Modeling inhomogeneous DNA replication kinetics.

Directory of Open Access Journals (Sweden)

Michel G Gauthier

Full Text Available In eukaryotic organisms, DNA replication is initiated at a series of chromosomal locations called origins, where replication forks are assembled proceeding bidirectionally to replicate the genome. The distribution and firing rate of these origins, in conjunction with the velocity at which forks progress, dictate the program of the replication process. Previous attempts at modeling DNA replication in eukaryotes have focused on cases where the firing rate and the velocity of replication forks are homogeneous, or uniform, across the genome. However, it is now known that there are large variations in origin activity along the genome and variations in fork velocities can also take place. Here, we generalize previous approaches to modeling replication, to allow for arbitrary spatial variation of initiation rates and fork velocities. We derive rate equations for left- and right-moving forks and for replication probability over time that can be solved numerically to obtain the mean-field replication program. This method accurately reproduces the results of DNA replication simulation. We also successfully adapted our approach to the inverse problem of fitting measurements of DNA replication performed on single DNA molecules. Since such measurements are performed on specified portion of the genome, the examined DNA molecules may be replicated by forks that originate either within the studied molecule or outside of it. This problem was solved by using an effective flux of incoming replication forks at the model boundaries to represent the origin activity outside the studied region. Using this approach, we show that reliable inferences can be made about the replication of specific portions of the genome even if the amount of data that can be obtained from single-molecule experiments is generally limited.

Hydrogen Field Test Standard: Laboratory and Field Performance

Science.gov (United States)

Pope, Jodie G.; Wright, John D.

2015-01-01

The National Institute of Standards and Technology (NIST) developed a prototype field test standard (FTS) that incorporates three test methods that could be used by state weights and measures inspectors to periodically verify the accuracy of retail hydrogen dispensers, much as gasoline dispensers are tested today. The three field test methods are: 1) gravimetric, 2) Pressure, Volume, Temperature (PVT), and 3) master meter. The FTS was tested in NIST's Transient Flow Facility with helium gas and in the field at a hydrogen dispenser location. All three methods agree within 0.57 % and 1.53 % for all test drafts of helium gas in the laboratory setting and of hydrogen gas in the field, respectively. The time required to perform six test drafts is similar for all three methods, ranging from 6 h for the gravimetric and master meter methods to 8 h for the PVT method. The laboratory tests show that 1) it is critical to wait for thermal equilibrium to achieve density measurements in the FTS that meet the desired uncertainty requirements for the PVT and master meter methods; in general, we found a wait time of 20 minutes introduces errors methods, respectively and 2) buoyancy corrections are important for the lowest uncertainty gravimetric measurements. The field tests show that sensor drift can become a largest component of uncertainty that is not present in the laboratory setting. The scale was calibrated after it was set up at the field location. Checks of the calibration throughout testing showed drift of 0.031 %. Calibration of the master meter and the pressure sensors prior to travel to the field location and upon return showed significant drifts in their calibrations; 0.14 % and up to 1.7 %, respectively. This highlights the need for better sensor selection and/or more robust sensor testing prior to putting into field service. All three test methods are capable of being successfully performed in the field and give equivalent answers if proper sensors without drift are

Replication stress-induced chromosome breakage is correlated with replication fork progression and is preceded by single-stranded DNA formation.

Science.gov (United States)

Feng, Wenyi; Di Rienzi, Sara C; Raghuraman, M K; Brewer, Bonita J

2011-10-01

Chromosome breakage as a result of replication stress has been hypothesized to be the direct consequence of defective replication fork progression, or "collapsed" replication forks. However, direct and genome-wide evidence that collapsed replication forks give rise to chromosome breakage is still lacking. Previously we showed that a yeast replication checkpoint mutant mec1-1, after transient exposure to replication impediment imposed by hydroxyurea (HU), failed to complete DNA replication, accumulated single-stranded DNA (ssDNA) at the replication forks, and fragmented its chromosomes. In this study, by following replication fork progression genome-wide via ssDNA detection and by direct mapping of chromosome breakage after HU exposure, we have tested the hypothesis that the chromosome breakage in mec1 cells occurs at collapsed replication forks. We demonstrate that sites of chromosome breakage indeed correlate with replication fork locations. Moreover, ssDNA can be detected prior to chromosome breakage, suggesting that ssDNA accumulation is the common precursor to double strand breaks at collapsed replication forks.

Effects of 60 Hz electromagnetic fields on early growth in three plant species and a replication of previous results

Energy Technology Data Exchange (ETDEWEB)

Davis, M.S. [Univ. of Sunderland (United Kingdom). Ecology Centre

1996-05-01

In an attempt to replicate the findings of Smith et al., seeds of Raphanus sativus L. (radish), Sinapsis alba L. (mustard), and Hordeum vulgare L. (barley) were grown for between 9 and 21 days in continuous electromagnetic fields (EMFs) at ion-cyclotron resonance conditions for stimulation of Ca{sup 2+} (B{sub H} = 78.3 {micro}T, B{sub HAC} = 40 {micro}T peak-peak at 60 Hz, B{sub v} = 0). On harvesting, radish showed results similar to those of Smith et al. Dry stem weight and plant height were both significantly greater (Mann-Whitney tests, Ps < 0.05) in EMF-exposed plants than in control plants in each EMF experiment. Wet root weight was significantly greater in EMF-exposed plants in two out of three experiments, as were dry leaf weight, dry whole weight, and stem diameter. Dry root weight, wet leaf weight, and wet whole weight were significantly greater in EMF-exposed plants in one of three experiments. All significant differences indicated an increase in weight or size in the EMF-exposed plants. In each of the sham experiments, no differences between exposed and control plants were evident. Mustard plants failed to respond to the EMFs in any of the plant parameters measured. In one experiment, barley similarly failed to respond; but in another showed significantly greater wet root weight and significantly smaller stem diameter and dry seed weight at the end of the experiment in exposed plants compared to control plants. Although these results give no clue about the underlying bioelectromagnetic mechanism, they demonstrate that, at least for one EMF-sensitive biosystem, results can be independently replicated in another laboratory. Such replication is crucial in establishing the validity of bioelectromagnetic science.

Repeatability study of replicate crash tests: A signal analysis approach.

Science.gov (United States)

Seppi, Jeremy; Toczyski, Jacek; Crandall, Jeff R; Kerrigan, Jason

2017-10-03

To provide an objective basis on which to evaluate the repeatability of vehicle crash test methods, a recently developed signal analysis method was used to evaluate correlation of sensor time history data between replicate vehicle crash tests. The goal of this study was to evaluate the repeatability of rollover crash tests performed with the Dynamic Rollover Test System (DRoTS) relative to other vehicle crash test methods. Test data from DRoTS tests, deceleration rollover sled (DRS) tests, frontal crash tests, frontal offset crash tests, small overlap crash tests, small overlap impact (SOI) crash tests, and oblique crash tests were obtained from the literature and publicly available databases (the NHTSA vehicle database and the Insurance Institute for Highway Safety TechData) to examine crash test repeatability. Signal analysis of the DRoTS tests showed that force and deformation time histories had good to excellent repeatability, whereas vehicle kinematics showed only fair repeatability due to the vehicle mounting method for one pair of tests and slightly dissimilar mass properties (2.2%) in a second pair of tests. Relative to the DRS, the DRoTS tests showed very similar or higher levels of repeatability in nearly all vehicle kinematic data signals with the exception of global X' (road direction of travel) velocity and displacement due to the functionality of the DRoTS fixture. Based on the average overall scoring metric of the dominant acceleration, DRoTS was found to be as repeatable as all other crash tests analyzed. Vertical force measures showed good repeatability and were on par with frontal crash barrier forces. Dynamic deformation measures showed good to excellent repeatability as opposed to poor repeatability seen in SOI and oblique deformation measures. Using the signal analysis method as outlined in this article, the DRoTS was shown to have the same or better repeatability of crash test methods used in government regulatory and consumer evaluation test

Transformation and Tumorigenicity Testing of Simian Cell Lines and Evaluation of Poliovirus Replication.

Directory of Open Access Journals (Sweden)

Silvia Dotti

Full Text Available The key role of cell cultures in different scientific fields is worldwide recognized, both as in vitro research models alternative to laboratory animals and substrates for biological production. However, many safety concerns rise from the use of animal/human cell lines that may be tumorigenic, leading to potential adverse contaminations in cell-derived biologicals. In order to evaluate the suitability of 13 different cell lines for Poliovirus vaccine production, safety and quality, in vitro/in vivo tumorigenicity and Poliovirus propagation properties were evaluated. Our results revealed that non-human primate cell lines CYNOM-K1, FRhK-4, 4MBr-5 and 4647 are free of tumorigenic features and represent highly susceptible substrates for attenuated Sabin Poliovirus strains. In particular, FRhK-4 and 4647 cell lines are characterized by a higher in vitro replication, resulting indicated for the use in large-scale production field.

A Successful Test of Parallel Replication Teams in Teaching Research Methods

Science.gov (United States)

Standing, Lionel G.; Astrologo, Lisa; Benbow, Felecia F.; Cyr-Gauthier, Chelsea S.; Williams, Charlotte A.

2016-01-01

This paper describes the novel use of parallel student teams from a research methods course to perform a replication study, and suggests that this approach offers pedagogical benefits for both students and teachers, as well as potentially contributing to a resolution of the replication crisis in psychology today. Four teams, of five undergraduates…

Reconstructing solar magnetic fields from historical observations: Testing the surface flux transport model

Science.gov (United States)

Virtanen, Iiro; Virtanen, Ilpo; Pevtsov, Alexei; Yeates, Anthony; Mursula, Kalevi

2017-04-01

We aim to use the surface flux transport model to simulate the long-term evolution of the photospheric magnetic field from historical observations. In this work we study the accuracy of the model and its sensitivity to uncertainties in its main parameters and the input data. We test the model by running simulations with different values of meridional circulation and supergranular diffusion parameters, and study how the flux distribution inside active regions and the initial magnetic field affect the simulation. We compare the results to assess how sensitive the simulation is to uncertainties in meridional circulation speed, supergranular diffusion and input data. We also compare the simulated magnetic field with observations. We find that there is generally good agreement between simulations and observations. While the model is not capable of replicating fine details of the magnetic field, the long-term evolution of the polar field is very similar in simulations and observations. Simulations typically yield a smoother evolution of polar fields than observations, that often include artificial variations due to observational limitations. We also find that the simulated field is fairly insensitive to uncertainties in model parameters or the input data. Due to the decay term included in the model the effects of the uncertainties are rather minor or temporary, lasting typically one solar cycle.

Direct Observation of Field and Temperature Induced Domain Replication in Dipolar Coupled Perpendicular Anisotropy Films

Energy Technology Data Exchange (ETDEWEB)

Hauet, T.; Gunther, C.M.; Pfau, B.; Eisebitt, S.; Fischer, P.; Rick, R. L.; Thiele, J.-U.; Hellwig, O.; Schabes, M.E.

2007-07-01

Dipolar interactions in a soft/Pd/hard [CoNi/Pd]{sub 30}/Pd/[Co/Pd]{sub 20} multilayer system, where a thick Pd layer between two ferromagnetic units prevents direct exchange coupling, are directly revealed by combining magnetometry and state-of-the-art layer resolving soft x-ray imaging techniques with sub-100-nm spatial resolution. The domains forming in the soft layer during external magnetic field reversal are found to match the domains previously trapped in the hard layer. The low Curie temperature of the soft layer allows varying its intrinsic parameters via temperature and thus studying the competition with dipolar fields due to the domains in the hard layer. Micromagnetic simulations elucidate the role of [CoNi/Pd] magnetization, exchange, and anisotropy in the duplication process. Finally, thermally driven domain replication in remanence during temperature cycling is demonstrated.

Using Replicates in Information Retrieval Evaluation.

Science.gov (United States)

Voorhees, Ellen M; Samarov, Daniel; Soboroff, Ian

2017-09-01

This article explores a method for more accurately estimating the main effect of the system in a typical test-collection-based evaluation of information retrieval systems, thus increasing the sensitivity of system comparisons. Randomly partitioning the test document collection allows for multiple tests of a given system and topic (replicates). Bootstrap ANOVA can use these replicates to extract system-topic interactions-something not possible without replicates-yielding a more precise value for the system effect and a narrower confidence interval around that value. Experiments using multiple TREC collections demonstrate that removing the topic-system interactions substantially reduces the confidence intervals around the system effect as well as increases the number of significant pairwise differences found. Further, the method is robust against small changes in the number of partitions used, against variability in the documents that constitute the partitions, and the measure of effectiveness used to quantify system effectiveness.

The Alleged Crisis and the Illusion of Exact Replication.

Science.gov (United States)

Stroebe, Wolfgang; Strack, Fritz

2014-01-01

There has been increasing criticism of the way psychologists conduct and analyze studies. These critiques as well as failures to replicate several high-profile studies have been used as justification to proclaim a "replication crisis" in psychology. Psychologists are encouraged to conduct more "exact" replications of published studies to assess the reproducibility of psychological research. This article argues that the alleged "crisis of replicability" is primarily due to an epistemological misunderstanding that emphasizes the phenomenon instead of its underlying mechanisms. As a consequence, a replicated phenomenon may not serve as a rigorous test of a theoretical hypothesis because identical operationalizations of variables in studies conducted at different times and with different subject populations might test different theoretical constructs. Therefore, we propose that for meaningful replications, attempts at reinstating the original circumstances are not sufficient. Instead, replicators must ascertain that conditions are realized that reflect the theoretical variable(s) manipulated (and/or measured) in the original study. © The Author(s) 2013.

Replication and robustness in developmental research.

Science.gov (United States)

Duncan, Greg J; Engel, Mimi; Claessens, Amy; Dowsett, Chantelle J

2014-11-01

Replications and robustness checks are key elements of the scientific method and a staple in many disciplines. However, leading journals in developmental psychology rarely include explicit replications of prior research conducted by different investigators, and few require authors to establish in their articles or online appendices that their key results are robust across estimation methods, data sets, and demographic subgroups. This article makes the case for prioritizing both explicit replications and, especially, within-study robustness checks in developmental psychology. It provides evidence on variation in effect sizes in developmental studies and documents strikingly different replication and robustness-checking practices in a sample of journals in developmental psychology and a sister behavioral science-applied economics. Our goal is not to show that any one behavioral science has a monopoly on best practices, but rather to show how journals from a related discipline address vital concerns of replication and generalizability shared by all social and behavioral sciences. We provide recommendations for promoting graduate training in replication and robustness-checking methods and for editorial policies that encourage these practices. Although some of our recommendations may shift the form and substance of developmental research articles, we argue that they would generate considerable scientific benefits for the field. (PsycINFO Database Record (c) 2014 APA, all rights reserved).

Structure of replicating intermediates of human herpesvirus type 6

International Nuclear Information System (INIS)

Severini, Alberto; Sevenhuysen, Claire; Garbutt, Michael; Tipples, Graham A.

2003-01-01

We have studied the structure of the replicative intermediates of human herpesvirus 6 (HHV-6) using pulsed-field gel electrophoresis, partial digestion, two-dimensional gel electrophoresis, and sedimentation centrifugation. The results show that DNA replication of HHV-6 produces head-to-tail concatemeric intermediates as well as approximately equal amounts of circular monomers or oligomers. Unlike the situation in herpes simplex virus, the intermediates of human herpesvirus 6 replication are not highly branched, suggesting a difference in the mechanism of replication or a lower frequency of homologous recombination in human herpesvirus 6 compared to herpes simplex virus

Test-potentiated learning: three independent replications, a disconfirmed hypothesis, and an unexpected boundary condition.

Science.gov (United States)

Wissman, Kathryn T; Rawson, Katherine A

2018-04-01

Arnold and McDermott [(2013). Test-potentiated learning: Distinguishing between direct and indirect effects of testing. Journal of Experimental Psychology: Learning, Memory, and Cognition, 39, 940-945] isolated the indirect effects of testing and concluded that encoding is enhanced to a greater extent following more versus fewer practice tests, referred to as test-potentiated learning. The current research provided further evidence for test-potentiated learning and evaluated the covert retrieval hypothesis as an alternative explanation for the observed effect. Learners initially studied foreign language word pairs and then completed either one or five practice tests before restudy occurred. Results of greatest interest concern performance on test trials following restudy for items that were not correctly recalled on the test trials that preceded restudy. Results replicate Arnold and McDermott (2013) by demonstrating that more versus fewer tests potentiate learning when trial time is limited. Results also provide strong evidence against the covert retrieval hypothesis concerning why the effect occurs (i.e., it does not reflect differential covert retrieval during pre-restudy trials). In addition, outcomes indicate that the magnitude of the test-potentiated learning effect decreases as trial length increases, revealing an unexpected boundary condition to test-potentiated learning.

A field test of a waste containment technology using a new generation of injectable barrier liquids

International Nuclear Information System (INIS)

Moridis, G.; Apps, J.; Persoff, P.; Myer, L.; Muller, S.; Pruess, K.; Yen, P.

1996-08-01

A first stage field injection of a new generation of barrier liquids was successfully completed. Two types of barrier liquids, colloidal silica (CS) and polysiloxane (PSX), were injected into heterogeneous unsaturated deposits of sand, silt, and gravel typical of many of the arid DOE cleanup sites and particularly analogous to the conditions of the Hanford Site. Successful injection by commercially available chemical grouting equipment and the tube-a-manchette technique was demonstrated. Excavation of the grout bulbs permitted visual evaluation of the soil permeation by the grout, as well as sample collection. Both grouts effectively permeated all of the formation. The PSX visually appeared to perform better, producing a more uniform and symmetric permeation regardless of heterogeneity, filling large as well as small pores and providing more structural strength than the CS. Numerical simulation of the injection tests incorporated a stochastic field to represent site heterogeneity and was able to replicate the general test behavior. Tiltmeters were used successfully to monitor surface displacements during grout injection

Evaluation of field test equipment for halide and DOP testing

International Nuclear Information System (INIS)

Schreiber, K.L.; Kovach, J.L.

1975-01-01

The Nucon Testing Services Department, field testing at power reactor sites, has performed tests using R-11, R-12, and R-112 in conjunction with gas chromatographs and direct reading halide detectors. The field operational experience with these detector systems, thus sensitivity, precision, and manner of field calibration, are presented. Laboratory experiments regarding 3 H-tagged methyl iodide for in place leak testing of adsorber systems indicate a low hazard, high reliability process for leak testing in facilities where atmospheric cross contamination occurs. (U.S.)

Toward a model for field-testing patient decision-support technologies: a qualitative field-testing study.

NARCIS (Netherlands)

Evans, R.; Elwyn, G.; Edwards, A.; Watson, E.; Austoker, J.; Grol, R.P.T.M.

2007-01-01

BACKGROUND: Field-testing is a quality assurance criterion in the development of patient decision-support technologies (PDSTs), as identified in the consensus statement of the International Patient Decision Aids Standards Collaboration. We incorporated field-testing into the development of a

Background field coils for the High Field Test Facility

International Nuclear Information System (INIS)

Zbasnik, J.P.; Cornish, D.N.; Scanlan, R.M.; Jewell, A.M.; Leber, R.L.; Rosdahl, A.R.; Chaplin, M.R.

1980-01-01

The High Field Test Facility (HFTF), presently under construction at LLNL, is a set of superconducting coils that will be used to test 1-m-o.d. coils of prototype conductors for fusion magnets in fields up to 12 T. The facility consists of two concentric sets of coils; the outer set is a stack of Nb-Ti solenoids, and the inner set is a pair of solenoids made of cryogenically-stabilized, multifilamentary Nb 3 Sn superconductor, developed for use in mirror-fusion magnets. The HFTF system is designed to be parted along the midplane to allow high-field conductors, under development for Tokamak fusion machines, to be inserted and tested. The background field coils were wound pancake-fashion, with cold-welded joints at both the inner and outer diameters. Turn-to-turn insulation was fabricated at LLNL from epoxy-fiberglass strip. The coils were assembled and tested in our 2-m-diam cryostat to verify their operation

The four-meter confrontation visual field test.

OpenAIRE

Kodsi, S R; Younge, B R

1992-01-01

The 4-m confrontation visual field test has been successfully used at the Mayo Clinic for many years in addition to the standard 0.5-m confrontation visual field test. The 4-m confrontation visual field test is a test of macular function and can identify small central or paracentral scotomas that the examiner may not find when the patient is tested only at 0.5 m. Also, macular sparing in homonymous hemianopias and quadrantanopias may be identified with the 4-m confrontation visual field test....

Distributional Replication

OpenAIRE

Beare, Brendan K.

2009-01-01

Suppose that X and Y are random variables. We define a replicating function to be a function f such that f(X) and Y have the same distribution. In general, the set of replicating functions for a given pair of random variables may be infinite. Suppose we have some objective function, or cost function, defined over the set of replicating functions, and we seek to estimate the replicating function with the lowest cost. We develop an approach to estimating the cheapest replicating function that i...

How many bootstrap replicates are necessary?

Science.gov (United States)

Pattengale, Nicholas D; Alipour, Masoud; Bininda-Emonds, Olaf R P; Moret, Bernard M E; Stamatakis, Alexandros

2010-03-01

Phylogenetic bootstrapping (BS) is a standard technique for inferring confidence values on phylogenetic trees that is based on reconstructing many trees from minor variations of the input data, trees called replicates. BS is used with all phylogenetic reconstruction approaches, but we focus here on one of the most popular, maximum likelihood (ML). Because ML inference is so computationally demanding, it has proved too expensive to date to assess the impact of the number of replicates used in BS on the relative accuracy of the support values. For the same reason, a rather small number (typically 100) of BS replicates are computed in real-world studies. Stamatakis et al. recently introduced a BS algorithm that is 1 to 2 orders of magnitude faster than previous techniques, while yielding qualitatively comparable support values, making an experimental study possible. In this article, we propose stopping criteria--that is, thresholds computed at runtime to determine when enough replicates have been generated--and we report on the first large-scale experimental study to assess the effect of the number of replicates on the quality of support values, including the performance of our proposed criteria. We run our tests on 17 diverse real-world DNA--single-gene as well as multi-gene--datasets, which include 125-2,554 taxa. We find that our stopping criteria typically stop computations after 100-500 replicates (although the most conservative criterion may continue for several thousand replicates) while producing support values that correlate at better than 99.5% with the reference values on the best ML trees. Significantly, we also find that the stopping criteria can recommend very different numbers of replicates for different datasets of comparable sizes. Our results are thus twofold: (i) they give the first experimental assessment of the effect of the number of BS replicates on the quality of support values returned through BS, and (ii) they validate our proposals for

The evolutionary ecology of molecular replicators.

Science.gov (United States)

Nee, Sean

2016-08-01

By reasonable criteria, life on the Earth consists mainly of molecular replicators. These include viruses, transposons, transpovirons, coviruses and many more, with continuous new discoveries like Sputnik Virophage. Their study is inherently multidisciplinary, spanning microbiology, genetics, immunology and evolutionary theory, and the current view is that taking a unified approach has great power and promise. We support this with a new, unified, model of their evolutionary ecology, using contemporary evolutionary theory coupling the Price equation with game theory, studying the consequences of the molecular replicators' promiscuous use of each others' gene products for their natural history and evolutionary ecology. Even at this simple expository level, we can make a firm prediction of a new class of replicators exploiting viruses such as lentiviruses like SIVs, a family which includes HIV: these have been explicitly stated in the primary literature to be non-existent. Closely connected to this departure is the view that multicellular organism immunology is more about the management of chronic infections rather than the elimination of acute ones and new understandings emerging are changing our view of the kind of theatre we ourselves provide for the evolutionary play of molecular replicators. This study adds molecular replicators to bacteria in the emerging field of sociomicrobiology.

Termination of DNA replication forks: "Breaking up is hard to do".

Science.gov (United States)

Bailey, Rachael; Priego Moreno, Sara; Gambus, Agnieszka

2015-01-01

To ensure duplication of the entire genome, eukaryotic DNA replication initiates from thousands of replication origins. The replication forks move through the chromatin until they encounter forks from neighboring origins. During replication fork termination forks converge, the replisomes disassemble and topoisomerase II resolves the daughter DNA molecules. If not resolved efficiently, terminating forks result in genomic instability through the formation of pathogenic structures. Our recent findings shed light onto the mechanism of replisome disassembly upon replication fork termination. We have shown that termination-specific polyubiquitylation of the replicative helicase component - Mcm7, leads to dissolution of the active helicase in a process dependent on the p97/VCP/Cdc48 segregase. The inhibition of terminating helicase disassembly resulted in a replication termination defect. In this extended view we present hypothetical models of replication fork termination and discuss remaining and emerging questions in the DNA replication termination field.

