST1 deficiency has recently been described in seven patients from three unrelated kindreds with profound T-cell deficiency and various viral and bacterial infections. The patient was also homozygous for a rare ERCC3 variation. Our findings broaden the clinical range of infections seen in MST1 deficiency and provide a new genetic etiology of susceptibility to EV-HPV infections. Together with the recent discovery of RHOH deficiency, they suggest that T cells are involved in the control of EV-HPVs, at least in some individuals.

Rapid and highly sensitive detection of Enterovirus 71 by using nanogold-enhanced electrochemical impedance spectroscopy

NASA Astrophysics Data System (ADS)

Li, Hsing-Yuan; Tseng, Shing-Hua; Cheng, Tsai-Mu; Chu, Hsueh-Liang; Lu, Yu-Ning; Wang, Fang-Yu; Tsai, Li-Yun; Shieh, Juo-Yu; Yang, Jyh-Yuan; Juan, Chien-Chang; Tu, Lung-Chen; Chang, Chia-Ching

2013-07-01

Enterovirus 71 (EV71) infection is an emerging infectious disease causing neurological complications and/or death within two to three days after the development of fever and rash. A low viral titre in clinical specimens makes the detection of EV71 difficult. Conventional approaches for detecting EV71 are time consuming, poorly sensitive, or complicated, and cannot be used effectively for clinical diagnosis. Furthermore, EV71 and Coxsackie virus A16 (CA16) may cross react in conventional assays. Therefore, a rapid, highly sensitive, specific, and user-friendly test is needed. We developed an EV71-specific nanogold-modified working electrode for electrochemical impedance spectroscopy in the detection of EV71. Our results show that EV71 can be distinguished from CA16, Herpes simplex virus, and lysozyme, with the modified nanogold electrode being able to detect EV71 in concentrations as low as 1 copy number/50 μl reaction volume, and the duration between sample preparation and detection being 11 min. This detection platform may have the potential for use in point-of-care diagnostics.

Anhydrous crystals of DNA bases are wide gap semiconductors.

PubMed

Maia, F F; Freire, V N; Caetano, E W S; Azevedo, D L; Sales, F A M; Albuquerque, E L

2011-05-07

We present the structural, electronic, and optical properties of anhydrous crystals of DNA nucleobases (guanine, adenine, cytosine, and thymine) found after DFT (Density Functional Theory) calculations within the local density approximation, as well as experimental measurements of optical absorption for powders of these crystals. Guanine and cytosine (adenine and thymine) anhydrous crystals are predicted from the DFT simulations to be direct (indirect) band gap semiconductors, with values 2.68 eV and 3.30 eV (2.83 eV and 3.22 eV), respectively, while the experimentally estimated band gaps we have measured are 3.83 eV and 3.84 eV (3.89 eV and 4.07 eV), in the same order. The electronic effective masses we have obtained at band extremes show that, at low temperatures, these crystals behave like wide gap semiconductors for electrons moving along the nucleobases stacking direction, while the hole transport are somewhat limited. Lastly, the calculated electronic dielectric functions of DNA nucleobases crystals in the parallel and perpendicular directions to the stacking planes exhibit a high degree of anisotropy (except cytosine), in agreement with published experimental results.

Hidden benefits of electric vehicles for addressing climate change.

PubMed

Li, Canbing; Cao, Yijia; Zhang, Mi; Wang, Jianhui; Liu, Jianguo; Shi, Haiqing; Geng, Yinghui

2015-03-19

There is an increasingly hot debate on whether the replacement of conventional vehicles (CVs) by electric vehicles (EVs) should be delayed or accelerated since EVs require higher cost and cause more pollution than CVs in the manufacturing process. Here we reveal two hidden benefits of EVs for addressing climate change to support the imperative acceleration of replacing CVs with EVs. As EVs emit much less heat than CVs within the same mileage, the replacement can mitigate urban heat island effect (UHIE) to reduce the energy consumption of air conditioners, benefitting local and global climates. To demonstrate these effects brought by the replacement of CVs by EVs, we take Beijing, China, as an example. EVs emit only 19.8% of the total heat emitted by CVs per mile. The replacement of CVs by EVs in 2012 could have mitigated the summer heat island intensity (HII) by about 0.94°C, reduced the amount of electricity consumed daily by air conditioners in buildings by 14.44 million kilowatt-hours (kWh), and reduced daily CO2 emissions by 10,686 tonnes.

Hidden benefits of electric vehicles for addressing climate change

DOE PAGES

Li, Canbing; Cao, Yijia; Zhang, Mi; ...

2015-03-19

There is an increasingly hot debate on whether the replacement of conventional vehicles (CVs) by electric vehicles (EVs) should be delayed or accelerated since EVs require higher cost and cause more pollution than CVs in the manufacturing process. Here we reveal two hidden benefits of EVs for addressing climate change to support the imperative acceleration of replacing CVs with EVs. As EVs emit much less heat than CVs within the same mileage, the replacement can mitigate urban heat island effect (UHIE) to reduce the energy consumption of air conditioners, benefitting local and global climates. To demonstrate these effects brought bymore » the replacement of CVs by EVs, we take Beijing, China, as an example. EVs emit only 19.8% of the total heat emitted by CVs per mile. The replacement of CVs by EVs in 2012 could have mitigated the summer heat island intensity (HII) by about 0.94°C, reduced the amount of electricity consumed daily by air conditioners in buildings by 14.44 million kilowatt-hours (kWh), and reduced daily CO₂ emissions by 10,686 tonnes.« less

Inherited MST1 Deficiency Underlies Susceptibility to EV-HPV Infections

PubMed Central

Crequer, Amandine; Picard, Capucine; Patin, Etienne; D’Amico, Aurelia; Abhyankar, Avinash; Munzer, Martine; Debré, Marianne; Zhang, Shen-Ying; de Saint-Basile, Geneviève; Fischer, Alain

2012-01-01

Epidermodysplasia verruciformis (EV) is characterized by persistent cutaneous lesions caused by a specific group of related human papillomavirus genotypes (EV-HPVs) in otherwise healthy individuals. Autosomal recessive (AR) EVER1 and EVER2 deficiencies account for two thirds of known cases of EV. AR RHOH deficiency has recently been described in two siblings with EV-HPV infections as well as other infectious and tumoral manifestations. We report here the whole-exome based discovery of AR MST1 deficiency in a 19-year-old patient with a T-cell deficiency associated with EV-HPV, bacterial and fungal infections. MST1 deficiency has recently been described in seven patients from three unrelated kindreds with profound T-cell deficiency and various viral and bacterial infections. The patient was also homozygous for a rare ERCC3 variation. Our findings broaden the clinical range of infections seen in MST1 deficiency and provide a new genetic etiology of susceptibility to EV-HPV infections. Together with the recent discovery of RHOH deficiency, they suggest that T cells are involved in the control of EV-HPVs, at least in some individuals. PMID:22952854

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

Li, Canbing; Cao, Yijia; Zhang, Mi

There is an increasingly hot debate on whether the replacement of conventional vehicles (CVs) by electric vehicles (EVs) should be delayed or accelerated since EVs require higher cost and cause more pollution than CVs in the manufacturing process. Here we reveal two hidden benefits of EVs for addressing climate change to support the imperative acceleration of replacing CVs with EVs. As EVs emit much less heat than CVs within the same mileage, the replacement can mitigate urban heat island effect (UHIE) to reduce the energy consumption of air conditioners, benefitting local and global climates. To demonstrate these effects brought bymore » the replacement of CVs by EVs, we take Beijing, China, as an example. EVs emit only 19.8% of the total heat emitted by CVs per mile. The replacement of CVs by EVs in 2012 could have mitigated the summer heat island intensity (HII) by about 0.94°C, reduced the amount of electricity consumed daily by air conditioners in buildings by 14.44 million kilowatt-hours (kWh), and reduced daily CO₂ emissions by 10,686 tonnes.« less

High sensitivity and label-free detection of Enterovirus 71 by nanogold modified electrochemical impedance spectroscopy

NASA Astrophysics Data System (ADS)

Wang, Fang-Yu; Li, Hsing-Yuan; Tseng, Shing-Hua; Cheng, Tsai-Mu; Chu, Hsueh-Liang; Yang, Jyh-Yuan; Chang, Chia-Ching

2013-03-01

Enterovirus 71 (EV71), which is the most fulminant and invasive species of enterovirus, can cause children neurologic complications and death within 2-3 days after fever and rash developed. Besides, EV71 has high sequence similarity with Coxsackie A 16 (CA16) that makes differential diagnosis difficult in clinic and laboratory. Since conventional viral diagnostic method cannot diagnose EV71 quickly and EV71 can transmit at low viral titer, the patients might delay in treatment. A quick, high sensitive, and high specific test for EV71 detection is pivotal. Electrochemical impedance spectroscopy (EIS) has been applied for detecting bio-molecules as biosensors recently. In this study, we try to build a detection platform for EV71 detection by nanogold modified EIS probe. The result shows that our probe can detect 3.6 VP1/50 μl (one EV71 particle has 60 VP1) in 3 minutes. The test can also distinguish EV71 from CA16 and lysozyme. Diagnosis of enterovirus 71 by electrochemical impedance spectroscopy has the potential to apply in clinic.

Pathogenic or Therapeutic Extracellular Vesicles in Rheumatic Diseases: Role of Mesenchymal Stem Cell-Derived Vesicles

PubMed Central

Cosenza, Stella; Ruiz, Maxime; Maumus, Marie; Jorgensen, Christian; Noël, Danièle

2017-01-01

Extracellular vesicles (EVs) are important mediators of cell-to-cell communication pathways via the transport of proteins, mRNA, miRNA and lipids. There are three main types of EVs, exosomes, microparticles and apoptotic bodies, which are classified according to their size and biogenesis. EVs are secreted by all cell types and their function reproduces that of the parental cell. They are involved in many biological processes that regulate tissue homeostasis and physiopathology of diseases. In rheumatic diseases, namely osteoarthritis (OA) and rheumatoid arthritis (RA), EVs have been isolated from synovial fluid and shown to play pathogenic roles contributing to progression of both diseases. By contrast, EVs may have therapeutic effect via the delivery of molecules that may stop disease evolution. In particular, EVs derived from mesenchymal stem cells (MSCs) reproduce the main functions of the parental cells and therefore represent the ideal type of EVs for modulating the course of either disease. The aim of this review is to discuss the role of EVs in OA and RA focusing on their potential pathogenic effect and possible therapeutic options. Special attention is given to MSCs and MSC-derived EVs for modulating OA and RA progression with the perspective of developing innovative therapeutic strategies. PMID:28441721

EV-Associated MMP9 in High-Grade Serous Ovarian Cancer Is Preferentially Localized to Annexin V-Binding EVs.

PubMed

Reiner, Agnes T; Tan, Sisareuth; Agreiter, Christiane; Auer, Katharina; Bachmayr-Heyda, Anna; Aust, Stefanie; Pecha, Nina; Mandorfer, Mattias; Pils, Dietmar; Brisson, Alain R; Zeillinger, Robert; Lim, Sai Kiang

2017-01-01

High-grade serous ovarian cancer (HGSOC) is the most aggressive type of ovarian cancer and is responsible for most deaths caused by gynecological cancers. Numerous candidate biomarkers were identified for this disease in the last decades, but most were not sensitive or specific enough for clinical applications. Hence, new biomarkers for HGSOC are urgently required. This study aimed to identify new markers by isolating different extracellular vesicle (EV) types from the ascites of ovarian cancer patients according to their affinities for lipid-binding proteins and analyzing their protein cargo. This approach circumvents the low signal-to-noise ratio when using biological fluids for biomarker discovery and the issue of contamination by large non-EV complexes. We isolated and analyzed three distinct EV populations from the ascites of patients with ovarian cancer or cirrhosis and observed that Annexin V-binding EVs have higher levels of matrix metalloproteinase 9 in malignant compared to portal-hypertensive ascites. As this protein was not detected in other EV populations, this study validates our approach of using different EV types for optimal biomarker discovery. Furthermore, MMP9 in Annexin V-binding EVs could be a HGSOC biomarker with enhanced specificity, because its identification requires detection of two distinct components, that is, lipid and protein.

Blood Volume: Importance and Adaptations to Exercise Training, Environmental Stresses and Trauma/Sickness

NASA Technical Reports Server (NTRS)

Sawka, Michael N.; Convertino, Victor A.; Eichner, E. Randy; Schnieder, Suzanne M.; Young, Andrew J.

2000-01-01

This paper reviews the influence of several perturbations (physical exercise, heat stress, terrestrial altitude, microgravity, and trauma/sickness) on adaptations of blood volume (BV), erythrocyte volume (EV), and plasma volume (PV). Exercise training can induced BV expansion; PV expansion usually occurs immediately, but EV expansion takes weeks. EV and PV expansion contribute to aerobic power improvements associated with exercise training. Repeated heat exposure induces PV expansion but does not alter EV. PV expansion does not improve thermoregulation, but EV expansion improves thermoregulation during exercise in the heat. Dehydration decreases PV (and increases plasma tonicity) which elevates heat strain and reduces exercise performance. High altitude exposure causes rapid (hours) plasma loss. During initial weeks at altitude, EV is unaffected, but a gradual expansion occurs with extended acclimatization. BV adjustments contribute, but are not key, to altitude acclimatization. Microgravity decreases PV and EV which contribute to orthostatic intolerance and decreased exercise capacity in astronauts. PV decreases may result from lower set points for total body water and central venous pressure, which EV decrease bay result form increased erythrocyte destruction. Trauma, renal disease, and chronic diseases cause anemia from hemorrhage and immune activation, which suppressions erythropoiesis. The re-establishment of EV is associated with healing, improved life quality, and exercise capabilities for these injured/sick persons.

Phospholipid composition of packed red blood cells and that of extracellular vesicles show a high resemblance and stability during storage.

PubMed

Laurén, Eva; Tigistu-Sahle, Feven; Valkonen, Sami; Westberg, Melissa; Valkeajärvi, Anne; Eronen, Juha; Siljander, Pia; Pettilä, Ville; Käkelä, Reijo; Laitinen, Saara; Kerkelä, Erja

2018-01-01

Red blood cells (RBCs) are stored up to 35-42days at 2-6°C in blood banks. During storage, the RBC membrane is challenged by energy depletion, decreasing pH, altered cation homeostasis, and oxidative stress, leading to several biochemical and morphological changes in RBCs and to shedding of extracellular vesicles (EVs) into the storage medium. These changes are collectively known as RBC storage lesions. EVs accumulate in stored RBC concentrates and are, thus, transfused into patients. The potency of EVs as bioactive effectors is largely acknowledged, and EVs in RBC concentrates are suspected to mediate some adverse effects of transfusion. Several studies have shown accumulation of lipid raft-associated proteins in RBC EVs during storage, whereas a comprehensive phospholipidomic study on RBCs and corresponding EVs during the clinical storage period is lacking. Our mass spectrometric and chromatographic study shows that RBCs maintain their major phospholipid (PL) content well during storage despite abundant vesiculation. The phospholipidomes were largely similar between RBCs and EVs. No accumulation of raft lipids in EVs was seen, suggesting that the primary mechanism of RBC vesiculation during storage might not be raft -based. Nonetheless, a slight tendency of EV PLs for shorter acyl chains was observed. Copyright © 2017 Elsevier B.V. All rights reserved.

Development of a sensitive and specific epitope-blocking ELISA for universal detection of antibodies to human enterovirus 71 strains.

PubMed

He, Fang; Kiener, Tanja K; Lim, Xiao Fang; Tan, Yunrui; Raj, Kattur Venkatachalam Ashok; Tang, Manli; Chow, Vincent T K; Chen, Qingfeng; Kwang, Jimmy

2013-01-01

Human Enterovirus 71 (EV71) is a common cause of hand, foot and mouth disease (HFMD) in young children. It is often associated with severe neurological diseases and mortalities in recent outbreaks across the Asia Pacific region. Currently, there is no efficient universal antibody test available to detect EV71 infections. In the present study, an epitope-blocking ELISA was developed to detect specific antibodies to human EV71 viruses in human or animal sera. The assay relies on a novel monoclonal antibody (Mab 1C6) that specifically binds to capsid proteins in whole EV71 viruses without any cross reaction to any EV71 capsid protein expressed alone. The sensitivity and specificity of the epitope-blocking ELISA for EV71 was evaluated and compared to microneutralization using immunized animal sera to multiple virus genotypes of EV71 and coxsackieviruses. Further, 200 serum sample from human individuals who were potentially infected with EV71 viruses were tested in both the blocking ELISA and microneutralization. Results indicated that antibodies to EV71 were readily detected in immunized animals or human sera by the epitope blocking ELISA whereas specimens with antibodies to other enteroviruses yielded negative results. This assay is not only simpler to perform but also shows higher sensitivity and specificity as compared to microneutralization. The epitope-blocking ELISA based on a unique Mab 1C6 provided highly sensitive and 100% specific detection of antibodies to human EV71 viruses in human sera.

Correlates of protection for inactivated enterovirus 71 vaccine: the analysis of immunological surrogate endpoints.

PubMed

Zhu, Wenbo; Jin, Pengfei; Li, Jing-Xin; Zhu, Feng-Cai; Liu, Pei

2017-09-01

Inactivated Enterovirus 71 (EV71) vaccines showed significant efficacy against the diseases associated with EV71 and a neutralizing antibody (NTAb) titer of 1:16-1:32 was suggested as the correlates of the vaccine protection. This paper aims to further estimate the immunological surrogate endpoints for the protection of inactivated EV71 vaccines and the effect factors. Pre-vaccination NTAb against EV71 at baseline (day 0), post-vaccination NTAb against EV71 at day 56, and the occurrence of laboratory-confirmed EV71-associated diseases during a 24-months follow-up period were collected from a phase 3 efficacy trial of an inactivated EV71 vaccine. We used the mixed-scaled logit model and the absolute sigmoid function by some extensions in continuous models to estimate the immunological surrogate endpoint for the EV71 vaccine protection, respectively. For children with a negative baseline of EV71 NTAb titers, an antibody level of 26.6 U/ml (1:30) was estimated to provide at least a 50% protection for 12 months, and an antibody level of 36.2 U/ml (1:42) may be needed to achieve a 50% protective level of the population for 24 months. Both the pre-vaccination NTAb level and the vaccine protective period could affect the estimation of the immunological surrogate for EV71 vaccine. A post-vaccination NTAb titer of 1:42 or more may be needed for long-term protection. NCT01508247.

Concentration-dependent protein loading of extracellular vesicles released by Histoplasma capsulatum after antibody treatment and its modulatory action upon macrophages

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

Baltazar, Ludmila Matos; Zamith-Miranda, Daniel; Burnet, Meagan C.

Here, diverse pathogenic fungi secrete extracellular vesicles (EV) that contain macromolecules, including virulence factors that can modulate the host immune response. We recently demonstrated that the binding of monoclonal antibodies (mAb) modulates how Histoplasma capsulatum load and releases its extracellular vesicles (EV). In the present paper, we addressed a concentration-dependent impact on the fungus’ EV loading and release with different mAb, as well as the pathophysiological role of these EV during the host-pathogen interaction. We found that the mAbs differentially regulate EV content in concentration-dependent and independent manners. Enzymatic assays demonstrated that laccase activity in EV from H. capsulatum opsonizedmore » with 6B7 was reduced, but urease activity was not altered. The uptake of H. capsulatum by macrophages pre-treated with EV, presented an antibody concentration-dependent phenotype. The intracellular killing of yeast cells was potently inhibited in macrophages pre-treated with EV from 7B6 (non-protective) mAb-opsonized H. capsulatum and this inhibition was associated with a decrease in the reactive-oxygen species generated by these macrophages. In summary, our findings show that opsonization quantitatively and qualitatively modifies H. capsulatum EV load and secretion leading to distinct effects on the host’s immune effector mechanisms, supporting the hypothesis that EV sorting and secretion are dynamic mechanisms for a fine-tuned response by fungal cells.« less

Concentration-dependent protein loading of extracellular vesicles released by Histoplasma capsulatum after antibody treatment and its modulatory action upon macrophages

DOE PAGES

Baltazar, Ludmila Matos; Zamith-Miranda, Daniel; Burnet, Meagan C.; ...

2018-05-23

Here, diverse pathogenic fungi secrete extracellular vesicles (EV) that contain macromolecules, including virulence factors that can modulate the host immune response. We recently demonstrated that the binding of monoclonal antibodies (mAb) modulates how Histoplasma capsulatum load and releases its extracellular vesicles (EV). In the present paper, we addressed a concentration-dependent impact on the fungus’ EV loading and release with different mAb, as well as the pathophysiological role of these EV during the host-pathogen interaction. We found that the mAbs differentially regulate EV content in concentration-dependent and independent manners. Enzymatic assays demonstrated that laccase activity in EV from H. capsulatum opsonizedmore » with 6B7 was reduced, but urease activity was not altered. The uptake of H. capsulatum by macrophages pre-treated with EV, presented an antibody concentration-dependent phenotype. The intracellular killing of yeast cells was potently inhibited in macrophages pre-treated with EV from 7B6 (non-protective) mAb-opsonized H. capsulatum and this inhibition was associated with a decrease in the reactive-oxygen species generated by these macrophages. In summary, our findings show that opsonization quantitatively and qualitatively modifies H. capsulatum EV load and secretion leading to distinct effects on the host’s immune effector mechanisms, supporting the hypothesis that EV sorting and secretion are dynamic mechanisms for a fine-tuned response by fungal cells.« less

Nanosized UCMSC-derived extracellular vesicles but not conditioned medium exclusively inhibit the inflammatory response of stimulated T cells: implications for nanomedicine.

PubMed

Monguió-Tortajada, Marta; Roura, Santiago; Gálvez-Montón, Carolina; Pujal, Josep Maria; Aran, Gemma; Sanjurjo, Lucía; Franquesa, Marcel la; Sarrias, Maria-Rosa; Bayes-Genis, Antoni; Borràs, Francesc E

2017-01-01

Undesired immune responses have drastically hampered outcomes after allogeneic organ transplantation and cell therapy, and also lead to inflammatory diseases and autoimmunity. Umbilical cord mesenchymal stem cells (UCMSCs) have powerful regenerative and immunomodulatory potential, and their secreted extracellular vesicles (EVs) are envisaged as a promising natural source of nanoparticles to increase outcomes in organ transplantation and control inflammatory diseases. However, poor EV preparations containing highly-abundant soluble proteins may mask genuine vesicular-associated functions and provide misleading data. Here, we used Size-Exclusion Chromatography (SEC) to successfully isolate EVs from UCMSCs-conditioned medium. These vesicles were defined as positive for CD9, CD63, CD73 and CD90, and their size and morphology characterized by NTA and cryo-EM. Their immunomodulatory potential was determined in polyclonal T cell proliferation assays, analysis of cytokine profiles and in the skewing of monocyte polarization. In sharp contrast to the non-EV containing fractions, to the complete conditioned medium and to ultracentrifuged pellet, SEC-purified EVs from UCMSCs inhibited T cell proliferation, resembling the effect of parental UCMSCs. Moreover, while SEC-EVs did not induce cytokine response, the non-EV fractions, conditioned medium and ultracentrifuged pellet promoted the secretion of pro-inflammatory cytokines by polyclonally stimulated T cells and supported Th17 polarization. In contrast, EVs did not induce monocyte polarization, but the non-EV fraction induced CD163 and CD206 expression and TNF-α production in monocytes. These findings increase the growing evidence confirming that EVs are an active component of MSC's paracrine immunosuppressive function and affirm their potential for therapeutics in nanomedicine. In addition, our results highlight the importance of well-purified and defined preparations of MSC-derived EVs to achieve the immunosuppressive effect.

Identification of point defects in HVPE-grown GaN by steady-state and time-resolved photoluminescence

NASA Astrophysics Data System (ADS)

Reshchikov, M. A.; Demchenko, D. O.; Usikov, A.; Helava, H.; Makarov, Yu.

2015-03-01

We have investigated point defects in GaN grown by HVPE by using steady-state and time-resolved photoluminescence (PL). Among the most common PL bands in this material are the red luminescence band with a maximum at 1.8 eV and a zero-phonon line (ZPL) at 2.36 eV (attributed to an unknown acceptor having an energy level 1.130 eV above the valence band), the blue luminescence band with a maximum at 2.9 eV (attributed to ZnGa), and the ultraviolet luminescence band with the main peak at 3.27 eV (related to an unknown shallow acceptor). In GaN with the highest quality, the dominant defect-related PL band at high excitation intensity is the green luminescence band with a maximum at about 2.4 eV. We attribute this band to transitions of electrons from the conduction band to the 0/+ level of the isolated CN defect. The yellow luminescence (YL) band, related to transitions via the -/0 level of the same defect, has a maximum at 2.1 eV. Another yellow luminescence band, which has similar shape but peaks at about 2.2 eV, is observed in less pure GaN samples and is attributed to the CNON complex. In semi-insulating GaN, the GL2 band with a maximum at 2.35 eV (attributed to VN) and the BL2 band with a maximum at 3.0 eV and the ZPL at 3.33 eV (attributed to a defect complex involving hydrogen) are observed. We also conclude that the gallium vacancy-related defects act as centers of nonradiative recombination.

Extracellular vesicles have variable dose-dependent effects on cultured draining cells in the eye.

PubMed

Tabak, Saray; Schreiber-Avissar, Sofia; Beit-Yannai, Elie

2018-03-01

The role of extracellular vesicles (EVs) as signal mediators has been described in many biological fields. How many EVs are needed to deliver the desired physiological signal is yet unclear. Using a normal trabecular meshwork (NTM) cell culture exposed to non-pigmented ciliary epithelium (NPCE)-derived EVs, a relevant model for studying the human ocular drainage system, we addressed the EVs dose-response effects on the Wnt signaling. The objective of the study was to investigate the dosing effects of NPCE-derived EVs on TM Wnt signaling. EVs were isolated by PEG 8000 method from NPCE and RPE cells (used as controls) conditioned media. Concentrations were determined by Tunable Resistive Pulse Sensing method. Various exosomes concentration were incubated with TM cells, for the determination of mRNA (β-Catenin, Axin2 and LEF1) and protein (β-Catenin, GSK-3β) expression using real-time quantitative PCR and Western blot, respectively. Exposure of NTM cells for 8 hrs to low EVs concentrations was associated with a significant decreased expression of β-Catenin, GSK-3β, as opposed to exposure to high exosomal concentrations. Pro-MMP9 and MMP9 activities were significantly enhanced in NTM cells treated with high EV concentrations of (X10) as compared to low EV concentrations of either NPCE- or RPE-derived EVs and to untreated control. Our data support the concept that EVs biological effects are concentration-dependent at their target site. Specifically in the present study, we described a general dose-response at the gene and MMPs activity and a different dose-response regarding key canonical Wnt proteins expression. © 2018 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Gastric Varices Bleed at Lower Portosystemic Pressure Gradients than Esophageal Varices.

PubMed

Morrison, Joseph D; Mendoza-Elias, Nasya; Lipnik, Andrew J; Lokken, R Peter; Bui, James T; Ray, Charles E; Gaba, Ron C

2018-05-01

To quantify and compare portosystemic pressure gradients (PSGs) between bleeding esophageal varices (EV) and gastric varices (GV). In a single-center, retrospective study, 149 patients with variceal bleeding (90 men, 59 women, mean age 52 y) with EV (n = 69; 46%) or GV (n = 80; 54%) were selected from 320 consecutive patients who underwent successful transjugular intrahepatic portosystemic shunt (TIPS) creation from 1998 to 2016. GV were subcategorized using the Sarin classification as gastroesophageal varices (GEV) (n = 57) or isolated gastric varices (IGV) (n = 23). PSG before TIPS was measured from the main portal vein to the right atrium. PSGs were compared across EV, GEV, and IGV groups using 1-way analysis of variance. Overall mean baseline PSG was 21 mm Hg ± 6. PSG was significantly higher in patients with EV versus GV (23 mm Hg vs 19 mm Hg; P < .001). Mean PSG was highest among EV (23 mm Hg) with lower PSGs identified for GEV (20 mm Hg) and IGV (16 mm Hg); this difference was statistically significant (P < .001). Among 95 acute bleeding cases, a similar pattern was evident (EV 23 mm Hg vs GEV mm Hg 20 vs IGV 17 mm Hg; P < .001). At baseline PSG < 12 mm Hg, 13% (3/23) of IGV bled versus 9% (5/57) of GEV and 3% (2/69) of EVs (P = .169). Mean final PSG after TIPS was 8 mm Hg (IGV 6 mm Hg vs EV and GEV 8 mm Hg; P = .005). GV bleed at lower PSGs than EV. EV, GEV, and IGV bleeding is associated with successively lower PSGs. These findings highlight distinct physiology, anatomy, and behavior of GV compared with EV. Copyright © 2017 SIR. Published by Elsevier Inc. All rights reserved.

Activation of TLR3 and its adaptor TICAM-1 increases miR-21 levels in extracellular vesicles released from human cells.

PubMed

Fukushima, Yoshimi; Okamoto, Masaaki; Ishikawa, Kana; Kouwaki, Takahisa; Tsukamoto, Hirotake; Oshiumi, Hiroyuki

2018-06-07

Pattern-recognition receptors (PRRs) recognizes viral RNAs and trigger the innate immune responses. Toll-like receptor 3 (TLR3), a PRR, recognizes viral double-stranded RNA (dsRNA) in endolysosomes, whereas cytoplasmic dsRNA is sensed by another PRR, MDA5. TLR3 and MDA5 utilize TICAM-1 and MAVS, respectively, to trigger the signal for inducing innate immune responses. Extracellular vesicles (EVs) include the exosomes and microvesicles; an accumulating body of evidence has shown that EVs delivers functional RNA, such as microRNAs (miRNAs), to other cells and thus mediate intercellular communications. Therefore, EVs carrying miRNAs affect innate immune responses in macrophages and dendritic cells. However, the mechanism underlying the regulation of miRNA levels in EVs remains unclear. To elucidate the mechanism, we sought to reveal the pathway that control miRNA expression levels in EVs. Here, we found that TLR3 stimulation increased miR-21 levels in EVs released from various types of human cells. Ectopic expression of the TLR3 adaptor, TICAM-1, increased miR-21 levels in EVs but not intracellular miR-21 levels, suggesting that TICAM-1 augmented sorting of miR-21 to EVs. In contrast, the MDA5 adaptor, MAVS, did not increase miR-21 levels in EVs. The siRNA for TICAM-1 reduced EV miR-21 levels after stimulation of TLR3. Collectively, our data indicate a novel role of the TLR3-TICAM-1 pathway in controlling miR-21 levels in EVs. Copyright © 2018 Elsevier Inc. All rights reserved.

Management of the 2014 Enterovirus 68 Outbreak at a Pediatric Tertiary Care Center.

PubMed

Schuster, Jennifer E; Newland, Jason G

2015-11-01

Enterovirus 68 (EV-D68) is an uncommonly recognized cause of acute respiratory tract infections. During the late summer of 2014, an international EV-D68 outbreak occurred. We review the steps of outbreak recognition and management in the context of 1 hospital's experience with the EV-D68 outbreak. We reviewed the role of Children's Mercy Hospital as one of the first hospitals to recognize the 2014 EV-D68 outbreak in the United States. The steps of outbreak management were applied to real-life examples as the outbreak unfolded at our hospital. Management of the 2014 EV-D68 outbreak was a multifaceted effort requiring close coordination with hospitals, local and state health departments, and the Centers for Disease Control and Prevention. The importance of clear and frequent communication is highlighted both intra- and interinstitutionally. Increased respiratory disease-related pediatric admissions at hospitals nationally were attributed to EV-D68. Outcomes for these children, including the association of EV-D68 with acute flaccid myelitis, remain under investigation. Following the steps of outbreak management is critical to providing optimal patient care and ensuring the health of the public. During the 2014 EV-D68 outbreak, close adherence to outbreak principles led to swift recognition of illness, rapid diagnostic measures, institution of appropriate therapies, and dissemination of information to health care providers and the public. Equally important was the subsequent identification of an increase in acute flaccid myelitis cases against the backdrop of an increase in EV-D68 detections nationally. Future prospective studies are needed to determine the true burden of EV-D68 disease, potential vaccines and therapeutics, and outcomes of children with EV-D68 infection. Copyright © 2015 Elsevier HS Journals, Inc. All rights reserved.

Deep level defects in Ge-doped (010) β-Ga2O3 layers grown by plasma-assisted molecular beam epitaxy

NASA Astrophysics Data System (ADS)

Farzana, Esmat; Ahmadi, Elaheh; Speck, James S.; Arehart, Aaron R.; Ringel, Steven A.

2018-04-01

Deep level defects were characterized in Ge-doped (010) β-Ga2O3 layers grown by plasma-assisted molecular beam epitaxy (PAMBE) using deep level optical spectroscopy (DLOS) and deep level transient (thermal) spectroscopy (DLTS) applied to Ni/β-Ga2O3:Ge (010) Schottky diodes that displayed Schottky barrier heights of 1.50 eV. DLOS revealed states at EC - 2.00 eV, EC - 3.25 eV, and EC - 4.37 eV with concentrations on the order of 1016 cm-3, and a lower concentration level at EC - 1.27 eV. In contrast to these states within the middle and lower parts of the bandgap probed by DLOS, DLTS measurements revealed much lower concentrations of states within the upper bandgap region at EC - 0.1 - 0.2 eV and EC - 0.98 eV. There was no evidence of the commonly observed trap state at ˜EC - 0.82 eV that has been reported to dominate the DLTS spectrum in substrate materials synthesized by melt-based growth methods such as edge defined film fed growth (EFG) and Czochralski methods [Zhang et al., Appl. Phys. Lett. 108, 052105 (2016) and Irmscher et al., J. Appl. Phys. 110, 063720 (2011)]. This strong sensitivity of defect incorporation on crystal growth method and conditions is unsurprising, which for PAMBE-grown β-Ga2O3:Ge manifests as a relatively "clean" upper part of the bandgap. However, the states at ˜EC - 0.98 eV, EC - 2.00 eV, and EC - 4.37 eV are reminiscent of similar findings from these earlier results on EFG-grown materials, suggesting that possible common sources might also be present irrespective of growth method.

Clinical severity of pediatric respiratory illness with enterovirus D68 compared with rhinovirus or other enterovirus genotypes.

PubMed

Mertz, Dominik; Alawfi, Abdulsalam; Pernica, Jeffrey M; Rutherford, Candy; Luinstra, Kathy; Smieja, Marek

2015-11-17

Enterovirus D68 (EV-D68) resulted in a reported increase in the number of children needing hospital or critical care admission because of respiratory insufficiency during 2014. It remains unclear, however, whether EV-D68 infections were more severe than rhinovirus or non-EV-D68 enterovirus infections. We evaluated consecutive children presenting to a pediatric hospital between Aug. 1 and Oct. 31, 2014, with positive nasopharyngeal swabs for rhinovirus or enterovirus that were sent automatically for EV-D68 testing. We compared characteristics and outcomes of patients with EV-D68 with those with rhinovirus or non-EV-D68 enterovirus in a matched cohort study. A total of 93/297 (31.3%) of rhinovirus or enterovirus samples tested positive for EV-D68, and it was possible to compare 87 matched pairs. Children with EV-D68 infection were more likely to have difficulty breathing (odds ratio [OR] 3.00, 95% confidence interval [CI] 1.47-6.14). There was no significant difference in admission to the critical care unit or death among children with EV-D68 infection compared with those with other rhinovirus or enterovirus infections (adjusted OR 1.47, 95% CI 0.61-3.52). Children with EV-D68 infection were more often admitted to hospital, but not significantly so (adjusted OR 2.29, 95% CI 0.96-5.46). Enterovirus D68 seems to be a more virulent pulmonary pathogen than rhinovirus or non-EV-D68 enterovirus, but we did not find a significant difference in death or need for critical care. © 2015 Canadian Medical Association or its licensors.

Human Rhinovirus 87 and Enterovirus 68 Represent a Unique Serotype with Rhinovirus and Enterovirus Features

PubMed Central

Blomqvist, Soile; Savolainen, Carita; Råman, Laura; Roivainen, Merja; Hovi, Tapani

2002-01-01

It has recently been reported that all but one of the 102 known serotypes of the genus Rhinovirus segregate into two genetic clusters (C. Savolainen, S. Blomqvist, M. N. Mulders, and T. Hovi, J. Gen. Virol. 83:333-340, 2002). The only exception is human rhinovirus 87 (HRV87). Here we demonstrate that HRV87 is genetically and antigenically highly similar to enterovirus 68 (EV68) and is related to EV70, the other member of human enterovirus group D. The partial nucleotide sequences of the 5′ untranslated region, capsid regions VP4/VP2 and VP1, and the 3D RNA polymerase gene of the HRV87 prototype strain F02-3607 Corn showed 97.3, 97.8, 95.2, and 95.9% identity to the corresponding regions of EV68 prototype strain Fermon. The amino acid identities were 100 and 98.1% for the products of the two capsid regions and 97.9% for 3D RNA polymerase. Antigenic cross-reaction between HRV87 and EV68 was indicated by microneutralization with monotypic antisera. Phylogenetic analysis showed definite clustering of HRV87 and EV68 with EV70 for all sequences examined. Both HRV87 and EV68 were shown to be acid sensitive by two different assays, while EV70 was acid resistant, which is typical of enteroviruses. The cytopathic effect induced by HRV87 or EV68 was inhibited by monoclonal antibodies to the decay-accelerating factor known to be the receptor of EV70. We conclude that HRV87 and EV68 are strains of the same picornavirus serotype presenting features of both rhinoviruses and enteroviruses. PMID:12409401

Immunodominant IgM and IgG Epitopes Recognized by Antibodies Induced in Enterovirus A71-Associated Hand, Foot and Mouth Disease Patients

PubMed Central

Aw-Yong, Kam Leng; Sam, I-Ching; Koh, Mia Tuang

2016-01-01

Enterovirus A71 (EV-A71) is one of the main causative agents of hand, foot and mouth disease (HFMD). Unlike other enteroviruses that cause HFMD, EV-A71 is more frequently associated with severe neurological complications and fatality. To date, no effective licensed antivirals are available to combat EV-A71 infection. Little is known about the immunogenicity of viral non-structural proteins in humans. Previous studies have mainly focused on characterization of epitopes of EV-A71 structural proteins by using immunized animal antisera. In this study, we have characterized human antibody responses against the structural and non-structural proteins of EV-A71. Each viral protein was cloned and expressed in either bacterial or mammalian systems, and tested with antisera by western blot. Results revealed that all structural proteins (VP1-4), and non-structural proteins 2A, 3C and 3D were targets of EV-A71 IgM, whereas EV-A71 IgG recognized all the structural and non-structural proteins. Sixty three synthetic peptides predicted to be immunogenic in silico were synthesized and used for the characterization of EV-A71 linear B-cell epitopes. In total, we identified 22 IgM and four IgG dominant epitopes. Synthetic peptide PEP27, corresponding to residues 142–156 of VP1, was identified as the EV-A71 IgM-specific immunodominant epitope. PEP23, mapped to VP1 41–55, was recognized as the EV-A71 IgG cross-reactive immunodominant epitope. The structural protein VP1 is the major immunodominant site targeted by anti-EV-A71 IgM and IgG antibodies, but epitopes against non-structural proteins were also detected. These data provide new understanding of the immune response to EV-A71 infection, which benefits the development of diagnostic tools, potential therapeutics and subunit vaccine candidates. PMID:27806091

Inhibition of enterovirus 71 (EV-71) infections by a novel antiviral peptide derived from EV-71 capsid protein VP1.

PubMed

Tan, Chee Wah; Chan, Yoke Fun; Sim, Kooi Mow; Tan, Eng Lee; Poh, Chit Laa

2012-01-01

Enterovirus 71 (EV-71) is the main causative agent of hand, foot and mouth disease (HFMD). In recent years, EV-71 infections were reported to cause high fatalities and severe neurological complications in Asia. Currently, no effective antiviral or vaccine is available to treat or prevent EV-71 infection. In this study, we have discovered a synthetic peptide which could be developed as a potential antiviral for inhibition of EV-71. Ninety five synthetic peptides (15-mers) overlapping the entire EV-71 capsid protein, VP1, were chemically synthesized and tested for antiviral properties against EV-71 in human Rhabdomyosarcoma (RD) cells. One peptide, SP40, was found to significantly reduce cytopathic effects of all representative EV-71 strains from genotypes A, B and C tested, with IC(50) values ranging from 6-9.3 µM in RD cells. The in vitro inhibitory effect of SP40 exhibited a dose dependent concentration corresponding to a decrease in infectious viral particles, total viral RNA and the levels of VP1 protein. The antiviral activity of SP40 peptide was not restricted to a specific cell line as inhibition of EV-71 was observed in RD, HeLa, HT-29 and Vero cells. Besides inhibition of EV-71, it also had antiviral activities against CV-A16 and poliovirus type 1 in cell culture. Mechanism of action studies suggested that the SP40 peptide was not virucidal but was able to block viral attachment to the RD cells. Substitutions of arginine and lysine residues with alanine in the SP40 peptide at positions R3A, R4A, K5A and R13A were found to significantly decrease antiviral activities, implying the importance of positively charged amino acids for the antiviral activities. The data demonstrated the potential and feasibility of SP40 as a broad spectrum antiviral agent against EV-71.

Hookworm Secreted Extracellular Vesicles Interact With Host Cells and Prevent Inducible Colitis in Mice

PubMed Central

Eichenberger, Ramon M.; Ryan, Stephanie; Jones, Linda; Buitrago, Geraldine; Polster, Ramona; Montes de Oca, Marcela; Zuvelek, Jennifer; Giacomin, Paul R.; Dent, Lindsay A.; Engwerda, Christian R.; Field, Matthew A.; Sotillo, Javier; Loukas, Alex

2018-01-01

Gastrointestinal (GI) parasites, hookworms in particular, have evolved to cause minimal harm to their hosts, allowing them to establish chronic infections. This is mediated by creating an immunoregulatory environment. Indeed, hookworms are such potent suppressors of inflammation that they have been used in clinical trials to treat inflammatory bowel diseases (IBD) and celiac disease. Since the recent description of helminths (worms) secreting extracellular vesicles (EVs), exosome-like EVs from different helminths have been characterized and their salient roles in parasite–host interactions have been highlighted. Here, we analyze EVs from the rodent parasite Nippostrongylus brasiliensis, which has been used as a model for human hookworm infection. N. brasiliensis EVs (Nb-EVs) are actively internalized by mouse gut organoids, indicating a role in driving parasitism. We used proteomics and RNA-Seq to profile the molecular composition of Nb-EVs. We identified 81 proteins, including proteins frequently present in exosomes (like tetraspanin, enolase, 14-3-3 protein, and heat shock proteins), and 27 sperm-coating protein-like extracellular proteins. RNA-Seq analysis revealed 52 miRNA species, many of which putatively map to mouse genes involved in regulation of inflammation. To determine whether GI nematode EVs had immunomodulatory properties, we assessed their potential to suppress GI inflammation in a mouse model of inducible chemical colitis. EVs from N. brasiliensis but not those from the whipworm Trichuris muris or control vesicles from grapes protected against colitic inflammation in the gut of mice that received a single intraperitoneal injection of EVs. Key cytokines associated with colitic pathology (IL-6, IL-1β, IFNγ, and IL-17a) were significantly suppressed in colon tissues from EV-treated mice. By contrast, high levels of the anti-inflammatory cytokine IL-10 were detected in Nb-EV-treated mice. Proteins and miRNAs contained within helminth EVs hold great potential application in development of drugs to treat helminth infections as well as chronic non-infectious diseases resulting from a dysregulated immune system, such as IBD. PMID:29760697

Structural Basis for Recognition of Human Enterovirus 71 by a Bivalent Broadly Neutralizing Monoclonal Antibody

PubMed Central

Ku, Zhiqiang; Zuo, Teng; Kong, Liangliang; Zhang, Chao; Shi, Jinping; Liu, Qingwei; Chen, Tan; Zhang, Yingyi; Jiang, Wen; Zhang, Linqi; Huang, Zhong; Cong, Yao

2016-01-01

Enterovirus 71 (EV71) is the main pathogen responsible for hand, foot and mouth disease with severe neurological complications and even death in young children. We have recently identified a highly potent anti-EV71 neutralizing monoclonal antibody, termed D5. Here we investigated the structural basis for recognition of EV71 by the antibody D5. Four three-dimensional structures of EV71 particles in complex with IgG or Fab of D5 were reconstructed by cryo-electron microscopy (cryo-EM) single particle analysis all at subnanometer resolutions. The most critical EV71 mature virion-Fab structure was resolved to a resolution of 4.8 Å, which is rare in cryo-EM studies of virus-antibody complex so far. The structures reveal a bivalent binding pattern of D5 antibody across the icosahedral 2-fold axis on mature virion, suggesting that D5 binding may rigidify virions to prevent their conformational changes required for subsequent RNA release. Moreover, we also identified that the complementary determining region 3 (CDR3) of D5 heavy chain directly interacts with the extremely conserved VP1 GH-loop of EV71, which was validated by biochemical and virological assays. We further showed that D5 is indeed able to neutralize a variety of EV71 genotypes and strains. Moreover, D5 could potently confer protection in a mouse model of EV71 infection. Since the conserved VP1 GH-loop is involved in EV71 binding with its uncoating receptor, the scavenger receptor class B, member 2 (SCARB2), the broadly neutralizing ability of D5 might attribute to its inhibition of EV71 from binding SCARB2. Altogether, our results elucidate the structural basis for the binding and neutralization of EV71 by the broadly neutralizing antibody D5, thereby enhancing our understanding of antibody-based protection against EV71 infection. PMID:26938634

Influence of processing conditions on the optical properties of chemically deposited zinc sulphide (ZnS) thin film

NASA Astrophysics Data System (ADS)

Igweoko, A. E.; Augustine, C.; Idenyi, N. E.; Okorie, B. A.; Anyaegbunam, F. N. C.

2018-03-01

In this paper, we present the influence of post deposition annealing and varying concentration on the optical properties of ZnS thin films fabricated by chemical bath deposition (CBD) at 65 °C from chemical baths comprising NH3/SC(NH2)2/ZnSO4 solutions at pH of about 10. The film samples were annealed at temperatures ranging from 373 K–473 K and the concentration of the film samples vary from 0.1 M–0.7 M. Post deposition annealing and concentration played an important role on the optical parameters investigated which includes absorbance, transmittance, reflectance, absorption coefficient, band gap, refractive index and extinction coefficient. The optical parameters were found to vary with post deposition annealing in one direction and concentration of Zn2+ in the reverse direction. For instance, post deposition annealing increases the band gap from 3.65 eV for as-deposited to 3.70 eV, 3.75 eV and 3.85 eV for annealed at 373 K, 423 K and 473 K respectively whereas concentration of Zn2+ decreases the band gap from 3.95 eV at 0.1 M to 3.90 eV, 3.85 eV and 3.80 eV at 0.3 M, 0.5 M and 0.7 M respectively. The fundamental absorption edge of ZnS thin films shifted toward the highest photon energies (blue shift) after annealing and shifted toward the lowest photon energies (red shift) with increasing Zn ions concentration. A linear relation between band gap energy and Urbach energy was found. After annealing, the Urbach energy increases form 3.10 eV to 3.50 eV and decreases from 3.40 eV to 3.10 eV at varying Zn2+ concentration. The property of wide band gap makes ZnS suitable for buffer layer of film solar cells, permitting more light especially the short wavelength light into absorber layer.

Imaging of extracellular vesicles derived from human bone marrow mesenchymal stem cells using fluorescent and magnetic labels.

PubMed

Dabrowska, Sylwia; Del Fattore, Andrea; Karnas, Elzbieta; Frontczak-Baniewicz, Malgorzata; Kozlowska, Hanna; Muraca, Maurizio; Janowski, Miroslaw; Lukomska, Barbara

2018-01-01

Mesenchymal stem cells have been shown therapeutic in various neurological disorders. Recent studies support the notion that the predominant mechanism by which MSCs act is through the release of extracellular vesicles (EVs). EVs seem to have similar therapeutic activity as their cellular counterparts and may represent an interesting alternative standalone therapy for various diseases. The aim of the study was to optimize the method of EV imaging to better understand therapeutic effects mediated by EVs. The fluorescent lipophilic stain PKH26 and superparamagnetic iron oxide nanoparticles conjugated with rhodamine (Molday ION Rhodamine B™) were used for the labeling of vesicles in human bone marrow MSCs (hBM-MSCs). The entire cycle from intracellular vesicles to EVs followed by their uptake by hBM-MSCs has been studied. The identity of vesicles has been proven by antibodies against: anti-CD9, -CD63, and -CD81 (tetraspanins). NanoSight particle tracking analysis (NTA), high-resolution flow cytometric analysis, transmission electron microscopy (TEM), ELYRA PS.1 super-resolution microscopy, and magnetic resonance imaging (MRI) were used for the characterization of vesicles. The PKH26 and Molday ION were exclusively localized in intracellular vesicles positively stained for EV markers: CD9, CD63, and CD81. The isolated EVs represent heterogeneous population of various sizes as confirmed by NTA. The TEM and MRI were capable to show successful labeling of EVs using ION. Co-culture of EVs with hBM-MSCs revealed their uptake by cells in vitro, as visualized by the co-localization of PKH26 or Molday ION with tetraspanins inside hBM-MSCs. PKH26 and Molday ION seem to be biocompatible with EVs, and the labeling did not interfere with the capability of EVs to re-enter hBM-MSCs during co-culture in vitro. Magnetic properties of IONs provide an additional advantage for the imaging of EV using TEM and MRI.

Production of EV71 vaccine candidates

PubMed Central

Chong, Pele; Hsieh, Shih-Yang; Liu, Chia-Chyi; Chou, Ai-Hsiang; Chang, Jui-Yuan; Wu, Suh-Chin; Liu, Shih-Jen; Chow, Yen-Hung; Su, Ih-Jen; Klein, Michel

2012-01-01

Enterovirus 71 (EV71) is now recognized as an emerging neurotropic virus in Asia and with Coxsackie virus (CV) it is the other major causative agent of hand-foot-mouth diseases (HFMD). Effective medications and/or prophylactic vaccines against HFMD are urgently needed. From a scientific (the feasibility of bioprocess, immunological responses and potency in animal challenge model) and business development (cost of goods) points of view, we in this review address and discuss the pros and cons of different EV71 vaccine candidates that have been produced and evaluated in animal models. Epitope-based synthetic peptide vaccine candidates containing residues 211–225 of VP1 formulated with Freund’s adjuvant (CFA/IFA) elicited low EV71 virus neutralizing antibody responses, but were protective in the suckling mouse challenge model. Among recombinant EV71 subunits (rVP1, rVP2 and rVP3) expressed in E. coli, purified and formulated with CFA/IFA, only VP1 elicited mouse antibody responses with measurable EV71-specific virus neutralization titers. Immunization of mice with either a DNA plasmid containing VP1 gene or VP1 expressed in Salmonella typhimurium also generated neutralizing antibody responses and protected animals against a live EV71 challenge. Recombinant EV71 virus-like particles (rVLP) produced from baculovirus formulated either with CFA/IFA or alum elicited good virus neutralization titers in both mice and non-human primates, and were found to be protective in the suckling mouse EV71 challenge model. Synthetic peptides or recombinant EV71 subunit vaccines (rVP1 and rVLP) formulated in alum were found to be poorly immunogenic in rabbits. Only formalin-inactivated (FI) EV71 virions formulated in alum elicited cross-neutralizing antibodies against different EV71 genotypes in mice, rabbits and non-human primates but induced weak neutralizing responses against CAV16. From a regulatory, economic and market acceptability standpoint, FI-EV71 virion vaccines are the most promising candidates and are currently being evaluated in human clinical trials. We further describe and analyze some new bioprocesses technologies that have great potential applications in EV71 vaccine development. This review also demonstrates the opportunities and challenges that the Asian vaccine industry faces today. PMID:22992566

Computational materials design of attractive Fermion system with large negative effective Ueff in the hole-doped Delafossite of CuAlO2, AgAlO2 and AuAlO2: Charge-excitation induced Ueff < 0

NASA Astrophysics Data System (ADS)

Nakanishi, A.; Fukushima, T.; Uede, H.; Katayama-Yoshida, H.