Chromatin Immunoprecipitation of Replication Factors Moving with the Replication Fork

OpenAIRE

Rapp, Jordan B.; Ansbach, Alison B.; Noguchi, Chiaki; Noguchi, Eishi

2009-01-01

Replication of chromosomes involves a variety of replication proteins including DNA polymerases, DNA helicases, and other accessory factors. Many of these proteins are known to localize at replication forks and travel with them as components of the replisome complex. Other proteins do not move with replication forks but still play an essential role in DNA replication. Therefore, in order to understand the mechanisms of DNA replication and its controls, it is important to examine localization ...

Open-field test site

Science.gov (United States)

Gyoda, Koichi; Shinozuka, Takashi

1995-06-01

An open-field test site with measurement equipment, a turn table, antenna positioners, and measurement auxiliary equipment was remodelled at the CRL north-site. This paper introduces the configuration, specifications and characteristics of this new open-field test site. Measured 3-m and 10-m site attenuations are in good agreement with theoretical values, and this means that this site is suitable for using 3-m and 10-m method EMI/EMC measurements. The site is expected to be effective for antenna measurement, antenna calibration, and studies on EMI/EMC measurement methods.

A field test of the effect of spiked ivermectin concentrations on the biodiversity of coprophagous dung insects in Switzerland.

Science.gov (United States)

Jochmann, Ralf; Lipkow, Erhard; Blanckenhorn, Wolf U

2016-08-01

Veterinary medical product residues can cause severe damage in the dung ecosystem. Depending on the manner of application and the time after treatment, the excreted concentration of a given pharmaceutical varies. The popular anthelmintic drug ivermectin can be applied to livestock in several different ways and is fecally excreted over a period of days to months after application. In a field experiment replicated in summer and autumn, the authors mixed 6 ivermectin concentrations plus a null control into fresh cow dung to assess the reaction of the dung insect community. Taxon richness of the insect dung fauna emerging from the dung, but not Hill diversity ((1) D) or the total number of individuals (abundance), decreased as ivermectin concentration increased. Corresponding declines in the number of emerging insects were found for most larger brachyceran flies and hymenopteran parasitoids, but not for most smaller nematoceran flies or beetles (except Hydrophilidae). Parallel pitfall traps recovered all major dung organism groups that emerged from the experimental dung, although at times in vastly different numbers. Ivermectin generally did not change the attractiveness of dung: differences in emergence therefore reflect differences in survival of coprophagous offspring of colonizing insects. Because sample size was limited to 6 replicates, the authors generally recommend more than 10 (seasonal) replicates and also testing higher concentrations than used in the present study as positive controls in future studies. Results accord with parallel experiments in which the substance was applied and passed through the cow's digestive system. In principle, therefore, the authors' experimental design is suitable for such higher-tier field tests of the response of the entire dung community to pharmaceutical residues, at least for ivermectin. Environ Toxicol Chem 2016;35:1947-1952. © 2015 SETAC. © 2015 SETAC.

Replication Catastrophe

DEFF Research Database (Denmark)

Toledo, Luis; Neelsen, Kai John; Lukas, Jiri

2017-01-01

Proliferating cells rely on the so-called DNA replication checkpoint to ensure orderly completion of genome duplication, and its malfunction may lead to catastrophic genome disruption, including unscheduled firing of replication origins, stalling and collapse of replication forks, massive DNA...... breakage, and, ultimately, cell death. Despite many years of intensive research into the molecular underpinnings of the eukaryotic replication checkpoint, the mechanisms underlying the dismal consequences of its failure remain enigmatic. A recent development offers a unifying model in which the replication...... checkpoint guards against global exhaustion of rate-limiting replication regulators. Here we discuss how such a mechanism can prevent catastrophic genome disruption and suggest how to harness this knowledge to advance therapeutic strategies to eliminate cancer cells that inherently proliferate under...

The use of statistical tools in field testing of putative effects of genetically modified plants on nontarget organisms.

Science.gov (United States)

Semenov, Alexander V; Elsas, Jan Dirk; Glandorf, Debora C M; Schilthuizen, Menno; Boer, Willem F

2013-08-01

To fulfill existing guidelines, applicants that aim to place their genetically modified (GM) insect-resistant crop plants on the market are required to provide data from field experiments that address the potential impacts of the GM plants on nontarget organisms (NTO's). Such data may be based on varied experimental designs. The recent EFSA guidance document for environmental risk assessment (2010) does not provide clear and structured suggestions that address the statistics of field trials on effects on NTO's. This review examines existing practices in GM plant field testing such as the way of randomization, replication, and pseudoreplication. Emphasis is placed on the importance of design features used for the field trials in which effects on NTO's are assessed. The importance of statistical power and the positive and negative aspects of various statistical models are discussed. Equivalence and difference testing are compared, and the importance of checking the distribution of experimental data is stressed to decide on the selection of the proper statistical model. While for continuous data (e.g., pH and temperature) classical statistical approaches - for example, analysis of variance (ANOVA) - are appropriate, for discontinuous data (counts) only generalized linear models (GLM) are shown to be efficient. There is no golden rule as to which statistical test is the most appropriate for any experimental situation. In particular, in experiments in which block designs are used and covariates play a role GLMs should be used. Generic advice is offered that will help in both the setting up of field testing and the interpretation and data analysis of the data obtained in this testing. The combination of decision trees and a checklist for field trials, which are provided, will help in the interpretation of the statistical analyses of field trials and to assess whether such analyses were correctly applied. We offer generic advice to risk assessors and applicants that will

Testing the Efficacy of a Tier 2 Mathematics Intervention: A Conceptual Replication Study

Science.gov (United States)

Doabler, Christian T.; Clarke, Ben; Kosty, Derek B.; Kurtz-Nelson, Evangeline; Fien, Hank; Smolkowski, Keith; Baker, Scott K.

2016-01-01

The purpose of this closely aligned conceptual replication study was to investigate the efficacy of a Tier 2 kindergarten mathematics intervention. The replication study differed from the initial randomized controlled trial on three important elements: geographical region, timing of the intervention, and instructional context of the…

Reconstructing solar magnetic fields from historical observations. II. Testing the surface flux transport model

Science.gov (United States)

Virtanen, I. O. I.; Virtanen, I. I.; Pevtsov, A. A.; Yeates, A.; Mursula, K.

2017-07-01

Aims: We aim to use the surface flux transport model to simulate the long-term evolution of the photospheric magnetic field from historical observations. In this work we study the accuracy of the model and its sensitivity to uncertainties in its main parameters and the input data. Methods: We tested the model by running simulations with different values of meridional circulation and supergranular diffusion parameters, and studied how the flux distribution inside active regions and the initial magnetic field affected the simulation. We compared the results to assess how sensitive the simulation is to uncertainties in meridional circulation speed, supergranular diffusion, and input data. We also compared the simulated magnetic field with observations. Results: We find that there is generally good agreement between simulations and observations. Although the model is not capable of replicating fine details of the magnetic field, the long-term evolution of the polar field is very similar in simulations and observations. Simulations typically yield a smoother evolution of polar fields than observations, which often include artificial variations due to observational limitations. We also find that the simulated field is fairly insensitive to uncertainties in model parameters or the input data. Due to the decay term included in the model the effects of the uncertainties are somewhat minor or temporary, lasting typically one solar cycle.

Materials Chemistry and Performance of Silicone-Based Replicating Compounds.

Energy Technology Data Exchange (ETDEWEB)

Brumbach, Michael T.; Mirabal, Alex James; Kalan, Michael; Trujillo, Ana B; Hale, Kevin

2014-11-01

Replicating compounds are used to cast reproductions of surface features on a variety of materials. Replicas allow for quantitative measurements and recordkeeping on parts that may otherwise be difficult to measure or maintain. In this study, the chemistry and replicating capability of several replicating compounds was investigated. Additionally, the residue remaining on material surfaces upon removal of replicas was quantified. Cleaning practices were tested for several different replicating compounds. For all replicating compounds investigated, a thin silicone residue was left by the replica. For some compounds, additional inorganic species could be identified in the residue. Simple solvent cleaning could remove some residue.

Evaluating the reproducibility of environmental radioactivity monitoring data through replicate sample analysis

International Nuclear Information System (INIS)

Lindeken, C.L.; White, J.H.; Silver, W.J.

1978-01-01

At the Lawrence Livermore Laboratory, about 10% of the sampling effort in the environmental monitoring program represents replicate sample collection. Replication of field samples was initiated as part of the quality assurance program for environmental monitoring to determine the reproducibility of environmental measurements. In the laboratory these replicates are processed along with routine samples. As all components of variance are included in analysis of such field samples, comparison of the analytical data from replicate analyses provides a basis for estimating the overall reproducibility of the measurements. The replication study indicates that the reproducibility of environmental radioactivity monitoring data is subject to considerably more variability than is indicated by the accompanying counting errors. The data are also compared with analyses of duplicate aliquots from a well mixed sample or with duplicate aliquots of samples with known radionuclide content. These comparisons show that most of the variability is associated with the collection and preparation of the sample rather than with the analytical procedures

Birth Order and Field Dependence-Independence: A Failure to Replicate

Science.gov (United States)

Finley, Gordon E.; Solla, Joseph

1975-01-01

The Children's Embedded Figures Test was individually administered to 116 Caucasian, middle class, second grade children. Results suggest that a child's early experience in a particular birth order position may not be related to the development of field dependence-independence in any unambiguous and simple fashion. (Author/ED)

Database Replication Prototype

OpenAIRE

Vandewall, R.

2000-01-01

This report describes the design of a Replication Framework that facilitates the implementation and com-parison of database replication techniques. Furthermore, it discusses the implementation of a Database Replication Prototype and compares the performance measurements of two replication techniques based on the Atomic Broadcast communication primitive: pessimistic active replication and optimistic active replication. The main contributions of this report can be split into four parts....

A Replication of the Internal Validity Structure of Three Major Teaching Rating Scales

Science.gov (United States)

Peters, Scott J.; Pereira, Nielsen

2017-01-01

Even as the importance of replication research has become more widely understood, the field of gifted education is almost completely devoid of replication studies. An area in which replication is a particular problem is in student identification research, since instrument validity is a necessary prerequisite for any sound psychometric decision. To…

Character relations and replication identities in 2d Conformal Field Theory

Energy Technology Data Exchange (ETDEWEB)

Bantay, P. [Institute for Theoretical Physics, Eötvös Loránd University,H-1117 Budapest, Pázmány P.s. 1/A (Hungary)

2016-10-05

We study replication identities satisfied by conformal characters of a 2D CFT, providing a natural framework for a physics interpretation of the famous Hauptmodul property of Monstrous Moonshine, and illustrate the underlying ideas in simple cases.

LHCb Data Replication During SC3

CERN Multimedia

Smith, A

2006-01-01

LHCb's participation in LCG's Service Challenge 3 involves testing the bulk data transfer infrastructure developed to allow high bandwidth distribution of data across the grid in accordance with the computing model. To enable reliable bulk replication of data, LHCb's DIRAC system has been integrated with gLite's File Transfer Service middleware component to make use of dedicated network links between LHCb computing centres. DIRAC's Data Management tools previously allowed the replication, registration and deletion of files on the grid. For SC3 supplementary functionality has been added to allow bulk replication of data (using FTS) and efficient mass registration to the LFC replica catalog.Provisional performance results have shown that the system developed can meet the expected data replication rate required by the computing model in 2007. This paper details the experience and results of integration and utilisation of DIRAC with the SC3 transfer machinery.

The Replication Recipe: What makes for a convincing replication?

NARCIS (Netherlands)

Brandt, M.J.; IJzerman, H.; Dijksterhuis, A.J.; Farach, F.J.; Geller, J.; Giner-Sorolla, R.; Grange, J.A.; Perugini, M.; Spies, J.R.; Veer, A. van 't

2014-01-01

Psychological scientists have recently started to reconsider the importance of close replications in building a cumulative knowledge base; however, there is no consensus about what constitutes a convincing close replication study. To facilitate convincing close replication attempts we have developed

The replication recipe : What makes for a convincing replication?

NARCIS (Netherlands)

Brandt, M.J.; IJzerman, H.; Dijksterhuis, Ap; Farach, Frank J.; Geller, Jason; Giner-Sorolla, Roger; Grange, James A.; Perugini, Marco; Spies, Jeffrey R.; van 't Veer, Anna

Psychological scientists have recently started to reconsider the importance of close replications in building a cumulative knowledge base; however, there is no consensus about what constitutes a convincing close replication study. To facilitate convincing close replication attempts we have developed

Useful field of view test.

Science.gov (United States)

Wood, Joanne M; Owsley, Cynthia

2014-01-01

The useful field of view test was developed to reflect the visual difficulties that older adults experience with everyday tasks. Importantly, the useful field of view test (UFOV) is one of the most extensively researched and promising predictor tests for a range of driving outcomes measures, including driving ability and crash risk as well as other everyday tasks. Currently available commercial versions of the test can be administered using personal computers; these measure the speed of visual processing for rapid detection and localization of targets under conditions of divided visual attention and in the presence and absence of visual clutter. The test is believed to assess higher-order cognitive abilities, but performance also relies on visual sensory function because in order for targets to be attended to, they must be visible. The format of the UFOV has been modified over the years; the original version estimated the spatial extent of useful field of view, while the latest version measures visual processing speed. While deficits in the useful field of view are associated with functional impairments in everyday activities in older adults, there is also emerging evidence from several research groups that improvements in visual processing speed can be achieved through training. These improvements have been shown to reduce crash risk, and can have a positive impact on health and functional well-being, with the potential to increase the mobility and hence the independence of older adults. © 2014 S. Karger AG, Basel

Registered Replication Report: Dijksterhuis and van Knippenberg (1998).

Science.gov (United States)

O'Donnell, Michael; Nelson, Leif D; Ackermann, Evi; Aczel, Balazs; Akhtar, Athfah; Aldrovandi, Silvio; Alshaif, Nasseem; Andringa, Ronald; Aveyard, Mark; Babincak, Peter; Balatekin, Nursena; Baldwin, Scott A; Banik, Gabriel; Baskin, Ernest; Bell, Raoul; Białobrzeska, Olga; Birt, Angie R; Boot, Walter R; Braithwaite, Scott R; Briggs, Jessie C; Buchner, Axel; Budd, Desiree; Budzik, Kathryn; Bullens, Lottie; Bulley, Richard L; Cannon, Peter R; Cantarero, Katarzyna; Cesario, Joseph; Chambers, Stephanie; Chartier, Christopher R; Chekroun, Peggy; Chong, Clara; Cleeremans, Axel; Coary, Sean P; Coulthard, Jacob; Cramwinckel, Florien M; Denson, Thomas F; Díaz-Lago, Marcos; DiDonato, Theresa E; Drummond, Aaron; Eberlen, Julia; Ebersbach, Titus; Edlund, John E; Finnigan, Katherine M; Fisher, Justin; Frankowska, Natalia; García-Sánchez, Efraín; Golom, Frank D; Graves, Andrew J; Greenberg, Kevin; Hanioti, Mando; Hansen, Heather A; Harder, Jenna A; Harrell, Erin R; Hartanto, Andree; Inzlicht, Michael; Johnson, David J; Karpinski, Andrew; Keller, Victor N; Klein, Olivier; Koppel, Lina; Krahmer, Emiel; Lantian, Anthony; Larson, Michael J; Légal, Jean-Baptiste; Lucas, Richard E; Lynott, Dermot; Magaldino, Corey M; Massar, Karlijn; McBee, Matthew T; McLatchie, Neil; Melia, Nadhilla; Mensink, Michael C; Mieth, Laura; Moore-Berg, Samantha; Neeser, Geraldine; Newell, Ben R; Noordewier, Marret K; Ali Özdoğru, Asil; Pantazi, Myrto; Parzuchowski, Michał; Peters, Kim; Philipp, Michael C; Pollmann, Monique M H; Rentzelas, Panagiotis; Rodríguez-Bailón, Rosa; Philipp Röer, Jan; Ropovik, Ivan; Roque, Nelson A; Rueda, Carolina; Rutjens, Bastiaan T; Sackett, Katey; Salamon, Janos; Sánchez-Rodríguez, Ángel; Saunders, Blair; Schaafsma, Juliette; Schulte-Mecklenbeck, Michael; Shanks, David R; Sherman, Martin F; Steele, Kenneth M; Steffens, Niklas K; Sun, Jessie; Susa, Kyle J; Szaszi, Barnabas; Szollosi, Aba; Tamayo, Ricardo M; Tinghög, Gustav; Tong, Yuk-Yue; Tweten, Carol; Vadillo, Miguel A; Valcarcel, Deisy; Van der Linden, Nicolas; van Elk, Michiel; van Harreveld, Frenk; Västfjäll, Daniel; Vazire, Simine; Verduyn, Philippe; Williams, Matt N; Willis, Guillermo B; Wood, Sarah E; Yang, Chunliang; Zerhouni, Oulmann; Zheng, Robert; Zrubka, Mark

2018-03-01

Dijksterhuis and van Knippenberg (1998) reported that participants primed with a category associated with intelligence ("professor") subsequently performed 13% better on a trivia test than participants primed with a category associated with a lack of intelligence ("soccer hooligans"). In two unpublished replications of this study designed to verify the appropriate testing procedures, Dijksterhuis, van Knippenberg, and Holland observed a smaller difference between conditions (2%-3%) as well as a gender difference: Men showed the effect (9.3% and 7.6%), but women did not (0.3% and -0.3%). The procedure used in those replications served as the basis for this multilab Registered Replication Report. A total of 40 laboratories collected data for this project, and 23 of these laboratories met all inclusion criteria. Here we report the meta-analytic results for those 23 direct replications (total N = 4,493), which tested whether performance on a 30-item general-knowledge trivia task differed between these two priming conditions (results of supplementary analyses of the data from all 40 labs, N = 6,454, are also reported). We observed no overall difference in trivia performance between participants primed with the "professor" category and those primed with the "hooligan" category (0.14%) and no moderation by gender.

Field testing a soil site field guide for Allegheny hardwoods

Science.gov (United States)

S.B. Jones

1991-01-01

A site quality evaluation decision model, developed for Allegheny hardwoods on the non-glaciated Allegheny Plateau of Pennsylvania and New York, was field tested by International Paper (IP) foresters and the author, on sites within the region of derivation and on glaciated sites north and west of the Wisconsin drift line. Results from the field testing are presented...

Open-field behavior of house mice selectively bred for high voluntary wheel-running.

Science.gov (United States)

Bronikowski, A M; Carter, P A; Swallow, J G; Girard, I A; Rhodes, J S; Garland, T

2001-05-01

Open-field behavioral assays are commonly used to test both locomotor activity and emotionality in rodents. We performed open-field tests on house mice (Mus domesticus) from four replicate lines genetically selected for high voluntary wheel-running for 22 generations and from four replicate random-bred control lines. Individual mice were recorded by video camera for 3 min in a 1-m2 open-field arena on 2 consecutive days. Mice from selected lines showed no statistical differences from control mice with respect to distance traveled, defecation, time spent in the interior, or average distance from the center of the arena during the trial. Thus, we found little evidence that open-field behavior, as traditionally defined, is genetically correlated with wheel-running behavior. This result is a useful converse test of classical studies that report no increased wheel-running in mice selected for increased open-field activity. However, mice from selected lines turned less in their travel paths than did control-line mice, and females from selected lines had slower travel times (longer latencies) to reach the wall. We discuss these results in the context of the historical open-field test and newly defined measures of open-field activity.

Prototype Engineered Barrier System Field Tests

International Nuclear Information System (INIS)

Ramirez, A.L.; Beatty, J.; Buscheck, T.A.

1989-01-01

This paper presents selected preliminary results obtained during the first 54 days of the Prototype Engineered Barrier System Field Tests (PEBSFT) that are being performed in G-Tunnel within the Nevada Test Site. The test described is a precursor to the Engineered Barrier Systems Field Tests (EBSFT). The EBSFT will consist of in situ tests of the geohydrologic and geochemical environment in the near field (within a few meters) of heaters emplaced in welded tuff to simulate the thermal effects of waste packages. The PEBSFTs are being conducted to evaluate the applicability of measurement techniques, numerical models, and procedures for future investigations that will be conducted in the Exploratory Shaft Facilities of the Yucca Mountain Project (YMP). The paper discusses the evolution of hydrothermal behavior during the prototype test, including rock temperatures, changes in rock moisture content, air permeability of fractures, gas pressures, and rock mass gas-phase humidity. 10 refs., 12 figs

Field testing of asphalt-emulsion radon-barrier system

International Nuclear Information System (INIS)

Hartley, J.N.; Freeman, H.D.; Baker, E.G.; Elmore, M.R.; Nelson, D.A.; Voss, C.F.; Koehmstedt, P.L.

1981-09-01

Three years of laboratory and field testing have demonstrated that asphalt emulsion seals are effective radon diffusion barriers. Both laboratory and field tests in 1979, 1980 and 1981 have shown that an asphalt emulsion seal can reduce radon fluxes by greater than 99.9%. The effective diffusion coefficient for the various asphalt emulsion admix seals averages about 10 -6 cm 2 /s. The 1981 joint field test is a culmination of all the technology developed to date for asphalt emulsion radon barrier systems. Preliminary results of this field test and the results of the 1980 field test are presented. 18 figures, 6 tables

Focus Article: Replication in Second Language Writing Research

Science.gov (United States)

Porte, Graeme; Richards, Keith

2012-01-01

This paper discusses the meaning and range of replication in L2 research from both quantitative and qualitative perspectives. In the first half of the paper, it will be argued that key quantitative studies need to be replicated to have their robustness and generalizability tested and that this is a requirement of scientific inquiry. Such research…

Topography measurements for determining the decay factors in surface replication

International Nuclear Information System (INIS)

Song, J; Zheng, A; Vorburger, T V; Rubert, P

2008-01-01

The electro-forming technique is used at National Institute of Standards and Technology (NIST) for the production of standard reference material (SRM) 2461 standard casings to support nationwide ballistics measurement traceability and measurement quality control in the US. In order to ensure that the SRM casings are produced with virtually the same surface topography, it is necessary to test the decay factors of the replication process. Twenty-six replica casings are replicated from the same master casing for the decay factor tests. The NIST topography measurement system is used for measurements and correlations of surface topography. The topography decays are quantified by the cross-correlation function maximum CCF max . Based on the test, it is expected that 256 SRM casings can be replicated from the same master with CCF max values higher than 95%

Replicative Intermediates of Human Papillomavirus Type 11 in Laryngeal Papillomas: Site of Replication Initiation and Direction of Replication

Science.gov (United States)

Auborn, K. J.; Little, R. D.; Platt, T. H. K.; Vaccariello, M. A.; Schildkraut, C. L.

1994-07-01

We have examined the structures of replication intermediates from the human papillomavirus type 11 genome in DNA extracted from papilloma lesions (laryngeal papillomas). The sites of replication initiation and termination utilized in vivo were mapped by using neutral/neutral and neutral/alkaline two-dimensional agarose gel electrophoresis methods. Initiation of replication was detected in or very close to the upstream regulatory region (URR; the noncoding, regulatory sequences upstream of the open reading frames in the papillomavirus genome). We also show that replication forks proceed bidirectionally from the origin and converge 180circ opposite the URR. These results demonstrate the feasibility of analysis of replication of viral genomes directly from infected tissue.

DNA replication and cancer: From dysfunctional replication origin activities to therapeutic opportunities.

Science.gov (United States)

Boyer, Anne-Sophie; Walter, David; Sørensen, Claus Storgaard

2016-06-01

A dividing cell has to duplicate its DNA precisely once during the cell cycle to preserve genome integrity avoiding the accumulation of genetic aberrations that promote diseases such as cancer. A large number of endogenous impacts can challenge DNA replication and cells harbor a battery of pathways to promote genome integrity during DNA replication. This includes suppressing new replication origin firing, stabilization of replicating forks, and the safe restart of forks to prevent any loss of genetic information. Here, we describe mechanisms by which oncogenes can interfere with DNA replication thereby causing DNA replication stress and genome instability. Further, we describe cellular and systemic responses to these insults with a focus on DNA replication restart pathways. Finally, we discuss the therapeutic potential of exploiting intrinsic replicative stress in cancer cells for targeted therapy. Copyright © 2016 Elsevier Ltd. All rights reserved.

Distribution of DNA replication proteins in Drosophila cells

Science.gov (United States)

Easwaran, Hariharan P; Leonhardt, Heinrich; Cardoso, M Cristina

2007-01-01

Background DNA replication in higher eukaryotic cells is organized in discrete subnuclear sites called replication foci (RF). During the S phase, most replication proteins assemble at the RF by interacting with PCNA via a PCNA binding domain (PBD). This has been shown to occur for many mammalian replication proteins, but it is not known whether this mechanism is conserved in evolution. Results Fluorescent fusions of mammalian replication proteins, Dnmt1, HsDNA Lig I and HsPCNA were analyzed for their ability to target to RF in Drosophila cells. Except for HsPCNA, none of the other proteins and their deletions showed any accumulation at RF in Drosophila cells. We hypothesized that in Drosophila cells there might be some other peptide sequence responsible for targeting proteins to RF. To test this, we identified the DmDNA Lig I and compared the protein sequence with HsDNA Lig I. The two orthologs shared the PBD suggesting a functionally conserved role for this domain in the Drosophila counterpart. A series of deletions of DmDNA Lig I were analyzed for their ability to accumulate at RF in Drosophila and mammalian cells. Surprisingly, no accumulation at RF was observed in Drosophila cells, while in mammalian cells DmDNA Lig I accumulated at RF via its PBD. Further, GFP fusions with the PBD domains from Dnmt1, HsDNA Lig I and DmDNA Lig I, were able to target to RF only in mammalian cells but not in Drosophila cells. Conclusion We show that S phase in Drosophila cells is characterized by formation of RF marked by PCNA like in mammalian cells. However, other than PCNA none of the replication proteins and their deletions tested here showed accumulation at RF in Drosophila cells while the same proteins and deletions are capable of accumulating at RF in mammalian cells. We hypothesize that unlike mammalian cells, in Drosophila cells, replication proteins do not form long-lasting interactions with the replication machinery, and rather perform their functions via very

The Acid Test for Biological Science: STAP Cells, Trust, and Replication.

Science.gov (United States)

Lancaster, Cheryl

2016-02-01

In January 2014, a letter and original research article were published in Nature describing a process whereby somatic mouse cells could be converted into stem cells by subjecting them to stress. These "stimulus-triggered acquisition of pluripotency" (STAP) cells were shown to be capable of contributing to all cell types of a developing embryo, and extra-embryonic tissues. The lead author of the publications, Haruko Obokata, became an overnight celebrity in Japan, where she was dubbed the new face of Japanese science. However, in the weeks that followed publication of the research, issues arose. Other laboratories and researchers (including authors on the original papers) found that they were unable to replicate Obokata et al.'s work. Closer scrutiny of the papers by the scientific community also suggested that there was manipulation of images that had been published, and Obokata was accused of misconduct. Those who should have been supervising her work (also her co-authors on the publications) were also heavily criticised. The STAP cell saga of 2014 is used as an example to highlight the importance of trust and replication in twenty-first century biological science. The role of trust in the scientific community is highlighted, and the effects on interactions between science and the public examined. Similarly, this essay aims to highlight the importance of replication, and how this is understood by researchers, the media, and the public. The expected behaviour of scientists in the twenty-first century is now more closely scrutinised.

USP7/HAUSP: A SUMO deubiquitinase at the heart of DNA replication.

Science.gov (United States)

Smits, Veronique A J; Freire, Raimundo

2016-09-01

DNA replication is both highly conserved and controlled. Problematic DNA replication can lead to genomic instability and therefore carcinogenesis. Numerous mechanisms work together to achieve this tight control and increasing evidence suggests that post-translational modifications (phosphorylation, ubiquitination, SUMOylation) of DNA replication proteins play a pivotal role in this process. Here we discuss such modifications in the light of a recent article that describes a novel role for the deubiquitinase (DUB) USP7/HAUSP in the control of DNA replication. USP7 achieves this function by an unusual and novel mechanism, namely deubiquitination of SUMOylated proteins at the replication fork, making USP7 also a SUMO DUB (SDUB). This work extends previous observations of increased levels of SUMO and low levels of ubiquitin at the on-going replication fork. Here, we discuss this novel study, its contribution to the DNA replication and genomic stability field and what questions arise from this work. © 2016 WILEY Periodicals, Inc.

Buildings, fields of activity, testing facilities

International Nuclear Information System (INIS)

1974-01-01

Since 1969 the activities of the Materialpruefungsanstalt Stuttgart (MPA) have grown quickly as planned, especially in the field of reactor safety research, which made it necessary to increase the staff to approximately 165 members, to supplement the machines and equipment and to extend the fields of activities occasioning a further departmental reorganization. At present the MPA has the following departments: 1. Teaching (materials testing, materials science and strength of materials) 2. Materials and Welding Technology 3. Materials Science and General Materials Testing with Tribology 4. Design and Strength 5. Creep and Fatigue Testing 6. Central Facilities 7. Vessel and Component Testing. (orig./RW) [de

A replicated climate change field experiment reveals rapid evolutionary response in an ecologically important soil invertebrate

DEFF Research Database (Denmark)

Bataillon, Thomas; Galtier, Nicolas; Bernard, Aurelien

2016-01-01

to climate change in a common annelid worm using a controlled replicated experiment where climatic conditions were manipulated in a natural setting. Analyzing the transcribed genome of 15 local populations, we found that about 12% of the genetic polymorphisms exhibit differences in allele frequencies......Whether species can respond evolutionarily to current climate change is crucial for the persistence of many species. Yet, very few studies have examined genetic responses to climate change in manipulated experiments carried out innatural field conditions. We examined the evolutionary response...... associated to changes in soil temperature and soil moisture. This shows an evolutionaryresponse to realistic climate change happening over short-time scale, and calls for incorporating evolution into modelspredicting future response of species to climate change. It also shows that designed climate change...

Adressing Replication and Model Uncertainty

DEFF Research Database (Denmark)

Ebersberger, Bernd; Galia, Fabrice; Laursen, Keld

innovation survey data for France, Germany and the UK, we conduct a ‘large-scale’ replication using the Bayesian averaging approach of classical estimators. Our method tests a wide range of determinants of innovation suggested in the prior literature, and establishes a robust set of findings on the variables...

Combining p-values in replicated single-case experiments with multivariate outcome.

Science.gov (United States)

Solmi, Francesca; Onghena, Patrick

2014-01-01

Interest in combining probabilities has a long history in the global statistical community. The first steps in this direction were taken by Ronald Fisher, who introduced the idea of combining p-values of independent tests to provide a global decision rule when multiple aspects of a given problem were of interest. An interesting approach to this idea of combining p-values is the one based on permutation theory. The methods belonging to this particular approach exploit the permutation distributions of the tests to be combined, and use a simple function to combine probabilities. Combining p-values finds a very interesting application in the analysis of replicated single-case experiments. In this field the focus, while comparing different treatments effects, is more articulated than when just looking at the means of the different populations. Moreover, it is often of interest to combine the results obtained on the single patients in order to get more global information about the phenomenon under study. This paper gives an overview of how the concept of combining p-values was conceived, and how it can be easily handled via permutation techniques. Finally, the method of combining p-values is applied to a simulated replicated single-case experiment, and a numerical illustration is presented.

Replication of avian influenza viruses in equine tracheal epithelium but not in horses

OpenAIRE

Chambers, Thomas M.; Balasuriya, Udeni B. R.; Reedy, Stephanie E.; Tiwari, Ashish

2013-01-01

We evaluated a hypothesis that horses are susceptible to avian influenza viruses by in vitro testing, using explanted equine tracheal epithelial cultures, and in vivo testing by aerosol inoculation of ponies. Results showed that several subtypes of avian influenza viruses detectably replicated in vitro. Three viruses with high in vitro replication competence were administered to ponies. None of the three demonstrably replicated or caused disease signs in ponies. While these results do not exh...

Identification of the determinants of efficient Pestivirus replication

DEFF Research Database (Denmark)

Risager, Peter Christian

, and in depth knowledge of the traits that determine the fitness of the virus in this regard are highly valuable. Recent advances in the field of molecular virology with methods to manipulate viral genomes have significantly helped to uncover these core mechanisms responsible for exploitation of the host......, BMC genomics). Manuscript II describes the generation of replicons that express two different types of luciferases (Rluc and Gluc), and their application as a tool for easy monitoring of replication competence (published paper, Journal of General Virology (94), 1739-1748). Manuscript III describes...... the properties of chimeric replicons and infectious clones that include a RNA dependent RNA polymerase (NS5B) from one of three different CSFV strains with distinct virulence properties. The entire NS5B proved to influence replication competence and key residues for replication competence was identified...

Greenness and school-wide test scores are not always positively associated – A replication of "linking student performance in Massachusetts elementary schools with the 'greenness' of school surroundings using remote sensing"

Science.gov (United States)

Matthew H.E.M. Browning; Ming Kuo; Sonya Sachdeva; Kangjae Lee; Lynne Westphal

2018-01-01

Recent studies find vegetation around schools correlates positively with student test scores. To test this relationship in schools with less green cover and more disadvantaged students, we replicated a leading study, using six years of NDVI-derived greenness data to predict school-level math and reading achievement in 404 Chicago public schools. A direct replication...

Repair replication in replicating and nonreplicating DNA after irradiation with uv light

Energy Technology Data Exchange (ETDEWEB)

Slor, H.; Cleaver, J.E.

1978-06-01

Ultraviolet light induces more pyrimidine dimers and more repair replication in DNA that replicates within 2 to 3 h of irradiation than in DNA that does not replicate during this period. This difference may be due to special conformational changes in DNA and chromatin that might be associated with semiconservative DNA replication.

Human Parvovirus B19 Utilizes Cellular DNA Replication Machinery for Viral DNA Replication.

Science.gov (United States)

Zou, Wei; Wang, Zekun; Xiong, Min; Chen, Aaron Yun; Xu, Peng; Ganaie, Safder S; Badawi, Yomna; Kleiboeker, Steve; Nishimune, Hiroshi; Ye, Shui Qing; Qiu, Jianming

2018-03-01

Human parvovirus B19 (B19V) infection of human erythroid progenitor cells (EPCs) induces a DNA damage response and cell cycle arrest at late S phase, which facilitates viral DNA replication. However, it is not clear exactly which cellular factors are employed by this single-stranded DNA virus. Here, we used microarrays to systematically analyze the dynamic transcriptome of EPCs infected with B19V. We found that DNA metabolism, DNA replication, DNA repair, DNA damage response, cell cycle, and cell cycle arrest pathways were significantly regulated after B19V infection. Confocal microscopy analyses revealed that most cellular DNA replication proteins were recruited to the centers of viral DNA replication, but not the DNA repair DNA polymerases. Our results suggest that DNA replication polymerase δ and polymerase α are responsible for B19V DNA replication by knocking down its expression in EPCs. We further showed that although RPA32 is essential for B19V DNA replication and the phosphorylated forms of RPA32 colocalized with the replicating viral genomes, RPA32 phosphorylation was not necessary for B19V DNA replication. Thus, this report provides evidence that B19V uses the cellular DNA replication machinery for viral DNA replication. IMPORTANCE Human parvovirus B19 (B19V) infection can cause transient aplastic crisis, persistent viremia, and pure red cell aplasia. In fetuses, B19V infection can result in nonimmune hydrops fetalis and fetal death. These clinical manifestations of B19V infection are a direct outcome of the death of human erythroid progenitors that host B19V replication. B19V infection induces a DNA damage response that is important for cell cycle arrest at late S phase. Here, we analyzed dynamic changes in cellular gene expression and found that DNA metabolic processes are tightly regulated during B19V infection. Although genes involved in cellular DNA replication were downregulated overall, the cellular DNA replication machinery was tightly

MOF Suppresses Replication Stress and Contributes to Resolution of Stalled Replication Forks.

Science.gov (United States)

Singh, Dharmendra Kumar; Pandita, Raj K; Singh, Mayank; Chakraborty, Sharmistha; Hambarde, Shashank; Ramnarain, Deepti; Charaka, Vijaya; Ahmed, Kazi Mokim; Hunt, Clayton R; Pandita, Tej K

2018-03-15

The human MOF (hMOF) protein belongs to the MYST family of histone acetyltransferases and plays a critical role in transcription and the DNA damage response. MOF is essential for cell proliferation; however, its role during replication and replicative stress is unknown. Here we demonstrate that cells depleted of MOF and under replicative stress induced by cisplatin, hydroxyurea, or camptothecin have reduced survival, a higher frequency of S-phase-specific chromosome damage, and increased R-loop formation. MOF depletion decreased replication fork speed and, when combined with replicative stress, also increased stalled replication forks as well as new origin firing. MOF interacted with PCNA, a key coordinator of replication and repair machinery at replication forks, and affected its ubiquitination and recruitment to the DNA damage site. Depletion of MOF, therefore, compromised the DNA damage repair response as evidenced by decreased Mre11, RPA70, Rad51, and PCNA focus formation, reduced DNA end resection, and decreased CHK1 phosphorylation in cells after exposure to hydroxyurea or cisplatin. These results support the argument that MOF plays an important role in suppressing replication stress induced by genotoxic agents at several stages during the DNA damage response. Copyright © 2018 American Society for Microbiology.

Database Replication

CERN Document Server

Kemme, Bettina

2010-01-01

Database replication is widely used for fault-tolerance, scalability and performance. The failure of one database replica does not stop the system from working as available replicas can take over the tasks of the failed replica. Scalability can be achieved by distributing the load across all replicas, and adding new replicas should the load increase. Finally, database replication can provide fast local access, even if clients are geographically distributed clients, if data copies are located close to clients. Despite its advantages, replication is not a straightforward technique to apply, and

SRS environmental technology development field test platform

International Nuclear Information System (INIS)

Riha, B.D.; Rossabi, J.; Eddy-Dilek, C.A.

1995-01-01

A critical and difficult step in the development and implementation of new technologies for environmental monitoring and characterization is successfully transferring these technologies to industry and government users for routine assessment and compliance activities. The Environmental Sciences Section of the DOE Savannah River Technology Center provides a forum for developers, potential users, and regulatory organizations to evaluate new technologies in comparison with baseline technologies in a well characterized field test bed. The principal objective of this project is to conduct comprehensive, objective field tests of monitoring and characterization technologies that are not currently used in EPA standard methods and evaluate their performance during actual operating conditions against baseline methods. This paper provides an overview of the field test site and a description of some of the technologies demonstrated at the site including their field applications

The Savannah River environmental technology field test platform

International Nuclear Information System (INIS)

Rossabi, J.; Riha, B.D.

1995-01-01

The principal goal in the development of new technologies for environmental monitoring and characterization is transferring them to organizations and individuals for use in site assessment and compliance monitoring. The Savannah River technology Center (SRTC) has been developing a program to rigorously field test promising environmental technologies that have not undergone EPA equivalency testing. The infrastructure and staff expertise developed as part of the activities of the Savannah River Integrated Demonstration Program allows field testing of technologies without the difficulties of providing remote field support. By providing a well-characterized site and a well-developed infrastructure, technologies are tested in actual field scenarios to determine their appropriate applications in environmental characterization and monitoring activities. The field tests provide regulatory organizations, potential industrial partners, and potential users with the opportunity to evaluate the technology's performance and its utility for implementation in environmental characterization and monitoring programs. This program has resulted in the successful implementation of several new technologies

X-irradiation affects all DNA replication intermediates when inhibiting replication initiation

International Nuclear Information System (INIS)

Loenn, U.; Karolinska Hospital, Stockholm

1982-01-01

When a human melanoma line was irradiated with 10 Gy, there was, after 30 to 60 min, a gradual reduction in the DNA replication rate. Ten to twelve hours after the irradiation, the DNA replication had returned to near normal rate. The results showed tht low dose-rate X-irradiation inhibits preferentially the formation of small DNA replication intermediates. There is no difference between the inhibition of these replication intermediates formed only in the irradiated cells and those formed also in untreated cells. (U.K.)

Chromatin Structure and Replication Origins: Determinants Of Chromosome Replication And Nuclear Organization

Science.gov (United States)

Smith, Owen K.; Aladjem, Mirit I.

2014-01-01

The DNA replication program is, in part, determined by the epigenetic landscape that governs local chromosome architecture and directs chromosome duplication. Replication must coordinate with other biochemical processes occurring concomitantly on chromatin, such as transcription and remodeling, to insure accurate duplication of both genetic and epigenetic features and to preserve genomic stability. The importance of genome architecture and chromatin looping in coordinating cellular processes on chromatin is illustrated by two recent sets of discoveries. First, chromatin-associated proteins that are not part of the core replication machinery were shown to affect the timing of DNA replication. These chromatin-associated proteins could be working in concert, or perhaps in competition, with the transcriptional machinery and with chromatin modifiers to determine the spatial and temporal organization of replication initiation events. Second, epigenetic interactions are mediated by DNA sequences that determine chromosomal replication. In this review we summarize recent findings and current models linking spatial and temporal regulation of the replication program with epigenetic signaling. We discuss these issues in the context of the genome’s three-dimensional structure with an emphasis on events occurring during the initiation of DNA replication. PMID:24905010

Prelife catalysts and replicators

OpenAIRE

Ohtsuki, Hisashi; Nowak, Martin A.

2009-01-01

Life is based on replication and evolution. But replication cannot be taken for granted. We must ask what there was prior to replication and evolution. How does evolution begin? We have proposed prelife as a generative system that produces information and diversity in the absence of replication. We model prelife as a binary soup of active monomers that form random polymers. ‘Prevolutionary’ dynamics can have mutation and selection prior to replication. Some sequences might have catalytic acti...

Data from Investigating Variation in Replicability: A “Many Labs” Replication Project

Directory of Open Access Journals (Sweden)

Richard A. Klein

2014-04-01

Full Text Available This dataset is from the Many Labs Replication Project in which 13 effects were replicated across 36 samples and over 6,000 participants. Data from the replications are included, along with demographic variables about the participants and contextual information about the environment in which the replication was conducted. Data were collected in-lab and online through a standardized procedure administered via an online link. The dataset is stored on the Open Science Framework website. These data could be used to further investigate the results of the included 13 effects or to study replication and generalizability more broadly.

Numerical simulations of capillary barrier field tests

International Nuclear Information System (INIS)

Morris, C.E.; Stormont, J.C.

1997-01-01

Numerical simulations of two capillary barrier systems tested in the field were conducted to determine if an unsaturated flow model could accurately represent the observed results. The field data was collected from two 7-m long, 1.2-m thick capillary barriers built on a 10% grade that were being tested to investigate their ability to laterally divert water downslope. One system had a homogeneous fine layer, while the fine soil of the second barrier was layered to increase its ability to laterally divert infiltrating moisture. The barriers were subjected first to constant infiltration while minimizing evaporative losses and then were exposed to ambient conditions. The continuous infiltration period of the field tests for the two barrier systems was modelled to determine the ability of an existing code to accurately represent capillary barrier behavior embodied in these two designs. Differences between the field test and the model data were found, but in general the simulations appeared to adequately reproduce the response of the test systems. Accounting for moisture retention hysteresis in the layered system will potentially lead to more accurate modelling results and is likely to be important when developing reasonable predictions of capillary barrier behavior

Entanglement replication in driven dissipative many-body systems.

Science.gov (United States)

Zippilli, S; Paternostro, M; Adesso, G; Illuminati, F

2013-01-25

We study the dissipative dynamics of two independent arrays of many-body systems, locally driven by a common entangled field. We show that in the steady state the entanglement of the driving field is reproduced in an arbitrarily large series of inter-array entangled pairs over all distances. Local nonclassical driving thus realizes a scale-free entanglement replication and long-distance entanglement distribution mechanism that has immediate bearing on the implementation of quantum communication networks.

Test fields cannot destroy extremal black holes

International Nuclear Information System (INIS)

Natário, José; Queimada, Leonel; Vicente, Rodrigo

2016-01-01

We prove that (possibly charged) test fields satisfying the null energy condition at the event horizon cannot overspin/overcharge extremal Kerr–Newman or Kerr–Newman–anti de Sitter black holes, that is, the weak cosmic censorship conjecture cannot be violated in the test field approximation. The argument relies on black hole thermodynamics (without assuming cosmic censorship), and does not depend on the precise nature of the fields. We also discuss generalizations of this result to other extremal black holes. (paper)

Genetic analysis without replications: Model evaluation and application in spring wheat

Science.gov (United States)

Genetic data collected from plant breeding and genetic studies may not be replicated in field designs even though field variation is present. In this study, we addressed this problem using spring wheat (Triticum eastivum L.) trial data collected from two locations. There were no intra-location repl...

Functions of Ubiquitin and SUMO in DNA Replication and Replication Stress

Science.gov (United States)

García-Rodríguez, Néstor; Wong, Ronald P.; Ulrich, Helle D.

2016-01-01

Complete and faithful duplication of its entire genetic material is one of the essential prerequisites for a proliferating cell to maintain genome stability. Yet, during replication DNA is particularly vulnerable to insults. On the one hand, lesions in replicating DNA frequently cause a stalling of the replication machinery, as most DNA polymerases cannot cope with defective templates. This situation is aggravated by the fact that strand separation in preparation for DNA synthesis prevents common repair mechanisms relying on strand complementarity, such as base and nucleotide excision repair, from working properly. On the other hand, the replication process itself subjects the DNA to a series of hazardous transformations, ranging from the exposure of single-stranded DNA to topological contortions and the generation of nicks and fragments, which all bear the risk of inducing genomic instability. Dealing with these problems requires rapid and flexible responses, for which posttranslational protein modifications that act independently of protein synthesis are particularly well suited. Hence, it is not surprising that members of the ubiquitin family, particularly ubiquitin itself and SUMO, feature prominently in controlling many of the defensive and restorative measures involved in the protection of DNA during replication. In this review we will discuss the contributions of ubiquitin and SUMO to genome maintenance specifically as they relate to DNA replication. We will consider cases where the modifiers act during regular, i.e., unperturbed stages of replication, such as initiation, fork progression, and termination, but also give an account of their functions in dealing with lesions, replication stalling and fork collapse. PMID:27242895

OPTIMAL STRATEGIES FOR REGIONAL CULTIVAR TESTING FOR VARIOUS CROPS

OpenAIRE

Kingwell, Ross S.

1999-01-01

Before a variety is released for adoption by farmers in a region it is subject to comparative field testing. An economic issue is how extensive and prolonged should be this field testing within the region; over how many locations, years and with what number of plot replications should testing occur? This paper addresses this issue for the main broadacre crops grown in Western Australia and shows through sensitivity analysis how robust are the findings for each crop type. Findings are contrast...

Replicating receptive fields of simple and complex cells in primary visual cortex in a neuronal network model with temporal and population sparseness and reliability.

Science.gov (United States)

Tanaka, Takuma; Aoyagi, Toshio; Kaneko, Takeshi

2012-10-01

We propose a new principle for replicating receptive field properties of neurons in the primary visual cortex. We derive a learning rule for a feedforward network, which maintains a low firing rate for the output neurons (resulting in temporal sparseness) and allows only a small subset of the neurons in the network to fire at any given time (resulting in population sparseness). Our learning rule also sets the firing rates of the output neurons at each time step to near-maximum or near-minimum levels, resulting in neuronal reliability. The learning rule is simple enough to be written in spatially and temporally local forms. After the learning stage is performed using input image patches of natural scenes, output neurons in the model network are found to exhibit simple-cell-like receptive field properties. When the output of these simple-cell-like neurons are input to another model layer using the same learning rule, the second-layer output neurons after learning become less sensitive to the phase of gratings than the simple-cell-like input neurons. In particular, some of the second-layer output neurons become completely phase invariant, owing to the convergence of the connections from first-layer neurons with similar orientation selectivity to second-layer neurons in the model network. We examine the parameter dependencies of the receptive field properties of the model neurons after learning and discuss their biological implications. We also show that the localized learning rule is consistent with experimental results concerning neuronal plasticity and can replicate the receptive fields of simple and complex cells.

Two Bayesian tests of the GLOMOsys Model.