2015-12-01

On the basis of general design rules for negative effective U(Ueff) systems by controlling purely-electronic and attractive Fermion mechanisms, we perform computational materials design (CMD®) for the negative Ueff system in hole-doped two-dimensional (2D) Delafossite CuAlO2, AgAlO2 and AuAlO2 by ab initio calculations with local density approximation (LDA) and self-interaction corrected-LDA (SIC-LDA). It is found that the large negative Ueff in the hole-doped attractive Fermion systems for CuAlO2 (UeffLDA = - 4.53 eV and UeffSIC-LDA = - 4.20 eV), AgAlO2 (UeffLDA = - 4.88 eV and UeffSIC-LDA = - 4.55 eV) and AuAlO2 (UeffLDA = - 4.14 eV and UeffSIC-LDA = - 3.55 eV). These values are 10 times larger than that in hole-doped three-dimensional (3D) CuFeS2 (Ueff = - 0.44 eV). For future calculations of Tc and phase diagram by quantum Monte Carlo simulations, we propose the negative Ueff Hubbard model with the anti-bonding single π-band model for CuAlO2, AgAlO2 and AuAlO2 using the mapped parameters obtained from ab initio electronic structure calculations. Based on the theory of negative Ueff Hubbard model (Noziéres and Schmitt-Rink, 1985), we discuss |Ueff| dependence of superconducting critical temperature (Tc) in the 2D Delafossite of CuAlO2, AgAlO2 and AuAlO2 and 3D Chalcopyrite of CuFeS2, which shows the interesting chemical trend, i.e., Tc increases exponentially (Tc ∝ exp [ - 1 / | Ueff | ]) in the weak coupling regime | Ueff(- 0.44 eV) | < W(∼ 2 eV) (where W is the band width of the negative Ueff Hubbard model) for the hole-doped CuFeS2, and then Tc goes through a maximum when | Ueff(- 4.88 eV , - 4.14 eV) | ∼ W(2.8 eV , 3.5 eV) for the hole-doped AgAlO2 and AuAlO2, and finally Tc decreases with increasing |Ueff| in the strong coupling regime, where | Ueff(- 4.53 eV) | > W(1.7 eV) , for the hole-doped CuAlO2.

EV-TRACK: transparent reporting and centralizing knowledge in extracellular vesicle research.

PubMed

Van Deun, Jan; Mestdagh, Pieter; Agostinis, Patrizia; Akay, Özden; Anand, Sushma; Anckaert, Jasper; Martinez, Zoraida Andreu; Baetens, Tine; Beghein, Els; Bertier, Laurence; Berx, Geert; Boere, Janneke; Boukouris, Stephanie; Bremer, Michel; Buschmann, Dominik; Byrd, James B; Casert, Clara; Cheng, Lesley; Cmoch, Anna; Daveloose, Delphine; De Smedt, Eva; Demirsoy, Seyma; Depoorter, Victoria; Dhondt, Bert; Driedonks, Tom A P; Dudek, Aleksandra; Elsharawy, Abdou; Floris, Ilaria; Foers, Andrew D; Gärtner, Kathrin; Garg, Abhishek D; Geeurickx, Edward; Gettemans, Jan; Ghazavi, Farzaneh; Giebel, Bernd; Kormelink, Tom Groot; Hancock, Grace; Helsmoortel, Hetty; Hill, Andrew F; Hyenne, Vincent; Kalra, Hina; Kim, David; Kowal, Joanna; Kraemer, Sandra; Leidinger, Petra; Leonelli, Carina; Liang, Yaxuan; Lippens, Lien; Liu, Shu; Lo Cicero, Alessandra; Martin, Shaun; Mathivanan, Suresh; Mathiyalagan, Prabhu; Matusek, Támas; Milani, Gloria; Monguió-Tortajada, Marta; Mus, Liselot M; Muth, Dillon C; Németh, Andrea; Nolte-'t Hoen, Esther N M; O'Driscoll, Lorraine; Palmulli, Roberta; Pfaffl, Michael W; Primdal-Bengtson, Bjarke; Romano, Erminia; Rousseau, Quentin; Sahoo, Susmita; Sampaio, Natalia; Samuel, Monisha; Scicluna, Benjamin; Soen, Bieke; Steels, Anneleen; Swinnen, Johannes V; Takatalo, Maarit; Thaminy, Safia; Théry, Clotilde; Tulkens, Joeri; Van Audenhove, Isabel; van der Grein, Susanne; Van Goethem, Alan; van Herwijnen, Martijn J; Van Niel, Guillaume; Van Roy, Nadine; Van Vliet, Alexander R; Vandamme, Niels; Vanhauwaert, Suzanne; Vergauwen, Glenn; Verweij, Frederik; Wallaert, Annelynn; Wauben, Marca; Witwer, Kenneth W; Zonneveld, Marijke I; De Wever, Olivier; Vandesompele, Jo; Hendrix, An

2017-02-28

We argue that the field of extracellular vesicle (EV) biology needs more transparent reporting to facilitate interpretation and replication of experiments. To achieve this, we describe EV-TRACK, a crowdsourcing knowledgebase (http://evtrack.org) that centralizes EV biology and methodology with the goal of stimulating authors, reviewers, editors and funders to put experimental guidelines into practice.

Distributed Solar Photovoltaics for Electric Vehicle Charging: Regulatory and Policy Considerations (Brochure)

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

Not Available

2014-09-01

Increasing demand for electric vehicle (EV) charging provides an opportunity for market expansion of distributed solar technology. A major barrier to the current deployment of solar technology for EV charging is a lack of clear information for policy makers, utilities and potential adopters. This paper introduces the pros and cons of EV charging during the day versus at night, summarizes the benefits and grid implications of combining solar and EV charging technologies, and offers some regulatory and policy options available to policy makers and regulators wanting to incentivize solar EV charging.

Cellular Engineering with Membrane Fusogenic Liposomes to Produce Functionalized Extracellular Vesicles.

PubMed

Lee, Junsung; Lee, Hyoungjin; Goh, Unbyeol; Kim, Jiyoung; Jeong, Moonkyoung; Lee, Jean; Park, Ji-Ho

2016-03-23

Engineering of extracellular vesicles (EVs) without affecting biological functions remains a challenge, limiting the broad applications of EVs in biomedicine. Here, we report a method to equip EVs with various functional agents, including fluorophores, drugs, lipids, and bio-orthogonal chemicals, in an efficient and controlled manner by engineering parental cells with membrane fusogenic liposomes, while keeping the EVs intact. As a demonstration of how this method can be applied, we prepared EVs containing azide-lipids, and conjugated them with targeting peptides using copper-free click chemistry to enhance targeting efficacy to cancer cells. We believe that this liposome-based cellular engineering method will find utility in studying the biological roles of EVs and delivering therapeutic agents through their innate pathway.

Do Extremely Violent Juveniles Respond Differently to Treatment?

PubMed Central

Asscher, Jessica J.; Deković, M.; Van den Akker, Alithe L.; Prins, Pier J. M.; Van der Laan, Peter H.

2016-01-01

This study increases knowledge on effectiveness of treatment for extremely violent (EV) youth by investigating their response to multisystemic therapy (MST). Using data of a randomized controlled trial on effectiveness of MST, we investigated differences in treatment response between EV youth and not extremely violent (NEV) youth. Pre- to post-treatment comparison indicated MST was equally effective for EV and NEV youth, whereas treatment as usual was not effective for either group. Growth curves of within-treatment changes indicated EV youth responded differently to MST than NEV youth. The within-treatment change was for EV youth non-linear: Initially, they show a deterioration; however, after one month, EV juveniles respond positively to MST, indicating longer lasting, intensive programs may be effective in treating extreme violence. PMID:27794135

CD30 on extracellular vesicles from malignant Hodgkin cells supports damaging of CD30 ligand-expressing bystander cells with Brentuximab-Vedotin, in vitro

PubMed Central

Hansen, Hinrich P.; Trad, Ahmad; Dams, Maria; Zigrino, Paola; Moss, Marcia; Tator, Maximilian; Schön, Gisela; Grenzi, Patricia C; Bachurski, Daniel; Aquino, Bruno; Dürkop, Horst; Reiners, Katrin S; von Bergwelt-Baildon, Michael; Hallek, Michael; Grötzinger, Joachim; Engert, Andreas; Leme, Adriana F Paes; von Strandmann, Elke Pogge

2016-01-01

The goal of targeted immunotherapy in cancer is to damage both malignant and tumor-supporting cells of the microenvironment but spare unaffected tissue. The malignant cells in classical Hodgkin lymphoma (cHL) selectively express CD30. They release this receptor on extracellular vesicles (EVs) for the tumor-supporting communication with CD30 ligand (CD30L)-positive bystander cells. Here, we investigated how CD30-positive EVs influence the efficacy of the CD30 antibody drug conjugate (ADC) Brentuximab Vedotin (SGN-35). The malignant cells and the EVs expressed the active sheddase ADAM10. ADAM10 cleaved and released the CD30 ectodomain (sCD30), causing a gradual depletion of SGN-35 binding sites on EVs and creating a soluble competitor of the ADC therapy. In a 3D semi-solid tumor microenvironment model, the EVs were retained in the matrix whereas sCD30 penetrated readily into the surrounding culture medium. This resulted in a lowered ratio of EV-associated CD30 (CD30EV) to sCD30 in the surrounding medium in comparison to non-embedded cultures. A low percentage of CD30EV was also detected in the plasma of cHL patients, supporting the clinical relevance of the model. The adherence of CD30EV but not sCD30 to CD30−/CD30L+ mast cells and eosinophils allowed the indirect binding of SGN-35. Moreover, SGN-35 damaged CD30-negative cells, provided they were loaded with CD30+ EVs. PMID:27105521

Detection of prostate cancer-specific transcripts in extracellular vesicles isolated from post-DRE urine

PubMed Central

Pellegrini, Kathryn L.; Patil, Dattatraya; Douglas, Kristen J.S.; Lee, Grace; Wehrmeyer, Kathryn; Torlak, Mersiha; Clark, Jeremy; Cooper, Colin S.; Moreno, Carlos S.; Sanda, Martin G.

2018-01-01

Background The measurement of gene expression in post-digital rectal examination (DRE) urine specimens provides a non-invasive method to determine a patient’s risk of prostate cancer. Many currently available assays use whole urine or cell pellets for the analysis of prostate cancer-associated genes, although the use of extracellular vesicles (EVs) has also recently been of interest. We investigated the expression of prostate-, kidney-, and bladder-specific transcripts and known prostate cancer biomarkers in urine EVs. Methods Cell pellets and EVs were recovered from post-DRE urine specimens, with the total RNA yield and quality determined by Bioanalyzer. The levels of prostate, kidney, and bladder-associated transcripts in EVs were assessed by TaqMan qPCR and targeted sequencing. Results RNA was more consistently recovered from the urine EV specimens, with over 80% of the patients demonstrating higher RNA yields in the EV fraction as compared to urine cell pellets. The median EV RNA yield of 36.4 ng was significantly higher than the median urine cell pellet RNA yield of 4.8 ng. Analysis of the post-DRE urine EVs indicated that prostate-specific transcripts were more abundant than kidney- or bladder-specific transcripts. Additionally, patients with prostate cancer had significantly higher levels of the prostate cancer-associated genes PCA3 and ERG. Conclusions Post-DRE urine EVs are a viable source of prostate-derived RNAs for biomarker discovery and prostate cancer status can be distinguished from analysis of these specimens. Continued analysis of urine EVs offers the potential discovery of novel biomarkers for pre-biopsy prostate cancer detection. PMID:28419548

Detection of prostate cancer-specific transcripts in extracellular vesicles isolated from post-DRE urine.

PubMed

Pellegrini, Kathryn L; Patil, Dattatraya; Douglas, Kristen J S; Lee, Grace; Wehrmeyer, Kathryn; Torlak, Mersiha; Clark, Jeremy; Cooper, Colin S; Moreno, Carlos S; Sanda, Martin G

2017-06-01

The measurement of gene expression in post-digital rectal examination (DRE) urine specimens provides a non-invasive method to determine a patient's risk of prostate cancer. Many currently available assays use whole urine or cell pellets for the analysis of prostate cancer-associated genes, although the use of extracellular vesicles (EVs) has also recently been of interest. We investigated the expression of prostate-, kidney-, and bladder-specific transcripts and known prostate cancer biomarkers in urine EVs. Cell pellets and EVs were recovered from post-DRE urine specimens, with the total RNA yield and quality determined by Bioanalyzer. The levels of prostate, kidney, and bladder-associated transcripts in EVs were assessed by TaqMan qPCR and targeted sequencing. RNA was more consistently recovered from the urine EV specimens, with over 80% of the patients demonstrating higher RNA yields in the EV fraction as compared to urine cell pellets. The median EV RNA yield of 36.4 ng was significantly higher than the median urine cell pellet RNA yield of 4.8 ng. Analysis of the post-DRE urine EVs indicated that prostate-specific transcripts were more abundant than kidney- or bladder-specific transcripts. Additionally, patients with prostate cancer had significantly higher levels of the prostate cancer-associated genes PCA3 and ERG. Post-DRE urine EVs are a viable source of prostate-derived RNAs for biomarker discovery and prostate cancer status can be distinguished from analysis of these specimens. Continued analysis of urine EVs offers the potential discovery of novel biomarkers for pre-biopsy prostate cancer detection. © 2017 Wiley Periodicals, Inc.

Mesenchymal Stromal/stem Cell-derived Extracellular Vesicles Promote Human Cartilage Regeneration In Vitro

PubMed Central

Vonk, Lucienne A.; van Dooremalen, Sanne F. J.; Liv, Nalan; Klumperman, Judith; Coffer, Paul J.; Saris, Daniël B.F.; Lorenowicz, Magdalena J.

2018-01-01

Osteoarthritis (OA) is a rheumatic disease leading to chronic pain and disability with no effective treatment available. Recently, allogeneic human mesenchymal stromal/stem cells (MSC) entered clinical trials as a novel therapy for OA. Increasing evidence suggests that therapeutic efficacy of MSC depends on paracrine signalling. Here we investigated the role of extracellular vesicles (EVs) secreted by human bone marrow derived MSC (BMMSC) in human OA cartilage repair. Methods: To test the effect of BMMSC-EVs on OA cartilage inflammation, TNF-alpha-stimulated OA chondrocyte monolayer cultures were treated with BMMSC-EVs and pro-inflammatory gene expression was measured by qRT-PCR after 48 h. To assess the impact of BMMSC-EVs on cartilage regeneration, BMMSC-EVs were added to the regeneration cultures of human OA chondrocytes, which were analyzed after 4 weeks for glycosaminoglycan content by 1,9-dimethylmethylene blue (DMMB) assay. Furthermore, paraffin sections of the regenerated tissue were stained for proteoglycans (safranin-O) and type II collagen (immunostaining). Results: We show that BMMSC-EVs inhibit the adverse effects of inflammatory mediators on cartilage homeostasis. When co-cultured with OA chondrocytes, BMMSC-EVs abrogated the TNF-alpha-mediated upregulation of COX2 and pro-inflammatory interleukins and inhibited TNF-alpha-induced collagenase activity. BMMSC-EVs also promoted cartilage regeneration in vitro. Addition of BMMSC-EVs to cultures of chondrocytes isolated from OA patients stimulated production of proteoglycans and type II collagen by these cells. Conclusion: Our data demonstrate that BMMSC-EVs can be important mediators of cartilage repair and hold great promise as a novel therapeutic for cartilage regeneration and osteoarthritis. PMID:29463990

[Expression of EV71-VP1, PSGL-1 and SCARB2 in Tissues of Infants with Brain Stem Encephalitis].

PubMed

Li, Ming; Kong, Xiao-ping; Liu, Hong; Cheng, Ling-xi; Huang, Jing-lu; Quan, Li; Wu, Fang-yu; Hao, Bo; Liu, Chao; Luo, Bin

2015-04-01

To understand the correlation of enterovirus 71 (EV71), P-selectin glycoprotein ligand-1 (PSGL-1), and scavenger receptor B2 (SCARB2) and to explore the possible pathway and mechanism of EV71 infection by observing the expression of EV71, PSGL-1 and SCARB2 in tissues of infants with brain stem encephalitis. The organs and tissues of infants with EV71-VP1 positivity in their brain stems were chosen. Expression and distribution of EV71-VP1, PSGL-1, and SCARB2 were detected and compared by immunohistochemistry. Strong staining of EV71 -VP1 was observed in the neuron, glial cells, the inflammatory cells of perivascular cuffing, parietal cells of the gastric fundus gland while alveolar macrophages, intestinal gland epithelium cells, mucosa lymphoid nodule and lymphocyte of palatine tonsil showed moderate staining and weak staining were displayed in mesenteric lymph nodes and lymphocyte of spleen. PSGL-1 expression was detected in parietal cells of the gastric fundus gland, tonsillar crypt squamous epithelium, alveolar macrophages and leukocytes in each tissue. SCARB2 expression was observed in all the above tissues except the intestines and spleen. The distribution of EV71 correlates with SCARB2 expression. SCARB2 plays an important role in virus infection and replication. Stomach may be an important site for EV71 replication.

How to Assess the External Validity and Model Validity of Therapeutic Trials: A Conceptual Approach to Systematic Review Methodology

PubMed Central

2014-01-01

Background. Evidence rankings do not consider equally internal (IV), external (EV), and model validity (MV) for clinical studies including complementary and alternative medicine/integrative medicine (CAM/IM) research. This paper describe this model and offers an EV assessment tool (EVAT©) for weighing studies according to EV and MV in addition to IV. Methods. An abbreviated systematic review methodology was employed to search, assemble, and evaluate the literature that has been published on EV/MV criteria. Standard databases were searched for keywords relating to EV, MV, and bias-scoring from inception to Jan 2013. Tools identified and concepts described were pooled to assemble a robust tool for evaluating these quality criteria. Results. This study assembled a streamlined, objective tool to incorporate for the evaluation of quality of EV/MV research that is more sensitive to CAM/IM research. Conclusion. Improved reporting on EV can help produce and provide information that will help guide policy makers, public health researchers, and other scientists in their selection, development, and improvement in their research-tested intervention. Overall, clinical studies with high EV have the potential to provide the most useful information about “real-world” consequences of health interventions. It is hoped that this novel tool which considers IV, EV, and MV on equal footing will better guide clinical decision making. PMID:24734111

Photoabsorption and S 2p photoionization of the SF6 molecule: resonances in the excitation energy range of 200-280 eV.

PubMed

Stener, M; Bolognesi, P; Coreno, M; O'Keeffe, P; Feyer, V; Fronzoni, G; Decleva, P; Avaldi, L; Kivimäki, A

2011-05-07

Photoabsorption and S 2p photoionization of the SF(6) molecule have been studied experimentally and theoretically in the excitation energy range up to 100 eV above the S 2p ionization potentials. In addition to the well-known 2t(2g) and 4e(g) shape resonances, the spin-orbit-resolved S 2p photoionization cross sections display two weak resonances between 200 and 210 eV, a wide resonance around 217 eV, a Fano-type resonance around 240 eV, and a second wide resonance around 260 eV. Calculations based on time-dependent density functional theory allow us to assign the 217-eV and 260-eV features to the shape resonances in S 2p photoionization. The Fano resonance is caused by the interference between the direct S 2p photoionization channel and the resonant channel that results from the participator decay of the S 2s(-1)6t(1u) excited state. The weak resonances below 210-eV photon energy, not predicted by theory, are tentatively suggested to originate from the coupling between S 2p shake-up photoionization and S 2p single-hole photoionization. The experimental and calculated angular anisotropy parameters for S 2p photoionization are in good agreement.

The impact of electric vehicles on the outlook of future energy system

NASA Astrophysics Data System (ADS)

Zhuk, A.; Buzoverov, E.

2018-02-01

Active promotion of electric vehicles (EVs) and technology of fast EV charging in the medium term may cause significant peak loads on the energy system, what necessitates making strategic decisions related to the development of generating capacities, distribution networks with EV charging infrastructure, and priorities in the development of battery electric vehicles and vehicles with electrochemical generators. The paper analyses one of the most significant aspects of joint development of electric transport system and energy system in the conditions of substantial growth of energy consumption by EVs. The assessments of per-unit-costs of operation and depreciation of EV power unit were made, taking into consideration the expenses of electric power supply. The calculations show that the choice of electricity buffering method for EV fast charging depends on the character of electricity infrastructure in the region where the electric transport is operating. In the conditions of high density of electricity network and a large number of EVs, the stationary storage facilities or the technology of distributed energy storage in EV batteries - vehicle-to-grid (V2G) technology may be used for buffering. In the conditions of low density and low capacity of electricity networks, the most economical solution could be usage of EVs with traction power units based on the combination of air-aluminum electrochemical generator and a buffer battery of small capacity.

Low-energy electron collisions with proline and pyrrolidine: A comparative study

NASA Astrophysics Data System (ADS)

Barbosa, Alessandra Souza; Freitas, Thiago Corrêa; Bettega, M. H. F.

2018-02-01

We present a comparative study on the calculated cross sections obtained for the elastic collisions of low-energy electrons with the amino acid proline (C5H9NO2) and its building block pyrrolidine (C4H9N). We employed the Schwinger multichannel method implemented with pseudopotentials to compute integral, differential, and momentum transfer cross sections in the static-exchange plus polarization approximation, for energies up to 15 eV. We report three shape resonances for proline at around 1.7 eV, 6.8 eV, and 10 eV and two shape resonances for pyrrolidine centered at 7 eV and 10.2 eV. The present resonance energies are compared with available experimental data on vertical attachment energies and dissociative electron attachment, where a good agreement is found. From the comparison of the present results with available calculated cross sections for the simplest carboxylic acid, formic acid (HCOOH), and from electronic structure calculations, we found that the first resonance of proline, at 1.7 eV, is due the presence of the carboxylic group, whereas the other two structures, at 6.8 eV and 10 eV, clearly arise from the pyrrolidine ring. A comparison between the differential cross sections for proline and pyrrolidine at some selected energies of the incident electron is also reported in this paper.

Evidence of bovine serum albumin-viologen herbicide binding interaction and associated structural modifications

NASA Astrophysics Data System (ADS)

Roy, Swarup; Saxena, Shailendra K.; Mishra, Suryakant; Yogi, Priyanka; Sagdeo, P. R.; Kumar, Rajesh

2017-07-01

The binding ability of viologen herbicide with bovine serum albumin (BSA) has been investigated to understand viologen associated hazards by investigating ethyl viologen's (EV) binding using various spectroscopies and in-silico molecular docking approaches. Apparent association constant (1.3 × 104 L/mol), calculated using UV-Vis spectra indicating a moderate complex formation between BSA and EV. A static mode of fluorescence quenching has been observed as evident from inverse temperature dependence of Stern-Volmer quenching constant which also confirms an EV-BSA complex formation. Emission and time resolved fluorescence studies reveal that the emission quenching of BSA with EV is initiated by static quenching mechanism. A moderately strong binding affinity between EV and BSA has been observed (binding constant value of 7.58 × 104 L/Mol) using fluorescence quenching titration, obtained at 298 K. Quantitative measurements of thermodynamic parameters like enthalpy and entropy changes clearly indicates hydrophobic force responsible for EV-BSA complex formation. The binding distance between EV and BSA was found to be 4.48 nm are involved in non-radiative energy transfer process. Furthermore, from the circular dichroism spectra it was observed that addition of EV is also found to change the secondary structure of BSA which leads to decrease in α-helix. Above mentioned results are found to be in consonance with molecular docking simulations and supports the EV-BSA binding.

7.87 eV Laser Desorption Postionization Mass Spectrometry of Adsorbed and Covalently Bound Bisphenol A Diglycidyl Methacrylate

PubMed Central

Zhou, Manshui; Wu, Chunping; Akhmetov, Artem; Edirisinghe, Praneeth D.; Drummond, James L.; Hanley, Luke

2007-01-01

Bisphenol A diglycidyl methacrylate (Bis-GMA) was adsorbed onto or covalently bound to a porous silicon oxide surface. Laser desorption 10.5 eV postionization mass spectrometry (LDPI-MS) was previously demonstrated for surface analysis of adsorbed and surface bound Bis-GMA, but signal to noise levels were low and ion fragmentation was extensive. 7.87 eV postionization using the fluorine laser was demonstrated here for Bis-GMA. However, signal levels remained low for LDPI-MS of Bis-GMA as its ionization potential was only ∼7.8 eV, near threshold for single photon ionization by the 7.87 eV fluorine laser. It is known that aromatic tagging of molecular analytes can lower the overall IP of the tagged molecular complex, allowing 7.87 eV single photon ionization. Therefore, Bis-GMA was also derivatized with several tags whose IPs were either below or above 7.87 eV: the tag with an IP below 7.87 eV enhanced single photon ionization while the tags with higher IPs did not. However, signal intensities were enhanced by resonant laser desorption for two of the derivatized Bis-GMAs. Intact ions of Bis-GMA and its derivatives were generally observed by 7.87 eV LDPI-MS, consistent with the formation of ions with relatively little internal energy upon threshold single photon ionization. PMID:17449273

Newt cells secrete extracellular vesicles with therapeutic bioactivity in mammalian cardiomyocytes.

PubMed

Middleton, Ryan C; Rogers, Russell G; De Couto, Geoffrey; Tseliou, Eleni; Luther, Kristin; Holewinski, Ronald; Soetkamp, Daniel; Van Eyk, Jennifer E; Antes, Travis J; Marbán, Eduardo

2018-01-01

Newts can regenerate amputated limbs and cardiac tissue, unlike mammals which lack broad regenerative capacity. Several signaling pathways involved in cell proliferation, differentiation and survival during newt tissue regeneration have been elucidated, however the factors that coordinate signaling between cells, as well as the conservation of these factors in other animals, are not well defined. Here we report that media conditioned by newt limb explant cells (A1 cells) protect mammalian cardiomyocytes from oxidative stress-induced apoptosis. The cytoprotective effect of A1-conditioned media was negated by exposing A1 cells to GW4869, which suppresses the generation of extracellular vesicles (EVs). A1-EVs are similar in diameter (~100-150 nm), structure, and share several membrane surface and cargo proteins with mammalian exosomes. However, isolated A1-EVs contain significantly higher levels of both RNA and protein per particle than mammalian EVs. Additionally, numerous cargo RNAs and proteins are unique to A1-EVs. Of particular note, A1-EVs contain numerous mRNAs encoding nuclear receptors, membrane ligands, as well as transcription factors. Mammalian cardiomyocytes treated with A1-EVs showed increased expression of genes in the PI3K/AKT pathway, a pivotal player in survival signaling. We conclude that newt cells secrete EVs with diverse, distinctive RNA and protein contents. Despite ~300 million years of evolutionary divergence between newts and mammals, newt EVs confer cytoprotective effects on mammalian cardiomyocytes.

Acoustic Enrichment of Extracellular Vesicles from Biological Fluids.

PubMed

Ku, Anson; Lim, Hooi Ching; Evander, Mikael; Lilja, Hans; Laurell, Thomas; Scheding, Stefan; Ceder, Yvonne

2018-06-11

Extracellular vesicles (EVs) have emerged as a rich source of biomarkers providing diagnostic and prognostic information in diseases such as cancer. Large-scale investigations into the contents of EVs in clinical cohorts are warranted, but a major obstacle is the lack of a rapid, reproducible, efficient, and low-cost methodology to enrich EVs. Here, we demonstrate the applicability of an automated acoustic-based technique to enrich EVs, termed acoustic trapping. Using this technology, we have successfully enriched EVs from cell culture conditioned media and urine and blood plasma from healthy volunteers. The acoustically trapped samples contained EVs ranging from exosomes to microvesicles in size and contained detectable levels of intravesicular microRNAs. Importantly, this method showed high reproducibility and yielded sufficient quantities of vesicles for downstream analysis. The enrichment could be obtained from a sample volume of 300 μL or less, an equivalent to 30 min of enrichment time, depending on the sensitivity of downstream analysis. Taken together, acoustic trapping provides a rapid, automated, low-volume compatible, and robust method to enrich EVs from biofluids. Thus, it may serve as a novel tool for EV enrichment from large number of samples in a clinical setting with minimum sample preparation.

Geographical and temporal differences in electric vehicle range due to cabin conditioning energy consumption

NASA Astrophysics Data System (ADS)

Kambly, Kiran; Bradley, Thomas H.

2015-02-01

Electric vehicles (EVs) are vehicles that are propelled by electric motors powered by rechargeable battery. They are generally asserted to have GHG emissions, driveability and life cycle cost benefits over conventional vehicles. Despite this, EVs face significant challenges due to their limited on-board energy storage capacity. In addition to providing energy for traction, the energy storage device operates HVAC systems for cabin conditioning. This results in reduced driving range. The factors such as local ambient temperature, local solar radiation, local humidity, duration and thermal soak have been identified to affect the cabin conditions. In this paper, the development of a detailed system-level approach to HVAC energy consumption in EVs as a function of transient environmental parameters is described. The resulting vehicle thermal comfort model is used to address several questions such as 1) How does day to day environmental conditions affect EV range? 2) How does frequency of EV range change geographically? 3) How does trip start time affect EV range? 4) Under what conditions does cabin preconditioning assist in increasing the EV range? 5) What percentage increase in EV range can be expected due to cabin preconditioning at a given location?

Molecular characterization of enteroviruses including a new type EV-C99 isolated from Xinjiang students in Shandong, China in 2011.

PubMed

Tao, Zexin; Yuan, Qun; Lin, Xiaojuan; Wang, Suting; Liu, Yao; Ji, Feng; Xiong, Ping; Cui, Ning; Song, Lizhi; Wang, Mei; Xu, Aiqiang

2014-10-09

The last case of infection with wild-type poliovirus indigenous to China was reported in 1994. In 2011, a poliomyelitis outbreak caused by imported wide-type poliovirus occurred in Xinjiang Uighur Autonomous Region. Here, we report the results of enterovirus (EV) isolation from Xinjiang students that returned to school in Shandong after summer vacation during this outbreak. Stool specimens from 376 students were collected and 10 EV strains were isolated including 4 polioviruses (All Sabin strains), 1 coxsackievirus (CV) A13, 3 CVA17 and 2 EV-C99. VP1 sequence analysis revealed these CVA13, CVA17 and EV-C99 strains had 71.3-81.8%, 76.5-84.6% and 74.2-82.9% nucleotide similarity with strains from other countries within a serotype, respectively. EV-C99 strains had 82.7-92.8% VP1 similarity with two previously reported Xinjiang strains. Complete genome analysis on EV-C99 strains revealed intra-serotypic genetic recombination events. These findings reflect great genetic divergence between Chinese strains and strains from other countries of the three types, and provide valuable information on monitoring EV transmission over long distance.

Computational materials design of negative effective U system in the hole-doped Delafossite of CuAlO2, AgAlO2 and AuAlO2

NASA Astrophysics Data System (ADS)

Nakanishi, Akitaka; Fukushima, Tetsuya; Uede, Hiroki; Katayama-Yoshida, Hiroshi

2015-03-01

In order to realize the super-high-TC superconductors (TC>1,000K) based on the general design rules for the negative Ueff system, we have performed computational materials design for theUeff<0 system in the hole-doped two-dimensional (2D) Delafossite CuAlO2, AgAlO2 and AuAlO2 from the first principles. We find the interesting chemical trend of TC in 2D and 3D systems; where the TC increases exponentially in the weak coupling regime (|Ueff (-0.44eV)|< W(2eV), W is the band width) for hole-doped CuFeS2, then the TC goes through a maximum when |Ueff (-4.88eV, -4.14eV)| ~ W (2.8eV, 3.5eV) for hole-doped AgAlO2 and AuAlO2, and the TC decreases with increasing |Ueff|in strong coupling regime, where |Ueff (-4.53eV)|> W(1.7eV) for hole-doped CuAlO2

The Sox2 promoter-driven CD63-GFP transgenic rat model allows tracking of neural stem cell-derived extracellular vesicles.

PubMed

Yoshimura, Aya; Adachi, Naoki; Matsuno, Hitomi; Kawamata, Masaki; Yoshioka, Yusuke; Kikuchi, Hisae; Odaka, Haruki; Numakawa, Tadahiro; Kunugi, Hiroshi; Ochiya, Takahiro; Tamai, Yoshitaka

2018-01-30

Extracellular vesicles (EVs) can modulate microenvironments by transferring biomolecules, including RNAs and proteins derived from releasing cells, to target cells. To understand the molecular mechanisms maintaining the neural stem cell (NSC) niche through EVs, a new transgenic (Tg) rat strain that can release human CD63-GFP-expressing EVs from the NSCs was established. Human CD63-GFP expression was controlled under the rat Sox2 promoter (Sox2/human CD63-GFP), and it was expressed in undifferentiated fetal brains. GFP signals were specifically observed in in vitro cultured NSCs obtained from embryonic brains of the Tg rats. We also demonstrated that embryonic NSC (eNSC)-derived EVs were labelled by human CD63-GFP. Furthermore, when we examined the transfer of EVs, eNSC-derived EVs were found to be incorporated into astrocytes and eNSCs, thus implying an EV-mediated communication between different cell types around NSCs. This new Sox2/human CD63-GFP Tg rat strain should provide resources to analyse the cell-to-cell communication via EVs in NSC microenvironments. © 2018. Published by The Company of Biologists Ltd.

Biological properties of extracellular vesicles and their physiological functions

PubMed Central

Yáñez-Mó, María; Siljander, Pia R.-M.; Andreu, Zoraida; Zavec, Apolonija Bedina; Borràs, Francesc E.; Buzas, Edit I.; Buzas, Krisztina; Casal, Enriqueta; Cappello, Francesco; Carvalho, Joana; Colás, Eva; Silva, Anabela Cordeiro-da; Fais, Stefano; Falcon-Perez, Juan M.; Ghobrial, Irene M.; Giebel, Bernd; Gimona, Mario; Graner, Michael; Gursel, Ihsan; Gursel, Mayda; Heegaard, Niels H. H.; Hendrix, An; Kierulf, Peter; Kokubun, Katsutoshi; Kosanovic, Maja; Kralj-Iglic, Veronika; Krämer-Albers, Eva-Maria; Laitinen, Saara; Lässer, Cecilia; Lener, Thomas; Ligeti, Erzsébet; Linē, Aija; Lipps, Georg; Llorente, Alicia; Lötvall, Jan; Manček-Keber, Mateja; Marcilla, Antonio; Mittelbrunn, Maria; Nazarenko, Irina; Hoen, Esther N.M. Nolte-‘t; Nyman, Tuula A.; O'Driscoll, Lorraine; Olivan, Mireia; Oliveira, Carla; Pállinger, Éva; del Portillo, Hernando A.; Reventós, Jaume; Rigau, Marina; Rohde, Eva; Sammar, Marei; Sánchez-Madrid, Francisco; Santarém, N.; Schallmoser, Katharina; Ostenfeld, Marie Stampe; Stoorvogel, Willem; Stukelj, Roman; Van der Grein, Susanne G.; Vasconcelos, M. Helena; Wauben, Marca H. M.; De Wever, Olivier

2015-01-01

In the past decade, extracellular vesicles (EVs) have been recognized as potent vehicles of intercellular communication, both in prokaryotes and eukaryotes. This is due to their capacity to transfer proteins, lipids and nucleic acids, thereby influencing various physiological and pathological functions of both recipient and parent cells. While intensive investigation has targeted the role of EVs in different pathological processes, for example, in cancer and autoimmune diseases, the EV-mediated maintenance of homeostasis and the regulation of physiological functions have remained less explored. Here, we provide a comprehensive overview of the current understanding of the physiological roles of EVs, which has been written by crowd-sourcing, drawing on the unique EV expertise of academia-based scientists, clinicians and industry based in 27 European countries, the United States and Australia. This review is intended to be of relevance to both researchers already working on EV biology and to newcomers who will encounter this universal cell biological system. Therefore, here we address the molecular contents and functions of EVs in various tissues and body fluids from cell systems to organs. We also review the physiological mechanisms of EVs in bacteria, lower eukaryotes and plants to highlight the functional uniformity of this emerging communication system. PMID:25979354

Biological properties of extracellular vesicles and their physiological functions.

PubMed

Yáñez-Mó, María; Siljander, Pia R-M; Andreu, Zoraida; Zavec, Apolonija Bedina; Borràs, Francesc E; Buzas, Edit I; Buzas, Krisztina; Casal, Enriqueta; Cappello, Francesco; Carvalho, Joana; Colás, Eva; Cordeiro-da Silva, Anabela; Fais, Stefano; Falcon-Perez, Juan M; Ghobrial, Irene M; Giebel, Bernd; Gimona, Mario; Graner, Michael; Gursel, Ihsan; Gursel, Mayda; Heegaard, Niels H H; Hendrix, An; Kierulf, Peter; Kokubun, Katsutoshi; Kosanovic, Maja; Kralj-Iglic, Veronika; Krämer-Albers, Eva-Maria; Laitinen, Saara; Lässer, Cecilia; Lener, Thomas; Ligeti, Erzsébet; Linē, Aija; Lipps, Georg; Llorente, Alicia; Lötvall, Jan; Manček-Keber, Mateja; Marcilla, Antonio; Mittelbrunn, Maria; Nazarenko, Irina; Nolte-'t Hoen, Esther N M; Nyman, Tuula A; O'Driscoll, Lorraine; Olivan, Mireia; Oliveira, Carla; Pállinger, Éva; Del Portillo, Hernando A; Reventós, Jaume; Rigau, Marina; Rohde, Eva; Sammar, Marei; Sánchez-Madrid, Francisco; Santarém, N; Schallmoser, Katharina; Ostenfeld, Marie Stampe; Stoorvogel, Willem; Stukelj, Roman; Van der Grein, Susanne G; Vasconcelos, M Helena; Wauben, Marca H M; De Wever, Olivier

2015-01-01

In the past decade, extracellular vesicles (EVs) have been recognized as potent vehicles of intercellular communication, both in prokaryotes and eukaryotes. This is due to their capacity to transfer proteins, lipids and nucleic acids, thereby influencing various physiological and pathological functions of both recipient and parent cells. While intensive investigation has targeted the role of EVs in different pathological processes, for example, in cancer and autoimmune diseases, the EV-mediated maintenance of homeostasis and the regulation of physiological functions have remained less explored. Here, we provide a comprehensive overview of the current understanding of the physiological roles of EVs, which has been written by crowd-sourcing, drawing on the unique EV expertise of academia-based scientists, clinicians and industry based in 27 European countries, the United States and Australia. This review is intended to be of relevance to both researchers already working on EV biology and to newcomers who will encounter this universal cell biological system. Therefore, here we address the molecular contents and functions of EVs in various tissues and body fluids from cell systems to organs. We also review the physiological mechanisms of EVs in bacteria, lower eukaryotes and plants to highlight the functional uniformity of this emerging communication system.

Inflammatory Stroke Extracellular Vesicles Induce Macrophage Activation.

PubMed

Couch, Yvonne; Akbar, Naveed; Davis, Simon; Fischer, Roman; Dickens, Alex M; Neuhaus, Ain A; Burgess, Annette I; Rothwell, Peter M; Buchan, Alastair M

2017-08-01

Extracellular vesicles (EVs) are protein-lipid complexes released from cells, as well as actively exocytosed, as part of normal physiology, but also during pathological processes such as those occurring during a stroke. Our aim was to determine the inflammatory potential of stroke EVs. EVs were quantified and analyzed in the sera of patients after an acute stroke (<24 hours; OXVASC [Oxford Vascular Study]). Isolated EV fractions were subjected to untargeted proteomic analysis by liquid chromatography mass-spectrometry/mass-spectrometry and then applied to macrophages in culture to investigate inflammatory gene expression. EV number, but not size, is significantly increased in stroke patients when compared to age-matched controls. Proteomic analysis reveals an overall increase in acute phase proteins, including C-reactive protein. EV fractions applied to monocyte-differentiated macrophage cultures induced inflammatory gene expression. Together these data show that EVs from stroke patients are proinflammatory in nature and are capable of inducing inflammation in immune cells. © 2017 American Heart Association, Inc.

VizieR Online Data Catalog: Wisconsin soft X-ray diffuse background all-sky Survey (McCammon+ 1983)

NASA Astrophysics Data System (ADS)

McCammon, D.; Burrows, D. N.; Sanders, W. T.; Kraushaar, W. L.

1997-10-01

The catalog contains all-sky survey of the soft X-ray diffuse background and the count-rate data from which the maps were made for the ten flights included in the survey. It contains 40 files in the machine-readable version and includes documentation and utility subroutines. The data files contain different band maps (B, C, M, M1, M2, I, J, 2-6 keV) in a 0 degree-centered Aitoff projection, in a 180-degree-centered Aitoff projection, in a north polar projection, and in a south polar projection. Lookup tables in the form of FITS images are provided for conversion between pixel coordinates and Galactic coordinates for the various projections. The bands are: B = 130-188eV C = 160-284eV M1 = 440-930eV M2 = 600-1100eV I = 770-1500eV J = 1100-2200eV 2-6keV = 1800-6300eV (51 data files).

The Seroprevalence and Seroincidence of Enterovirus71 Infection in Infants and Children in Ho Chi Minh City, Viet Nam

PubMed Central

Tran, Chau Bich Nguyen; Nguyen, Hieu Trong; Phan, Ha Thanh Thi; Tran, Ngoc Van; Wills, Bridget; Farrar, Jeremy; Santangelo, Joseph D.; Simmons, Cameron P.

2011-01-01

Enterovirus 71 (EV71)-associated hand, foot and mouth disease has emerged as a serious public health problem in South East Asia over the last decade. To better understand the prevalence of EV71 infection, we determined EV71 seroprevalence and seroincidence amongst healthy infants and children in Ho Chi Minh City, Viet Nam. In a cohort of 200 newborns, 55% of cord blood samples contained EV71 neutralizing antibodies and these decayed to undetectable levels by 6 months of age in 98% of infants. The EV71 neutralizing antibody seroconversion rate was 5.6% in the first year and 14% in the second year of life. In children 5–15 yrs of age, seroprevalence of EV71 neutralizing antibodies was 84% and in cord blood it was 55%. Taken together, these data suggest EV71 force of infection is high and highlights the need for more research into its epidemiology and pathogenesis in high disease burden countries. PMID:21765891

Extracellular vesicle-mediated transfer of processed and functional RNY5 RNA

PubMed Central

Chakrabortty, Sudipto K.; Prakash, Ashwin; Nechooshtan, Gal; Hearn, Stephen; Gingeras, Thomas R.

2015-01-01

Extracellular vesicles (EVs) have been proposed as a means to promote intercellular communication. We show that when human primary cells are exposed to cancer cell EVs, rapid cell death of the primary cells is observed, while cancer cells treated with primary or cancer cell EVs do not display this response. The active agents that trigger cell death are 29- to 31-nucleotide (nt) or 22- to 23-nt processed fragments of an 83-nt primary transcript of the human RNY5 gene that are highly likely to be formed within the EVs. Primary cells treated with either cancer cell EVs, deproteinized total RNA from either primary or cancer cell EVs, or synthetic versions of 31- and 23-nt fragments trigger rapid cell death in a dose-dependent manner. The transfer of processed RNY5 fragments through EVs may reflect a novel strategy used by cancer cells toward the establishment of a favorable microenvironment for their proliferation and invasion. PMID:26392588

Detection and genotyping of enteroviruses in cerebrospinal fluid in patients in Victoria, Australia, 2007-2013.

PubMed

Papadakis, Georgina; Chibo, Doris; Druce, Julian; Catton, Michael; Birch, Chris

2014-09-01

Genotyping by VP1 fragment polymerase chain reaction (PCR) and nucleic acid sequencing to detect enterovirus (EV) genotypes was performed directly on 729 EV PCR positive cerebrospinal fluid (CSF) samples collected between 2007 and 2012 from Victorian hospital inpatients. The overall genotype identification rate from CSF-positive material was 43%. The four most common genotypes identified were Echovirus 6 (24%), Echovirus 30 (17%), Echovirus 25 (10%), and Coxsackievirus A9 (10%), together comprising 61% of all EVs typed. The seasonal distribution of all EVs identified followed the recognized pattern of mainly summer epidemics. Three of the four predominant genotypes were present in each of the 6 years in which the study was conducted, with 20 other EV genotypes also detected, often in only a single year. Genotyping of EVs directly in CSF is faster, simpler and more sensitive than traditional virus neutralization assays performed on EV positive samples. © 2014 Wiley Periodicals, Inc.

Predicting Rosseland mean opacities of Aluminum using a difference converging method

NASA Astrophysics Data System (ADS)

Sun, Weiguo; Fan, Zhixiang; Zhang, Yi; Fu, Jia; Hu, Shide; Zhang, Lulu; Fan, Qunchao

2018-01-01

A difference converging method for opacity (DCMo) calculation is suggested to predict opacities and transmissions of metal plasmas at other photon energies that were not measured. The applications of the DCMo to the opacity and transmission calculations of Aluminum plasmas, in the spectral range of 73.5eV ≤ hν ≤ 245eV with Te = 20eV, and of 1475eV ≤ hν ≤ 1540eV with Te = 19eV, show that: 1) the DCMo correctly reproduce all important absorption structures of the Al plasmas and their corresponding intensities; 2) the agreement between the experimental and the DCMo transmissions is notably better than that between the experimental and some other computational transmissions like the ones obtained using OPAL and ATOMIC models.

Marcasite revisited: Optical absorption gap at room temperature

NASA Astrophysics Data System (ADS)

Sánchez, C.; Flores, E.; Barawi, M.; Clamagirand, J. M.; Ares, J. R.; Ferrer, I. J.

2016-03-01

Jagadeesh and Seehra published in 1980 that the marcasite band gap energy is 0.34 eV. However, recent calculations and experimental approximations accomplished by several research groups point out that the marcasite band gap energy should be quite similar to that of pyrite (of the order of 0.8-1.0 eV). By using diffuse reflectance spectroscopy (DRS) we have determined that marcasite has no optical absorption gap at photon energies 0.06 ≤ hν ≤ 0.75 eV and that it has two well defined optical transitions at ~ 0.9 eV and ~ 2.2 eV quite similar to those of pyrite. Marcasite optical absorption gap appears to be Eg ≅ 0.83 ± 0.02 eV and it is due to an allowed indirect transition.

Time-resolved SERS for characterizing extracellular vesicles

NASA Astrophysics Data System (ADS)

Rojalin, Tatu; Saari, Heikki; Somersalo, Petter; Laitinen, Saara; Turunen, Mikko; Viitala, Tapani; Wachsmann-Hogiu, Sebastian; Smith, Zachary J.; Yliperttula, Marjo

2017-02-01

The aim of this work is to develop a platform for characterizing extracellular vesicles (EV) by using gold-polymer nanopillar SERS arrays simultaneously circumventing the photoluminescence-related disadvantages of Raman with a time-resolved approach. EVs are rich of biochemical information reporting of, for example, diseased state of the biological system. Currently, straightforward, label-free and fast EV characterization methods with low sample consumption are warranted. In this study, SERS spectra of red blood cell and platelet derived EVs were successfully measured and their biochemical contents analyzed using multivariate data analysis techniques. The developed platform could be conveniently used for EV analytics in general.

Potentials and capabilities of the Extracellular Vesicle (EV) Array.

PubMed

Jørgensen, Malene Møller; Bæk, Rikke; Varming, Kim

2015-01-01

Extracellular vesicles (EVs) and exosomes are difficult to enrich or purify from biofluids, hence quantification and phenotyping of these are tedious and inaccurate. The multiplexed, highly sensitive and high-throughput platform of the EV Array presented by Jørgensen et al., (J Extracell Vesicles, 2013; 2: 10) has been refined regarding the capabilities of the method for characterization and molecular profiling of EV surface markers. Here, we present an extended microarray platform to detect and phenotype plasma-derived EVs (optimized for exosomes) for up to 60 antigens without any enrichment or purification prior to analysis.

Co-circulation of multiple subtypes of enterovirus A71 (EV- A71) genotype C, including novel recombinants characterised by use of whole genome sequencing (WGS), Denmark 2016.

PubMed

Midgley, Sofie E; Nielsen, Astrid G; Trebbien, Ramona; Poulsen, Mille W; Andersen, Peter H; Fischer, Thea K

2017-06-29

In Europe, enterovirus A71 (EV-A71) has primarily been associated with sporadic cases of neurological disease. The recent emergence of new genotypes and larger outbreaks with severely ill patients demonstrates a potential for the spread of new, highly pathogenic EV-A71 strains. Detection and characterisation of these new emerging EV variants is challenging as standard EV assays may not be adequate, necessitating the use of whole genome analysis. This article is copyright of The Authors, 2017.

Effects of a chitin binding vicilin from Erythrina velutina seeds on bean bruchid pests (Callosobruchus maculatus and Zabrotes subfasciatus).

PubMed

Teixeira, Fabiano M; Oliveira, Adeliana S; Macedo, Leonardo L P; Santos, Elizeu A; de Sales, Mauricio P

2008-01-01

Erythrina velutina vicilin, EvV, is a dimeric glycoprotein with Mr of 124.6 kDa. EvV was tested for anti-insect activity against bean bruchid larvae. EvV had LD(50) of 0.10% and ED(50) of 0.14% for Z. subfasciatus and LD(50) of 0.26% and ED(50) of 0.19% for C. maculatus. EvV was not digested by bean larvae enzymes until 12 h of incubation, and at 24 h EvV was more resistant to Z. subfasciatus enzymes.

Electronic voting to encourage interactive lectures: a randomised trial

PubMed Central

2007-01-01

Background Electronic Voting Systems have been used for education in a variety of disciplines. Outcomes from these studies have been mixed. Because results from these studies have been mixed, we examined whether an EVS system could enhance a lecture's effect on educational outcomes. Methods A cohort of 127 Year 5 medical students at the University of Adelaide was stratified by gender, residency status and academic record then randomised into 2 groups of 64 and 63 students. Each group received consecutive 40-minute lectures on two clinical topics. One group received the EVS for both topics. The other group received traditional teaching only. Evaluation was undertaken with two, 15-question multiple-choice questionnaires (MCQ) assessing knowledge and problem solving and undertaken as a written paper immediately before and after the lectures and repeated online 8–12 weeks later. Standardised institutional student questionnaires were completed for each lecture and independent observers assessed student behaviour during the lectures. Lecturer's opinions were assessed by a questionnaire developed for this study. Results Two-thirds of students randomised to EVS and 59% of students randomised to traditional lectures attended. One-half of the students in the EVS group and 41% in the traditional group completed all questionnaires. There was no difference in MCQ scores between EVS and traditional lectures (p = 0.785). The cervical cancer lectures showed higher student ranking in favour of EVS in all parameters. The breast cancer lectures showed higher ranking in favour of traditional lectures in 5 of 7 parameters (p < 0.001). The observed higher-order lecturer-students interactions were increased in the EVS lecture for one lecturer and reduced for the other. Both lecturers felt that the EVS lectures were difficult to prepare, that they were able to keep to time in the traditional lectures, that the educational value of both lecture styles was similar, and that they were neutral-to-slightly favourably disposed to continue with the EVS technology. The 2 lecturers disagreed regarding the ease of preparation of the traditional lecture, their ability to keep to time in the EVS lecture, and personal satisfaction with the EVS lecture. The lecturers felt that EVS encouraged student participation and helped identify where students were having difficulty. Conclusion In this setting, EVS technology used in large group lectures did not offer significant advantages over the more traditional lecture format. PMID:17655773

Genetic characterization of enterovirus strains identified in Hand, Foot and Mouth Disease (HFMD): Emergence of B1c, C1 subgenotypes, E2 sublineage of CVA16, EV71 and CVA6 strains in India.