Science.gov (United States)

Field, Sarahanne M; Wagenmakers, Eric-Jan; Newell, Ben R; Zeelenberg, René; van Ravenzwaaij, Don

2016-12-01

Priming is arguably one of the key phenomena in contemporary social psychology. Recent retractions and failed replication attempts have led to a division in the field between proponents and skeptics and have reinforced the importance of confirming certain priming effects through replication. In this study, we describe the results of 2 preregistered replication attempts of 1 experiment by Förster and Denzler (2012). In both experiments, participants first processed letters either globally or locally, then were tested using a typicality rating task. Bayes factor hypothesis tests were conducted for both experiments: Experiment 1 (N = 100) yielded an indecisive Bayes factor of 1.38, indicating that the in-lab data are 1.38 times more likely to have occurred under the null hypothesis than under the alternative. Experiment 2 (N = 908) yielded a Bayes factor of 10.84, indicating strong support for the null hypothesis that global priming does not affect participants' mean typicality ratings. The failure to replicate this priming effect challenges existing support for the GLOMO sys model. (PsycINFO Database Record (c) 2016 APA, all rights reserved).

NACSA Charter School Replication Guide: The Spectrum of Replication Options. Authorizing Matters. Replication Brief 1

Science.gov (United States)

O'Neill, Paul

2010-01-01

One of the most important and high-profile issues in public education reform today is the replication of successful public charter school programs. With more than 5,000 failing public schools in the United States, there is a tremendous need for strong alternatives for parents and students. Replicating successful charter school models is an…

Relationship between mitochondrial gene rearrangements and stability of the origin of light strand replication

Directory of Open Access Journals (Sweden)

Miguel M. Fonseca

2008-01-01

Full Text Available Mitochondrial gene rearrangements are much more frequent in vertebrates than initially thought. It has been suggested that the origin of light strand replication could have an important role in the process of gene rearrangements, but this hypothesis has never been tested before. We used amphibians to test the correlation between light-strand replication origin thermodynamic stability and the occurrence of gene rearrangements. The two variables were correlated in a non-phylogenetic approach, but when tested in a phylogenetically based comparative method the correlation was not significant, although species with unstable light-strand replication origins were much more likely to have undergone gene rearrangements. This indicates that within amphibians there are stable and unstable phylogenetic groups regarding mitochondrial gene order. The species analyzed showed variability in the thermodynamic stability of the secondary structure, in the length of its stem and loop, and several species did not present the 5’-GCCGG-3’ motif reported to be necessary for efficient mitochondrial DNA replication. Future studies should focus on the role of the light-strand replication origin in mitochondrial DNA replication and gene rearrangements mechanisms.

Replicative phenotyping adds value to genotypic resistance testing in heavily pre-treated HIV-infected individuals - the Swiss HIV Cohort Study

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Martinetti Gladys

2011-01-01

Full Text Available Abstract Background Replicative phenotypic HIV resistance testing (rPRT uses recombinant infectious virus to measure viral replication in the presence of antiretroviral drugs. Due to its high sensitivity of detection of viral minorities and its dissecting power for complex viral resistance patterns and mixed virus populations rPRT might help to improve HIV resistance diagnostics, particularly for patients with multiple drug failures. The aim was to investigate whether the addition of rPRT to genotypic resistance testing (GRT compared to GRT alone is beneficial for obtaining a virological response in heavily pre-treated HIV-infected patients. Methods Patients with resistance tests between 2002 and 2006 were followed within the Swiss HIV Cohort Study (SHCS. We assessed patients' virological success after their antiretroviral therapy was switched following resistance testing. Multilevel logistic regression models with SHCS centre as a random effect were used to investigate the association between the type of resistance test and virological response (HIV-1 RNA Results Of 1158 individuals with resistance tests 221 with GRT+rPRT and 937 with GRT were eligible for analysis. Overall virological response rates were 85.1% for GRT+rPRT and 81.4% for GRT. In the subgroup of patients with >2 previous failures, the odds ratio (OR for virological response of GRT+rPRT compared to GRT was 1.45 (95% CI 1.00-2.09. Multivariate analyses indicate a significant improvement with GRT+rPRT compared to GRT alone (OR 1.68, 95% CI 1.31-2.15. Conclusions In heavily pre-treated patients rPRT-based resistance information adds benefit, contributing to a higher rate of treatment success.

DNA Replication in Engineered Escherichia coli Genomes with Extra Replication Origins.

Science.gov (United States)

Milbredt, Sarah; Farmani, Neda; Sobetzko, Patrick; Waldminghaus, Torsten

2016-10-21

The standard outline of bacterial genomes is a single circular chromosome with a single replication origin. From the bioengineering perspective, it appears attractive to extend this basic setup. Bacteria with split chromosomes or multiple replication origins have been successfully constructed in the last few years. The characteristics of these engineered strains will largely depend on the respective DNA replication patterns. However, the DNA replication has not been investigated systematically in engineered bacteria with multiple origins or split replicons. Here we fill this gap by studying a set of strains consisting of (i) E. coli strains with an extra copy of the native replication origin (oriC), (ii) E. coli strains with an extra copy of the replication origin from the secondary chromosome of Vibrio cholerae (oriII), and (iii) a strain in which the E. coli chromosome is split into two linear replicons. A combination of flow cytometry, microarray-based comparative genomic hybridization (CGH), and modeling revealed silencing of extra oriC copies and differential timing of ectopic oriII copies compared to the native oriC. The results were used to derive construction rules for future multiorigin and multireplicon projects.

On Scalability and Replicability of Smart Grid Projects—A Case Study

Directory of Open Access Journals (Sweden)

Lukas Sigrist

2016-03-01

Full Text Available This paper studies the scalability and replicability of smart grid projects. Currently, most smart grid projects are still in the R&D or demonstration phases. The full roll-out of the tested solutions requires a suitable degree of scalability and replicability to prevent project demonstrators from remaining local experimental exercises. Scalability and replicability are the preliminary requisites to perform scaling-up and replication successfully; therefore, scalability and replicability allow for or at least reduce barriers for the growth and reuse of the results of project demonstrators. The paper proposes factors that influence and condition a project’s scalability and replicability. These factors involve technical, economic, regulatory and stakeholder acceptance related aspects, and they describe requirements for scalability and replicability. In order to assess and evaluate the identified scalability and replicability factors, data has been collected from European and national smart grid projects by means of a survey, reflecting the projects’ view and results. The evaluation of the factors allows quantifying the status quo of on-going projects with respect to the scalability and replicability, i.e., they provide a feedback on to what extent projects take into account these factors and on whether the projects’ results and solutions are actually scalable and replicable.

Field testing at the Climax Stock on the Nevada Test Site: spent fuel test and radionuclide migration experiments

International Nuclear Information System (INIS)

Ballou, L.B.; Isherwood, D.J.; Patrick, W.C.

1982-01-01

Two field tests in the Climax Stock are being conducted. The Climax Stock, a granitic instrusive, has been administratively excluded from consideration as a full-scale repository site. However, it provides a readily available facility for field testing with high-level radioactive materials at a depth (420 m) approaching that of a repository. The major test activity in the 1980 fiscal year has been initiation of the Spent Fuel Test-Climax (SFT-C). This test, which was authorized in June 1978, is designed to evaluate the generic feasibility of geologic storage and retrievability of commercial power reactor spent fuel assemblies in a granitic medium. In addition, the test is configured and instrumented to provide thermal and thermomechanical response data that will be relevant to the design of a repository in hard crystalline rock. The other field activity in the Climax Stock is a radionuclide migration test. It combines a series of field and laboratory migration experiments with the use of existing hydrologic models for pretest predictions and data interpretation. Goals of this project are to develop: (1) field measurement techniques for radionuclide migration studies in a hydrologic regime where the controlling mechanism is fracture permeability; (2) field test data on radionuclide migration; and (3) a comparison of laboratory- and field-measured retardation factors. This radionuclide migration test, which was authorized in the middle of the 1980 fiscal year, is in the preliminary design phase. The detailed program plan was prepared and subjected to formal peer review in August. In September/October researchers conducted preliminary flow tests with water in selected near-vertical fractures intersected by small horizontal boreholes. These tests were needed to establish the range of pressures, flow rates, and other operating parameters to be used in conducting the nuclide migration tests. 21 references, 14 figures, 1 table

Hydroxyurea-Induced Replication Stress

Directory of Open Access Journals (Sweden)

Kenza Lahkim Bennani-Belhaj

2010-01-01

Full Text Available Bloom's syndrome (BS displays one of the strongest known correlations between chromosomal instability and a high risk of cancer at an early age. BS cells combine a reduced average fork velocity with constitutive endogenous replication stress. However, the response of BS cells to replication stress induced by hydroxyurea (HU, which strongly slows the progression of replication forks, remains unclear due to publication of conflicting results. Using two different cellular models of BS, we showed that BLM deficiency is not associated with sensitivity to HU, in terms of clonogenic survival, DSB generation, and SCE induction. We suggest that surviving BLM-deficient cells are selected on the basis of their ability to deal with an endogenous replication stress induced by replication fork slowing, resulting in insensitivity to HU-induced replication stress.

Deep Borehole Field Test Laboratory and Borehole Testing Strategy

Energy Technology Data Exchange (ETDEWEB)

Kuhlman, Kristopher L. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Brady, Patrick V. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); MacKinnon, Robert J. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Heath, Jason E. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Herrick, Courtney G. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Jensen, Richard P. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Gardner, W. Payton [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Sevougian, S. David [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Bryan, Charles R. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Jang, Je-Hun [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Stein, Emily R. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Bauer, Stephen J. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Daley, Tom [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Freifeld, Barry M. [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Birkholzer, Jens [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Spane, Frank A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

2016-09-19

Deep Borehole Disposal (DBD) of high-level radioactive wastes has been considered an option for geological isolation for many years (Hess et al. 1957). Recent advances in drilling technology have decreased costs and increased reliability for large-diameter (i.e., ≥50 cm [19.7”]) boreholes to depths of several kilometers (Beswick 2008; Beswick et al. 2014). These advances have therefore also increased the feasibility of the DBD concept (Brady et al. 2009; Cornwall 2015), and the current field test design will demonstrate the DBD concept and these advances. The US Department of Energy (DOE) Strategy for the Management and Disposal of Used Nuclear Fuel and High-Level Radioactive Waste (DOE 2013) specifically recommended developing a research and development plan for DBD. DOE sought input or expression of interest from States, local communities, individuals, private groups, academia, or any other stakeholders willing to host a Deep Borehole Field Test (DBFT). The DBFT includes drilling two boreholes nominally 200m [656’] apart to approximately 5 km [16,400’] total depth, in a region where crystalline basement is expected to begin at less than 2 km depth [6,560’]. The characterization borehole (CB) is the smaller-diameter borehole (i.e., 21.6 cm [8.5”] diameter at total depth), and will be drilled first. The geologic, hydrogeologic, geochemical, geomechanical and thermal testing will take place in the CB. The field test borehole (FTB) is the larger-diameter borehole (i.e., 43.2 cm [17”] diameter at total depth). Surface handling and borehole emplacement of test package will be demonstrated using the FTB to evaluate engineering feasibility and safety of disposal operations (SNL 2016).

Testing the family replication-model through Bsup(O)-Bsup(-O) mixing

International Nuclear Information System (INIS)

Datta, A.; Pati, J.C.

1985-07-01

It is observed that the family-replication idea, proposed in the context of a minimal preon-model, necessarily implies a maximal mixing (i.e. ΔM>>GAMMA) either in the Bsub(s)sup(O)-B-barsub(s)sup(O) or the Bsub(d)sup(O)-B-barsub(d)sup(O) system, in contrast to the standard model. (author)

Role of energy systems in two intermittent field tests in women field hockey players

NARCIS (Netherlands)

Lemmink, Koen A. P. M.; Visscher, Susan H.

The energetics of 2 field tests that reflect physical performance in intermittent sports (i.e., the Interval Shuttle Sprint Test [ISST] and the Interval Shuttle Run Test [ISRT]) were examined in 21 women field hockey players. The ISST required the players to perform 10 shuttle sprints starting every

Performance Test of CCTV in a Test Field

Energy Technology Data Exchange (ETDEWEB)

Seo, Hyung Min [Korea Institute of Nuclear Nonproliferation and Control, Daejeon (Korea, Republic of)

2011-10-15

On April 12-13, 2010, US President Obama hosted a Nuclear Security Summit in Washington, DC, to enhance international cooperation to prevent nuclear terrorism, an issue which he has identified as the most immediate and extreme threat to global security. The Summit focused on the security of nuclear materials, nonproliferation, disarmament, and peaceful nuclear energy. At the summit, the Republic of Korea was chosen as the host of the next Summit in 2012. This series of events reflects the growing global interest on 'Nuclear Security' and as the host country of the next Nuclear Summit it is the time for Korea to strengthen the physical protection regime for nuclear facilities as a first step of securing its nuclear security capability. KINAC has been operating Test field as a mean of preparing solid backup data for reviewing and revising DBT (Design Basis Threat) and to test components of the conventional physical protection system. CCTV is a key component which is used worldwide for the assessment measure of alarms. In terms of performance test of CCTV, there are several elements such as image quality, coverage and mechanical features (speed of zoom-in-out, capture, angle shift etc.). Speaking of image quality acquired by the CCTV, the quality is subject to resolution, monitor specification, camera housing, camera mounting and lightening. Thus it is clear that performance tests on image quality should consider those factors and vary the factors respectively in order to verify the influence and the interaction among those. Nevertheless due to the restrictions of the current Test field, this paper focuses on the image quality through resolution test under the various lightening conditions

Reproducibility and replicability of rodent phenotyping in preclinical studies

NARCIS (Netherlands)

Kafkafi, Neri; Agassi, Joseph; Chesler, Elissa J.; Crabbe, John C.; Crusio, Wim E.; Eilam, David; Gerlai, Robert; Golani, Ilan; Gomez-Marin, Alex; Heller, Ruth; Iraqi, Fuad; Jaljuli, Iman; Karp, Natasha A.; Morgan, Hugh; Nicholson, George; Pfaff, Donald W.; Richter, S. Helene; Stark, Philip B.; Stiedl, Oliver; Stodden, Victoria; Tarantino, Lisa M.; Tucci, Valter; Valdar, William; Williams, Robert W.; Würbel, Hanno; Benjamini, Yoav

The scientific community is increasingly concerned with the proportion of published “discoveries” that are not replicated in subsequent studies. The field of rodent behavioral phenotyping was one of the first to raise this concern, and to relate it to other methodological issues: the complex

Chromatin Controls DNA Replication Origin Selection, Lagging-Strand Synthesis, and Replication Fork Rates.

Science.gov (United States)

Kurat, Christoph F; Yeeles, Joseph T P; Patel, Harshil; Early, Anne; Diffley, John F X

2017-01-05

The integrity of eukaryotic genomes requires rapid and regulated chromatin replication. How this is accomplished is still poorly understood. Using purified yeast replication proteins and fully chromatinized templates, we have reconstituted this process in vitro. We show that chromatin enforces DNA replication origin specificity by preventing non-specific MCM helicase loading. Helicase activation occurs efficiently in the context of chromatin, but subsequent replisome progression requires the histone chaperone FACT (facilitates chromatin transcription). The FACT-associated Nhp6 protein, the nucleosome remodelers INO80 or ISW1A, and the lysine acetyltransferases Gcn5 and Esa1 each contribute separately to maximum DNA synthesis rates. Chromatin promotes the regular priming of lagging-strand DNA synthesis by facilitating DNA polymerase α function at replication forks. Finally, nucleosomes disrupted during replication are efficiently re-assembled into regular arrays on nascent DNA. Our work defines the minimum requirements for chromatin replication in vitro and shows how multiple chromatin factors might modulate replication fork rates in vivo. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

Applied field test procedures on petroleum release sites

International Nuclear Information System (INIS)

Gilbert, G.; Nichols, L.

1995-01-01

The effective remediation of petroleum contaminated soils and ground water is a significant issue for Williams Pipe Line Co. (Williams): costing $6.8 million in 1994. It is in the best interest, then, for Williams to adopt approaches and apply technologies that will be both cost-effective and comply with regulations. Williams has found the use of soil vapor extraction (SVE) and air sparging (AS) field test procedures at the onset of a petroleum release investigation/remediation accomplish these goals. This paper focuses on the application of AS/SVE as the preferred technology to a specific type of remediation: refined petroleum products. In situ field tests are used prior to designing a full-scale remedial system to first validate or disprove initial assumptions on applicability of the technology. During the field test, remedial system design parameters are also collected to tailor the design and operation of a full-scale system to site specific conditions: minimizing cost and optimizing effectiveness. In situ field tests should be designed and operated to simulate as close as possible the operation of a full-scale remedial system. The procedures of an in situ field test will be presented. The results of numerous field tests and the associated costs will also be evaluated and compared to full-scale remedial systems and total project costs to demonstrate overall effectiveness. There are many advantages of As/SVE technologies over conventional fluid extraction or SVE systems alone. However, the primary advantage is the ability to simultaneously reduce volatile and biodegradable compound concentrations in the phreatic, capillary fringe, and unsaturated zones

Mechanisms of DNA replication termination.

Science.gov (United States)

Dewar, James M; Walter, Johannes C

2017-08-01

Genome duplication is carried out by pairs of replication forks that assemble at origins of replication and then move in opposite directions. DNA replication ends when converging replication forks meet. During this process, which is known as replication termination, DNA synthesis is completed, the replication machinery is disassembled and daughter molecules are resolved. In this Review, we outline the steps that are likely to be common to replication termination in most organisms, namely, fork convergence, synthesis completion, replisome disassembly and decatenation. We briefly review the mechanism of termination in the bacterium Escherichia coli and in simian virus 40 (SV40) and also focus on recent advances in eukaryotic replication termination. In particular, we discuss the recently discovered E3 ubiquitin ligases that control replisome disassembly in yeast and higher eukaryotes, and how their activity is regulated to avoid genome instability.

Rolling replication of UV-irradiated duplex DNA in the phi X174 replicative-form----single-strand replication system in vitro

International Nuclear Information System (INIS)

Shavitt, O.; Livneh, Z.

1989-01-01

Cloning of the phi X174 viral origin of replication into phage M13mp8 produced an M13-phi X174 chimera, the DNA of which directed efficient replicative-form----single-strand rolling replication in vitro. This replication assay was performed with purified phi X174-encoded gene A protein, Escherichia coli rep helicase, single-stranded DNA-binding protein, and DNA polymerase III holoenzyme. The nicking of replicative-form I (RFI) DNA by gene A protein was essentially unaffected by the presence of UV lesions in the DNA. However, unwinding of UV-irradiated DNA by the rep helicase was inhibited twofold as compared with unwinding of the unirradiated substrate. UV irradiation of the substrate DNA caused a strong inhibition in its ability to direct DNA synthesis. However, even DNA preparations that contained as many as 10 photodimers per molecule still supported the synthesis of progeny full-length single-stranded DNA. The appearance of full-length radiolabeled products implied at least two full rounds of replication, since the first round released the unlabeled plus viral strand of the duplex DNA. Pretreatment of the UV-irradiated DNA substrate with purified pyrimidine dimer endonuclease from Micrococcus luteus, which converted photodimer-containing supercoiled RFI DNA into relaxed, nicked RFII DNA and thus prevented its replication, reduced DNA synthesis by 70%. Analysis of radiolabeled replication products by agarose gel electrophoresis followed by autoradiography revealed that this decrease was due to a reduction in the synthesis of progeny full-length single-stranded DNA. This implies that 70 to 80% of the full-length DNA products produced in this system were synthesized on molecules that carried photodimers

Mammalian RAD52 Functions in Break-Induced Replication Repair of Collapsed DNA Replication Forks

DEFF Research Database (Denmark)

Sotiriou, Sotirios K; Kamileri, Irene; Lugli, Natalia

2016-01-01

Human cancers are characterized by the presence of oncogene-induced DNA replication stress (DRS), making them dependent on repair pathways such as break-induced replication (BIR) for damaged DNA replication forks. To better understand BIR, we performed a targeted siRNA screen for genes whose...... RAD52 facilitates repair of collapsed DNA replication forks in cancer cells....

Field Testing of Cryogenic Carbon Capture

Energy Technology Data Exchange (ETDEWEB)

Sayre, Aaron [Sustainable Energy Solutions, LLC; Frankman, Dave [Sustainable Energy Solutions, LLC; Baxter, Andrew [Sustainable Energy Solutions, LLC; Stitt, Kyler [Sustainable Energy Solutions, LLC; Baxter, Larry [Sustainable Energy Solutions, LLC; Brigham Young Univ., Provo, UT (United States)

2017-07-17

Sustainable Energy Solutions has been developing Cryogenic Carbon Capture™ (CCC) since 2008. In that time two processes have been developed, the External Cooling Loop and Compressed Flue Gas Cryogenic Carbon Capture processes (CCC ECL™ and CCC CFG™ respectively). The CCC ECL™ process has been scaled up to a 1TPD CO2 system. In this process the flue gas is cooled by an external refrigerant loop. SES has tested CCC ECL™ on real flue gas slip streams from subbituminous coal, bituminous coal, biomass, natural gas, shredded tires, and municipal waste fuels at field sites that include utility power stations, heating plants, cement kilns, and pilot-scale research reactors. The CO2 concentrations from these tests ranged from 5 to 22% on a dry basis. CO2 capture ranged from 95-99+% during these tests. Several other condensable species were also captured including NO2, SO2 and PMxx at 95+%. NO was also captured at a modest rate. The CCC CFG™ process has been scaled up to a .25 ton per day system. This system has been tested on real flue gas streams including subbituminous coal, bituminous coal and natural gas at field sites that include utility power stations, heating plants, and pilot-scale research reactors. CO2 concentrations for these tests ranged from 5 to 15% on a dry basis. CO2 capture ranged from 95-99+% during these tests. Several other condensable species were also captured including NO2, SO2 and PMxx at 95+%. NO was also captured at 90+%. Hg capture was also verified and the resulting effluent from CCC CFG™ was below a 1ppt concentration. This paper will focus on discussion of the capabilities of CCC, the results of field testing and the future steps surrounding the development of this technology.

REPLICATION TOOL AND METHOD OF PROVIDING A REPLICATION TOOL

DEFF Research Database (Denmark)

2016-01-01

The invention relates to a replication tool (1, 1a, 1b) for producing a part (4) with a microscale textured replica surface (5a, 5b, 5c, 5d). The replication tool (1, 1a, 1b) comprises a tool surface (2a, 2b) defining a general shape of the item. The tool surface (2a, 2b) comprises a microscale...... energy directors on flange portions thereof uses the replication tool (1, 1a, 1b) to form an item (4) with a general shape as defined by the tool surface (2a, 2b). The formed item (4) comprises a microscale textured replica surface (5a, 5b, 5c, 5d) with a lateral arrangement of polydisperse microscale...

Recommendations for Replication Research in Special Education: A Framework of Systematic, Conceptual Replications

Science.gov (United States)

Coyne, Michael D.; Cook, Bryan G.; Therrien, William J.

2016-01-01

Special education researchers conduct studies that can be considered replications. However, they do not often refer to them as replication studies. The purpose of this article is to consider the potential benefits of conceptualizing special education intervention research within a framework of systematic, conceptual replication. Specifically, we…

DNA replication and cancer

DEFF Research Database (Denmark)

Boyer, Anne-Sophie; Walter, David; Sørensen, Claus Storgaard

2016-01-01

A dividing cell has to duplicate its DNA precisely once during the cell cycle to preserve genome integrity avoiding the accumulation of genetic aberrations that promote diseases such as cancer. A large number of endogenous impacts can challenge DNA replication and cells harbor a battery of pathways...... causing DNA replication stress and genome instability. Further, we describe cellular and systemic responses to these insults with a focus on DNA replication restart pathways. Finally, we discuss the therapeutic potential of exploiting intrinsic replicative stress in cancer cells for targeted therapy....

Sequential accelerated tests: Improving the correlation of accelerated tests to module performance in the field

Science.gov (United States)

Felder, Thomas; Gambogi, William; Stika, Katherine; Yu, Bao-Ling; Bradley, Alex; Hu, Hongjie; Garreau-Iles, Lucie; Trout, T. John

2016-09-01

DuPont has been working steadily to develop accelerated backsheet tests that correlate with solar panels observations in the field. This report updates efforts in sequential testing. Single exposure tests are more commonly used and can be completed more quickly, and certain tests provide helpful predictions of certain backsheet failure modes. DuPont recommendations for single exposure tests are based on 25-year exposure levels for UV and humidity/temperature, and form a good basis for sequential test development. We recommend a sequential exposure of damp heat followed by UV then repetitions of thermal cycling and UVA. This sequence preserves 25-year exposure levels for humidity/temperature and UV, and correlates well with a large body of field observations. Measurements can be taken at intervals in the test, although the full test runs 10 months. A second, shorter sequential test based on damp heat and thermal cycling tests mechanical durability and correlates with loss of mechanical properties seen in the field. Ongoing work is directed toward shorter sequential tests that preserve good correlation to field data.

Barriers and facilitators to replicating an evidence-based palliative care model.

Science.gov (United States)

Davis, E Maxwell; Jamison, Paula; Brumley, Richard; Enguídanos, Susan

2006-01-01

Recognition of the difficulties involved in replicating evidence- based interventions is well documented in the literature within the medical field. Promising research findings are often not translated into practice, and if they are, there is a significant time gap between study conclusion and practice adoption. The purpose of this article is to describe the barriers and facilitators encountered by two managed care organizations while replicating an evidence-based end of life in-home palliative care model. Using Diffusion of Innovation Theory as a theoretical framework, results from focus groups and interviews with the project's clinical, administrative and research teams are presented and recommendations made for improving translational efforts. The process of replicating the end of life in-home palliative care model clearly illustrated the key elements required for successfully diffusing innovation. These key elements include marketing and communication, leadership, organizational support and training and mentorship. This qualitative process study provides clear, real world perspectives of the myriad of challenges encountered in replicating an evidence-based project.

Late-replicating X-chromosome: replication patterns in mammalian females

Directory of Open Access Journals (Sweden)

Tunin Karen

2002-01-01

Full Text Available The GTG-banding and 5-BrdU incorporation patterns of the late-replicating X-chromosome were studied in female dogs and cattle, and compared to human female patterns. The replication patterns of the short arm of the X-chromosomes did not show any difference between human, dog and cattle females. As to the long arm, some bands showed differences among the three studied species regarding the replication kinetics pattern. These differences were observed in a restricted region of the X-chromosome, delimited by Xq11 -> q25 in humans, by Xq1 -> q8 in dogs, and by Xq12 -> q32 in cattle. In an attempt to find out if these differences in the replication kinetics could be a reflection of differences in the localization of genes in that region of the X-chromosome, we used the probe for the human androgen receptor gene (AR localized at Xq12, which is in the region where we observed differences among the three studied species. We did not, however, observe hybridization signals. Our study goes on, using other human probes for genes located in the region Xq11 -> Xq25.

Faithful replication of grating patterns in polymer through electrohydrodynamic instabilities

International Nuclear Information System (INIS)

Li, H; Yu, W; Wang, T; Zhang, H; Cao, Y; Abraham, E; Desmulliez, M P Y

2014-01-01

Electrohydrodynamic instability patterning (EHDIP) as an alternative patterning method has attracted a great deal of attention over the past decade. This article demonstrates the faithful transfer of patterns with a high aspect ratio onto a polymer film via electrohydrodynamic instabilities for a given patterned grating mask. We perform a simple mathematical analysis to determine the influence of process parameters on the pressure difference ▵P. Through numerical simulation, it is demonstrated that thick films subject to large electric fields are essential to realize this faithful replication. In particular, the influence of the material properties of the polymer on pattern replication is discussed in detail. It is found that, to achieve the smaller periodic patterns with a higher resolution, film with a larger value of the dielectric constant and smaller value of the surface tension should be chosen. In addition, an ideal replication of the mask pattern with a short evolution time is possible by reducing the viscosity of the polymer liquid. Finally, the experiments of the pattern replication with and without defects are demonstrated to compare with the numerical simulation results. It is found that experiments are in good agreement with the simulation results and prove that the numerical simulation method provides an effective way to predict faithful replication. (paper)

A Replication by Any Other Name: A Systematic Review of Replicative Intervention Studies

Science.gov (United States)

Cook, Bryan G.; Collins, Lauren W.; Cook, Sara C.; Cook, Lysandra

2016-01-01

Replication research is essential to scientific knowledge. Reviews of replication studies often electronically search for "replicat*" as a textword, which does not identify studies that replicate previous research but do not self-identify as such. We examined whether the 83 intervention studies published in six non-categorical research…

Identifying significant temporal variation in time course microarray data without replicates

Directory of Open Access Journals (Sweden)

Porter Weston

2009-03-01

Full Text Available Abstract Background An important component of time course microarray studies is the identification of genes that demonstrate significant time-dependent variation in their expression levels. Until recently, available methods for performing such significance tests required replicates of individual time points. This paper describes a replicate-free method that was developed as part of a study of the estrous cycle in the rat mammary gland in which no replicate data was collected. Results A temporal test statistic is proposed that is based on the degree to which data are smoothed when fit by a spline function. An algorithm is presented that uses this test statistic together with a false discovery rate method to identify genes whose expression profiles exhibit significant temporal variation. The algorithm is tested on simulated data, and is compared with another recently published replicate-free method. The simulated data consists both of genes with known temporal dependencies, and genes from a null distribution. The proposed algorithm identifies a larger percentage of the time-dependent genes for a given false discovery rate. Use of the algorithm in a study of the estrous cycle in the rat mammary gland resulted in the identification of genes exhibiting distinct circadian variation. These results were confirmed in follow-up laboratory experiments. Conclusion The proposed algorithm provides a new approach for identifying expression profiles with significant temporal variation without relying on replicates. When compared with a recently published algorithm on simulated data, the proposed algorithm appears to identify a larger percentage of time-dependent genes for a given false discovery rate. The development of the algorithm was instrumental in revealing the presence of circadian variation in the virgin rat mammary gland during the estrous cycle.

Registered Replication Report

DEFF Research Database (Denmark)

Bouwmeester, S.; Verkoeijen, P. P.J.L.; Aczel, B.

2017-01-01

and colleagues. The results of studies using time pressure have been mixed, with some replication attempts observing similar patterns (e.g., Rand et al., 2014) and others observing null effects (e.g., Tinghög et al., 2013; Verkoeijen & Bouwmeester, 2014). This Registered Replication Report (RRR) assessed...... the size and variability of the effect of time pressure on cooperative decisions by combining 21 separate, preregistered replications of the critical conditions from Study 7 of the original article (Rand et al., 2012). The primary planned analysis used data from all participants who were randomly assigned...

Recovery from Proactive Semantic Interference and MRI Volume: A Replication and Extension Study.

Science.gov (United States)

Loewenstein, David A; Curiel, Rosie E; DeKosky, Steven; Rosselli, Monica; Bauer, Russell; Grieg-Custo, Maria; Penate, Ailyn; Li, Chunfei; Lizagarra, Gabriel; Golde, Todd; Adjouadi, Malek; Duara, Ranjan

2017-01-01

The rise in incidence of Alzheimer's disease (AD) has led to efforts to advance early detection of the disease during its preclinical stages. To achieve this, the field needs to develop more sensitive cognitive tests that relate to biological markers of disease pathology. Failure to recover from proactive interference (frPSI) is one such cognitive marker that is associated with volumetric reductions in the hippocampus, precuneus, and other AD-prone regions, and to amyloid load in the brain. The current study attempted to replicate and extend our previous findings that frPSI is a sensitive marker of early AD, and related to a unique pattern of volumetric loss in AD prone areas. Three different memory measures were examined relative to volumetric loss and cortical thickness among 45 participants with amnestic mild cognitive impairment. frPSI was uniquely associated with reduced volumes in the hippocampus (r = 0.50) precuneus (r = 0.41), and other AD prone regions, replicating previous findings. Strong associations between frPSI and lower entorhinal cortex volumes and cortical thickness (r≥0.60) and precuneus (r = 0.50) were also observed. Unique and strong associations between volumetric reductions and frPSI as observed by Loewenstein and colleagues were replicated. Together with cortical thickness findings, these results indicate that frPSI is worthy of further study as a sensitive and early cognitive marker of AD.

Combustion Safety Simplified Test Protocol Field Study

Energy Technology Data Exchange (ETDEWEB)

Brand, L. [Gas Technology Inst., Des Plaines, IL (United States); Cautley, D. [Gas Technology Inst., Des Plaines, IL (United States); Bohac, D. [Gas Technology Inst., Des Plaines, IL (United States); Francisco, P. [Gas Technology Inst., Des Plaines, IL (United States); Shen, L. [Gas Technology Inst., Des Plaines, IL (United States); Gloss, S. [Gas Technology Inst., Des Plaines, IL (United States)

2015-11-01

Combustions safety is an important step in the process of upgrading homes for energy efficiency. There are several approaches used by field practitioners, but researchers have indicated that the test procedures in use are complex to implement and provide too many false positives. Field failures often mean that the house is not upgraded until after remediation or not at all, if not include in the program. In this report the PARR and NorthernSTAR DOE Building America Teams provide a simplified test procedure that is easier to implement and should produce fewer false positives. A survey of state weatherization agencies on combustion safety issues, details of a field data collection instrumentation package, summary of data collected over seven months, data analysis and results are included. The project team collected field data on 11 houses in 2015.

International Expansion through Flexible Replication

DEFF Research Database (Denmark)

Jonsson, Anna; Foss, Nicolai Juul

2011-01-01

Business organizations may expand internationally by replicating a part of their value chain, such as a sales and marketing format, in other countries. However, little is known regarding how such “international replicators” build a format for replication, or how they can adjust it in order to ada......, etc.) are replicated in a uniform manner across stores, and change only very slowly (if at all) in response to learning (“flexible replication”). We conclude by discussing the factors that influence the approach to replication adopted by an international replicator.......Business organizations may expand internationally by replicating a part of their value chain, such as a sales and marketing format, in other countries. However, little is known regarding how such “international replicators” build a format for replication, or how they can adjust it in order to adapt...

Chromosomal Replication Complexity: A Novel DNA Metrics and Genome Instability Factor.

Directory of Open Access Journals (Sweden)

Andrei Kuzminov

2016-10-01

Full Text Available As the ratio of the copy number of the most replicated to the unreplicated regions in the same chromosome, the definition of chromosomal replication complexity (CRC appears to leave little room for variation, being either two during S-phase or one otherwise. However, bacteria dividing faster than they replicate their chromosome spike CRC to four and even eight. A recent experimental inquiry about the limits of CRC in Escherichia coli revealed two major reasons to avoid elevating it further: (i increased chromosomal fragmentation and (ii complications with subsequent double-strand break repair. Remarkably, examples of stable elevated CRC in eukaryotic chromosomes are well known under various terms like "differential replication," "underreplication," "DNA puffs," "onion-skin replication," or "re-replication" and highlight the phenomenon of static replication fork (sRF. To accurately describe the resulting "amplification by overinitiation," I propose a new term: "replification" (subchromosomal overreplication. In both prokaryotes and eukaryotes, replification, via sRF processing, causes double-strand DNA breaks and, with their repair elevating chromosomal rearrangements, represents a novel genome instability factor. I suggest how static replication bubbles could be stabilized and speculate that some tandem duplications represent such persistent static bubbles. Moreover, I propose how static replication bubbles could be transformed into tandem duplications, double minutes, or inverted triplications. Possible experimental tests of these models are discussed.

Success-Breeds-Success in Collective Political Behavior: Evidence from a Field Experiment

NARCIS (Netherlands)

Van De Rijt, Arnout; Akin, Idil; Willer, Robb; Feinberg, Matthew

2016-01-01

Scholars have proposed that the emergence of political movements is highly pathdependent, such that early mobilization successes may lead to disproportionately greater eventual success. This article replicates a unique field experiment testing for positive feedback in internet petition signing (van

Sterol Binding by the Tombusviral Replication Proteins Is Essential for Replication in Yeast and Plants.

Science.gov (United States)

Xu, Kai; Nagy, Peter D

2017-04-01

Membranous structures derived from various organelles are important for replication of plus-stranded RNA viruses. Although the important roles of co-opted host proteins in RNA virus replication have been appreciated for a decade, the equally important functions of cellular lipids in virus replication have been gaining full attention only recently. Previous work with Tomato bushy stunt tombusvirus (TBSV) in model host yeast has revealed essential roles for phosphatidylethanolamine and sterols in viral replication. To further our understanding of the role of sterols in tombusvirus replication, in this work we showed that the TBSV p33 and p92 replication proteins could bind to sterols in vitro The sterol binding by p33 is supported by cholesterol recognition/interaction amino acid consensus (CRAC) and CARC-like sequences within the two transmembrane domains of p33. Mutagenesis of the critical Y amino acids within the CRAC and CARC sequences blocked TBSV replication in yeast and plant cells. We also showed the enrichment of sterols in the detergent-resistant membrane (DRM) fractions obtained from yeast and plant cells replicating TBSV. The DRMs could support viral RNA synthesis on both the endogenous and exogenous templates. A lipidomic approach showed the lack of enhancement of sterol levels in yeast and plant cells replicating TBSV. The data support the notion that the TBSV replication proteins are associated with sterol-rich detergent-resistant membranes in yeast and plant cells. Together, the results obtained in this study and the previously published results support the local enrichment of sterols around the viral replication proteins that is critical for TBSV replication. IMPORTANCE One intriguing aspect of viral infections is their dependence on efficient subcellular assembly platforms serving replication, virion assembly, or virus egress via budding out of infected cells. These assembly platforms might involve sterol-rich membrane microdomains, which are

Mcm10 regulates DNA replication elongation by stimulating the CMG replicative helicase.

Science.gov (United States)

Lõoke, Marko; Maloney, Michael F; Bell, Stephen P

2017-02-01

Activation of the Mcm2-7 replicative DNA helicase is the committed step in eukaryotic DNA replication initiation. Although Mcm2-7 activation requires binding of the helicase-activating proteins Cdc45 and GINS (forming the CMG complex), an additional protein, Mcm10, drives initial origin DNA unwinding by an unknown mechanism. We show that Mcm10 binds a conserved motif located between the oligonucleotide/oligosaccharide fold (OB-fold) and A subdomain of Mcm2. Although buried in the interface between these domains in Mcm2-7 structures, mutations predicted to separate the domains and expose this motif restore growth to conditional-lethal MCM10 mutant cells. We found that, in addition to stimulating initial DNA unwinding, Mcm10 stabilizes Cdc45 and GINS association with Mcm2-7 and stimulates replication elongation in vivo and in vitro. Furthermore, we identified a lethal allele of MCM10 that stimulates initial DNA unwinding but is defective in replication elongation and CMG binding. Our findings expand the roles of Mcm10 during DNA replication and suggest a new model for Mcm10 function as an activator of the CMG complex throughout DNA replication. © 2017 Lõoke et al.; Published by Cold Spring Harbor Laboratory Press.

Challenges in reproducibility of genetic association studies: lessons learned from the obesity field.

Science.gov (United States)

Li, A; Meyre, D

2013-04-01

A robust replication of initial genetic association findings has proved to be difficult in human complex diseases and more specifically in the obesity field. An obvious cause of non-replication in genetic association studies is the initial report of a false positive result, which can be explained by a non-heritable phenotype, insufficient sample size, improper correction for multiple testing, population stratification, technical biases, insufficient quality control or inappropriate statistical analyses. Replication may, however, be challenging even when the original study describes a true positive association. The reasons include underpowered replication samples, gene × gene, gene × environment interactions, genetic and phenotypic heterogeneity and subjective interpretation of data. In this review, we address classic pitfalls in genetic association studies and provide guidelines for proper discovery and replication genetic association studies with a specific focus on obesity.

A prototype tap test imaging system: Initial field test results

Science.gov (United States)

Peters, J. J.; Barnard, D. J.; Hudelson, N. A.; Simpson, T. S.; Hsu, D. K.

2000-05-01

This paper describes a simple, field-worthy tap test imaging system that gives quantitative information about the size, shape, and severity of defects and damages. The system consists of an accelerometer, electronic circuits for conditioning the signal and measuring the impact duration, a laptop PC and data acquisition and processing software. The images are generated manually by tapping on a grid printed on a plastic sheet laid over the part's surface. A mechanized scanner is currently under development. The prototype has produced images for a variety of aircraft composite and metal honeycomb structures containing flaws, damages, and repairs. Images of the local contact stiffness, deduced from the impact duration using a spring model, revealed quantitatively the stiffness reduction due to flaws and damages, as well as the stiffness enhancement due to substructures. The system has been field tested on commercial and military aircraft as well as rotor blades and engine decks on helicopters. Field test results will be shown and the operation of the system will be demonstrated.—This material is based upon work supported by the Federal Aviation Administration under Contract #DTFA03-98-D-00008, Delivery Order No. IA016 and performed at Iowa State University's Center for NDE as part of the Center for Aviation Systems Reliability program.

A Blood Test for Alzheimer's Disease: Progress, Challenges, and Recommendations.

Science.gov (United States)

Kiddle, Steven J; Voyle, Nicola; Dobson, Richard J B

2018-03-29

Ever since the discovery of APOEɛ4 around 25 years ago, researchers have been excited about the potential of a blood test for Alzheimer's disease (AD). Since then researchers have looked for genetic, protein, metabolite, and/or gene expression markers of AD and related phenotypes. However, no blood test for AD is yet being used in the clinical setting. We first review the trends and challenges in AD blood biomarker research, before giving our personal recommendations to help researchers overcome these challenges. While some degree of consistency and replication has been seen across independent studies, several high-profile studies have seemingly failed to replicate. Partly due to academic incentives, there is a reluctance in the field to report predictive ability, to publish negative findings, and to independently replicate the work of others. If this can be addressed, then we will know sooner whether a blood test for AD or related phenotypes with clinical utility can be developed.

Automated particulate sampler field test model operations guide

Energy Technology Data Exchange (ETDEWEB)

Bowyer, S.M.; Miley, H.S.

1996-10-01

The Automated Particulate Sampler Field Test Model Operations Guide is a collection of documents which provides a complete picture of the Automated Particulate Sampler (APS) and the Field Test in which it was evaluated. The Pacific Northwest National Laboratory (PNNL) Automated Particulate Sampler was developed for the purpose of radionuclide particulate monitoring for use under the Comprehensive Test Ban Treaty (CTBT). Its design was directed by anticipated requirements of small size, low power consumption, low noise level, fully automatic operation, and most predominantly the sensitivity requirements of the Conference on Disarmament Working Paper 224 (CDWP224). This guide is intended to serve as both a reference document for the APS and to provide detailed instructions on how to operate the sampler. This document provides a complete description of the APS Field Test Model and all the activity related to its evaluation and progression.

Fracture behavior of low-density replicated aluminum alloy foams

NARCIS (Netherlands)

Amsterdam, E.; Goodall, R.; Mortensen, A.; Onck, P. R.; De Hosson, J. Th. M.

2008-01-01

Tensile tests have been performed on replicated aluminum alloy foams of relative density between 4.5% and 8%. During the test the electrical resistance was measured with a four-point set-up and the displacements along the gage section were measured using a digital image correlation (DIC) technique.

Remote field eddy current testing

International Nuclear Information System (INIS)

Cheong, Y. M.; Jung, H. K.; Huh, H.; Lee, Y. S.; Shim, C. M.

2001-03-01

The state-of-art technology of the remote field eddy current, which is actively developed as an electromagnetic non-destructive testing tool for ferromagnetic tubes, is described. The historical background and recent R and D activities of remote-field eddy current technology are explained including the theoretical development of remote field eddy current, such as analytical and numerical approach, and the results of finite element analysis. The influencing factors for actual applications, such as the effect of frequency, magnetic permeability, receiving sensitivity, and difficulties of detection and classification of defects are also described. Finally, two examples of actual application, 1) the gap measurement between pressure tubes and calandria tube in CANDU reactor and, 2) the detection of defects in the ferromagnetic heat exchanger tubes, are described. The future research efforts are also included

Replication initiatives will not salvage the trustworthiness of psychology.

Science.gov (United States)

Coyne, James C

2016-05-31

Replication initiatives in psychology continue to gather considerable attention from far outside the field, as well as controversy from within. Some accomplishments of these initiatives are noted, but this article focuses on why they do not provide a general solution for what ails psychology. There are inherent limitations to mass replications ever being conducted in many areas of psychology, both in terms of their practicality and their prospects for improving the science. Unnecessary compromises were built into the ground rules for design and publication of the Open Science Collaboration: Psychology that undermine its effectiveness. Some ground rules could actually be flipped into guidance for how not to conduct replications. Greater adherence to best publication practices, transparency in the design and publishing of research, strengthening of independent post-publication peer review and firmer enforcement of rules about data sharing and declarations of conflict of interest would make many replications unnecessary. Yet, it has been difficult to move beyond simple endorsement of these measures to consistent implementation. Given the strong institutional support for questionable publication practices, progress will depend on effective individual and collective use of social media to expose lapses and demand reform. Some recent incidents highlight the necessity of this.

A serine palmitoyltransferase inhibitor blocks hepatitis C virus replication in human hepatocytes.

Science.gov (United States)

Katsume, Asao; Tokunaga, Yuko; Hirata, Yuichi; Munakata, Tsubasa; Saito, Makoto; Hayashi, Hitohisa; Okamoto, Koichi; Ohmori, Yusuke; Kusanagi, Isamu; Fujiwara, Shinya; Tsukuda, Takuo; Aoki, Yuko; Klumpp, Klaus; Tsukiyama-Kohara, Kyoko; El-Gohary, Ahmed; Sudoh, Masayuki; Kohara, Michinori

2013-10-01

Host cell lipid rafts form a scaffold required for replication of hepatitis C virus (HCV). Serine palmitoyltransferases (SPTs) produce sphingolipids, which are essential components of the lipid rafts that associate with HCV nonstructural proteins. Prevention of the de novo synthesis of sphingolipids by an SPT inhibitor disrupts the HCV replication complex and thereby inhibits HCV replication. We investigated the ability of the SPT inhibitor NA808 to prevent HCV replication in cells and mice. We tested the ability of NA808 to inhibit SPT's enzymatic activity in FLR3-1 replicon cells. We used a replicon system to select for HCV variants that became resistant to NA808 at concentrations 4- to 6-fold the 50% inhibitory concentration, after 14 rounds of cell passage. We assessed the ability of NA808 or telaprevir to inhibit replication of HCV genotypes 1a, 1b, 2a, 3a, and 4a in mice with humanized livers (transplanted with human hepatocytes). NA808 was injected intravenously, with or without pegylated interferon alfa-2a and HCV polymerase and/or protease inhibitors. NA808 prevented HCV replication via noncompetitive inhibition of SPT; no resistance mutations developed. NA808 prevented replication of all HCV genotypes tested in mice with humanized livers. Intravenous NA808 significantly reduced viral load in the mice and had synergistic effects with pegylated interferon alfa-2a and HCV polymerase and protease inhibitors. The SPT inhibitor NA808 prevents replication of HCV genotypes 1a, 1b, 2a, 3a, and 4a in cultured hepatocytes and in mice with humanized livers. It might be developed for treatment of HCV infection or used in combination with pegylated interferon alfa-2a or HCV polymerase or protease inhibitors. Copyright © 2013 AGA Institute. Published by Elsevier Inc. All rights reserved.

Who Needs Replication?