PubMed

Ganorkar, Nital N; Patil, Pooja R; Tikute, Sanjay S; Gopalkrishna, Varanasi

2017-10-01

Hand, Foot and Mouth disease (HFMD) is a common childhood disease and caused due to Enterovirus-A (EV-A), EV-B and EV-C species worldwide. Cases of HFMD were reported from, Ahmedabad (Gujarat, 2012) and Pune (Maharashtra, 2013-2014) in India. The present study highlights the identification of EV strains (CVA16, CVA6, CVA4 and Echo12), characterization of subgenotypes of CVA16, CVA6 strains during 2012-14 and CVA16, CVA6, EV71 strains reported from the earlier study (2009-10) in HFMD cases from India. A total 158 clinical specimens collected from 64 HFMD cases (2012-2014) were included in the study. EV detection was carried out by 5'NCR based RT-PCR, molecular typing and subgenotyping was by VP1/2A junction or VP1, full VP1 gene amplification respectively followed by phylogenetic analysis. The present study reports 63.92% (101/158) EV positivity by RT-PCR. Ninety four of the 101 (93.06%) EV positive strains were amplified by VP1/2A junction or VP1 regions. Sequence analysis revealed the presence of CVA16 (61.7%), CVA6 (34.04%), CVA4 and Echo12 (4.3%). A total of 114 EV positive strains were genotyped using full and partial VP1 region. All CVA16 Indian strains (n=70) clustered with rarely reported B1c subgenotype, CVA6 (n=43) and EV71 (n=1) strains clustered with sub-lineage E2 and C1 subgenotypes respectively. In summary, the study reports genetic characterization of CVA16, CVA6, CVA4 and Echo12 strains in HFMD cases from India. Circulation of B1c subgenotype of CVA16, E2 sub-lineage of CVA6 and C1 subgenotype of EV 71 strains in HFMD cases were reported for the first time from India. This study helps to understand the genotype distribution, genetic diversity of EV strains associated with HFMD from Eastern, Western and Southern regions in India. Copyright © 2017 Elsevier B.V. All rights reserved.

Co-Circulation and Genomic Recombination of Coxsackievirus A16 and Enterovirus 71 during a Large Outbreak of Hand, Foot, and Mouth Disease in Central China

PubMed Central

Liu, Weiyong; Wu, Shimin; Xiong, Ying; Li, Tongya; Wen, Zhou; Yan, Mingzhe; Qin, Kai; Liu, Yingle; Wu, Jianguo

2014-01-01

A total of 1844 patients with hand, foot, and mouth disease (HFMD), most of them were children of age 1–3-year-old, in Central China were hospitalized from 2011 to 2012. Among them, 422 were infected with coxsackievirus A16 (CVA16), 334 were infected with enterovirus 71 (EV71), 38 were co-infected with EV71 and CVA16, and 35 were infected with other enteroviruses. Molecular epidemiology analysis revealed that EV71 and CVA16 were detected year-round, but EV71 circulated mainly in July and CVA16 circulated predominantly in November, and incidence of HFMD was reduced in January and February and increased in March. Clinical data showed that hyperglycemia and neurologic complications were significantly higher in EV71-infected patients, while upper respiratory tract infection and C-reactive protein were significantly higher in CVA16-associated patients. 124 EV71 and 80 CVA16 strains were isolated, among them 56 and 68 EV71 strains were C4a and C4b, while 25 and 55 CVA16 strains were B1a and B1b, respectively. Similarity plots and bootscan analyses based on entire genomic sequences revealed that the three C4a sub-genotype EV71 strains were recombinant with C4b sub-genotype EV71 in 2B–2C region, and the three CVA16 strains were recombinant with EV71 in 2A–2B region. Thus, CVA16 and EV71 were the major causative agents in a large HFMD outbreak in Central China. HFMD incidence was high for children among household contact and was detected year-round, but outbreak was seasonal dependent. CVA16 B1b and EV71 C4b reemerged and caused a large epidemic in China after a quiet period of many years. Moreover, EV71 and CVA16 were co-circulated during the outbreak, which may have contributed to the genomic recombination between the pathogens. It should gain more attention as there may be an upward trend in co-circulation of the two pathogens globally and the new role recombination plays in the emergence of new enterovirus variants. PMID:24776922

Association of Tic Disorders and Enterovirus Infection: A Nationwide Population-Based Study.

PubMed

Tsai, Ching-Shu; Yang, Yao-Hsu; Huang, Kuo-You; Lee, Yena; McIntyre, Roger S; Chen, Vincent Chin-Hung

2016-04-01

There has been growing interest in the association between infectious disease and mental disorders, but an association between enterovirus (EV) infection and tic disorders has not been sufficiently explored. Herein, we aim to investigate the association between EV infection and incidence of tic disorders in a nationwide population-based sample using Taiwan's National Health Insurance Research Database. We identified individuals aged ≤18 years prior to 2005 with an inpatient diagnosis of EV infection and/or history of EV infection. Tic disorder was operationalized using International Classification of Disease, Revision 9, Clinical Modification (ICD-9-CM) codes 307.20-307.23. A total of 47,998 individuals with history of EV infection were compared to 47,998 sex-, age-, and urbanization-matched controls on incidence of tic disorders. The mean ± standard deviation follow-up period for all subjects was 9.7 ± 3.6 years; the mean latency period between initial EV infection and incident diagnosis of tic disorder diagnosis was 5.4 ± 2.8 years. EV infection was significantly associated with greater incidence of tic disorders (hazard ratio [HR] = 1.24, 95% CI: 1.07-1.45). When subgrouped on the basis of central nervous system (CNS) involvement, EV infection with CNS involvement was not significantly associated with greater incidence of tic disorders when compared to controls (HR = 1.25, 95% CI: 0.64-2.43); EV infection without CNS involvement was significantly associated greater incidence of tic disorders when compared to controls (HR = 1.24, 95% CI: 1.07-1.45). In addition, hospitalization for an EV infection did not increase the hazard for greater incidence of tic disorders (HR = 1.32, 95% CI: 1.04-1.67 with hospitalization and 1.22, 95% CI: 1.04-1.44 without hospitalization). EV infection is temporally associated with incidence of tic disorders. Our observations add to the growing body of literature implicating immune-inflammatory system in the pathoetiology of brain disorders in a subpopulation of individuals and serve as a clarion call for surveillance of symptoms suggestive of tic disorders in individuals with history of EV infection.

Extracellular vesicles in urine of women with but not without kidney stones manifest patterns similar to men: a case control study.

PubMed

Jayachandran, Muthuvel; Lugo, Ghiara; Heiling, Hillary; Miller, Virginia M; Rule, Andrew D; Lieske, John C

2015-01-01

The lifetime incidence of kidney stones is about two times greater in men compared to women. Extracellular vesicles (EVs) shed from activated cells are present in the urine and may reflect or even mediate renal physiology and/or pathology. This study was designed to standardize methodology to characterize urinary EVs by digital flow cytometry and to identify possible sex differences in EVs in persons with and without their first symptomatic kidney stones. Twenty-four-hour urine collections were obtained from persons presenting with their first kidney stone episode (n = 50 women, 60 men; age 19-76 years) and sex- and age-matched controls from the general population (n = 24 women, 36 men). Standardization: Size of EV was variable within all groups. EV positivity was verified with two fluorophores for surface phosphatidylserine and/or using two different protein markers specific for renal-specific cells. The number of phosphatidylserine- and exosome marker-positive EVs did not correlate with urine osmolality and were similar in fresh vs. frozen and between two sequential urine collections from the same individual. Sex differences: Urine from women controls contained greater (P < 0.05) numbers of EVs positive for phosphatidylserine, exosomes, inflammatory factors and adhesion molecules, and cell-specific markers from different segments of the nephron, renal pelvis, and bladder compared to control men. In contrast, urine from women with kidney stones contained significantly (P < 0.05) lower numbers of EVs derived from podocytes, parietal cells, proximal convoluted tubule, thin and thick loop of Henle, distal tubule, collecting duct, renal pelvis, and bladder compared to control women and contained similar quantities of these types of EVs in men with and without kidney stones. There were also no sex differences in EVs positive for cell adhesion (E-cadherin and inter-cellular adhesion molecule-1 [ICAM-1]) molecules. Unlike women who do not have kidney stones, EVs in urine from women with nephrolithiasis are similar to men with and without kidney stones. Thus, EVs may mediate or reflect aspects of kidney stone pathogenesis and perhaps provide clues regarding sex differences in kidney stone incidence rates.

Unusual Features of Vaccinia Virus Extracellular Virion Form Neutralization Resistance Revealed in Human Antibody Responses to the Smallpox Vaccine

PubMed Central

Benhnia, Mohammed Rafii-El-Idrissi; Maybeno, Matthew; Blum, David; Aguilar-Sino, Rowena; Matho, Michael; Meng, Xiangzhi; Head, Steven; Felgner, Philip L.; Zajonc, Dirk M.; Koriazova, Lilia; Kato, Shinichiro; Burton, Dennis R.; Xiang, Yan; Crowe, James E.; Peters, Bjoern

2013-01-01

The extracellular virion form (EV) of vaccinia virus (VACV) is essential for viral pathogenesis and is difficult to neutralize with antibodies. Why this is the case and how the smallpox vaccine overcomes this challenge remain incompletely understood. We previously showed that high concentrations of anti-B5 antibodies are insufficient to directly neutralize EV (M. R. Benhnia, et al., J. Virol. 83:1201–1215, 2009). This allowed for at least two possible interpretations: covering the EV surface is insufficient for neutralization, or there are insufficient copies of B5 to allow anti-B5 IgG to cover the whole surface of EV and another viral receptor protein remains active. We endeavored to test these possibilities, focusing on the antibody responses elicited by immunization against smallpox. We tested whether human monoclonal antibodies (MAbs) against the three major EV antigens, B5, A33, and A56, could individually or together neutralize EV. While anti-B5 or anti-A33 (but not anti-A56) MAbs of appropriate isotypes were capable of neutralizing EV in the presence of complement, a mixture of anti-B5, anti-A33, and anti-A56 MAbs was incapable of directly neutralizing EV, even at high concentrations. This remained true when neutralizing the IHD-J strain, which lacks a functional version of the fourth and final known EV surface protein, A34. These immunological data are consistent with the possibility that viral proteins may not be the active component of the EV surface for target cell binding and infectivity. We conclude that the protection afforded by the smallpox vaccine anti-EV response is predominantly mediated not by direct neutralization but by isotype-dependent effector functions, such as complement recruitment for antibodies targeting B5 and A33. PMID:23152530

Human Neural Stem Cell Extracellular Vesicles Improve Recovery in a Porcine Model of Ischemic Stroke

PubMed Central

Webb, Robin L.; Kaiser, Erin E.; Jurgielewicz, Brian J.; Spellicy, Samantha; Scoville, Shelley L.; Thompson, Tyler A.; Swetenburg, Raymond L.; Hess, David C.; West, Franklin D.

2018-01-01

Background and Purpose— Recent work from our group suggests that human neural stem cell–derived extracellular vesicle (NSC EV) treatment improves both tissue and sensorimotor function in a preclinical thromboembolic mouse model of stroke. In this study, NSC EVs were evaluated in a pig ischemic stroke model, where clinically relevant end points were used to assess recovery in a more translational large animal model. Methods— Ischemic stroke was induced by permanent middle cerebral artery occlusion (MCAO), and either NSC EV or PBS treatment was administered intravenously at 2, 14, and 24 hours post-MCAO. NSC EV effects on tissue level recovery were evaluated via magnetic resonance imaging at 1 and 84 days post-MCAO. Effects on functional recovery were also assessed through longitudinal behavior and gait analysis testing. Results— NSC EV treatment was neuroprotective and led to significant improvements at the tissue and functional levels in stroked pigs. NSC EV treatment eliminated intracranial hemorrhage in ischemic lesions in NSC EV pigs (0 of 7) versus control pigs (7 of 8). NSC EV–treated pigs exhibited a significant decrease in cerebral lesion volume and decreased brain swelling relative to control pigs 1-day post-MCAO. NSC EVs significantly reduced edema in treated pigs relative to control pigs, as assessed by improved diffusivity through apparent diffusion coefficient maps. NSC EVs preserved white matter integrity with increased corpus callosum fractional anisotropy values 84 days post-MCAO. Behavior and mobility improvements paralleled structural changes as NSC EV–treated pigs exhibited improved outcomes, including increased exploratory behavior and faster restoration of spatiotemporal gait parameters. Conclusions— This study demonstrated for the first time that in a large animal model novel NSC EVs significantly improved neural tissue preservation and functional levels post-MCAO, suggesting NSC EVs may be a paradigm changing stroke therapeutic. PMID:29650593

Changes in the pattern of plasma extracellular vesicles after severe trauma

PubMed Central

Kuravi, Sahithi J.; Yates, Clara M.; Foster, Mark; Hampson, Peter; Watson, Chris; Midwinter, Mark

2017-01-01

Background Extracellular vesicles (EV) released into the circulation after traumatic injury may influence complications. We thus evaluated the numbers of EV in plasma over 28 days after trauma and evaluated their pro-coagulant and inflammatory effects. Methods and findings 37 patients suffering trauma with an injury severity score >15 were studied along with 24 healthy controls. Plasma samples were isolated by double centrifugation (2000g 20min; 13000g 2min) from blood collected from within an hour up to 28 days after injury. Plasma EV were counted and sized using nanoparticle tracking analysis (NTA); counts and cellular origins were also determined by flow cytometry (FC) using cell-specific markers. Functional effects were tested in a procoagulant phospholipid assay and in flow-based, leukocyte adhesion assay after endothelial cells (EC) were treated with EV. We found that EV concentrations measured by NTA were significantly increased in trauma patients compared to healthy controls, and remained elevated over days. In addition, or FC showed that patients with trauma had higher numbers of EV derived from platelets (CD41+), leukocytes (CD45+) and endothelial EC (CD144+). The increases were evident throughout the 28-day follow-up. However, the FC count represented <1% of the count detected by NTA, and only 1–2% of EV identified using NTA had a diameter >400nm. The procoagulant phospholipid activity assay showed that patient plasma accelerated coagulation on day 1 and day 3 after trauma, with coagulation times correlated with EV counts. Furthermore, treatment of EC for 24 hours with plasma containing EV tended to increase the recruitment of peripheral flowing blood mononuclear cells. Conclusions EV counted by FC represent a small sub-population of the total load detected by NTA. Both methods however indicate a significant increase in plasma EV after severe traumatic injury that have pro-coagulant and pro-inflammatory effects that may influence outcomes. PMID:28837705

Ratio of platelet count/spleen diameter predicted the presence of esophageal varices in patients with schistosomiasis liver cirrhosis.

PubMed

Xu, Xiao-Dan; Xu, Chun-Fang; Dai, Jian-Jun; Qian, Jian-Qing; Pin, Xun

2016-05-01

To examine the platelet count (PC)/spleen diameter (SD) ratio in predicting the presence of esophageal varices (EV) in patients with schistosomiasis liver cirrhosis. A total of 95 consecutive patients with EV induced by schistosomiasis liver cirrhosis were enrolled in this trial. A total of 141 schistosomiasis liver cirrhosis patients without EV were enrolled as controls. All patients were diagnosed by endoscopy. Demographic, laboratory, and Doppler ultrasound parameters were collected and analyzed. Binary logistic regression analysis was carried out to identify independent risk factors associated with EV occurrence. Receiver operating curves were generated to obtain the PC/SD ratio cutoff values for the optimal sensitivity and specificity with respect to EV. The accuracy was increased in diagnosing for EV using the ratio of PC/SD compared with the SD alone [area under the curve: 0.891 95% confidence interval (CI): 0.844-0.928 vs. 0.764 95% CI: 0.705-0.817; P<0.01]. The optimal cutoff value was 1004, with a 77.1% (95% CI: 67.9-84.8%) positive-predictive value and an 89.3% (95% CI: 82.7-94.0%) negative-predictive value. Using a cutoff of 1004, it was determined that 117/141 (83.0%) patients without EV could avoid undergoing unnecessary endoscopy, whereas 14/95 (14.7%) patients with EV would be misdiagnosed. In contrast, when the ratio was set at 909, the positive-predictive and negative-predictive values were 79.5% (95% CI: 69.5-87.4%) and 83.1% (95% CI: 76.1-88.8%), respectively. A ratio of 909 would accurately predict the absence of EV in 123/141 (87.2%) patients; however, 24/95 (25.3%) patients with EV would miss the necessary screening endoscopy. The ratio of PC/SD was a useful marker in predicting the presence of EV in patients with schistosomiasis liver cirrhosis.

Non-Singular Modeling of Rigid Manipulators.

DTIC Science & Technology

1986-12-01

SOJQCE 09 FONDING NoMBERS PROG-RAM PRO,ECT -ASK v1 W K .. Ni ELEMEFNT NO NO NO I ACCES;C)1 NO N ON:-zlVTLvIP ’"fPFILINC OFPTRCI1) TAIT rT( Q ; E RSO...AUTHORIS r eamrred. khavvq’_ .33 " t0 QFOQ 13t)’ E COIRED TO4 DATE OF REPORT lYcir MonfhP Day) S P.%t~r (,),%T S I ; . - A T 0 N& (C)SAI. CODES I i...tr he po--,nt of singul -ar-ity. One solution is to s:e:-, a --,mmamu: t me at r oosit:ocn or to avoid t>i:c’ . alt :- e ::.: n t-.te r *solution is to

Proteomic analysis reveals different composition of extracellular vesicles released by two Trypanosoma cruzi strains associated with their distinct interaction with host cells.

PubMed

Ribeiro, Kleber Silva; Vasconcellos, Camilla Ioshida; Soares, Rodrigo Pedro; Mendes, Maria Tays; Ellis, Cameron C; Aguilera-Flores, Marcela; de Almeida, Igor Correia; Schenkman, Sergio; Iwai, Leo Kei; Torrecilhas, Ana Claudia

2018-01-01

Trypanosoma cruzi , the aetiologic agent of Chagas disease, releases vesicles containing a wide range of surface molecules known to affect the host immunological responses and the cellular infectivity. Here, we compared the secretome of two distinct strains (Y and YuYu) of T. cruzi , which were previously shown to differentially modulate host innate and acquired immune responses. Tissue culture-derived trypomastigotes of both strains secreted extracellular vesicles (EVs), as demonstrated by electron scanning microscopy. EVs were purified by exclusion chromatography or ultracentrifugation and quantitated using nanoparticle tracking analysis. Trypomastigotes from YuYu strain released higher number of EVs than those from Y strain, enriched with virulence factors trans -sialidase (TS) and cruzipain. Proteomic analysis confirmed the increased abundance of proteins coded by the TS gene family, mucin-like glycoproteins, and some typical exosomal proteins in the YuYu strain, which also showed considerable differences between purified EVs and vesicle-free fraction as compared to the Y strain. To evaluate whether such differences were related to parasite infectivity, J774 macrophages and LLC-MK2 kidney cells were preincubated with purified EVs from both strains and then infected with Y strain trypomastigotes. EVs released by YuYu strain caused a lower infection but higher intracellular proliferation in J774 macrophages than EVs from Y strain. In contrast, YuYu strain-derived EVs caused higher infection of LLC-MK2 cells than Y strain-derived EVs. In conclusion, quantitative and qualitative differences in EVs and secreted proteins from different T. cruzi strains may correlate with infectivity/virulence during the host-parasite interaction.

Proteomic analysis reveals different composition of extracellular vesicles released by two Trypanosoma cruzi strains associated with their distinct interaction with host cells

PubMed Central

Ribeiro, Kleber Silva; Vasconcellos, Camilla Ioshida; Soares, Rodrigo Pedro; Ellis, Cameron C.; Aguilera-Flores, Marcela; de Almeida, Igor Correia

2018-01-01

ABSTRACT Trypanosoma cruzi, the aetiologic agent of Chagas disease, releases vesicles containing a wide range of surface molecules known to affect the host immunological responses and the cellular infectivity. Here, we compared the secretome of two distinct strains (Y and YuYu) of T. cruzi, which were previously shown to differentially modulate host innate and acquired immune responses. Tissue culture-derived trypomastigotes of both strains secreted extracellular vesicles (EVs), as demonstrated by electron scanning microscopy. EVs were purified by exclusion chromatography or ultracentrifugation and quantitated using nanoparticle tracking analysis. Trypomastigotes from YuYu strain released higher number of EVs than those from Y strain, enriched with virulence factors trans-sialidase (TS) and cruzipain. Proteomic analysis confirmed the increased abundance of proteins coded by the TS gene family, mucin-like glycoproteins, and some typical exosomal proteins in the YuYu strain, which also showed considerable differences between purified EVs and vesicle-free fraction as compared to the Y strain. To evaluate whether such differences were related to parasite infectivity, J774 macrophages and LLC-MK2 kidney cells were preincubated with purified EVs from both strains and then infected with Y strain trypomastigotes. EVs released by YuYu strain caused a lower infection but higher intracellular proliferation in J774 macrophages than EVs from Y strain. In contrast, YuYu strain-derived EVs caused higher infection of LLC-MK2 cells than Y strain-derived EVs. In conclusion, quantitative and qualitative differences in EVs and secreted proteins from different T. cruzi strains may correlate with infectivity/virulence during the host–parasite interaction. PMID:29696081

Extracellular vesicles released from mesenchymal stromal cells stimulate bone growth in osteogenesis imperfecta.

PubMed

Otsuru, Satoru; Desbourdes, Laura; Guess, Adam J; Hofmann, Ted J; Relation, Theresa; Kaito, Takashi; Dominici, Massimo; Iwamoto, Masahiro; Horwitz, Edwin M

2018-01-01

Systemic infusion of mesenchymal stromal cells (MSCs) has been shown to induce acute acceleration of growth velocity in children with osteogenesis imperfecta (OI) despite minimal engraftment of infused MSCs in bones. Using an animal model of OI we have previously shown that MSC infusion stimulates chondrocyte proliferation in the growth plate and that this enhanced proliferation is also observed with infusion of MSC conditioned medium in lieu of MSCs, suggesting that bone growth is due to trophic effects of MSCs. Here we sought to identify the trophic factor secreted by MSCs that mediates this therapeutic activity. To examine whether extracellular vesicles (EVs) released from MSCs have therapeutic activity, EVs were isolated from MSC conditioned medium by ultracentrifugation. To further characterize the trophic factor, RNA or microRNA (miRNA) within EVs was depleted by either ribonuclease (RNase) treatment or suppressing miRNA biogenesis in MSCs. The functional activity of these modified EVs was evaluated using an in vitro chondrocyte proliferation assay. Finally, bone growth was evaluated in an animal model of OI treated with EVs. We found that infusion of MSC-derived EVs stimulated chondrocyte proliferation in the growth plate, resulting in improved bone growth in a mouse model of OI. However, infusion of neither RNase-treated EVs nor miRNA-depleted EVs enhanced chondrocyte proliferation. MSCs exert therapeutic effects in OI by secreting EVs containing miRNA, and EV therapy has the potential to become a novel cell-free therapy for OI that will overcome some of the current limitations in MSC therapy. Copyright © 2017 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

Defect annealing in electron-irradiated boron-doped silicon

NASA Astrophysics Data System (ADS)

Awadelkarim, O. O.; Chen, W. M.; Weman, H.; Monemar, B.

1990-01-01

Defects introduced by room-temperature electron irradiation and subsequent annealing in boron-doped silicon are studied by means of deep-level transient spectroscopy, photoluminescence, and optical detection of magnetic resonance (ODMR) techniques. ODMR reveals a thermally induced paramagnetic (S=(1/2) defect center that is produced following annealing at 400 °C. The center possesses a C3v point-group symmetry with the trigonal axis along <111>. Detailed analysis of the ODMR line shapes indicates the involvement of a silicon atom in the defect center. It appears from the results that boron is either another possible defect component or an essential catalyst for the defect formation. The occurrence of the ODMR signal together with a luminescence band peaking at 0.80 eV is independent of oxygen or carbon contents in the samples. The band does not belong to the center observed by ODMR; however, a decrease in its intensity, under resonance conditions in the ODMR center, is explained in terms of carrier recombination, capture, or energy-transfer processes involving this center. Annealing studies on a metastable hole trap observed at Ev+0.12 eV (Ev being the top of the valence band) establish the trap assignment to a carbon-interstitial-carbon-substitutional pair. The introduction of postannealing traps observed at Ev+0.07 eV, Ev+0.45 eV, and Ec-0.59 eV (Ec being the conduction-band edge) is found to be boron dependent. Isothermal formation of the centers responsible for these traps are observed, and none of the traps appears to be related to either the center observed by ODMR or the 0.80-eV band.