Science.gov (United States)

Porte, Graeme

2013-01-01

In this paper, the editor of a recent Cambridge University Press book on research methods discusses replicating previous key studies to throw more light on their reliability and generalizability. Replication research is presented as an accepted method of validating previous research by providing comparability between the original and replicated…

Mechanisms of bacterial DNA replication restart

Science.gov (United States)

Windgassen, Tricia A; Wessel, Sarah R; Bhattacharyya, Basudeb

2018-01-01

Abstract Multi-protein DNA replication complexes called replisomes perform the essential process of copying cellular genetic information prior to cell division. Under ideal conditions, replisomes dissociate only after the entire genome has been duplicated. However, DNA replication rarely occurs without interruptions that can dislodge replisomes from DNA. Such events produce incompletely replicated chromosomes that, if left unrepaired, prevent the segregation of full genomes to daughter cells. To mitigate this threat, cells have evolved ‘DNA replication restart’ pathways that have been best defined in bacteria. Replication restart requires recognition and remodeling of abandoned replication forks by DNA replication restart proteins followed by reloading of the replicative DNA helicase, which subsequently directs assembly of the remaining replisome subunits. This review summarizes our current understanding of the mechanisms underlying replication restart and the proteins that drive the process in Escherichia coli (PriA, PriB, PriC and DnaT). PMID:29202195

Testing of Photomultiplier Tubes in a Magnetic Field

Science.gov (United States)

Waldron, Zachary; A1 Collaboration

2016-09-01

The A1 collaboration at MAMI in Mainz, Germany has designed a neutron detector that can be used in experiments to measure the electric form factor of the neutron. They will measure elastic scattering from the neutron, using the polarized electron beam from MAMI at A1's experimental hall. The detector will be composed of two walls of staggered scintillator bars which will be read out by photomultiplier tubes (PMT), connected to both ends of each scintillator via light guides. The experiment requires a magnetic field with strength of 1 Tesla, 2m away from the first scintillator wall. The resulting fringe field is sufficient to disrupt the PMTs, despite the addition of Mu Metal shielding. The effects of the fringe field on these PMTs was tested to optimize the amplification of the PMTs. A Helmholtz Coil was designed to generate a controlled magnetic field with equivalent strength to the field that the PMTs will encounter. The PMTs were read out using a multi-channel analyzer, were tested at various angles relative to the magnetic field in order to determine the optimal orientation to minimize signal disruption. Tests were also performed to determine: the neutron detector response to cosmic radiation; and the best method for measuring a magnetic field's strength in two dimensions. National Science Foundation Grant No. IIA-1358175.

Why threefold-replication of families?

Science.gov (United States)

Fitzpatrick, Gerald L.

1998-04-01

In spite of the many successes of the standard model of particle physics, the observed proliferation of matter-fields, in the form of ``replicated'' generations or families, is a major unsolved problem. In this paper, I explore some of the algebraic, geometric and physical consequences of a new organizing principle for fundamental fermions (quarks and leptons)(Gerald L. Fitzpatrick, phThe Family Problem--New Internal Algebraic and Geometric Regularities), Nova Scientific Press, Issaquah, Washington, 1997. Read more about this book (ISBN 0--9655695--0--0) and its subject matter at: http://www.tp.umu.se/TIPTOP and/or amazon.com>http://www.amazon.com.. The essence of the new organizing principle is the idea that the standard-model concept of scalar fermion numbers f can be generalized. In particular, a ``generalized fermion number,'' which consists of a 2× 2 matrix F that ``acts'' on an internal 2-space, instead of spacetime, is taken to describe certain internal properties of fundamental fermions. This generalization automatically introduces internal degrees of freedom that ``explain,'' among other things, family replication and the number (three) of families observed in nature.

Analysis of JC virus DNA replication using a quantitative and high-throughput assay

Science.gov (United States)

Shin, Jong; Phelan, Paul J.; Chhum, Panharith; Bashkenova, Nazym; Yim, Sung; Parker, Robert; Gagnon, David; Gjoerup, Ole; Archambault, Jacques; Bullock, Peter A.

2015-01-01

Progressive Multifocal Leukoencephalopathy (PML) is caused by lytic replication of JC virus (JCV) in specific cells of the central nervous system. Like other polyomaviruses, JCV encodes a large T-antigen helicase needed for replication of the viral DNA. Here, we report the development of a luciferase-based, quantitative and high-throughput assay of JCV DNA replication in C33A cells, which, unlike the glial cell lines Hs 683 and U87, accumulate high levels of nuclear T-ag needed for robust replication. Using this assay, we investigated the requirement for different domains of T-ag, and for specific sequences within and flanking the viral origin, in JCV DNA replication. Beyond providing validation of the assay, these studies revealed an important stimulatory role of the transcription factor NF1 in JCV DNA replication. Finally, we show that the assay can be used for inhibitor testing, highlighting its value for the identification of antiviral drugs targeting JCV DNA replication. PMID:25155200

Some Tests on Heather Field Moraine Clay

DEFF Research Database (Denmark)

Jørgensen, Mogens B.; Jacobsen, Moust

This report deals with oedometer tests on three samples of moraine clay from the Heather Field in the English part of the North Sea. The tests have been carried out in the very unelastic apparatus used in Denmark and with special test procedures differing from the ones used elsewhere. In Denmark...... Moraine Clay covers a large part of the surface, and it has therefore been investigated extensively in the field and in the laboratories during the last 25 years. It is to day - from a geotechnical point of view - the best known clay in Denmark. It could therefore be of some interest to compare...... the English North Sea moraine clays with the corresponding Danish Moraine Clays. The Danish test procedures are explained in details and some comments are given in the hope that they may not be banalities all of them....

What Should Researchers Expect When They Replicate Studies? A Statistical View of Replicability in Psychological Science.

Science.gov (United States)

Patil, Prasad; Peng, Roger D; Leek, Jeffrey T

2016-07-01

A recent study of the replicability of key psychological findings is a major contribution toward understanding the human side of the scientific process. Despite the careful and nuanced analysis reported, the simple narrative disseminated by the mass, social, and scientific media was that in only 36% of the studies were the original results replicated. In the current study, however, we showed that 77% of the replication effect sizes reported were within a 95% prediction interval calculated using the original effect size. Our analysis suggests two critical issues in understanding replication of psychological studies. First, researchers' intuitive expectations for what a replication should show do not always match with statistical estimates of replication. Second, when the results of original studies are very imprecise, they create wide prediction intervals-and a broad range of replication effects that are consistent with the original estimates. This may lead to effects that replicate successfully, in that replication results are consistent with statistical expectations, but do not provide much information about the size (or existence) of the true effect. In this light, the results of the Reproducibility Project: Psychology can be viewed as statistically consistent with what one might expect when performing a large-scale replication experiment. © The Author(s) 2016.

Field tests on migration of TRU-nuclide, (2). Migration test for engineered barrier materials in aerated soil

International Nuclear Information System (INIS)

Maeda, Toshikatsu; Tanaka, Tadao; Mukai, Masayuki

2003-01-01

Field tests on migration of radionuclides for engineered barrier materials such as bentonite and cementitious materials were performed. The tests were run under both wet conditions with artificial rainfall and dry conditions with natural rainfall. Laboratory experiments such as batch adsorption tests were also conducted to analyze the result of field test. The results of field tests agreed with the predicted moisture conditions and the migration behaviors observed at the laboratory experiment that is reported so far. For bentonite material, the movements of the tracer were calculated using known information such as the results of batch sorption tests and migration mechanism. Comparing the result of field test and calculations, it is suggested that tracer migration behavior in bentonite material in field can be evaluated quantitatively by the known migration mechanism and the results of laboratory experiments such as batch sorption test. (author)

Prospective elementary teachers' conceptions of multidigit number: exemplifying a replication framework for mathematics education

Science.gov (United States)

Jacobson, Erik; Simpson, Amber

2018-04-01

Replication studies play a critical role in scientific accumulation of knowledge, yet replication studies in mathematics education are rare. In this study, the authors replicated Thanheiser's (Educational Studies in Mathematics 75:241-251, 2010) study of prospective elementary teachers' conceptions of multidigit number and examined the main claim that most elementary pre-service teachers think about digits incorrectly at least some of the time. Results indicated no statistically significant difference in the distribution of conceptions between the original and replication samples and, moreover, no statistically significant differences in the distribution of sub-conceptions among prospective teachers with the most common conception. These results suggest confidence is warranted both in the generality of the main claim and in the utility of the conceptions framework for describing prospective elementary teachers' conceptions of multidigit number. The report further contributes a framework for replication of mathematics education research adapted from the field of psychology.

Field tests on migration of TRU-nuclide, (1). General introduction

International Nuclear Information System (INIS)

Ogawa, Hiromichi; Tanaka, Tadao; Mukai, Masayuki

2003-01-01

The field migration test using TRU nuclide was carried out as a cooperative research project between JAERI (Japan Atomic Energy Research Institute) and CIRP (China Institute for Radiation Protection). This report introduced the out-line of the field migration test and described the outline of the series of 'Field Test on Migration of TRU-nuclide' and main results as a summary report. (author)

Preirradiation of host (monkey) cells mitigates the effects of UV upon simian virus 40 DNA replication

International Nuclear Information System (INIS)

Scaria, A.; Edenberg, H.J.

1987-01-01

The authors examined the effects of preirradiation of host (monkey) cells upon the replication of UV-damaged SV40. Control cells and cells preirradiated with low fluences of UV were infected with undamaged SV40, and the immediate effects of a subsequent irradiation were determined. UV inhibited total SV40 DNA synthesis in both preirradiated and control cells, but the extent of inhibition was less in the preirradiated cells. A test fluence of 60 J/m 2 to SV40 replicating in preirradiated cells reduced synthesis only as much as a test fluence of 25 J/m 2 in control cells. The fraction of recently replicated SV40 molecules that re-entered the replication pool and subsequently completed one round of replication in the first 2 h after UV was also decreased less in the preirradiated cells. Thus preirradiation of the host cell mitigates the immediate inhibitory effects of a subsequent UV exposure upon SV40 replication. (Auth.)

Timing, coordination, and rhythm: Acrobatics at the DNA replication fork

KAUST Repository

Hamdan, Samir

2010-04-09

In DNA replication, the antiparallel nature of the parental duplex imposes certain constraints on the activity of the DNA polymerases that synthesize new DNA. The leading-strand polymerase advances in a continuous fashion, but the lagging-strand polymerase is forced to restart at short intervals. In several prokaryotic systems studied so far, this problem is solved by the formation of a loop in the lagging strand of the replication fork to reorient the lagging-strand DNA polymerase so that it advances in parallel with the leading-strand polymerase. The replication loop grows and shrinks during each cycle of Okazaki fragment synthesis. The timing of Okazaki fragment synthesis and loop formation is determined by a subtle interplay of enzymatic activities at the fork. Recent developments in single-molecule techniques have enabled the direct observation of these processes and have greatly contributed to a better understanding of the dynamic nature of the replication fork. Here, we will review recent experimental advances, present the current models, and discuss some of the exciting developments in the field. 2010 by The American Society for Biochemistry and Molecular Biology, Inc.

Timing, coordination, and rhythm: Acrobatics at the DNA replication fork

KAUST Repository

Hamdan, Samir; van Oijen, Antoine M.

2010-01-01

In DNA replication, the antiparallel nature of the parental duplex imposes certain constraints on the activity of the DNA polymerases that synthesize new DNA. The leading-strand polymerase advances in a continuous fashion, but the lagging-strand polymerase is forced to restart at short intervals. In several prokaryotic systems studied so far, this problem is solved by the formation of a loop in the lagging strand of the replication fork to reorient the lagging-strand DNA polymerase so that it advances in parallel with the leading-strand polymerase. The replication loop grows and shrinks during each cycle of Okazaki fragment synthesis. The timing of Okazaki fragment synthesis and loop formation is determined by a subtle interplay of enzymatic activities at the fork. Recent developments in single-molecule techniques have enabled the direct observation of these processes and have greatly contributed to a better understanding of the dynamic nature of the replication fork. Here, we will review recent experimental advances, present the current models, and discuss some of the exciting developments in the field. 2010 by The American Society for Biochemistry and Molecular Biology, Inc.

Statistical correction of the Winner's Curse explains replication variability in quantitative trait genome-wide association studies.

Directory of Open Access Journals (Sweden)

Cameron Palmer

2017-07-01

Full Text Available Genome-wide association studies (GWAS have identified hundreds of SNPs responsible for variation in human quantitative traits. However, genome-wide-significant associations often fail to replicate across independent cohorts, in apparent inconsistency with their apparent strong effects in discovery cohorts. This limited success of replication raises pervasive questions about the utility of the GWAS field. We identify all 332 studies of quantitative traits from the NHGRI-EBI GWAS Database with attempted replication. We find that the majority of studies provide insufficient data to evaluate replication rates. The remaining papers replicate significantly worse than expected (p < 10-14, even when adjusting for regression-to-the-mean of effect size between discovery- and replication-cohorts termed the Winner's Curse (p < 10-16. We show this is due in part to misreporting replication cohort-size as a maximum number, rather than per-locus one. In 39 studies accurately reporting per-locus cohort-size for attempted replication of 707 loci in samples with similar ancestry, replication rate matched expectation (predicted 458, observed 457, p = 0.94. In contrast, ancestry differences between replication and discovery (13 studies, 385 loci cause the most highly-powered decile of loci to replicate worse than expected, due to difference in linkage disequilibrium.

DNA Replication Profiling Using Deep Sequencing.

Science.gov (United States)

Saayman, Xanita; Ramos-Pérez, Cristina; Brown, Grant W

2018-01-01

Profiling of DNA replication during progression through S phase allows a quantitative snap-shot of replication origin usage and DNA replication fork progression. We present a method for using deep sequencing data to profile DNA replication in S. cerevisiae.

Prototype Engineered Barrier System Field Test (PEBSFT)

International Nuclear Information System (INIS)

Ramirez, A.L.; Buscheck, T.; Carlson, R.; Daily, W.; Lee, K.; Lin, Wunan; Mao, Nai-hsien; Ueng, Tzou-Shin; Wang, H.; Watwood, D.

1991-08-01

This final report represents a summary of data and interpretations obtained from the Prototype Engineered Barrier System Field Test (PEBSFT) performed in G-Tunnel within the Nevada Test Site. The PEBSFT was conducted to evaluate the applicability of measurement techniques, numerical models, and procedures developed for future field tests that will be conducted in the Exploratory Studies Facilities (ESF) at Yucca Mountain. The primary objective of the test was to provide a basis for determining whether tests planned for the ESF have the potential to be successful. Chapter 1 on high frequency electromagnetic tomography discusses the rock mass electromagnetic permittivity and attenuation rate changes that were measured to characterize the water distribution in the near field of a simulated waste container. The data are used to obtain quantitative estimates of how the moisture content in the rock mass changes during heating and to infer properties of the spatial variability of water distribution, leading to conclusions about the role of fractures in the system. Chapter 2 discusses the changes in rock moisture content detected by the neutron logging probe. Chapter 3 permeability tests discusses the characterization of the in-situ permeability of the fractured tuff around the borehole. The air permeability testing apparatus, the testing procedures, and the data analysis are presented. Chapter 4 describes the moisture collection system installed in the heater borehole to trap and measure the moisture volumes. Chapter 5 describes relative humidity measurements made with the thermocouple psychrometer and capacitance sensors. Chapter 6 discusses gas pressure measurements in the G-Tunnel, addressing the calibration and installation of piezoresistive-gaged transducers. Chapter 7 describes the calibration and installation of thermocouples for temperature measurements. Chapter 8 discusses the results of the PEBSFT

Replication, randomization, and treatment design concepts for on-farm research

Science.gov (United States)

For most agronomists, randomization and replication are fundamental concepts that have a nearly sacred or spiritual status. They are an integral part of nearly all of our field-based activities. Some on-farm research falls into this category, simply because it is driven and designed by researchers w...

Induction of UV-resistant DNA replication in Escherichia coli: Induced stable DNA replication as an SOS function

International Nuclear Information System (INIS)

Kogoma, T.; Torrey, T.A.; Connaughton, M.J.

1979-01-01

The striking similarity between the treatments that induce SOS functions and those that result in stable DNA replication (continuous DNA replication in the absence of protein synthesis) prompted us to examine the possibility of stable DNA replication being a recA + lexA + -dependent SOS function. In addition to the treatments previously reported, ultraviolet (UV) irradiation or treatment with mitomycin C was also found to induce stable DNA replication. The thermal treatment of tif-1 strains did not result in detectable levels of stable DNA replication, but nalidixic acid readily induced the activity in these strains. The induction of stable DNA replication with nalidixic acid was severely suppressed in tif-1 lex A mutant strains. The inhibitory activity of lexA3 was negated by the presence of the spr-5l mutation, an intragenic suppressor of lexA3. Induced stable DNA replication was found to be considerably more resistant to UV irradiation than normal replication both in a uvr A6 strain and a uvr + strain. The UV-resistant replication occurred mostly in the semiconservative manner. The possible roles of stable DNA replication in repair of damaged DNA are discussed. (orig.)

Chromatin replication and epigenome maintenance

DEFF Research Database (Denmark)

Alabert, Constance; Groth, Anja

2012-01-01

Stability and function of eukaryotic genomes are closely linked to chromatin structure and organization. During cell division the entire genome must be accurately replicated and the chromatin landscape reproduced on new DNA. Chromatin and nuclear structure influence where and when DNA replication...... initiates, whereas the replication process itself disrupts chromatin and challenges established patterns of genome regulation. Specialized replication-coupled mechanisms assemble new DNA into chromatin, but epigenome maintenance is a continuous process taking place throughout the cell cycle. If DNA...

36 CFR 910.64 - Replication.

Science.gov (United States)

2010-07-01

... 36 Parks, Forests, and Public Property 3 2010-07-01 2010-07-01 false Replication. 910.64 Section 910.64 Parks, Forests, and Public Property PENNSYLVANIA AVENUE DEVELOPMENT CORPORATION GENERAL... DEVELOPMENT AREA Glossary of Terms § 910.64 Replication. Replication means the process of using modern methods...

Field test and mathematical modeling of bioremediation of an oil-contaminated soil. Part 1: Field test

International Nuclear Information System (INIS)

Li, K.Y.; Xu, T.; Colapret, J.A.; Cawley, W.A.; Bonner, J.S.; Ernest, A.; Verramachaneni, P.B.

1994-01-01

A fire-wall area (about 270 ft x 310 ft) with the Bunker C oil contaminated soil was selected for the bioremediation field test. This fire-wall area was separated into 18 plots by dirt dikes to test 6 bioremediation methods with three tests of each method. The six treatment methods were: (a) aeration with basic nutrients and indigenous organisms (BNIO); (b) aeration with basic nutrients and inoculation from a refinery wastewater treatment facility (BNSIWT); (c) aeration with an oleophilic fertilizer and indigenous organisms (INIPOL); (d) aeration with basic nutrients and biosurfactant organisms (EPA Seal Beach consortia) (EPA); (e) aeration with proprietary nutrients and organisms (PRO); and (f) aeration only for active control (CONTROL). This field test was conducted for 91 days. In general the oil contents in 18 plots were reduced, but the results showed significant fluctuations. A statistical method was used to examine if the oil reductions of six methods were the results from the random error of sampling and sample analysis or biodegradation. The results of the statistical analysis showed that oil reduction was concluded from all but the plots of PRO. From the data analysis, it may be concluded that the oil reduction rate in these studies is controlled by oil transfer from soil into the aqueous solution. An example of calculation was used to illustrate this conclusion

Eukaryotic DNA Replication Fork.

Science.gov (United States)

Burgers, Peter M J; Kunkel, Thomas A

2017-06-20

This review focuses on the biogenesis and composition of the eukaryotic DNA replication fork, with an emphasis on the enzymes that synthesize DNA and repair discontinuities on the lagging strand of the replication fork. Physical and genetic methodologies aimed at understanding these processes are discussed. The preponderance of evidence supports a model in which DNA polymerase ε (Pol ε) carries out the bulk of leading strand DNA synthesis at an undisturbed replication fork. DNA polymerases α and δ carry out the initiation of Okazaki fragment synthesis and its elongation and maturation, respectively. This review also discusses alternative proposals, including cellular processes during which alternative forks may be utilized, and new biochemical studies with purified proteins that are aimed at reconstituting leading and lagging strand DNA synthesis separately and as an integrated replication fork.

The progression of replication forks at natural replication barriers in live bacteria

NARCIS (Netherlands)

Moolman, M.C.; Tiruvadi Krishnan, S; Kerssemakers, J.W.J.; de Leeuw, R.; Lorent, V.J.F.; Sherratt, David J.; Dekker, N.H.

2016-01-01

Protein-DNA complexes are one of the principal barriers the replisome encounters during replication. One such barrier is the Tus-ter complex, which is a direction dependent barrier for replication fork progression. The details concerning the dynamics of the replisome when encountering these

Analysis of JC virus DNA replication using a quantitative and high-throughput assay

International Nuclear Information System (INIS)

Shin, Jong; Phelan, Paul J.; Chhum, Panharith; Bashkenova, Nazym; Yim, Sung; Parker, Robert; Gagnon, David; Gjoerup, Ole; Archambault, Jacques; Bullock, Peter A.

2014-01-01

Progressive Multifocal Leukoencephalopathy (PML) is caused by lytic replication of JC virus (JCV) in specific cells of the central nervous system. Like other polyomaviruses, JCV encodes a large T-antigen helicase needed for replication of the viral DNA. Here, we report the development of a luciferase-based, quantitative and high-throughput assay of JCV DNA replication in C33A cells, which, unlike the glial cell lines Hs 683 and U87, accumulate high levels of nuclear T-ag needed for robust replication. Using this assay, we investigated the requirement for different domains of T-ag, and for specific sequences within and flanking the viral origin, in JCV DNA replication. Beyond providing validation of the assay, these studies revealed an important stimulatory role of the transcription factor NF1 in JCV DNA replication. Finally, we show that the assay can be used for inhibitor testing, highlighting its value for the identification of antiviral drugs targeting JCV DNA replication. - Highlights: • Development of a high-throughput screening assay for JCV DNA replication using C33A cells. • Evidence that T-ag fails to accumulate in the nuclei of established glioma cell lines. • Evidence that NF-1 directly promotes JCV DNA replication in C33A cells. • Proof-of-concept that the HTS assay can be used to identify pharmacological inhibitor of JCV DNA replication

Analysis of JC virus DNA replication using a quantitative and high-throughput assay

Energy Technology Data Exchange (ETDEWEB)

Shin, Jong; Phelan, Paul J.; Chhum, Panharith; Bashkenova, Nazym; Yim, Sung; Parker, Robert [Department of Developmental, Molecular and Chemical Biology, Tufts University School of Medicine, Boston, MA 02111 (United States); Gagnon, David [Institut de Recherches Cliniques de Montreal (IRCM), 110 Pine Avenue West, Montreal, Quebec, Canada H2W 1R7 (Canada); Department of Biochemistry and Molecular Medicine, Université de Montréal, Montréal, Quebec (Canada); Gjoerup, Ole [Molecular Oncology Research Institute, Tufts Medical Center, Boston, MA 02111 (United States); Archambault, Jacques [Institut de Recherches Cliniques de Montreal (IRCM), 110 Pine Avenue West, Montreal, Quebec, Canada H2W 1R7 (Canada); Department of Biochemistry and Molecular Medicine, Université de Montréal, Montréal, Quebec (Canada); Bullock, Peter A., E-mail: Peter.Bullock@tufts.edu [Department of Developmental, Molecular and Chemical Biology, Tufts University School of Medicine, Boston, MA 02111 (United States)

2014-11-15

Progressive Multifocal Leukoencephalopathy (PML) is caused by lytic replication of JC virus (JCV) in specific cells of the central nervous system. Like other polyomaviruses, JCV encodes a large T-antigen helicase needed for replication of the viral DNA. Here, we report the development of a luciferase-based, quantitative and high-throughput assay of JCV DNA replication in C33A cells, which, unlike the glial cell lines Hs 683 and U87, accumulate high levels of nuclear T-ag needed for robust replication. Using this assay, we investigated the requirement for different domains of T-ag, and for specific sequences within and flanking the viral origin, in JCV DNA replication. Beyond providing validation of the assay, these studies revealed an important stimulatory role of the transcription factor NF1 in JCV DNA replication. Finally, we show that the assay can be used for inhibitor testing, highlighting its value for the identification of antiviral drugs targeting JCV DNA replication. - Highlights: • Development of a high-throughput screening assay for JCV DNA replication using C33A cells. • Evidence that T-ag fails to accumulate in the nuclei of established glioma cell lines. • Evidence that NF-1 directly promotes JCV DNA replication in C33A cells. • Proof-of-concept that the HTS assay can be used to identify pharmacological inhibitor of JCV DNA replication.

The Importance of Replication in Measurement Research: Using Curriculum-Based Measures with Postsecondary Students with Developmental Disabilities

Science.gov (United States)

Hosp, John L.; Ford, Jeremy W.; Huddle, Sally M.; Hensley, Kiersten K.

2018-01-01

Replication is a foundation of the development of a knowledge base in an evidence-based field such as education. This study includes two direct replications of Hosp, Hensley, Huddle, and Ford which found evidence of criterion-related validity of curriculum-based measurement (CBM) for reading and mathematics with postsecondary students with…

Flock House virus subgenomic RNA3 is replicated and its replication correlates with transactivation of RNA2

International Nuclear Information System (INIS)

Eckerle, Lance D.; Albarino, Cesar G.; Ball, L. Andrew.