The compatibility of inactivated-Enterovirus 71 vaccination with Coxsackievirus A16 and Poliovirus immunizations in humans and animals

PubMed Central

Mao, Qunying; Wang, Yiping; Shao, Jie; Ying, Zhifang; Gao, Fan; Yao, Xin; Li, Changgui; Ye, Qiang; Xu, Miao; Li, Rongcheng; Zhu, Fengcai; Liang, Zhenglun

2015-01-01

Enterovirus 71 (EV71) is the key pathogen for Hand, Foot, and Mouth Disease (HFMD) and can result in severe neurological complications and death among young children. Three inactivated-EV71 vaccines have gone through phase III clinical trials and have demonstrated good safety and efficacy. These vaccines will benefit young children under the threat of severe HFMD. However, the potential immunization-related compatibility for different enterovirus vaccines remains unclear, making it hard to include the EV71 vaccine in Expanded Program on Immunization (EPI). Here, we measured the neutralizing antibodies (NTAbs) against EV71, Coxsackievirus A16 (CA16) and Poliovirus from infants enrolled in those EV71 vaccine clinical trials. The results indicated that the levels of NTAb GMTs for EV71 increased significantly in all 3 vaccine groups (high, middle and low dosages, respectively) post-vaccination. Seroconversion ratios and Geometric mean fold increase were significantly higher in the vaccine groups (≥7/9 and 8.9～228.1) than in the placebo group (≤1/10 and 0.8～1.7, P < 0.05). But no similar NTAb response trends were found in CA16 and 3 types of Poliovirus. The decrease of 3 types of Poliovirus NTAb GMTs and an increase of CA16 GMTs post-EV71-vaccination were found in vaccine and placebo groups. Further animal study on CA16 and poliovirus vaccine co-immunization or pre-immunization with EV71 vaccine in mice indicated that there was no NTAb cross-activity between EV71 and CA16/Poliovirus. Our research showed that inactivated-EV71 vaccine has good specific-neutralizing capacity and can be included in EPI. PMID:25715318

Ab Initio Study of Aluminium Impurity and Interstitial-Substitutional Complexes in Ge Using a Hybrid Functional (HSE)

NASA Astrophysics Data System (ADS)

Igumbor, E.; Mapasha, R. E.; Meyer, W. E.

2017-07-01

The results of an ab initio modelling of aluminium substitutional impurity ({\\hbox {Al}}_Ge), aluminium interstitial in Ge [{\\hbox {I}}_Al for the tetrahedral (T) and hexagonal (H) configurations] and aluminium interstitial-substitutional pairs in Ge ({\\hbox {I}}_Al{\\hbox {Al}}_Ge) are presented. For all calculations, the hybrid functional of Heyd, Scuseria, and Ernzerhof in the framework of density functional theory was used. Defects formation energies, charge state transition levels and minimum energy configurations of the {\\hbox {Al}}_Ge, {\\hbox {I}}_Al and {\\hbox {I}}_Al{\\hbox {Al}}_Ge were obtained for -2, -1, 0, +1 and +2 charge states. The calculated formation energy shows that for the neutral charge state, the {\\hbox {I}}_Al is energetically more favourable in the T than the H configuration. The {\\hbox {I}}_Al{\\hbox {Al}}_Ge forms with formation energies of -2.37 eV and -2.32 eV, when the interstitial atom is at the T and H sites, respectively. The {\\hbox {I}}_Al{\\hbox {Al}}_Ge is energetically more favourable when the interstitial atom is at the T site with a binding energy of 0.8 eV. The {\\hbox {I}}_Al in the T configuration, induced a deep donor (+2/+1) level at EV+0.23 eV and the {\\hbox {Al}}_Ge induced a single acceptor level (0/-1) at EV+0.14 eV in the band gap of Ge. The {\\hbox {I}}_Al{\\hbox {Al}}_Ge induced double-donor levels are at E_V+0.06 and E_V+0.12 eV, when the interstitial atom is at the T and H sites, respectively. The {\\hbox {I}}_Al and {\\hbox {I}}_Al{\\hbox {Al}}_Ge exhibit properties of charge state-controlled metastability.

Optical properties of tetragonal and nanoscale BiFeO3

NASA Astrophysics Data System (ADS)

Chen, P.; Xu, X. S.; Musfeldt, J. L.; Santulli, A. C.; Koenigsmann, C.; Wong, S. S.; Podraza, N. J.; Melville, A.; Vlahos, E.; Gopalan, V.; Schlom, D. G.; Ramesh, R.

2010-03-01

We measured the optical properties of tetragonal thin film and nanoscale rhombohedral BiFeO3 in the range from near infrared to the near ultraviolet. The absorption spectrum in the tetragonal film is overall blue-shifted compared with that of the rhombohedral BiFeO3 film. It shows an absorption onset near 2.25 eV, a direct 3.1 eV band gap, and charge transfer excitations that are ˜0.4 eV higher than those of the rhombohedral counterpart. In the nanoparticles, the band gap decreases from 2.7 eV to ˜2.3 eV, and the well-known 3.2 and 4.5 eV charge transfer excitations split into multiplets. We discuss these results in terms of structural strain, surface strain, and local symmetry breaking.

INTERACTION OF INTERSTITIAL CLUSTERS WITH RHENIUM, OSMIUM, AND TANTALUM IN TUNGSTEN

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

Setyawan, Wahyu; Nandipati, Giridhar; Kurtz, Richard J.

2016-09-01

In the previous semi annual report, we explored the stability of interstitial clusters in W up to size seven. In this report, we study the binding of those clusters to Re, Os, and Ta atoms. For each cluster size, the three most stable configurations are considered to average the binding property. The average binding energy to a Re decreases from 0.79 eV for a size-1 cluster (a [111] dumbbell) to 0.65 eV for a size-7 cluster. For Os, the binding decreases from 1.61 eV for a [111] dumbbell to 1.34 eV for a size-7 cluster. Tantalum is repulsive to interstitialmore » clusters with binding energy ranges from -0.61 eV for a [111] dumbbell to -0.5 eV for a size-7 cluster.« less

Electric Vehicles Charging Scheduling Strategy Considering the Uncertainty of Photovoltaic Output

NASA Astrophysics Data System (ADS)

Wei, Xiangxiang; Su, Su; Yue, Yunli; Wang, Wei; He, Luobin; Li, Hao; Ota, Yutaka

2017-05-01

The rapid development of electric vehicles and distributed generation bring new challenges to security and economic operation of the power system, so the collaborative research of the EVs and the distributed generation have important significance in distribution network. Under this background, an EVs charging scheduling strategy considering the uncertainty of photovoltaic(PV) output is proposed. The characteristics of EVs charging are analysed first. A PV output prediction method is proposed with a PV database then. On this basis, an EVs charging scheduling strategy is proposed with the goal to satisfy EVs users’ charging willingness and decrease the power loss in distribution network. The case study proves that the proposed PV output prediction method can predict the PV output accurately and the EVs charging scheduling strategy can reduce the power loss and stabilize the fluctuation of the load in distributed network.

Extracellular vesicle-mediated transfer of donor genomic DNA to recipient cells is a novel mechanism for genetic influence between cells

PubMed Central

Cai, Jin; Han, Yu; Ren, Hongmei; Chen, Caiyu; He, Duofen; Zhou, Lin; Eisner, Gilbert M.; Asico, Laureano D.; Jose, Pedro A.; Zeng, Chunyu

2013-01-01

Extracellular vesicles (EVs) carry signals within or at their limiting membranes, providing a mechanism by which cells can exchange more complex information than what was previously thought. In addition to mRNAs and microRNAs, there are DNA fragments in EVs. Solexa sequencing indicated the presence of at least 16434 genomic DNA (gDNA) fragments in the EVs from human plasma. Immunofluorescence study showed direct evidence that acridine orange-stained EV DNAs could be transferred into the cells and localize to and inside the nuclear membrane. However, whether the transferred EV DNAs are functional or not is not clear. We found that EV gDNAs could be homologously or heterologously transferred from donor cells to recipient cells, and increase gDNA-coding mRNA, protein expression, and function (e.g. AT1 receptor). An endogenous promoter of the AT1 receptor, NF-κB, could be recruited to the transferred DNAs in the nucleus, and increase the transcription of AT1 receptor in the recipient cells. Moreover, the transferred EV gDNAs have pathophysiological significance. BCR/ABL hybrid gene, involved in the pathogenesis of chronic myeloid leukemia, could be transferred from K562 EVs to HEK293 cells or neutrophils. Our present study shows that the gDNAs transferred from EVs to cells have physiological significance, not only to increase the gDNA-coding mRNA and protein levels, but also to influence function in recipient cells. PMID:23580760

Extracellular vesicles and their synthetic analogues in aging and age-associated brain diseases.

PubMed

Smith, J A; Leonardi, T; Huang, B; Iraci, N; Vega, B; Pluchino, S

2015-04-01

Multicellular organisms rely upon diverse and complex intercellular communications networks for a myriad of physiological processes. Disruption of these processes is implicated in the onset and propagation of disease and disorder, including the mechanisms of senescence at both cellular and organismal levels. In recent years, secreted extracellular vesicles (EVs) have been identified as a particularly novel vector by which cell-to-cell communications are enacted. EVs actively and specifically traffic bioactive proteins, nucleic acids, and metabolites between cells at local and systemic levels, modulating cellular responses in a bidirectional manner under both homeostatic and pathological conditions. EVs are being implicated not only in the generic aging process, but also as vehicles of pathology in a number of age-related diseases, including cancer and neurodegenerative and disease. Thus, circulating EVs-or specific EV cargoes-are being utilised as putative biomarkers of disease. On the other hand, EVs, as targeted intercellular shuttles of multipotent bioactive payloads, have demonstrated promising therapeutic properties, which can potentially be modulated and enhanced through cellular engineering. Furthermore, there is considerable interest in employing nanomedicinal approaches to mimic the putative therapeutic properties of EVs by employing synthetic analogues for targeted drug delivery. Herein we describe what is known about the origin and nature of EVs and subsequently review their putative roles in biology and medicine (including the use of synthetic EV analogues), with a particular focus on their role in aging and age-related brain diseases.

Inhibition of oncogenic epidermal growth factor receptor kinase triggers release of exosome-like extracellular vesicles and impacts their phosphoprotein and DNA content.

PubMed

Montermini, Laura; Meehan, Brian; Garnier, Delphine; Lee, Wan Jin; Lee, Tae Hoon; Guha, Abhijit; Al-Nedawi, Khalid; Rak, Janusz

2015-10-02

Cancer cells emit extracellular vesicles (EVs) containing unique molecular signatures. Here, we report that the oncogenic EGF receptor (EGFR) and its inhibitors reprogram phosphoproteomes and cargo of tumor cell-derived EVs. Thus, phosphorylated EGFR (P-EGFR) and several other receptor tyrosine kinases can be detected in EVs purified from plasma of tumor-bearing mice and from conditioned media of cultured cancer cells. Treatment of EGFR-driven tumor cells with second generation EGFR kinase inhibitors (EKIs), including CI-1033 and PF-00299804 but not with anti-EGFR antibody (Cetuximab) or etoposide, triggers a burst in emission of exosome-like EVs containing EGFR, P-EGFR, and genomic DNA (exo-gDNA). The EV release can be attenuated by treatment with inhibitors of exosome biogenesis (GW4869) and caspase pathways (ZVAD). The content of P-EGFR isoforms (Tyr-845, Tyr-1068, and Tyr-1173), ERK, and AKT varies between cells and their corresponding EVs and as a function of EKI treatment. Immunocapture experiments reveal the presence of EGFR and exo-gDNA within the same EV population following EKI treatment. These findings suggest that targeted agents may induce cancer cells to change the EV emission profiles reflective of drug-related therapeutic stress. We suggest that EV-based assays may serve as companion diagnostics for targeted anticancer agents. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

Fibronectin on extracellular vesicles from microvascular endothelial cells is involved in the vesicle uptake into oligodendrocyte precursor cells

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

Osawa, Sho; Department of Molecular and Cellular Neurobiology, Gunma University Graduate School of Medicine, 3-39-22 Showa-machi, Maebashi, Gunma 371-8511; Kurachi, Masashi

We previously reported transplantation of brain microvascular endothelial cells (MVECs) into cerebral white matter infarction model improved the animal's behavioral outcome by increasing the number of oligodendrocyte precursor cells (OPCs). We also revealed extracellular vesicles (EVs) derived from MVECs promoted survival and proliferation of OPCs in vitro. In this study, we investigated the mechanism how EVs derived from MVECs contribute to OPC survival and proliferation. Protein mass spectrometry and enzyme-linked immunosorbent assay revealed fibronectin was abundant on the surface of EVs from MVECs. As fibronectin has been reported to promote OPC survival and proliferation via integrin signaling pathway, we blocked themore » binding between fibronectin and integrins using RGD sequence mimics. Blocking the binding, however, did not attenuate the survival and proliferation promoting effect of EVs on OPCs. Flow cytometric and imaging analyses revealed fibronectin on EVs mediates their internalization into OPCs by its binding to heparan sulfate proteoglycan on OPCs. OPC survival and proliferation promoted by EVs were attenuated by blocking the internalization of EVs into OPCs. These lines of evidence suggest that fibronectin on EVs mediates their internalization into OPCs, and the cargo of EVs promotes survival and proliferation of OPCs, independent of integrin signaling pathway. - Highlights: • Fibronectin exists on the surface of extracellular vesicles from endothelial cells. • Integrin signaling is not involved in effects of extracellular vesicles on OPCs. • Fibronectin on the surface of extracellular vesicles mediates their uptake into OPCs.« less

Formulation and immunological evaluation of a trivalent vaccine comprising emulsified submicron particles and inactivated virions of H5N1/EV71/JEV

PubMed Central

Lin, Chih-Wei; Chang, Ching-Yun; Chen, Wei-Lin; Lin, Shih-Chang; Liao, Chien-Chun; Chang, Jui-Yuan; Liu, Chia-Chyi; Hu, Alan Yung-Chih; Lu, Tsung-Chun; Chou, Ai-Hsiang; Wu, Suh-Chin; Chong, Pele; Huang, Ming-Hsi

2013-01-01

Combination vaccines can reduce the number of injections and simplify the immunization schedule required to prevent different diseases. Here we assessed the immunogenicity in a mouse model of a vaccine composition comprising inactivated influenza viruses (H5N1/H1N1), enterovirus 71 (EV71), and/or Japanese encephalitis virus (JEV) and investigated whether the vaccine formulations can overcome the immunologic interference between the individual vaccine components. We demonstrated that the antigenic competition happens between H5N1/H1N1 or H5N1/EV71 inactivated virions when the vaccine combinations either formulated with Alum suspensions or without adjuvant. In the presence of PELC emulsified particles, EV71-specific immune responses before and after incorporating H5N1 virus into EV71 vaccine were detected of no significant difference; in addition, H5N1- and EV71-specific immune responses were found at the same level when H5N1/EV71/JEV consolidating into combination vaccine. Emulsified vaccine formulation was represented as a potential tool that is found to reduce the number of injections required to prevent multiple infectious strains causing the same disease (H5N1/H1N1) and/or that protect against different diseases (H5N1/EV71). Combination vaccines can also include a third component to protect against H5N1/EV71/JEV at the same time. PMID:23838466

Intraspecific variation of Centruroides sculpturatus scorpion venom from two regions of Arizona.

PubMed

Carcamo-Noriega, Edson Norberto; Olamendi-Portugal, Timoteo; Restano-Cassulini, Rita; Rowe, Ashlee; Uribe-Romero, Selene Jocelyn; Becerril, Baltazar; Possani, Lourival Domingos

2018-01-15

This study investigated geographic variability in the venom of Centruroides sculpturatus scorpions from different biotopes. Venom from scorpions collected from two different regions in Arizona; Santa Rita Foothills (SR) and Yarnell (Yar) were analyzed. We found differences between venoms, mainly in the two most abundant peptides; SR (CsEv2e and CsEv1f) and Yar (CsEv2 and CsEv1c) identified as natural variants of CsEv1 and CsEv2. Sequence analyses of these peptides revealed conservative amino acid changes between variants, which may underlie biological activity against arthropods. A third peptide (CsEv6) was highly abundant in the Yar venom compared to the SR venom. CsEv6 is a 67 amino acid peptide with 8 cysteines. CsEv6 did not exhibit toxicity to the three animal models tested. However, both venoms shared similarities in peptides that are predicted to deter predators. For example, both venoms expressed CsEI (lethal to chick) in similar abundance, while CsEd and CsEM1a (toxic to mammals) displayed only moderate variation in their abundance. Electrophysiological evaluation of CsEd and CsEM1a showed that both toxins act on the human sodium-channel subtype 1.6 (hNav 1.6). Complete sequencing revealed that both toxins are structurally similar to beta-toxins isolated from different Centruroides species that also target hNav 1.6. Copyright © 2017 Elsevier Inc. All rights reserved.

Detection and proteomic characterization of extracellular vesicles in human pancreatic juice.

PubMed

Osteikoetxea, Xabier; Benke, Márton; Rodriguez, Marta; Pálóczi, Krisztina; Sódar, Barbara W; Szvicsek, Zsuzsanna; Szabó-Taylor, Katalin; Vukman, Krisztina V; Kittel, Ágnes; Wiener, Zoltán; Vékey, Károly; Harsányi, László; Szűcs, Ákos; Turiák, Lilla; Buzás, Edit I

2018-04-30

The prognosis of patients with pancreatic cancer has remained virtually unchanged with a high mortality rate compared to other types of cancers. An earlier detection would provide a time window of opportunity for treatment and prevention of deaths. In the present study we investigated extracellular vesicle (EV)-associated potential biomarkers for pancreatic cancer by directly assessing EV size-based subpopulations in pancreatic juice samples of patients with chronic pancreatitis or pancreatic cancer. In addition, we also studied blood plasma and pancreatic cancer cell line-derived EVs. Comparative proteomic analysis was performed of 102 EV preparations from human pancreatic juices, blood, and pancreatic cancer cell lines Capan-1 and MIA PaCa-2. EV preparations were also characterized by electron microscopy, tunable resistive pulse sensing, and flow cytometry. Here we describe the presence of EVs in human pancreatic juice samples. Pancreatic juice EV-associated proteins that we identified as possible candidate markers for pancreatic cancer included mucins, such as MUC1, MUC4, MUC5AC, MUC6 and MUC16, CFTR, and MDR1 proteins. These candidate biomarkers could also be detected by flow cytometry in EVs found in pancreatic juice and those secreted by pancreatic cancer cell lines. Together our data show that detection and characterization of EVs directly in pancreatic juice is feasible and may prove to be a valuable source of potential biomarkers of pancreatic cancer. Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

Effects of Acute Aerobic Exercise on Rats Serum Extracellular Vesicles Diameter, Concentration and Small RNAs Content.

PubMed

Oliveira, Getúlio P; Porto, William F; Palu, Cintia C; Pereira, Lydyane M; Petriz, Bernardo; Almeida, Jeeser A; Viana, Juliane; Filho, Nezio N A; Franco, Octavio L; Pereira, Rinaldo W

2018-01-01

Physical exercise stimulates organs, mainly the skeletal muscle, to release a broad range of molecules, recently dubbed exerkines. Among them, RNAs, such as miRNAs, piRNAs, and tRNAs loaded in extracellular vesicles (EVs) have the potential to play a significant role in the way muscle and other organs communicate to translate exercise into health. Low, moderate and high intensity treadmill protocols were applied to rat groups, aiming to investigate the impact of exercise on serum EVs and their associated small RNA molecules. Transmission electron microscopy, resistive pulse sensing, and western blotting were used to investigate EVs morphology, size distribution, concentration and EVs marker proteins. Small RNA libraries from EVs RNA were sequenced. Exercise did not change EVs size, while increased EVs concentration. Twelve miRNAs were found differentially expressed after exercise: rno-miR-128-3p, 103-3p, 330-5p, 148a-3p, 191a-5p, 10b-5p, 93-5p, 25-3p, 142-5p, 3068-3p, 142-3p, and 410-3p. No piRNA was found differentially expressed, and one tRNA, trna8336, was found down-regulated after exercise. The differentially expressed miRNAs were predicted to target genes involved in the MAPK pathway. A single bout of exercise impacts EVs and their small RNA load, reinforcing the need for a more detailed investigation into EVs and their load as mediators of health-promoting exercise.

Thermal stability of deep level defects induced by high energy proton irradiation in n-type GaN

NASA Astrophysics Data System (ADS)

Zhang, Z.; Farzana, E.; Sun, W. Y.; Chen, J.; Zhang, E. X.; Fleetwood, D. M.; Schrimpf, R. D.; McSkimming, B.; Kyle, E. C. H.; Speck, J. S.; Arehart, A. R.; Ringel, S. A.

2015-10-01

The impact of annealing of proton irradiation-induced defects in n-type GaN devices has been systematically investigated using deep level transient and optical spectroscopies. Moderate temperature annealing (>200-250 °C) causes significant reduction in the concentration of nearly all irradiation-induced traps. While the decreased concentration of previously identified N and Ga vacancy related levels at EC - 0.13 eV, 0.16 eV, and 2.50 eV generally followed a first-order reaction model with activation energies matching theoretical values for NI and VGa diffusion, irradiation-induced traps at EC - 0.72 eV, 1.25 eV, and 3.28 eV all decrease in concentration in a gradual manner, suggesting a more complex reduction mechanism. Slight increases in concentration are observed for the N-vacancy related levels at EC - 0.20 eV and 0.25 eV, which may be due to the reconfiguration of other N-vacancy related defects. Finally, the observed reduction in concentrations of the states at EC - 1.25 and EC - 3.28 eV as a function of annealing temperature closely tracks the detailed recovery behavior of the background carrier concentration as a function of annealing temperature. As a result, it is suggested that these two levels are likely to be responsible for the underlying carrier compensation effect that causes the observation of carrier removal in proton-irradiated n-GaN.

The Current Status of the Disease Caused by Enterovirus 71 Infections: Epidemiology, Pathogenesis, Molecular Epidemiology, and Vaccine Development.

PubMed

Chang, Ping-Chin; Chen, Shou-Chien; Chen, Kow-Tong

2016-09-09

Enterovirus 71 (EV71) infections have a major public health impact in the Asia-Pacific region. We reviewed the epidemiology, pathogenesis, and molecular epidemiology of EV71 infection as well as EV71 vaccine development. Previous studies were found using the search terms "enterovirus 71" and "epidemiology" or "pathogenesis" or "molecular epidemiology" or "vaccine" in Medline and PubMed. Articles that were not published in the English language, manuscripts without an abstract, and opinion articles were excluded from the review. The reported epidemiology of cases caused by EV71 infection varied from country to country; seasonal variations in incidence were observed. Most cases of EV71 infection that resulted in hospitalization for complications occurred in children less than five years old. The brainstem was the most likely major target of EV71 infection. The emergence of the EV71 epidemic in the Asia-Pacific region has been associated with the circulation of different genetic lineages (genotypes B3, B4, C1, C2, and C4) that appear to be undergoing rapid evolutionary changes. The relationship between the gene structure of the EV71 virus and the factors that ensure its survival, circulation, and evasion of immunity is still unknown. EV71 infection has emerged as an important global public health problem. Vaccine development, including the development of inactivated whole-virus live attenuated, subviral particles, and DNA vaccines, has been progressing.

Fair Energy Scheduling for Vehicle-to-Grid Networks Using Adaptive Dynamic Programming.

PubMed

Xie, Shengli; Zhong, Weifeng; Xie, Kan; Yu, Rong; Zhang, Yan

2016-08-01

Research on the smart grid is being given enormous supports worldwide due to its great significance in solving environmental and energy crises. Electric vehicles (EVs), which are powered by clean energy, are adopted increasingly year by year. It is predictable that the huge charge load caused by high EV penetration will have a considerable impact on the reliability of the smart grid. Therefore, fair energy scheduling for EV charge and discharge is proposed in this paper. By using the vehicle-to-grid technology, the scheduler controls the electricity loads of EVs considering fairness in the residential distribution network. We propose contribution-based fairness, in which EVs with high contributions have high priorities to obtain charge energy. The contribution value is defined by both the charge/discharge energy and the timing of the action. EVs can achieve higher contribution values when discharging during the load peak hours. However, charging during this time will decrease the contribution values seriously. We formulate the fair energy scheduling problem as an infinite-horizon Markov decision process. The methodology of adaptive dynamic programming is employed to maximize the long-term fairness by processing online network training. The numerical results illustrate that the proposed EV energy scheduling is able to mitigate and flatten the peak load in the distribution network. Furthermore, contribution-based fairness achieves a fast recovery of EV batteries that have deeply discharged and guarantee fairness in the full charge time of all EVs.

Molecular and epidemiological study of enterovirus D68 in Taiwan.

PubMed

Huang, Yuan-Pin; Lin, Tsuey-Li; Lin, Ting-Han; Wu, Ho-Sheng

2017-08-01

As an immunofluorescence assay for enterovirus D68 (EV-D68) is not available in the enteroviruses surveillance network in Taiwan, EV-D68 may be the actual pathogen of untypeable enterovirus-suspected isolates. The untypeable isolates collected from 2007 through 2014 were identified by nucleic acid amplification-based methods and sequencing of the VP1 region to analyze the phylogeny and epidemiology of EV-D68 in Taiwan. Twenty-nine EV-D68 isolates were sequenced, including 15 Cluster 3 and 14 Cluster 1 viruses. Approximately 41% of the patients were children under 5 years of age and their infections peaked in August. The ratio of male to female patients was 1.5 and 3.67 for Cluster 3 and Cluster 1, respectively. Fever and respiratory symptoms were commonly reported in EV-D68-infected patients. The results of phylogenetic analyses showed that EV-D68 isolates between 2007 and 2014 belonged to different clusters and existed for years, indicating that endemic circulation of EV-D68 existed in Taiwan. This study showed that EV-D68 has been endemic in Taiwan for some years despite a small number of positive cases. The continuous monitoring and efforts towards the improvement of diagnostic techniques are required to complete the surveillance system. This study provided the genetic and epidemiological information which could contribute to understanding the etiology and epidemiology of EV-D68. Copyright © 2015. Published by Elsevier B.V.