2003-01-01

The nodavirus Flock House virus has a bipartite genome composed of RNAs 1 and 2, which encode the catalytic component of the RNA-dependent RNA polymerase (RdRp) and the capsid protein precursor, respectively. In addition to catalyzing replication of the viral genome, the RdRp also transcribes from RNA1 a subgenomic RNA3, which is both required for and suppressed by RNA2 replication. Here, we show that in the absence of RNA1 replication, FHV RdRp replicated positive-sense RNA3 transcripts fully and copied negative-sense RNA3 transcripts into positive strands. The two nonstructural proteins encoded by RNA3 were dispensable for replication, but sequences in the 3'-terminal 58 nucleotides were required. RNA3 variants that failed to replicate also failed to transactivate RNA2. These results imply that RNA3 is naturally produced both by transcription from RNA1 and by subsequent RNA1-independent replication and that RNA3 replication may be necessary for transactivation of RNA2

[New visual field testing possibilities (a preliminary report)].

Science.gov (United States)

Erichev, V P; Ermolaev, A P; Antonov, A A; Grigoryan, G L; Kosova, D V

2018-01-01

There are currently no portable mobile perimeters that allow visual field testing outside ophthalmologist's examination rooms. To develop a mobile perimetry technique based on use of a virtual reality headset (VR). The study involved 26 patients (30 eyes) with II-III stage primary open-angle glaucoma (POAG) with compensated IOP. Perimetry was performed for each patient twice - on Humphrey analyzer (test 30-2, 76 points) and employing similar strategy on a perimeter integrated into VR headset (Total Vision, Russia). Visual field testing was performed with an interval from 1 hour to 3 days. The results were comparatively analyzed. Patients tolerated the examination well. Comparative analysis of preliminary perimetry results obtained with both methods showed high degree of identity, so the results were concluded to be comparable. By visually isolating the wearer, VR headset achieves elimination of distractions and stable light conditions for visual field testing. The headset-perimeter is compact, mobile, easily transportable, can be used in the work of visiting medical teams and for examination at home.

How (and Whether) to Teach Undergraduates about the Replication Crisis in Psychological Science

Science.gov (United States)

Chopik, William J.; Bremner, Ryan H.; Defever, Andrew M.; Keller, Victor N.

2018-01-01

Over the past 10 years, crises surrounding replication, fraud, and best practices in research methods have dominated discussions in the field of psychology. However, no research exists examining how to communicate these issues to undergraduates and what effect this has on their attitudes toward the field. We developed and validated a 1-hr lecture…

Superconductor shields test chamber from ambient magnetic fields

Science.gov (United States)

Hildebrandt, A. F.

1965-01-01

Shielding a test chamber for magnetic components enables it to maintain a constant, low magnetic field. The chamber is shielded from ambient magnetic fields by a lead foil cylinder maintained in a superconducting state by liquid helium.

Sultan - forced flow, high field test facility

International Nuclear Information System (INIS)

Horvath, I.; Vecsey, G.; Weymuth, P.; Zellweger, J.

1981-01-01

Three European laboratories: CNEN (Frascati, I) ECN (Petten, NL) and SIN (Villigen, CH) decided to coordinate their development efforts and to install a common high field forced flow test facility at Villigen Switzerland. The test facility SULTAN (Supraleiter Testanlage) is presently under construction. As a first step, an 8T/1m bore solenoid with cryogenic periphery will be ready in 1981. The cryogenic system, data acquisition system and power supplies which are contributed by SIN are described. Experimental feasibilities, including cooling, and instrumentation are reviewed. Progress of components and facility construction is described. Planned extension of the background field up to 12T by insert coils is outlined. 5 refs

Test-particle motion in Einstein's unified field theory. I. General theory and application to neutral test particles

International Nuclear Information System (INIS)

Johnson, C.R.

1985-01-01

We develop a method for finding the exact equations of structure and motion of multipole test particles in Einstein's unified field theory: the theory of the nonsymmetric field. The method is also applicable to Einstein's gravitational theory. Particles are represented by singularities in the field. The method is covariant at each step of the analysis. We also apply the method and find both in Einstein's unified field theory and in Einstein's gravitational theory the equations of structure and motion of neutral pole-dipole test particles possessing no electromagnetic multipole moments. In the case of Einstein's gravitational theory the results are the well-known equations of structure and motion of a neutral pole-dipole test particle in a given background gravitational field. In the case of Einstein's unified field theory the results are the same, providing we identify a certain symmetric second-rank tensor field appearing in Einstein's theory with the metric and gravitational field. We therefore discover not only the equations of structure and motion of a neutral test particle in Einstein's unified field theory, but we also discover what field in Einstein's theory plays the role of metric and gravitational field

Test field for airborne laser scanning in Finland

Science.gov (United States)

Ahokas, E.; Kaartinen, H.; Kukko, A.; Litkey, P.

2014-11-01

Airborne laser scanning (ALS) is a widely spread operational measurement tool for obtaining 3D coordinates of the ground surface. There is a need for calibrating the ALS system and a test field for ALS was established at the end of 2013. The test field is situated in the city of Lahti, about 100 km to the north of Helsinki. The size of the area is approximately 3.5 km × 3.2 km. Reference data was collected with a mobile laser scanning (MLS) system assembled on a car roof. Some streets were measured both ways and most of them in one driving direction only. The MLS system of the Finnish Geodetic Institute (FGI) consists of a navigation system (NovAtel SPAN GNSS-IMU) and a laser scanner (FARO Focus3D 120). In addition to the MLS measurements more than 800 reference points were measured using a Trimble R8 VRS-GNSS system. Reference points are along the streets, on parking lots, and white pedestrian crossing line corners which can be used as reference targets. The National Land Survey of Finland has already used this test field this spring for calibrating their Leica ALS-70 scanner. Especially it was easier to determine the encoder scale factor parameter using this test field. Accuracy analysis of the MLS points showed that the point height RMSE is 2.8 cm and standard deviation is 2.6 cm. Our purpose is to measure both more MLS data and more reference points in the test field area to get a better spatial coverage. Calibration flight heights are planned to be 1000 m and 2500 m above ground level. A cross pattern, southwest-northeast and northwest-southeast, will be flown both in opposite directions.

Field testing plan for unsaturated zone monitoring and field studies

International Nuclear Information System (INIS)

Young, M.H.; Wierenga, P.J.; Warrick, A.W.

1996-10-01

The University of Arizona, in cooperation with the Bureau of Economic Geology at The University of Texas at Austin, and Stephens and Associates in Albuquerque, New Mexico has developed a field testing plan for evaluating subsurface monitoring systems. The U.S. Nuclear Regulatory Commission has requested development of these testing plans for low-level radioactive waste disposal sites (LLW) and for monitoring at decommissioned facilities designated under the open-quotes Site Decommissioning Management Planclose quotes (SDMP). The tests are conducted on a 50 m by 50 m plot on the University of Arizona's Maricopa Agricultural Center. Within the 50 m by 50 m plot one finds: (1) an instrumented buried trench, (2) monitoring islands similar to those proposed for the Ward Valley, California LLW Facility, (3) deep borehole monitoring sites, (4) gaseous transport monitoring, and (5) locations for testing non-invasive geophysical measurement techniques. The various subplot areas are instrumented with commercially available instruments such as neutron probes, time domain reflectometry probes, tensiometers, psychrometers, heat dissipation sensors, thermocouples, solution samplers, and cross-hole geophysics electrodes. Measurement depths vary from ground surface to 15 m. The data from the controlled flow and transport experiments, conducted over the plot, will be used to develop an integrated approach to long-term monitoring of the vadose zone at waste disposal sites. The data will also be used to test field-scale flow and transport models. This report describes in detail the design of the experiment and the methodology proposed for evaluating the data

Replication Rate, Framing, and Format Affect Attitudes and Decisions about Science Claims.

Science.gov (United States)

Barnes, Ralph M; Tobin, Stephanie J; Johnston, Heather M; MacKenzie, Noah; Taglang, Chelsea M

2016-01-01

A series of five experiments examined how the evaluation of a scientific finding was influenced by information about the number of studies that had successfully replicated the initial finding. The experiments also tested the impact of frame (negative, positive) and numeric format (percentage, natural frequency) on the evaluation of scientific findings. In Experiments 1 through 4, an attitude difference score served as the dependent measure, while a measure of choice served as the dependent measure in Experiment 5. Results from a diverse sample of 188 non-institutionalized U.S. adults (Experiment 2) and 730 undergraduate college students (Experiments 1, 3, and 4) indicated that attitudes became more positive as the replication rate increased and attitudes were more positive when the replication information was framed positively. The results also indicate that the manner in which replication rate was framed had a greater impact on attitude than the replication rate itself. The large effect for frame was attenuated somewhat when information about replication was presented in the form of natural frequencies rather than percentages. A fifth study employing 662 undergraduate college students in a task in which choice served as the dependent measure confirmed the framing effect and replicated the replication rate effect in the positive frame condition, but provided no evidence that the use of natural frequencies diminished the effect.

Replication Rate, Framing, and Format Affect Attitudes and Decisions about Science Claims

Directory of Open Access Journals (Sweden)

Ralph M. Barnes

2016-11-01

Full Text Available A series of five experiments examined how the evaluation of a scientific finding was influenced by information about the number of studies that had successfully replicated the initial finding. The experiments also tested the impact of frame (negative, positive and numeric format (percentage, natural frequency on the evaluation of scientific findings. In Experiments 1 through 4, an attitude difference score served as the dependent measure, while a measure of choice served as the dependent measure in Experiment 5. Results from a diverse sample of 188 non-institutionalized U.S. adults (Experiment 2 and 730 undergraduate college students (Experiments 1, 3, and 4 indicated that attitudes became more positive as the replication rate increased and attitudes were more positive when the replication information was framed positively. The results also indicate that the manner in which replication rate was framed had a greater impact on attitude than the replication rate itself. The large effect for frame was attenuated somewhat when information about replication was presented in the form of natural frequencies rather than percentages. A fifth study employing 662 undergraduate college students in a task in which choice served as the dependent measure confirmed the framing effect and replicated the replication rate effect in the positive frame condition, but provided no evidence that the use of natural frequencies diminished the effect.

Replication Rate, Framing, and Format Affect Attitudes and Decisions about Science Claims

Science.gov (United States)

Barnes, Ralph M.; Tobin, Stephanie J.; Johnston, Heather M.; MacKenzie, Noah; Taglang, Chelsea M.

2016-01-01

A series of five experiments examined how the evaluation of a scientific finding was influenced by information about the number of studies that had successfully replicated the initial finding. The experiments also tested the impact of frame (negative, positive) and numeric format (percentage, natural frequency) on the evaluation of scientific findings. In Experiments 1 through 4, an attitude difference score served as the dependent measure, while a measure of choice served as the dependent measure in Experiment 5. Results from a diverse sample of 188 non-institutionalized U.S. adults (Experiment 2) and 730 undergraduate college students (Experiments 1, 3, and 4) indicated that attitudes became more positive as the replication rate increased and attitudes were more positive when the replication information was framed positively. The results also indicate that the manner in which replication rate was framed had a greater impact on attitude than the replication rate itself. The large effect for frame was attenuated somewhat when information about replication was presented in the form of natural frequencies rather than percentages. A fifth study employing 662 undergraduate college students in a task in which choice served as the dependent measure confirmed the framing effect and replicated the replication rate effect in the positive frame condition, but provided no evidence that the use of natural frequencies diminished the effect. PMID:27920743

DNA Replication Control During Drosophila Development: Insights into the Onset of S Phase, Replication Initiation, and Fork Progression

Science.gov (United States)

Hua, Brian L.; Orr-Weaver, Terry L.

2017-01-01

Proper control of DNA replication is critical to ensure genomic integrity during cell proliferation. In addition, differential regulation of the DNA replication program during development can change gene copy number to influence cell size and gene expression. Drosophila melanogaster serves as a powerful organism to study the developmental control of DNA replication in various cell cycle contexts in a variety of differentiated cell and tissue types. Additionally, Drosophila has provided several developmentally regulated replication models to dissect the molecular mechanisms that underlie replication-based copy number changes in the genome, which include differential underreplication and gene amplification. Here, we review key findings and our current understanding of the developmental control of DNA replication in the contexts of the archetypal replication program as well as of underreplication and differential gene amplification. We focus on the use of these latter two replication systems to delineate many of the molecular mechanisms that underlie the developmental control of replication initiation and fork elongation. PMID:28874453

Replicable in vivo physiological and behavioral phenotypes of the Shank3B null mutant mouse model of autism.

Science.gov (United States)

Dhamne, Sameer C; Silverman, Jill L; Super, Chloe E; Lammers, Stephen H T; Hameed, Mustafa Q; Modi, Meera E; Copping, Nycole A; Pride, Michael C; Smith, Daniel G; Rotenberg, Alexander; Crawley, Jacqueline N; Sahin, Mustafa

2017-01-01

Autism spectrum disorder (ASD) is a clinically and biologically heterogeneous condition characterized by social, repetitive, and sensory behavioral abnormalities. No treatments are approved for the core diagnostic symptoms of ASD. To enable the earliest stages of therapeutic discovery and development for ASD, robust and reproducible behavioral phenotypes and biological markers are essential to establish in preclinical animal models. The goal of this study was to identify electroencephalographic (EEG) and behavioral phenotypes that are replicable between independent cohorts in a mouse model of ASD. The larger goal of our strategy is to empower the preclinical biomedical ASD research field by generating robust and reproducible behavioral and physiological phenotypes in animal models of ASD, for the characterization of mechanistic underpinnings of ASD-relevant phenotypes, and to ensure reliability for the discovery of novel therapeutics. Genetic disruption of the SHANK3 gene, a scaffolding protein involved in the stability of the postsynaptic density in excitatory synapses, is thought to be responsible for a relatively large number of cases of ASD. Therefore, we have thoroughly characterized the robustness of ASD-relevant behavioral phenotypes in two cohorts, and for the first time quantified translational EEG activity in Shank3B null mutant mice. In vivo physiology and behavioral assays were conducted in two independently bred and tested full cohorts of Shank3B null mutant ( Shank3B KO) and wildtype littermate control (WT) mice. EEG was recorded via wireless implanted telemeters for 7 days of baseline followed by 20 min of recording following pentylenetetrazol (PTZ) challenge. Behaviors relevant to the diagnostic and associated symptoms of ASD were tested on a battery of established behavioral tests. Assays were designed to reproduce and expand on the original behavioral characterization of Shank3B KO mice. Two or more corroborative tests were conducted within each

Qualifying tests for TRIAM-1M superconducting toroidal magnetic field coil

Energy Technology Data Exchange (ETDEWEB)

Nakanura, Yukio; Hiraki, Naoji; Nakamura, Kazuo; Tanaka, Masayoshi; Nagao, Akihiro; Kawasaki, Shoji; Itoh, Satoshi

1984-09-01

In the strong toroidal magnetic field experimental facility ''TRIAM-1M'' currently under construction, construction of the superconducting toroidal magnetic field coil and the following qualifying tests conducted on the full-scale superconducting toroidal magnetic field coil actually fabricated are described: (1) coil excitation test, (2) superconducting stability test, (3) external magnetic field application test, and (4) high-speed excitation test. On the basis of these test results, stability was evaluated of the superconducting coil being operated in the tokamak device. In normal tokamak operation, there occurs no normal conduction transition. At the time of plasma disruption, though this transition takes place in part of the coil, the superconducting state is immediately restored. By its electromagnetic force analysis, the superconducting coil is also stable in structure.

Comparative Field Tests of Pressurised Rover Prototypes

Science.gov (United States)

Mann, G. A.; Wood, N. B.; Clarke, J. D.; Piechochinski, S.; Bamsey, M.; Laing, J. H.

The conceptual designs, interior layouts and operational performances of three pressurised rover prototypes - Aonia, ARES and Everest - were field tested during a recent simulation at the Mars Desert Research Station in Utah. A human factors experiment, in which the same crew of three executed the same simulated science mission in each of the three vehicles, yielded comparative data on the capacity of each vehicle to safely and comfortably carry explorers away from the main base, enter and exit the vehicle in spacesuits, perform science tasks in the field, and manage geological and biological samples. As well as offering recommendations for design improvements for specific vehicles, the results suggest that a conventional Sports Utility Vehicle (SUV) would not be suitable for analog field work; that a pressurised docking tunnel to the main habitat is essential; that better provisions for spacesuit storage are required; and that a crew consisting of one driver/navigator and two field science crew specialists may be optimal. From a field operations viewpoint, a recurring conflict between rover and habitat crews at the time of return to the habitat was observed. An analysis of these incidents leads to proposed refinements of operational protocols, specific crew training for rover returns and again points to the need for a pressurised docking tunnel. Sound field testing, circulating of results, and building the lessons learned into new vehicles is advocated as a way of producing ever higher fidelity rover analogues.

The progression of replication forks at natural replication barriers in live bacteria.

Science.gov (United States)

Moolman, M Charl; Tiruvadi Krishnan, Sriram; Kerssemakers, Jacob W J; de Leeuw, Roy; Lorent, Vincent; Sherratt, David J; Dekker, Nynke H

2016-07-27

Protein-DNA complexes are one of the principal barriers the replisome encounters during replication. One such barrier is the Tus-ter complex, which is a direction dependent barrier for replication fork progression. The details concerning the dynamics of the replisome when encountering these Tus-ter barriers in the cell are poorly understood. By performing quantitative fluorescence microscopy with microfuidics, we investigate the effect on the replisome when encountering these barriers in live Escherichia coli cells. We make use of an E. coli variant that includes only an ectopic origin of replication that is positioned such that one of the two replisomes encounters a Tus-ter barrier before the other replisome. This enables us to single out the effect of encountering a Tus-ter roadblock on an individual replisome. We demonstrate that the replisome remains stably bound after encountering a Tus-ter complex from the non-permissive direction. Furthermore, the replisome is only transiently blocked, and continues replication beyond the barrier. Additionally, we demonstrate that these barriers affect sister chromosome segregation by visualizing specific chromosomal loci in the presence and absence of the Tus protein. These observations demonstrate the resilience of the replication fork to natural barriers and the sensitivity of chromosome alignment to fork progression. © The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

Field-test programs of borehole plugs in southeastern New Mexico

International Nuclear Information System (INIS)

Christensen, C.L.; Peterson, E.W.

1981-01-01

This paper gives a general overview of the repository-sealing field test effort being conducted by Sandia National Laboratories in support of the Waste Isolation Pilot Plant in southeast New Mexico. Summary descriptions of supporting activities, such as performance assessment and plugging materials development, are included to create the connection between modeling and laboratory activities as they relate to field results. Results of tests on a portion of a 17-year-old plug (Plug 217) recovered from a mine horizon and the Bell Canyon Test, in which a cement plug was emplaced to isolate a naturally pressurized aquifer, are given. Conclusions from these field plugging tests are included

Phosphorylation of Large T Antigen Regulates Merkel Cell Polyomavirus Replication

International Nuclear Information System (INIS)

Diaz, Jason; Wang, Xin; Tsang, Sabrina H.; Jiao, Jing; You, Jianxin

2014-01-01

Merkel Cell Polyomavirus (MCPyV) was recently discovered as a novel human polyomavirus that is associated with ~80% of Merkel Cell Carcinomas. The Large Tumor antigen (LT) is an early viral protein which has a variety of functions, including manipulation of the cell cycle and initiating viral DNA replication. Phosphorylation plays a critical regulatory role for polyomavirus LT proteins, but no investigation of MCPyV LT phosphorylation has been performed to date. In this report mass spectrometry analysis reveals three unique phosphorylation sites: T271, T297 and T299. In vivo replication assays confirm that phosphorylation of T271 does not play a role in viral replication, while modification at T297 and T299 have dramatic and opposing effects on LT’s ability to initiate replication from the viral origin. We test these mutants for their ability to bind, unwind, and act as a functional helicase at the viral origin. These studies provide a framework for understanding how phosphorylation of LT may dynamically regulate viral replication. Although the natural host cell of MCPyV has not yet been established, this work provides a foundation for understanding how LT activity is regulated and provides tools for better exploring this regulation in both natural host cells and Merkel cells

Phosphorylation of Large T Antigen Regulates Merkel Cell Polyomavirus Replication

Energy Technology Data Exchange (ETDEWEB)

Diaz, Jason; Wang, Xin; Tsang, Sabrina H. [Department of Microbiology, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104 (United States); Jiao, Jing [Department of Pathology and Laboratory Medicine, Children’s Hospital of Philadelphia, Philadelphia, PA 19104 (United States); You, Jianxin, E-mail: jianyou@mail.med.upenn.edu [Department of Microbiology, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104 (United States)

2014-07-08

Merkel Cell Polyomavirus (MCPyV) was recently discovered as a novel human polyomavirus that is associated with ~80% of Merkel Cell Carcinomas. The Large Tumor antigen (LT) is an early viral protein which has a variety of functions, including manipulation of the cell cycle and initiating viral DNA replication. Phosphorylation plays a critical regulatory role for polyomavirus LT proteins, but no investigation of MCPyV LT phosphorylation has been performed to date. In this report mass spectrometry analysis reveals three unique phosphorylation sites: T271, T297 and T299. In vivo replication assays confirm that phosphorylation of T271 does not play a role in viral replication, while modification at T297 and T299 have dramatic and opposing effects on LT’s ability to initiate replication from the viral origin. We test these mutants for their ability to bind, unwind, and act as a functional helicase at the viral origin. These studies provide a framework for understanding how phosphorylation of LT may dynamically regulate viral replication. Although the natural host cell of MCPyV has not yet been established, this work provides a foundation for understanding how LT activity is regulated and provides tools for better exploring this regulation in both natural host cells and Merkel cells.

FUELS IN SOIL TEST KIT: FIELD USE OF DIESEL DOG SOIL TEST KITS

Energy Technology Data Exchange (ETDEWEB)

Susan S. Sorini; John F. Schabron; Joseph F. Rovani, Jr.

2002-09-30

Western Research Institute (WRI) has developed a new commercial product ready for technology transfer, the Diesel Dog{reg_sign} Portable Soil Test Kit, for performing analysis of fuel-contaminated soils in the field. The technology consists of a method developed by WRI (U.S. Patents 5,561,065 and 5,976,883) and hardware developed by WRI that allows the method to be performed in the field (patent pending). The method is very simple and does not require the use of highly toxic reagents. The aromatic components in a soil extract are measured by absorption at 254 nm with a field-portable photometer. WRI added significant value to the technology by taking the method through the American Society for Testing and Materials (ASTM) approval and validation processes. The method is designated as ASTM Method D 5831-96, Standard Test Method for Screening Fuels in Soils. This ASTM designation allows the method to be used for federal compliance activities. In June 2001, the Diesel Dog technology won an American Chemical Society Regional Industrial Innovations Award. To gain field experience with the new technology, Diesel Dog kits have been used for a variety of site evaluation and cleanup activities. Information gained from these activities has led to improvements in hardware configurations and additional insight into correlating Diesel Dog results with results from laboratory methods. The Wyoming Department of Environmental Quality (DEQ) used Diesel Dog Soil Test Kits to guide cleanups at a variety of sites throughout the state. ENSR, of Acton, Massachusetts, used a Diesel Dog Portable Soil Test Kit to evaluate sites in the Virgin Islands and Georgia. ChemTrack and the U.S. Army Corps of Engineers successfully used a test kit to guide excavation at an abandoned FAA fuel-contaminated site near Fairbanks, Alaska. Barenco, Inc. is using a Diesel Dog Portable Soil Test Kit for site evaluations in Canada. A small spill of diesel fuel was cleaned up in Laramie, Wyoming using a Diesel

Replication of micro and nano surface geometries

DEFF Research Database (Denmark)

Hansen, Hans Nørgaard; Hocken, R.J.; Tosello, Guido

2011-01-01

The paper describes the state-of-the-art in replication of surface texture and topography at micro and nano scale. The description includes replication of surfaces in polymers, metals and glass. Three different main technological areas enabled by surface replication processes are presented......: manufacture of net-shape micro/nano surfaces, tooling (i.e. master making), and surface quality control (metrology, inspection). Replication processes and methods as well as the metrology of surfaces to determine the degree of replication are presented and classified. Examples from various application areas...... are given including replication for surface texture measurements, surface roughness standards, manufacture of micro and nano structured functional surfaces, replicated surfaces for optical applications (e.g. optical gratings), and process chains based on combinations of repeated surface replication steps....