The Current Status of the Disease Caused by Enterovirus 71 Infections: Epidemiology, Pathogenesis, Molecular Epidemiology, and Vaccine Development

PubMed Central

Chang, Ping-Chin; Chen, Shou-Chien; Chen, Kow-Tong

2016-01-01

Enterovirus 71 (EV71) infections have a major public health impact in the Asia-Pacific region. We reviewed the epidemiology, pathogenesis, and molecular epidemiology of EV71 infection as well as EV71 vaccine development. Previous studies were found using the search terms “enterovirus 71” and “epidemiology” or “pathogenesis” or “molecular epidemiology” or “vaccine” in Medline and PubMed. Articles that were not published in the English language, manuscripts without an abstract, and opinion articles were excluded from the review. The reported epidemiology of cases caused by EV71 infection varied from country to country; seasonal variations in incidence were observed. Most cases of EV71 infection that resulted in hospitalization for complications occurred in children less than five years old. The brainstem was the most likely major target of EV71 infection. The emergence of the EV71 epidemic in the Asia-Pacific region has been associated with the circulation of different genetic lineages (genotypes B3, B4, C1, C2, and C4) that appear to be undergoing rapid evolutionary changes. The relationship between the gene structure of the EV71 virus and the factors that ensure its survival, circulation, and evasion of immunity is still unknown. EV71 infection has emerged as an important global public health problem. Vaccine development, including the development of inactivated whole-virus live attenuated, subviral particles, and DNA vaccines, has been progressing. PMID:27618078

Evaluation and Validation (E&V) Team Public Report. Volume 1.

DTIC Science & Technology

1984-11-30

Components. .......... K-109 *Table K-6. Possible Approaches to Integrating Methodology Components Across Life Cycle Phases. .........K-109 SECTION I...provide a detailed and organized approach to the development of technology which will be used as a basis for the E&V of APSEs. The E&V Plan which is...A-4 1.2 Background ....... .................. A-6 2. SCOPE ...... ....................... A-8 3. E&V TECHNICAL APPROACH

Electron-impact coherence parameters for 41 P 1 excitation of zinc

NASA Astrophysics Data System (ADS)

Piwiński, Mariusz; Kłosowski, Łukasz; Chwirot, Stanisław; Fursa, Dmitry V.; Bray, Igor; Das, Tapasi; Srivastava, Rajesh

2018-04-01

We present electron-impact coherence parameters (EICP) for electron-impact excitation of 41 P 1 state of zinc atoms for collision energies 40 eV and 60 eV. The experimental results are presented together with convergent close-coupling and relativistic distorted-wave approximation theoretical predictions. The results are compared and discussed with EICP data for collision energies 80 eV and 100 eV.

Impact of polyphenols on extracellular vesicle levels and effects and their properties as tools for drug delivery for nutrition and health.

PubMed

Soleti, Raffaella; Andriantsitohaina, Ramaroson; Martinez, M Carmen

2018-04-15

Polyphenols are found in plant-derived foods and beverages and display numerous protective effects against cancers, cardiovascular, metabolic and neurodegenerative diseases. Extracellular vesicles (EVs), microparticles, exosomes, and apoptotic bodies, originated by different cell types are emerging as a novel mean of cell-to-cell communication in physiology and pathology and represent a new way to convey fundamental information between cells. Polyphenols can act on signaling pathways that interfere with the biogenesis of EVs. Thus, they are able to control EV release from cells and their content and therefore their functional properties. Both EVs and polyphenols are therapeutic tools that can be used against several diseases. In this context, the combination of both tools can increase their therapeutic potential. Three types of strategies can be used: (i) plants are able to produce EVs that encapsulate natural components from vegetables, polyphenols for instance, (ii) mammalian cells can be treated with polyphenols and the subsequent EVs produced are enriched in these components, and (iii) EVs from mammalian cells can be uploaded with polyphenols. We review the novel aspects of the interplay between polyphenols and EVs that could trigger and improve the health benefits in cancer, cardiovascular, metabolic and neurodegenerative diseases. Copyright © 2018 Elsevier Inc. All rights reserved.

[Cardiovascular risk study in patients with renin-angiotensin system blockade by means of the proteone of circulating extracellular vesicles].

PubMed

de la Cuesta, F; Baldan-Martin, M; Mourino-Alvarez, L; Sastre-Oliva, T; Alvarez-Llamas, G; Gonzalez-Calero, L; Ruiz-Hurtado, G; Segura, J; Vivanco, F; Ruilope, L M; Barderas, M G

2016-01-01

Extracellular vesicles (EVs) are released to the bloodstream by certain cell types due to transport, activation and cell death processes. Blood count of EVs from platelet and endothelial origin has been proved to be a cardiovascular risk biomarker. Thus, EVs proteome might reflect the underlying cellular processes in hypertensive patients with albuminuria. Protein content of circulating EVs was analyzed by liquid chromatography coupled to mass spectrometry. EVs were isolated by an ultracentrifugation protocol optimized in order to avoid contamination by blood plasma proteins. Purity of the isolated fraction was verified by electronic and confocal microscopy, and by flow cytometry. We hereby show a method to isolate circulating EVs from hypertensive patients with/without albuminuria with high yield and purity. Besides, we provide a reference proteome of the EVs of these patients, composed of 2,463 proteins, and prove that the proteins carried by these vesicles are associated with crucial processes involved in the inherent cardiovascular risk. The proteome of circulating EVs is an interesting source of indicators in the evaluation of cardiovascular risk in hypertensive patients with renin-angiotensin system blockage. Copyright © 2015 SEHLELHA. Published by Elsevier España, S.L.U. All rights reserved.

Naturally enveloped AAV vectors for shielding neutralizing antibodies and robust gene delivery in vivo

PubMed Central

György, Bence; Fitzpatrick, Zachary; Crommentuijn, Matheus HW; Mu, Dakai; Maguire, Casey A.

2014-01-01

Recently adeno-associated virus (AAV) became the first clinically approved gene therapy product in the western world. To develop AAV for future clinical application in a widespread patient base, particularly in therapies which require intravenous (i.v.) administration of vector, the virus must be able to evade pre-existing antibodies to the wild type virus. Here we demonstrate that in mice, AAV vectors associated with extracellular vesicles (EVs) can evade human anti-AAV neutralizing antibodies. We observed different antibody evasion and gene transfer abilities with populations of EVs isolated by different centrifugal forces. EV-associated AAV vector (ev-AAV) was up to 136-fold more resistant over a range of neutralizing antibody concentrations relative to standard AAV vector in vitro. Importantly in mice, at a concentration of passively transferred human antibodies which decreased i.v. administered standard AAV transduction of brain by 80%, transduction of ev-AAV transduction was not reduced and was 4,000-fold higher. Finally, we show that expressing a brain targeting peptide on the EV surface allowed significant enhancement of transduction compared to untargeted ev-AAV. Using ev-AAV represents an effective, clinically relevant approach to evade human neutralizing anti-AAV antibodies after systemic administration of vector. PMID:24917028

Quantifying EV battery end-of-life through analysis of travel needs with vehicle powertrain models

NASA Astrophysics Data System (ADS)

Saxena, Samveg; Le Floch, Caroline; MacDonald, Jason; Moura, Scott

2015-05-01

Electric vehicles enable clean and efficient transportation, however concerns about range anxiety and battery degradation hinder EV adoption. The common definition for battery end-of-life is when 70-80% of original energy capacity remains, however little analysis is available to support this retirement threshold. By applying detailed physics-based models of EVs with data on how drivers use their cars, we show that EV batteries continue to meet daily travel needs of drivers well beyond capacity fade of 80% remaining energy storage capacity. Further, we show that EV batteries with substantial energy capacity fade continue to provide sufficient buffer charge for unexpected trips with long distances. We show that enabling charging in more locations, even if only with 120 V wall outlets, prolongs useful life of EV batteries. Battery power fade is also examined and we show EVs meet performance requirements even down to 30% remaining power capacity. Our findings show that defining battery retirement at 70-80% remaining capacity is inaccurate. Battery retirement should instead be governed by when batteries no longer satisfy daily travel needs of a driver. Using this alternative retirement metric, we present results on the fraction of EV batteries that may be retired with different levels of energy capacity fade.

Quantifying EV battery end-of-life through analysis of travel needs with vehicle powertrain models

DOE PAGES

Saxena, Samveg; Le Floch, Caroline; MacDonald, Jason; ...

2015-05-15

Electric vehicles enable clean and efficient transportation; however, concerns about range anxiety and battery degradation hinder EV adoption. The common definition for battery end-of-life is when 70-80% of original energy capacity remain;, however, little analysis is available to support this retirement threshold. By applying detailed physics-based models of EVs with data on how drivers use their cars, we show that EV batteries continue to meet daily travel needs of drivers well beyond capacity fade of 80% remaining energy storage capacity. Further, we show that EV batteries with substantial energy capacity fade continue to provide sufficient buffer charge for unexpected tripsmore » with long distances. We show that enabling charging in more locations, even if only with 120 V wall outlets, prolongs useful life of EV batteries. Battery power fade is also examined and we show EVs meet performance requirements even down to 30% remaining power capacity. Our findings show that defining battery retirement at 70-80% remaining capacity is inaccurate. Battery retirement should instead be governed by when batteries no longer satisfy daily travel needs of a driver. Using this alternative retirement metric, we present results on the fraction of EV batteries that may be retired with different levels of energy capacity fade.« less

Quantifying EV battery end-of-life through analysis of travel needs with vehicle powertrain models

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

Saxena, Samveg; Le Floch, Caroline; MacDonald, Jason

Electric vehicles enable clean and efficient transportation; however, concerns about range anxiety and battery degradation hinder EV adoption. The common definition for battery end-of-life is when 70-80% of original energy capacity remain;, however, little analysis is available to support this retirement threshold. By applying detailed physics-based models of EVs with data on how drivers use their cars, we show that EV batteries continue to meet daily travel needs of drivers well beyond capacity fade of 80% remaining energy storage capacity. Further, we show that EV batteries with substantial energy capacity fade continue to provide sufficient buffer charge for unexpected tripsmore » with long distances. We show that enabling charging in more locations, even if only with 120 V wall outlets, prolongs useful life of EV batteries. Battery power fade is also examined and we show EVs meet performance requirements even down to 30% remaining power capacity. Our findings show that defining battery retirement at 70-80% remaining capacity is inaccurate. Battery retirement should instead be governed by when batteries no longer satisfy daily travel needs of a driver. Using this alternative retirement metric, we present results on the fraction of EV batteries that may be retired with different levels of energy capacity fade.« less

Enterovirus-71 genotype C isolated in Peru between 2006 and 2009.

PubMed

Huaman, Jose L; Carrion, Gladys; Ampuero, Julia S; Ocaña, Victor; Laguna-Torres, V Alberto; Hontz, Robert D

2016-12-01

Enterovirus-71 (EV71) was first isolated in California, United States in 1969, belongs to the genus Enterovirus, family Picornaviridae. Although infection normally causes mild, often undiagnosed illness, it can cause central nervous system infections that could turn fatal. Based on VP1 gene analysis, EV71 has been classified into six separate genotypes. Although the molecular epidemiology of EV71 has been well described via studies originating from Asia and Europe, it is mostly unknown in South America. From our study, four EV71 isolates from Peru were characterized using phylogenetic methods to determine their relationship with known reference strains. These four Peruvian EV71 isolates from between 2006 and 2009 were analyzed by RT-PCR using primers capable of amplifying the entire VP1 gene. Reference strains representing all six known genotypes were used to determine any recognizable phylogenetic relationships. In fact, all of our isolates clustered together within the genotype C1 lineage- separate from Asian, European, North American, and Australian strains. We present evidence that EV71 genotype C1 exists in Peru, and this is the first such report documenting EV71 genotype C1 circulating in South America. Gathering additional isolates will help elucidate a more complete global epidemiological picture of EV71 infections. Published by Elsevier B.V.

Environmental Justice Aspects of Exposure to PM2.5 Emissions from Electric Vehicle Use in China.

PubMed

Ji, Shuguang; Cherry, Christopher R; Zhou, Wenjun; Sawhney, Rapinder; Wu, Ye; Cai, Siyi; Wang, Shuxiao; Marshall, Julian D

2015-12-15

Plug-in electric vehicles (EVs) in China aim to improve sustainability and reduce environmental health impacts of transport emissions. Urban use of EVs rather than conventional vehicles shifts transportation's air pollutant emissions from urban areas (tailpipes) to predominantly rural areas (power plants), changing the geographic distribution of health impacts. We model PM2.5-related health impacts attributable to urban EV use for 34 major cities. Our investigation focuses on environmental justice (EJ) by comparing pollutant inhalation versus income among impacted counties. We find that EVs could increase EJ challenge in China: most (~77%, range: 41-96%) emission inhalation attributable to urban EVs use is distributed to predominately rural communities whose incomes are on average lower than the cities where EVs are used. Results vary dramatically across cities depending on urban income and geography. Discriminant analysis reveals that counties with low income and high inhalation of urban EV emissions have comparatively higher agricultural employment rates, higher mortality rates, more children in the population, and lower education levels. We find that low-emission electricity sources such as renewable energy can help mitigate EJ issues raised here. Findings here are not unique to EVs, but instead are relevant for nearly all electricity-consuming technologies in urban areas.

Observation of CH A 2X 2Πr and 2Σ--->X 2Πr emissions in gas-phase collisions of fast O(3P) atoms with acetylene

NASA Astrophysics Data System (ADS)

Orient, O. J.; Chutjian, A.; Murad, E.

1995-03-01

Optical emissions in single-collision, beam-beam reactions of fast (3-22-eV translational energy) O(3P) atoms with C2H2 have been measured in the wavelength range 300-850 nm. Two features were observed, one with a peak wavelength at 431 nm, corresponding to the CH A 2X 2Πr transition, and a second weaker emission in the range 380-400 nm corresponding to the B 2Σ--->X 2Πr transition. Both the A-->X and B-->X emissions were fit to a synthetic spectrum of CH(A) at a vibrational temperature Tv of 10 000 K (0.86 eV) and a rotational temperature Tr of approximately 5000 K (0.43 eV); and CH(B) to Tv=2500 K (0.22 eV) and Tr=1000 K (0.09 eV). The energy threshold for the A-->X emission was measured to be 7.3+/-0.4 eV (lab) or 4.5+/-0.2 eV (c.m.). This agrees with the energy threshold of 7.36 eV (lab) for the reaction O(3P)+C2H2-->CH(A)+HCO.

Lowering Low-Density Lipoprotein Particles in Plasma Using Dextran Sulphate Co-Precipitates Procoagulant Extracellular Vesicles.

PubMed

Wang, Jiong-Wei; Zhang, Ya-Nan; Sze, Siu Kwan; van de Weg, Sander M; Vernooij, Flora; Schoneveld, Arjan H; Tan, Sock-Hwee; Versteeg, Henri H; Timmers, Leo; Lam, Carolyn S P; de Kleijn, Dominique P V

2017-12-29

Plasma extracellular vesicles (EVs) are lipid membrane vesicles involved in several biological processes including coagulation. Both coagulation and lipid metabolism are strongly associated with cardiovascular events. Lowering very-low- and low-density lipoprotein ((V)LDL) particles via dextran sulphate LDL apheresis also removes coagulation proteins. It remains unknown, however, how coagulation proteins are removed in apheresis. We hypothesize that plasma EVs that contain high levels of coagulation proteins are concomitantly removed with (V)LDL particles by dextran sulphate apheresis. For this, we precipitated (V)LDL particles from human plasma with dextran sulphate and analyzed the abundance of coagulation proteins and EVs in the precipitate. Coagulation pathway proteins, as demonstrated by proteomics and a bead-based immunoassay, were over-represented in the (V)LDL precipitate. In this precipitate, both bilayer EVs and monolayer (V)LDL particles were observed by electron microscopy. Separation of EVs from (V)LDL particles using density gradient centrifugation revealed that almost all coagulation proteins were present in the EVs and not in the (V)LDL particles. These EVs also showed a strong procoagulant activity. Our study suggests that dextran sulphate used in LDL apheresis may remove procoagulant EVs concomitantly with (V)LDL particles, leading to a loss of coagulation proteins from the blood.

Choices and changes: Eccles' Expectancy-Value model and upper-secondary school students' longitudinal reflections about their choice of a STEM education

NASA Astrophysics Data System (ADS)

Lykkegaard, Eva; Ulriksen, Lars

2016-03-01

During the past 30 years, Eccles' comprehensive social-psychological Expectancy-Value Model of Motivated Behavioural Choices (EV-MBC model) has been proven suitable for studying educational choices related to Science, Technology, Engineering and/or Mathematics (STEM). The reflections of 15 students in their last year in upper-secondary school concerning their choice of tertiary education were examined using quantitative EV-MBC surveys and repeated qualitative interviews. This article presents the analyses of three cases in detail. The analytical focus was whether the factors indicated in the EV-MBC model could be used to detect significant changes in the students' educational choice processes. An important finding was that the quantitative EV-MBC surveys and the qualitative interviews gave quite different results concerning the students' considerations about the choice of tertiary education, and that significant changes in the students' reflections were not captured by the factors of the EV-MBC model. This questions the validity of the EV-MBC surveys. Moreover, the quantitative factors from the EV-MBC model did not sufficiently explain students' dynamical educational choice processes where students in parallel considered several different potential educational trajectories. We therefore call for further studies of the EV-MBC model's use in describing longitudinal choice processes and especially in investigating significant changes.

Extracellular vesicles in Alzheimer's disease: friends or foes? Focus on aβ-vesicle interaction.

PubMed

Joshi, Pooja; Benussi, Luisa; Furlan, Roberto; Ghidoni, Roberta; Verderio, Claudia

2015-03-03

The intercellular transfer of amyloid-β (Aβ) and tau proteins has received increasing attention in Alzheimer's disease (AD). Among other transfer modes, Aβ and tau dissemination has been suggested to occur through release of Extracellular Vesicles (EVs), which may facilitate delivery of pathogenic proteins over large distances. Recent evidence indicates that EVs carry on their surface, specific molecules which bind to extracellular Aβ, opening the possibility that EVs may also influence Aβ assembly and synaptotoxicity. In this review we focus on studies which investigated the impact of EVs in Aβ-mediated neurodegeneration and showed either detrimental or protective role for EVs in the pathology.

Electron heated target temperature measurements in petawatt laser experiments based on extreme ultraviolet imaging and spectroscopy.

PubMed

Ma, T; Beg, F N; MacPhee, A G; Chung, H-K; Key, M H; Mackinnon, A J; Patel, P K; Hatchett, S; Akli, K U; Stephens, R B; Chen, C D; Freeman, R R; Link, A; Offermann, D T; Ovchinnikov, V; Van Woerkom, L D

2008-10-01

Three independent methods (extreme ultraviolet spectroscopy, imaging at 68 and 256 eV) have been used to measure planar target rear surface plasma temperature due to heating by hot electrons. The hot electrons are produced by ultraintense laser-plasma interactions using the 150 J, 0.5 ps Titan laser. Soft x-ray spectroscopy in the 50-400 eV region and imaging at the 68 and 256 eV photon energies give a planar deuterated carbon target rear surface pre-expansion temperature in the 125-150 eV range, with the rear plasma plume averaging a temperature approximately 74 eV.

Power control apparatus and methods for electric vehicles

DOEpatents

Gadh, Rajit; Chung, Ching-Yen; Chu, Chi-Cheng; Qiu, Li

2016-03-22

Electric vehicle (EV) charging apparatus and methods are described which allow the sharing of charge current between multiple vehicles connected to a single source of charging energy. In addition, this charge sharing can be performed in a grid-friendly manner by lowering current supplied to EVs when necessary in order to satisfy the needs of the grid, or building operator. The apparatus and methods can be integrated into charging stations or can be implemented with a middle-man approach in which a multiple EV charging box, which includes an EV emulator and multiple pilot signal generation circuits, is coupled to a single EV charge station.

A novel Enterovirus 96 circulating in China causes hand, foot, and mouth disease.

PubMed

Xu, Yi; Sun, Yisuo; Ma, Jinmin; Zhou, Shuru; Fang, Wei; Ye, Jiawei; Tan, Limei; Ji, Jingkai; Luo, Dan; Li, Liqiang; Li, Jiandong; Fang, Chunxiao; Pei, Na; Shi, Shuo; Liu, Xin; Jiang, Hui; Gong, Sitang; Xu, Xun

2017-06-01

Enterovirus 96 (EV-96) is a recently described member of the species Enterovirus C and is associated with paralysis and myelitis. In this study, using metagenomic sequencing, we identified a new enterovirus 96 strain (EV-96-SZ/GD/CHN/2014) as the sole pathogen causing hand, foot, and mouth disease (HFMD). A genomic comparison showed that EV-96-SZ/GD/CHN/2014 is most similar to the EV-96-05517 strain (85% identity), which has also been detected in Guangdong Province. This is the first time that metagenomic sequencing has been used to identify an EV-96 strain shown to be associated with HFMD.

Hand, Foot, and Mouth Disease in China: Modeling Epidemic Dynamics of Enterovirus Serotypes and Implications for Vaccination

PubMed Central

Takahashi, Saki; Liao, Qiaohong; Van Boeckel, Thomas P.; Xing, Weijia; Sun, Junling; Hsiao, Victor Y.; Metcalf, C. Jessica E.; Chang, Zhaorui; Liu, Fengfeng; Zhang, Jing; Wu, Joseph T.; Cowling, Benjamin J.; Leung, Gabriel M.; Farrar, Jeremy J.; van Doorn, H. Rogier; Grenfell, Bryan T.; Yu, Hongjie

2016-01-01

Background Hand, foot, and mouth disease (HFMD) is a common childhood illness caused by serotypes of the Enterovirus A species in the genus Enterovirus of the Picornaviridae family. The disease has had a substantial burden throughout East and Southeast Asia over the past 15 y. China reported 9 million cases of HFMD between 2008 and 2013, with the two serotypes Enterovirus A71 (EV-A71) and Coxsackievirus A16 (CV-A16) being responsible for the majority of these cases. Three recent phase 3 clinical trials showed that inactivated monovalent EV-A71 vaccines manufactured in China were highly efficacious against HFMD associated with EV-A71, but offered no protection against HFMD caused by CV-A16. To better inform vaccination policy, we used mathematical models to evaluate the effect of prospective vaccination against EV-A71-associated HFMD and the potential risk of serotype replacement by CV-A16. We also extended the model to address the co-circulation, and implications for vaccination, of additional non-EV-A71, non-CV-A16 serotypes of enterovirus. Methods and Findings Weekly reports of HFMD incidence from 31 provinces in Mainland China from 1 January 2009 to 31 December 2013 were used to fit multi-serotype time series susceptible–infected–recovered (TSIR) epidemic models. We obtained good model fit for the two-serotype TSIR with cross-protection, capturing the seasonality and geographic heterogeneity of province-level transmission, with strong correlation between the observed and simulated epidemic series. The national estimate of the basic reproduction number, R 0, weighted by provincial population size, was 26.63 for EV-A71 (interquartile range [IQR]: 23.14, 30.40) and 27.13 for CV-A16 (IQR: 23.15, 31.34), with considerable variation between provinces (however, predictions about the overall impact of vaccination were robust to this variation). EV-A71 incidence was projected to decrease monotonically with higher coverage rates of EV-A71 vaccination. Across provinces, CV-A16 incidence in the post-EV-A71-vaccination period remained either comparable to or only slightly increased from levels prior to vaccination. The duration and strength of cross-protection following infection with EV-A71 or CV-A16 was estimated to be 9.95 wk (95% confidence interval [CI]: 3.31, 23.40) in 68% of the population (95% CI: 37%, 96%). Our predictions are limited by the necessarily short and under-sampled time series and the possible circulation of unidentified serotypes, but, nonetheless, sensitivity analyses indicate that our results are robust in predicting that the vaccine should drastically reduce incidence of EV-A71 without a substantial competitive release of CV-A16. Conclusions The ability of our models to capture the observed epidemic cycles suggests that herd immunity is driving the epidemic dynamics caused by the multiple serotypes of enterovirus. Our results predict that the EV-A71 and CV-A16 serotypes provide a temporary immunizing effect against each other. Achieving high coverage rates of EV-A71 vaccination would be necessary to eliminate the ongoing transmission of EV-A71, but serotype replacement by CV-A16 following EV-A71 vaccination is likely to be transient and minor compared to the corresponding reduction in the burden of EV-A71-associated HFMD. Therefore, a mass EV-A71 vaccination program of infants and young children should provide significant benefits in terms of a reduction in overall HFMD burden. PMID:26882540

Hand, Foot, and Mouth Disease in China: Modeling Epidemic Dynamics of Enterovirus Serotypes and Implications for Vaccination.

PubMed

Takahashi, Saki; Liao, Qiaohong; Van Boeckel, Thomas P; Xing, Weijia; Sun, Junling; Hsiao, Victor Y; Metcalf, C Jessica E; Chang, Zhaorui; Liu, Fengfeng; Zhang, Jing; Wu, Joseph T; Cowling, Benjamin J; Leung, Gabriel M; Farrar, Jeremy J; van Doorn, H Rogier; Grenfell, Bryan T; Yu, Hongjie

2016-02-01

Hand, foot, and mouth disease (HFMD) is a common childhood illness caused by serotypes of the Enterovirus A species in the genus Enterovirus of the Picornaviridae family. The disease has had a substantial burden throughout East and Southeast Asia over the past 15 y. China reported 9 million cases of HFMD between 2008 and 2013, with the two serotypes Enterovirus A71 (EV-A71) and Coxsackievirus A16 (CV-A16) being responsible for the majority of these cases. Three recent phase 3 clinical trials showed that inactivated monovalent EV-A71 vaccines manufactured in China were highly efficacious against HFMD associated with EV-A71, but offered no protection against HFMD caused by CV-A16. To better inform vaccination policy, we used mathematical models to evaluate the effect of prospective vaccination against EV-A71-associated HFMD and the potential risk of serotype replacement by CV-A16. We also extended the model to address the co-circulation, and implications for vaccination, of additional non-EV-A71, non-CV-A16 serotypes of enterovirus. Weekly reports of HFMD incidence from 31 provinces in Mainland China from 1 January 2009 to 31 December 2013 were used to fit multi-serotype time series susceptible-infected-recovered (TSIR) epidemic models. We obtained good model fit for the two-serotype TSIR with cross-protection, capturing the seasonality and geographic heterogeneity of province-level transmission, with strong correlation between the observed and simulated epidemic series. The national estimate of the basic reproduction number, R0, weighted by provincial population size, was 26.63 for EV-A71 (interquartile range [IQR]: 23.14, 30.40) and 27.13 for CV-A16 (IQR: 23.15, 31.34), with considerable variation between provinces (however, predictions about the overall impact of vaccination were robust to this variation). EV-A71 incidence was projected to decrease monotonically with higher coverage rates of EV-A71 vaccination. Across provinces, CV-A16 incidence in the post-EV-A71-vaccination period remained either comparable to or only slightly increased from levels prior to vaccination. The duration and strength of cross-protection following infection with EV-A71 or CV-A16 was estimated to be 9.95 wk (95% confidence interval [CI]: 3.31, 23.40) in 68% of the population (95% CI: 37%, 96%). Our predictions are limited by the necessarily short and under-sampled time series and the possible circulation of unidentified serotypes, but, nonetheless, sensitivity analyses indicate that our results are robust in predicting that the vaccine should drastically reduce incidence of EV-A71 without a substantial competitive release of CV-A16. The ability of our models to capture the observed epidemic cycles suggests that herd immunity is driving the epidemic dynamics caused by the multiple serotypes of enterovirus. Our results predict that the EV-A71 and CV-A16 serotypes provide a temporary immunizing effect against each other. Achieving high coverage rates of EV-A71 vaccination would be necessary to eliminate the ongoing transmission of EV-A71, but serotype replacement by CV-A16 following EV-A71 vaccination is likely to be transient and minor compared to the corresponding reduction in the burden of EV-A71-associated HFMD. Therefore, a mass EV-A71 vaccination program of infants and young children should provide significant benefits in terms of a reduction in overall HFMD burden.

Nuclear transport of cancer extracellular vesicle-derived biomaterials through nuclear envelope invagination-associated late endosomes.

PubMed

Rappa, Germana; Santos, Mark F; Green, Toni M; Karbanová, Jana; Hassler, Justin; Bai, Yongsheng; Barsky, Sanford H; Corbeil, Denis; Lorico, Aurelio

2017-02-28

Extracellular membrane vesicles (EVs) function as vehicles of intercellular communication, but how the biomaterials they carry reach the target site in recipient cells is an open question. We report that subdomains of Rab7+ late endosomes and nuclear envelope invaginations come together to create a sub-nuclear compartment, where biomaterials associated with CD9+ EVs are delivered. EV-derived biomaterials were also found in the nuclei of host cells. The inhibition of nuclear import and export pathways abrogated the nuclear localization of EV-derived biomaterials or led to their accumulation therein, respectively, suggesting that their translocation is dependent on nuclear pores. Nuclear envelope invagination-associated late endosomes were observed in ex vivo biopsies in both breast carcinoma and associated stromal cells. The transcriptome of stromal cells exposed to cancer cell-derived CD9+ EVs revealed that the regulation of eleven genes, notably those involved in inflammation, relies on the nuclear translocation of EV-derived biomaterials. Our findings uncover a new cellular pathway used by EVs to reach nuclear compartment.

[The early diagnosis value of EV 71 IgM class antibodies in the hand, foot and mouth disease].

PubMed

Zhao, Jing; Xu, Jun; Chen, Wei-wei; Li, Yong-li; Tang, Yan; Li, Jia; Wang, Hai-bin; Guo, Tong-sheng; Zhao, Min; Li, Bo-an; Mao, Yuan-li

2011-04-01

Assessment of detection of IgM antibodies for human enterovirus 71 (EV 71) in early diagnosis for the hand, foot and mouth disease (HFMD). The sera and throat swabs from 38 patients which were clinical diagnosis as HFMD, were continuous daily collected in our hospital in 2010. These specimens were detected by EV 71 IgM antibodies assay, real time RT-PCR methods for EV 71 and Enterovirus. Among 38 HFMD patients, the cumulative positive rates of EV 71 IgM antibodies were: 60.5% on day 1, 71.1% on day 2, 81.5% in the first 3-4 days, 92.1% on day 5, 92.1% on day 6, and the positive rate of nucleic acid detected by the real time RT-PCR for EV 71 and Enterovirus were 60.5%, 73.6%. The positive rate of EV 71 IgM antibodies in the hand, foot and mouth disease just can occur on day 1, and reach to peak on day 5, which can be used as one of indicators of early diagnosis of hand, foot and mouth disease.

Emerging Roles for Extracellular Vesicles in Tissue Engineering and Regenerative Medicine

PubMed Central

Lamichhane, Tek N.; Sokic, Sonja; Schardt, John S.; Raiker, Rahul S.; Lin, Jennifer W.

2015-01-01

Extracellular vesicles (EVs)—comprising a heterogeneous population of cell-derived lipid vesicles including exosomes, microvesicles, and others—have recently emerged as both mediators of intercellular information transfer in numerous biological systems and vehicles for drug delivery. In both roles, EVs have immense potential to impact tissue engineering and regenerative medicine applications. For example, the therapeutic effects of several progenitor and stem cell-based therapies have been attributed primarily to EVs secreted by these cells, and EVs have been recently reported to play direct roles in injury-induced tissue regeneration processes in multiple physiological systems. In addition, EVs have been utilized for targeted drug delivery in regenerative applications and possess unique potential to be harnessed as patient-derived drug delivery vehicles for personalized medicine. This review discusses EVs in the context of tissue repair and regeneration, including their utilization as drug carriers and their crucial role in cell-based therapies. Furthermore, the article highlights the growing need for bioengineers to understand, consider, and ultimately design and specifically control the activity of EVs to maximize the efficacy of tissue engineering and regenerative therapies. PMID:24957510

Band alignments at Ga2O3 heterojunction interfaces with Si and Ge

NASA Astrophysics Data System (ADS)

Gibbon, J. T.; Jones, L.; Roberts, J. W.; Althobaiti, M.; Chalker, P. R.; Mitrovic, Ivona Z.; Dhanak, V. R.

2018-06-01

Amorphous Ga2O3 thin films were deposited on p-type (111) and (100) surfaces of silicon and (100) germanium by atomic layer deposition (ALD). X-ray photoelectron spectroscopy (XPS) was used to investigate the band alignments at the interfaces using the Kraut Method. The valence band offsets were determined to be 3.49± 0.08 eV and 3.47± 0.08 eV with Si(111) and Si(100) respectively and 3.51eV± 0.08 eV with Ge(100). Inverse photoemission spectroscopy (IPES) was used to investigate the conduction band of a thick Ga2O3 film and the band gap of the film was determined to be 4.63±0.14 eV. The conduction band offsets were found to be 0.03 eV and 0.05eV with Si(111) and Si(100) respectively, and 0.45eV with Ge(100). The results indicate that the heterojunctions of Ga2O3 with Si(100), Si(111) and Ge(100) are all type I heterojunctions.

Band alignment of atomic layer deposited SiO2 and HfSiO4 with (\\bar{2}01) β-Ga2O3

NASA Astrophysics Data System (ADS)

Carey, Patrick H., IV; Ren, Fan; Hays, David C.; Gila, Brent P.; Pearton, Stephen J.; Jang, Soohwan; Kuramata, Akito

2017-07-01

The valence band offset at both SiO2/β-Ga2O3 and HfSiO4/β-Ga2O3 heterointerfaces was measured using X-ray photoelectron spectroscopy. Both dielectrics were deposited by atomic layer deposition (ALD) onto single-crystal β-Ga2O3. The bandgaps of the materials were determined by reflection electron energy loss spectroscopy as 4.6 eV for Ga2O3, 8.7 eV for Al2O3 and 7.0 eV for HfSiO4. The valence band offset was determined to be 1.23 ± 0.20 eV (straddling gap, type I alignment) for ALD SiO2 on β-Ga2O3 and 0.02 ± 0.003 eV (also type I alignment) for HfSiO4. The respective conduction band offsets were 2.87 ± 0.70 eV for ALD SiO2 and 2.38 ± 0.50 eV for HfSiO4, respectively.

Neonatal Subventricular Zone Neural Stem Cells Release Extracellular Vesicles that Act as a Microglial Morphogen.

PubMed

Morton, Mary C; Neckles, Victoria N; Seluzicki, Caitlin M; Holmberg, Jennie C; Feliciano, David M

2018-04-03

Subventricular zone (SVZ) neural stem cells (NSCs) are the cornerstone of the perinatal neurogenic niche. Microglia are immune cells of the nervous system that are enriched in the neonatal SVZ. Although microglia regulate NSCs, the extent to which this interaction is bi-directional is unclear. Extracellular vesicles (EVs) are cell-derived particles that encase miRNA and proteins. Here, we demonstrate that SVZ NSCs generate and release EVs. Neonatal electroporated fluorescent EV fusion proteins were released by NSCs and subsequently cleared from the SVZ. EVs were preferentially targeted to microglia. Small RNA sequencing identified miRNAs within the EVs that regulate microglia physiology and morphology. EVs induced a transition to a CD11b/Iba1 non-stellate microglial morphology. The transition accompanied a microglial transcriptional state characterized by Let-7-regulated cytokine release and a negative feedback loop that controlled NSC proliferation. These findings implicate an NSC-EV-microglia axis and provide insight to normal and pathophysiological brain development. Copyright © 2018 The Author(s). Published by Elsevier Inc. All rights reserved.

Focus on Extracellular Vesicles: Physiological Role and Signalling Properties of Extracellular Membrane Vesicles

PubMed Central

Iraci, Nunzio; Leonardi, Tommaso; Gessler, Florian; Vega, Beatriz; Pluchino, Stefano

2016-01-01

Extracellular vesicles (EVs) are a heterogeneous population of secreted membrane vesicles, with distinct biogenesis routes, biophysical properties and different functions both in physiological conditions and in disease. The release of EVs is a widespread biological process, which is conserved across species. In recent years, numerous studies have demonstrated that several bioactive molecules are trafficked with(in) EVs, such as microRNAs, mRNAs, proteins and lipids. The understanding of their final impact on the biology of specific target cells remains matter of intense debate in the field. Also, EVs have attracted great interest as potential novel cell-free therapeutics. Here we describe the proposed physiological and pathological functions of EVs, with a particular focus on their molecular content. Also, we discuss the advances in the knowledge of the mechanisms regulating the secretion of EV-associated molecules and the specific pathways activated upon interaction with the target cell, highlighting the role of EVs in the context of the immune system and as mediators of the intercellular signalling in the brain. PMID:26861302

Electric vehicle (EV) storage supply chain risk and the energy market: A micro and macroeconomic risk management approach

NASA Astrophysics Data System (ADS)

Aguilar, Susanna D.

As a cost effective storage technology for renewable energy sources, Electric Vehicles can be integrated into energy grids. Integration must be optimized to ascertain that renewable energy is available through storage when demand exists so that cost of electricity is minimized. Optimization models can address economic risks associated with the EV supply chain- particularly the volatility in availability and cost of critical materials used in the manufacturing of EV motors and batteries. Supply chain risk can reflect itself in a shortage of storage, which can increase the price of electricity. We propose a micro-and macroeconomic framework for managing supply chain risk through utilization of a cost optimization model in combination with risk management strategies at the microeconomic and macroeconomic level. The study demonstrates how risk from the EVs vehicle critical material supply chain affects manufacturers, smart grid performance, and energy markets qualitatively and quantitatively. Our results illustrate how risk in the EV supply chain affects EV availability and the cost of ancillary services, and how EV critical material supply chain risk can be mitigated through managerial strategies and policy.

Current status of BL-2B at photon factory

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

Nambu, Akira, E-mail: akia.nambu.tw@hitachi.com; Ueda, Kazuhiro; Horiba, Koji

A new soft x-ray beamline BL-2B at Photon Factory of High Energy Accelerator Research Organization (KEK-PF) covers energy range from vacuum ultraviolet (30 eV) to soft x-ray (4000 eV). This wide energy range could be achieved by employing two undulators and two monochromators. Two different energy range undulators were installed tandem to a 9-meter straight section of PF storage ring. The 1{sup st} undulator is for VUV (30 eV) to SX (280 eV), while the other one is for SX (280 eV) to HX (4000 eV). It is also necessary to be equipped with two different monochrometors for energy abovemore » and under 2000 eV; grating monochrometor and double crystal monochrometor. One of the main purposes of this bemaline is spectroscopic study of light elements contained in several functional materials. The beamline is designed for photoemission spectroscopy (PES), X-ray absorption fine structure (XAFS) and other types of experiments. The performance of the new beamline is reported and typical examples of its application to material science are demonstrated.« less

Enterovirus meningitis in Tunisia (Monastir, Mahdia, 2011-2013): identification of virus variants cocirculating in France.

PubMed

Othman, Ines; Volle, Romain; Elargoubi, Aida; Guediche, Mohamed Neji; Chakroun, Mohamed; Sfar, Mohamed Tahar; Pereira, Bruno; Peigue-Lafeuille, Hélène; Aouni, Mahjoub; Archimbaud, Christine; Bailly, Jean-Luc

2016-02-01

Acute enterovirus (EV) meningitis is a frequent cause of hospitalisation, and over 100 EV serotypes may be involved. A total of 215 patients of all ages with meningitis signs were investigated in 2 Tunisian hospitals. Their cerebrospinal fluid (CSF) was analysed retrospectively for EVs with a TaqMan real-time RT-qPCR. The virus strains were typed, and their evolutionary relationships were determined by Bayesian phylogenetic methods. An EV genome was detected in 21/215 patients (9.8%). The CSF viral loads ranged from 3.27 to 5.63 log10 genome copies/mL. The strains were identified in 13/21 patients and assigned to EV-B types. Viruses identified in Tunisian patients were genetically related to variants detected in France. The viral loads were similar in Tunisian and French patients for most EV types. The phylogenetic data and viral loads determined in Tunisian and French patients suggest that close EV variants were involved in aseptic meningitis in the 2 countries over a same period. Copyright © 2016 Elsevier Inc. All rights reserved.

Improved Methods of Producing and Administering Extracellular Vesicles | Poster

Cancer.gov

An efficient method of producing purified extracellular vesicles (EVs), in conjunction with a method that blocks liver macrophages from clearing EVs from the body, has produced promising results for the use of EVs in cancer therapy.

Mechanisms and functions of extracellular vesicle release in vivo-What we can learn from flies and worms.

PubMed

Beer, Katharina B; Wehman, Ann Marie

2017-03-04

Cells from bacteria to man release extracellular vesicles (EVs) that contain signaling molecules like proteins, lipids, and nucleic acids. The content, formation, and signaling roles of these conserved vesicles are diverse, but the physiological relevance of EV signaling in vivo is still debated. Studies in classical genetic model organisms like C. elegans and Drosophila have begun to reveal the developmental and behavioral roles for EVs. In this review, we discuss the emerging evidence for the in vivo signaling roles of EVs. Significant effort has also been made to understand the mechanisms behind the formation and release of EVs, specifically of exosomes derived from exocytosis of multivesicular bodies and of microvesicles derived from plasma membrane budding called ectocytosis. In this review, we detail the impact of flies and worms on understanding the proteins and lipids involved in EV biogenesis and highlight the open questions in the field.

Study of chemical shift in Kα, Kβ1,3 and Kβ// X-ray emission lines of 37Rb compounds with WDXRF

NASA Astrophysics Data System (ADS)

Kainth, Harpreet Singh; Singh, Ranjit; Singh, Tejbir; Mehta, D.; Shahi, J. S.; Kumar, Sanjeev

2018-05-01

The positive and negative chemical shifts in Kα, Kβ1,3 and Kβ// X-ray emission lines of rubidium compounds were measured with high resolution WDXRF spectrometer. The measured energy shifts in Kα emission lines ranges from -2.95 eV to -3.64 eV, Kβ1,3 emission lines ranges from 1.16 eV to 1.32 eV and Kβ// emission lines ranges from 1.31 eV to 4.36 eV respectively. In the present work, it has been found that chemical shift in Kβ// X-ray emission lines were found to be larger than Kα and Kβ1,3 X-ray emission lines. To find the cause of chemical shift, various factors like effective charge, line intensity ratio, bond length and electro-negativity were calculated and correlated with the chemical shift.

EV Charging Algorithm Implementation with User Price Preference

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

Wang, Bin; Hu, Boyang; Qiu, Charlie

2015-02-17

in this paper, we propose and implement a smart Electric Vehicle (EV) charging algorithm to control the EV charging infrastructures according to users’ price preferences. EVSE (Electric Vehicle Supply Equipment), equipped with bidirectional communication devices and smart meters, can be remotely monitored by the proposed charging algorithm applied to EV control center and mobile app. On the server side, ARIMA model is utilized to fit historical charging load data and perform day-ahead prediction. A pricing strategy with energy bidding policy is proposed and implemented to generate a charging price list to be broadcasted to EV users through mobile app. Onmore » the user side, EV drivers can submit their price preferences and daily travel schedules to negotiate with Control Center to consume the expected energy and minimize charging cost simultaneously. The proposed algorithm is tested and validated through the experimental implementations in UCLA parking lots.« less

An electric vehicle driving behavior model in the traffic system with a wireless charging lane

NASA Astrophysics Data System (ADS)

He, Jia; Huang, Hai-Jun; Yang, Hai; Tang, Tie-Qiao

2017-09-01

In this paper, a car-following model is proposed to study each EV's (electric vehicle) motion behavior near the WCL (wireless charging lane) and a lane-changing rule is designed to describe the EV's lane-changing behavior. Then, the car-following model and lane-changing rule are used to explore each EV's micro driving behavior in a two-lane system with a WCL. Finally, the impacts of the WCL on each EV's motion behavior are investigated. The numerical results show that each EV should run slowly on the WCL if it needs charge of electricity, that the EV's lane-changing behavior has great effects on the whole system, that the delay time caused by the WCL turns more prominent when the traffic turns heavy, and that lane-changing frequently occurs near the WCL (especially at the downstream of the WCL).

Compact, Interactive Electric Vehicle Charger: Gallium-Nitride Switch Technology for Bi-directional Battery-to-Grid Charger Applications

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

None

2010-10-01

ADEPT Project: HRL Laboratories is using gallium nitride (GaN) semiconductors to create battery chargers for electric vehicles (EVs) that are more compact and efficient than traditional EV chargers. Reducing the size and weight of the battery charger is important because it would help improve the overall performance of the EV. GaN semiconductors process electricity faster than the silicon semiconductors used in most conventional EV battery chargers. These high-speed semiconductors can be paired with lighter-weight electrical circuit components, which helps decrease the overall weight of the EV battery charger. HRL Laboratories is combining the performance advantages of GaN semiconductors with anmore » innovative, interactive battery-to-grid energy distribution design. This design would support 2-way power flow, enabling EV battery chargers to not only draw energy from the power grid, but also store and feed energy back into it.« less

Extracellular Vesicles: How the External and Internal Environment Can Shape Cell-To-Cell Communication.

PubMed

Neven, Kristof Y; Nawrot, Tim S; Bollati, Valentina

2017-03-01

To summarize the scientific evidence regarding the effects of environmental exposures on extracellular vesicle (EV) release and their contents. As environmental exposures might influence the aging phenotype in a very strict way, we will also report the role of EVs in the biological aging process. EV research is a new and quickly developing field. With many investigations conducted so far, only a limited number of studies have explored the potential role EVs play in the response and adaptation to environmental stimuli. The investigations available to date have identified several exposures or lifestyle factors able to modify EV trafficking including air pollutants, cigarette smoke, alcohol, obesity, nutrition, physical exercise, and oxidative stress. EVs are a very promising tool, as biological fluids are easily obtainable biological media that, if successful in identifying early alterations induced by the environment and predictive of disease, would be amenable to use for potential future preventive and diagnostic applications.

Low dimensional CH3NH3PbBr3 cubes for persistent luminescence: Energy variation of electron excitation

NASA Astrophysics Data System (ADS)

Besral, N.; Paul, T.; Thakur, S.; Sarkar, S.; Sardar, K.; Chanda, K.; Das, A.; Chattopadhyay, K. K.

2018-04-01

The impact of varying electron beam voltage upon room temperature CL (cathodoluminescence) properties of crystalline organic-inorganic lead halide perovskite CH3NH3PbBr3 (Methylammonium lead tribromide) microcubes have been studied. CH3NH3PbBr3 microcubes were synthesized at room temperature by a very straight forward wet chemical route. After preliminary characterizations like XRD (X-ray diffraction), FESEM (Field emission scanning electron microscopy), UV-Vis spectroscopy, CL study at three different beam voltages i.e. 5 kV, 10 kV and 15 kV respectively was performed at room temperature. Prominent emission signals were obtained with emission peaks at 2.190 eV (FWHM 0.120 eV), 2.222 eV (FWHM 0.108 eV) and 2.242 eV (FWHM 0.095 eV) for electron beam voltages 5 kV, 10 kV and 15 kV respectively.

The cosmic ray spectrum and composition measured by KASCADE-Grande between 1016 eV and 1018 eV

NASA Astrophysics Data System (ADS)

Bertaina, M.; Apel, W. D.; Arteaga-Velázquez, J. C.; Bekk, K.; Blümer, J.; Bozdog, H.; Brancus, I. M.; Cantoni, E.; Chiavassa, A.; Cossavella, F.; Daumiller, K.; de Souza, V.; Di Pierro, F.; Doll, P.; Engel, R.; Engler, J.; Fuchs, B.; Fuhrmann, D.; Gherghel-Lascu, A.; Gils, H. J.; Glasstetter, R.; Grupen, C.; Haungs, A.; Heck, D.; Hörandel, J. R.; Huber, D.; Huege, T.; Kampert, K.-H.; Kang, D.; Klages, H. O.; Link, K.; Łuczak, P.; Mathes, H. J.; Mayer, H. J.; Milke, J.; Mitrica, B.; Morello, C.; Oehlschläger, J.; Ostapchenko, S.; Palmieri, N.; Petcu, M.; Pierog, T.; Rebel, H.; Roth, M.; Schieler, H.; Schoo, S.; Schröder, F. G.; Sima, O.; Toma, G.; Trinchero, G. C.; Ulrich, H.; Weindl, A.; Wochele, J.; Zabierowski, J.

2014-11-01

The shape and composition of the primary spectrum of cosmic rays are key elements to understand the origin, acceleration and propagation of the Galactic cosmic rays. Besides the well known knee and ankle features, the recent results of KASCADE-Grande indicate that the measured energy spectrum exhibits also a less pronounced but still clear deviation from a single power law between the knee and the ankle, with a spectral hardening at 2 × 1016 eV and a steepening at 1017 eV. The average mass composition gets heavier after the knee till 1017 eV where a bending of the heavy component is observed. An indication of a hardening of the light component just above 1017 eV has been measured as well. In this paper the major results obtained so far by the KASCADE-Grande experiment are reviewed.

Extracellular vesicles are independent metabolic units with asparaginase activity

PubMed Central

Leonardi, Tommaso; Costa, Ana S. H.; Cossetti, Chiara; Peruzzotti-Jametti, Luca; Bernstock, Joshua D.; Saini, Harpreet K.; Gelati, Maurizio; Vescovi, Angelo Luigi; Bastos, Carlos; Faria, Nuno; Occhipinti, Luigi G.; Enright, Anton J.; Frezza, Christian; Pluchino, Stefano

2017-01-01

Extracellular vesicles (EVs) are membrane particles involved in the exchange of a broad range of bioactive molecules between cells and the microenvironment. While it has been shown that cells can traffic metabolic enzymes via EVs much remains to be elucidated with regard to their intrinsic metabolic activity. Accordingly, herein we assessed the ability of neural stem/progenitor cell (NSC)-derived EVs to consume and produce metabolites. Both our metabolomics and functional analyses revealed that EVs harbour L-asparaginase activity catalysed by the enzyme Asparaginase-like protein 1 (Asrgl1). Critically, we show that Asrgl1 activity is selective for asparagine and is devoid of glutaminase activity. We found that mouse and human NSC-derived EVs traffic ASRGL1. Our results demonstrate for the first time that NSC EVs function as independent, extracellular metabolic units able to modify the concentrations of critical nutrients, with the potential to affect the physiology of their microenvironment. PMID:28671681

Effects of excess oxygen on the 4.5-6.3 eV absorption spectra of oxygen-rich high purity silica

NASA Astrophysics Data System (ADS)

Magruder, R. H.; Robinson, S. J.

2016-05-01

Type III silica samples were implanted with O using a multi-energy process that produced a layer of constant concentration to within ±5% beginning ∼80 nm from the surface and extending to ∼640 nm below the surfaces of the samples. The concentrations of excess oxygen in the layer ranged from 0.035 to ∼2.1at.%. In these samples we show that E‧ centers and NBOHCs, as well as the normal cadre of ODC (II) centers, were suppressed, and the optical absorption from 4.7 to 6.4 eV was primarily due to oxygen excess defects. Using Gaussian fitting techniques to examine the optical difference spectra, we have been able to identify four defect centers that are related to excess oxygen defect bands at 4.76 eV, 5.42 eV, 5.75 eV and 6.25 eV.

Therapeutic Applications of Extracellular Vesicles: Clinical Promise and Open Questions

PubMed Central

Breakefield, Xandra O.; Leonard, Joshua N.

2015-01-01

This review provides an updated perspective on rapidly proliferating efforts to harness extracellular vesicles (EVs) for therapeutic applications. We summarize current knowledge, emerging strategies, and open questions pertaining to clinical potential and translation. Potentially useful EVs comprise diverse products of various cell types and species. EV components may also be combined with liposomes and nanoparticles to facilitate manufacturing as well as product safety and evaluation. Potential therapeutic cargoes include RNA, proteins, and drugs. Strategic issues considered herein include choice of therapeutic agent, means of loading cargoes into EVs, promotion of EV stability, tissue targeting, and functional delivery of cargo to recipient cells. Some applications may harness natural EV properties, such as immune modulation, regeneration promotion, and pathogen suppression. These properties can be enhanced or customized to enable a wide range of therapeutic applications, including vaccination, improvement of pregnancy outcome, and treatment of autoimmune disease, cancer, and tissue injury. PMID:25292428