A New Replication Norm for Psychology

Directory of Open Access Journals (Sweden)

Etienne P LeBel

2015-10-01

Full Text Available In recent years, there has been a growing concern regarding the replicability of findings in psychology, including a mounting number of prominent findings that have failed to replicate via high-powered independent replication attempts. In the face of this replicability “crisis of confidence”, several initiatives have been implemented to increase the reliability of empirical findings. In the current article, I propose a new replication norm that aims to further boost the dependability of findings in psychology. Paralleling the extant social norm that researchers should peer review about three times as many articles that they themselves publish per year, the new replication norm states that researchers should aim to independently replicate important findings in their own research areas in proportion to the number of original studies they themselves publish per year (e.g., a 4:1 original-to-replication studies ratio. I argue this simple approach could significantly advance our science by increasing the reliability and cumulative nature of our empirical knowledge base, accelerating our theoretical understanding of psychological phenomena, instilling a focus on quality rather than quantity, and by facilitating our transformation toward a research culture where executing and reporting independent direct replications is viewed as an ordinary part of the research process. To help promote the new norm, I delineate (1 how each of the major constituencies of the research process (i.e., funders, journals, professional societies, departments, and individual researchers can incentivize replications and promote the new norm and (2 any obstacles each constituency faces in supporting the new norm.

IMOM Field Test Study and Accuracy Verification

National Research Council Canada - National Science Library

Levien, Fred

1998-01-01

.... It was desired to obtain flight test data for both TAMPS and IMOM in order to compare their ability to accurately predict the effects of Radar Terrain Masking (RTM). It was initially planned to have NPS compare predictive data from both of these systems and then do analysis of how they compared to actual field test data.

Influence of the magnetic field profile on ITER conductor testing

International Nuclear Information System (INIS)

Nijhuis, A; Ilyin, Y; Kate, H H J ten

2006-01-01

We performed simulations with the numerical CUDI-CICC code on a typical short ITER (International Thermonuclear Experimental Reactor) conductor test sample of dual leg configuration, as usually tested in the SULTAN test facility, and made a comparison with the new EFDA-Dipole test facility offering a larger applied DC field region. The new EFDA-Dipole test facility, designed for short sample testing of conductors for ITER, has a homogeneous high field region of 1.2 m, while in the SULTAN facility this region is three times shorter. The inevitable non-uniformity of the current distribution in the cable, introduced by the joints at both ends, has a degrading effect on voltage-current (VI) and voltage-temperature (VT) characteristics, particularly for these short samples. This can easily result in an underestimation or overestimation of the actual conductor performance. A longer applied DC high field region along a conductor suppresses the current non-uniformity by increasing the overall longitudinal cable electric field when reaching the current sharing mode. The numerical interpretation study presented here gives a quantitative analysis for a relevant practical case of a test of a short sample poloidal field coil insert (PFCI) conductor in SULTAN. The simulation includes the results of current distribution analysis from self-field measurements with Hall sensor arrays, current sharing measurements and inter-petal resistance measurements. The outcome of the simulations confirms that the current uniformity improves with a longer high field region but the 'measured' VI transition is barely affected, though the local peak voltages become somewhat suppressed. It appears that the location of the high field region and voltage taps has practically no influence on the VI curve as long as the transverse voltage components are adequately cancelled. In particular, for a thin conduit wall, the voltage taps should be connected to the conduit in the form of an (open) azimuthally

Centromere replication timing determines different forms of genomic instability in Saccharomyces cerevisiae checkpoint mutants during replication stress.

Science.gov (United States)

Feng, Wenyi; Bachant, Jeff; Collingwood, David; Raghuraman, M K; Brewer, Bonita J

2009-12-01

Yeast replication checkpoint mutants lose viability following transient exposure to hydroxyurea, a replication-impeding drug. In an effort to understand the basis for this lethality, we discovered that different events are responsible for inviability in checkpoint-deficient cells harboring mutations in the mec1 and rad53 genes. By monitoring genomewide replication dynamics of cells exposed to hydroxyurea, we show that cells with a checkpoint deficient allele of RAD53, rad53K227A, fail to duplicate centromeres. Following removal of the drug, however, rad53K227A cells recover substantial DNA replication, including replication through centromeres. Despite this recovery, the rad53K227A mutant fails to achieve biorientation of sister centromeres during recovery from hydroxyurea, leading to secondary activation of the spindle assembly checkpoint (SAC), aneuploidy, and lethal chromosome segregation errors. We demonstrate that cell lethality from this segregation defect could be partially remedied by reinforcing bipolar attachment. In contrast, cells with the mec1-1 sml1-1 mutations suffer from severely impaired replication resumption upon removal of hydroxyurea. mec1-1 sml1-1 cells can, however, duplicate at least some of their centromeres and achieve bipolar attachment, leading to abortive segregation and fragmentation of incompletely replicated chromosomes. Our results highlight the importance of replicating yeast centromeres early and reveal different mechanisms of cell death due to differences in replication fork progression.

Cutthroat trout virus as a surrogate in vitro infection model for testing inhibitors of hepatitis E virus replication

Science.gov (United States)

Debing, Yannick; Winton, James; Neyts, Johan; Dallmeier, Kai

2013-01-01

Hepatitis E virus (HEV) is one of the most important causes of acute hepatitis worldwide. Although most infections are self-limiting, mortality is particularly high in pregnant women. Chronic infections can occur in transplant and other immune-compromised patients. Successful treatment of chronic hepatitis E has been reported with ribavirin and pegylated interferon-alpha, however severe side effects were observed. We employed the cutthroat trout virus (CTV), a non-pathogenic fish virus with remarkable similarities to HEV, as a potential surrogate for HEV and established an antiviral assay against this virus using the Chinook salmon embryo (CHSE-214) cell line. Ribavirin and the respective trout interferon were found to efficiently inhibit CTV replication. Other known broad-spectrum inhibitors of RNA virus replication such as the nucleoside analog 2′-C-methylcytidine resulted only in a moderate antiviral activity. In its natural fish host, CTV levels largely fluctuate during the reproductive cycle with the virus detected mainly during spawning. We wondered whether this aspect of CTV infection may serve as a surrogate model for the peculiar pathogenesis of HEV in pregnant women. To that end the effect of three sex steroids on in vitro CTV replication was evaluated. Whereas progesterone resulted in marked inhibition of virus replication, testosterone and 17β-estradiol stimulated viral growth. Our data thus indicate that CTV may serve as a surrogate model for HEV, both for antiviral experiments and studies on the replication biology of the Hepeviridae.

Registered Replication Report: Testing Disruptive Effects of Irrelevant Speech on Visual-Spatial Working Memory

Directory of Open Access Journals (Sweden)

Tatiana Kvetnaya

2018-04-01

Full Text Available A Partial Replication of “Functional Equivalence of Verbal and Spatial Information in Serial Short-Term Memory (Jones, Farrand, Stuart, & Morris, 1995; Experiment 4” The irrelevant speech effect (ISE—the phenomenon that background speech impairs serial recall of visually presented material—has been widely used for examining the structure of short-term memory. In Experiment 4, Jones, Farrand, Stuart, and Morris (1995 employed the ISE to demonstrate that impairment of performance is determined by the changing-state characteristics of the material, rather than its modality of origin. The present study directly replicated the spatial condition of Experiment 4 with 'N' = 40 German participants. In contrast to the original findings, no main effect of sound type was observed, 'F'(2, 78 = 0.81, 'p' = .450, η2'p' = .02. The absence of an ISE in the spatial domain does not support the changing state hypothesis.

The Inherent Asymmetry of DNA Replication.

Science.gov (United States)

Snedeker, Jonathan; Wooten, Matthew; Chen, Xin

2017-10-06

Semiconservative DNA replication has provided an elegant solution to the fundamental problem of how life is able to proliferate in a way that allows cells, organisms, and populations to survive and replicate many times over. Somewhat lost, however, in our admiration for this mechanism is an appreciation for the asymmetries that occur in the process of DNA replication. As we discuss in this review, these asymmetries arise as a consequence of the structure of the DNA molecule and the enzymatic mechanism of DNA synthesis. Increasing evidence suggests that asymmetries in DNA replication are able to play a central role in the processes of adaptation and evolution by shaping the mutagenic landscape of cells. Additionally, in eukaryotes, recent work has demonstrated that the inherent asymmetries in DNA replication may play an important role in the process of chromatin replication. As chromatin plays an essential role in defining cell identity, asymmetries generated during the process of DNA replication may play critical roles in cell fate decisions related to patterning and development.

BinQuasi: a peak detection method for ChIP-sequencing data with biological replicates.

Science.gov (United States)

Goren, Emily; Liu, Peng; Wang, Chao; Wang, Chong

2018-04-19

ChIP-seq experiments that are aimed at detecting DNA-protein interactions require biological replication to draw inferential conclusions, however there is no current consensus on how to analyze ChIP-seq data with biological replicates. Very few methodologies exist for the joint analysis of replicated ChIP-seq data, with approaches ranging from combining the results of analyzing replicates individually to joint modeling of all replicates. Combining the results of individual replicates analyzed separately can lead to reduced peak classification performance compared to joint modeling. Currently available methods for joint analysis may fail to control the false discovery rate at the nominal level. We propose BinQuasi, a peak caller for replicated ChIP-seq data, that jointly models biological replicates using a generalized linear model framework and employs a one-sided quasi-likelihood ratio test to detect peaks. When applied to simulated data and real datasets, BinQuasi performs favorably compared to existing methods, including better control of false discovery rate than existing joint modeling approaches. BinQuasi offers a flexible approach to joint modeling of replicated ChIP-seq data which is preferable to combining the results of replicates analyzed individually. Source code is freely available for download at https://cran.r-project.org/package=BinQuasi, implemented in R. pliu@iastate.edu or egoren@iastate.edu. Supplementary material is available at Bioinformatics online.

Deep Borehole Field Test Research Activities at LBNL

Energy Technology Data Exchange (ETDEWEB)

Dobson, Patrick [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Tsang, Chin-Fu [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Kneafsey, Timothy [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Borglin, Sharon [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Piceno, Yvette [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Andersen, Gary [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Nakagawa, Seiji [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Nihei, Kurt [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Rutqvist, Jonny [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Doughty, Christine [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Reagan, Matthew [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)

2016-08-19

The goal of the U.S. Department of Energy Used Fuel Disposition’s (UFD) Deep Borehole Field Test is to drill two 5 km large-diameter boreholes: a characterization borehole with a bottom-hole diameter of 8.5 inches and a field test borehole with a bottom-hole diameter of 17 inches. These boreholes will be used to demonstrate the ability to drill such holes in crystalline rocks, effectively characterize the bedrock repository system using geophysical, geochemical, and hydrological techniques, and emplace and retrieve test waste packages. These studies will be used to test the deep borehole disposal concept, which requires a hydrologically isolated environment characterized by low permeability, stable fluid density, reducing fluid chemistry conditions, and an effective borehole seal. During FY16, Lawrence Berkeley National Laboratory scientists conducted a number of research studies to support the UFD Deep Borehole Field Test effort. This work included providing supporting data for the Los Alamos National Laboratory geologic framework model for the proposed deep borehole site, conducting an analog study using an extensive suite of geoscience data and samples from a deep (2.5 km) research borehole in Sweden, conducting laboratory experiments and coupled process modeling related to borehole seals, and developing a suite of potential techniques that could be applied to the characterization and monitoring of the deep borehole environment. The results of these studies are presented in this report.

Deep Borehole Field Test Research Activities at LBNL

International Nuclear Information System (INIS)

Dobson, Patrick; Tsang, Chin-Fu; Kneafsey, Timothy; Borglin, Sharon; Piceno, Yvette; Andersen, Gary; Nakagawa, Seiji; Nihei, Kurt; Rutqvist, Jonny; Doughty, Christine; Reagan, Matthew

2016-01-01

The goal of the U.S. Department of Energy Used Fuel Disposition's (UFD) Deep Borehole Field Test is to drill two 5 km large-diameter boreholes: a characterization borehole with a bottom-hole diameter of 8.5 inches and a field test borehole with a bottom-hole diameter of 17 inches. These boreholes will be used to demonstrate the ability to drill such holes in crystalline rocks, effectively characterize the bedrock repository system using geophysical, geochemical, and hydrological techniques, and emplace and retrieve test waste packages. These studies will be used to test the deep borehole disposal concept, which requires a hydrologically isolated environment characterized by low permeability, stable fluid density, reducing fluid chemistry conditions, and an effective borehole seal. During FY16, Lawrence Berkeley National Laboratory scientists conducted a number of research studies to support the UFD Deep Borehole Field Test effort. This work included providing supporting data for the Los Alamos National Laboratory geologic framework model for the proposed deep borehole site, conducting an analog study using an extensive suite of geoscience data and samples from a deep (2.5 km) research borehole in Sweden, conducting laboratory experiments and coupled process modeling related to borehole seals, and developing a suite of potential techniques that could be applied to the characterization and monitoring of the deep borehole environment. The results of these studies are presented in this report.

Replication dynamics of the yeast genome.

Science.gov (United States)

Raghuraman, M K; Winzeler, E A; Collingwood, D; Hunt, S; Wodicka, L; Conway, A; Lockhart, D J; Davis, R W; Brewer, B J; Fangman, W L

2001-10-05

Oligonucleotide microarrays were used to map the detailed topography of chromosome replication in the budding yeast Saccharomyces cerevisiae. The times of replication of thousands of sites across the genome were determined by hybridizing replicated and unreplicated DNAs, isolated at different times in S phase, to the microarrays. Origin activations take place continuously throughout S phase but with most firings near mid-S phase. Rates of replication fork movement vary greatly from region to region in the genome. The two ends of each of the 16 chromosomes are highly correlated in their times of replication. This microarray approach is readily applicable to other organisms, including humans.

IN SITU FIELD TESTING OF PROCESSES

International Nuclear Information System (INIS)

YANG, J.S.Y.

2004-01-01

The purpose of this scientific analysis report is to update and document the data and subsequent analyses from ambient field-testing activities performed in underground drifts and surface-based boreholes through unsaturated zone (UZ) tuff rock units. In situ testing, monitoring, and associated laboratory studies are conducted to directly assess and evaluate the waste emplacement environment and the natural barriers to radionuclide transport at Yucca Mountain. This scientific analysis report supports and provides data to UZ flow and transport model reports, which in turn contribute to the Total System Performance Assessment (TSPA) of Yucca Mountain, an important document for the license application (LA). The objectives of ambient field-testing activities are described in Section 1.1. This report is the third revision (REV 03), which supercedes2. The scientific analysis of data for inputs to model calibration and validation as documented in2 were developed in accordance with the Technical Work Plan (TWP) ''Technical Work Plan for: Performance Assessment Unsaturated Zone'' (BSC 2004 [DIRS 167969]). This revision was developed in accordance with the ''Technical Work Plan for: Unsaturated Zone Flow Analysis and Model Report Integration'' (BSC 2004 [DIRS 169654], Section 1.2.4) for better integrated, consistent, transparent, traceable, and more complete documentation in this scientific analysis report and associated UZ flow and transport model reports. No additional testing or analyses were performed as part of this revision. The list of relevant acceptance criteria is provided by ''Technical Work Plan for: Unsaturated Zone Flow Analysis and Model Report Integration'' (BSC 2004 [DIRS 169654]), Table 3-1. Additional deviations from the TWP regarding the features, events, and processes (FEPs) list are discussed in Section 1.3. Documentation in this report includes descriptions of how, and under what conditions, the tests were conducted. The descriptions and analyses

Deep Borehole Field Test Conceptual Design Report

Energy Technology Data Exchange (ETDEWEB)

Hardin, Ernest L. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

2016-09-30

This report documents conceptual design development for the Deep Borehole Field Test (DBFT), including test packages (simulated waste packages, not containing waste) and a system for demonstrating emplacement and retrieval of those packages in the planned Field Test Borehole (FTB). For the DBFT to have demonstration value, it must be based on conceptualization of a deep borehole disposal (DBD) system. This document therefore identifies key options for a DBD system, describes an updated reference DBD concept, and derives a recommended concept for the DBFT demonstration. The objective of the DBFT is to confirm the safety and feasibility of the DBD concept for long-term isolation of radioactive waste. The conceptual design described in this report will demonstrate equipment and operations for safe waste handling and downhole emplacement of test packages, while contributing to an evaluation of the overall safety and practicality of the DBD concept. The DBFT also includes drilling and downhole characterization investigations that are described elsewhere (see Section 1). Importantly, no radioactive waste will be used in the DBFT, nor will the DBFT site be used for disposal of any type of waste. The foremost performance objective for conduct of the DBFT is to demonstrate safe operations in all aspects of the test.

High-Resolution Replication Profiles Define the Stochastic Nature of Genome Replication Initiation and Termination

Directory of Open Access Journals (Sweden)

Michelle Hawkins

2013-11-01

Full Text Available Eukaryotic genome replication is stochastic, and each cell uses a different cohort of replication origins. We demonstrate that interpreting high-resolution Saccharomyces cerevisiae genome replication data with a mathematical model allows quantification of the stochastic nature of genome replication, including the efficiency of each origin and the distribution of termination events. Single-cell measurements support the inferred values for stochastic origin activation time. A strain, in which three origins were inactivated, confirmed that the distribution of termination events is primarily dictated by the stochastic activation time of origins. Cell-to-cell variability in origin activity ensures that termination events are widely distributed across virtually the whole genome. We propose that the heterogeneity in origin usage contributes to genome stability by limiting potentially deleterious events from accumulating at particular loci.

DNA replication and post-replication repair in U.V.-sensitive mouse neuroblastoma cells

International Nuclear Information System (INIS)

Lavin, M.F.; McCombe, P.; Kidson, C.

1976-01-01

Mouse neuroblastoma cells differentiated when grown in the absence of serum; differentiation was reversed on the addition of serum. Differentiated cells were more sensitive to U.V.-radiation than proliferating cells. Whereas addition of serum to differentiated neuroblastoma cells normally resulted in immediate, synchronous entry into S phase, irradiation just before the addition of serum resulted in a long delay in the onset of DNA replication. During this lag period, incorporated 3 H-thymidine appeared in the light density region of CsCl gradients, reflecting either repair synthesis or abortive replication. Post-replication repair (gap-filling) was found to be present in proliferating cells and at certain times in differentiated cells. It is suggested that the sensitivity of differentiated neuroblastoma cells to U.V.-radiation may have been due to ineffective post-replication repair or to deficiencies in more than one repair mechanism, with reduction in repair capacity beyond a critical threshold. (author)

Success and failure in replication of genotype-phenotype associations: How does replication help in understanding the genetic basis of phenotypic variation in outbred populations?

Science.gov (United States)

Schielzeth, Holger; Rios Villamil, Alejandro; Burri, Reto

2018-03-25

Recent developments in sequencing technologies have facilitated genomewide mapping of phenotypic variation in natural populations. Such mapping efforts face a number of challenges potentially leading to low reproducibility. However, reproducible research forms the basis of scientific progress. We here discuss the options for replication and the reasons for potential nonreproducibility. We then review the evidence for reproducible quantitative trait loci (QTL) with a focus on natural animal populations. Existing case studies of replication fall into three categories: (i) traits that have been mapped to major effect loci (including chromosomal inversion and supergenes) by independent research teams; (ii) QTL fine-mapped in discovery populations; and (iii) attempts to replicate QTL across multiple populations. Major effect loci, in particular those associated with inversions, have been successfully replicated in several cases within and across populations. Beyond such major effect variants, replication has been more successful within than across populations, suggesting that QTL discovered in natural populations may often be population-specific. This suggests that biological causes (differences in linkage patterns, allele frequencies or context-dependencies of QTL) contribute to nonreproducibility. Evidence from other fields, notably animal breeding and QTL mapping in humans, suggests that a significant fraction of QTL is indeed reproducible in direction and magnitude at least within populations. However, there is also a large number of QTL that cannot be easily reproduced. We put forward that more studies should explicitly address the causes and context-dependencies of QTL signals, in particular to disentangle linkage differences, allele frequency differences and gene-by-environment interactions as biological causes of nonreproducibility of QTL, especially between populations. © 2018 John Wiley & Sons Ltd.

Effectiveness of strategies to increase the validity of findings from association studies: size vs. replication

Directory of Open Access Journals (Sweden)

Kallischnigg Gerd

2010-05-01

Full Text Available Abstract Background The capacity of multiple comparisons to produce false positive findings in genetic association studies is abundantly clear. To address this issue, the concept of false positive report probability (FPRP measures "the probability of no true association between a genetic variant and disease given a statistically significant finding". This concept involves the notion of prior probability of an association between a genetic variant and a disease, making it difficult to achieve acceptable levels for the FPRP when the prior probability is low. Increasing the sample size is of limited efficiency to improve the situation. Methods To further clarify this problem, the concept of true report probability (TRP is introduced by analogy to the positive predictive value (PPV of diagnostic testing. The approach is extended to consider the effects of replication studies. The formula for the TRP after k replication studies is mathematically derived and shown to be only dependent on prior probability, alpha, power, and number of replication studies. Results Case-control association studies are used to illustrate the TRP concept for replication strategies. Based on power considerations, a relationship is derived between TRP after k replication studies and sample size of each individual study. That relationship enables study designers optimization of study plans. Further, it is demonstrated that replication is efficient in increasing the TRP even in the case of low prior probability of an association and without requiring very large sample sizes for each individual study. Conclusions True report probability is a comprehensive and straightforward concept for assessing the validity of positive statistical testing results in association studies. By its extension to replication strategies it can be demonstrated in a transparent manner that replication is highly effective in distinguishing spurious from true associations. Based on the generalized TRP

Can Coloring Mandalas Reduce Anxiety? A Replication Study

Science.gov (United States)

van der Vennet, Renee; Serice, Susan

2012-01-01

This experimental study replicated Curry and Kasser's (2005) research that tested whether coloring a mandala would reduce anxiety. After inducing an anxious mood via a writing activity, participants were randomly assigned to three groups that colored either on a mandala design, on a plaid design, or on a blank paper. Anxiety level was measured…

DATABASE REPLICATION IN HETEROGENOUS PLATFORM

OpenAIRE

Hendro Nindito; Evaristus Didik Madyatmadja; Albert Verasius Dian Sano

2014-01-01

The application of diverse database technologies in enterprises today is increasingly a common practice. To provide high availability and survavibality of real-time information, a database replication technology that has capability to replicate databases under heterogenous platforms is required. The purpose of this research is to find the technology with such capability. In this research, the data source is stored in MSSQL database server running on Windows. The data will be replicated to MyS...

Overcoming natural replication barriers: differential helicase requirements.

Science.gov (United States)

Anand, Ranjith P; Shah, Kartik A; Niu, Hengyao; Sung, Patrick; Mirkin, Sergei M; Freudenreich, Catherine H

2012-02-01

DNA sequences that form secondary structures or bind protein complexes are known barriers to replication and potential inducers of genome instability. In order to determine which helicases facilitate DNA replication across these barriers, we analyzed fork progression through them in wild-type and mutant yeast cells, using 2-dimensional gel-electrophoretic analysis of the replication intermediates. We show that the Srs2 protein facilitates replication of hairpin-forming CGG/CCG repeats and prevents chromosome fragility at the repeat, whereas it does not affect replication of G-quadruplex forming sequences or a protein-bound repeat. Srs2 helicase activity is required for hairpin unwinding and fork progression. Also, the PCNA binding domain of Srs2 is required for its in vivo role of replication through hairpins. In contrast, the absence of Sgs1 or Pif1 helicases did not inhibit replication through structural barriers, though Pif1 did facilitate replication of a telomeric protein barrier. Interestingly, replication through a protein barrier but not a DNA structure barrier was modulated by nucleotide pool levels, illuminating a different mechanism by which cells can regulate fork progression through protein-mediated stall sites. Our analyses reveal fundamental differences in the replication of DNA structural versus protein barriers, with Srs2 helicase activity exclusively required for fork progression through hairpin structures.

Cutthroat trout virus as a surrogate in vitro infection model for testing inhibitors of hepatitis E virus replication.

Science.gov (United States)

Debing, Yannick; Winton, James; Neyts, Johan; Dallmeier, Kai

2013-10-01

Hepatitis E virus (HEV) is one of the most important causes of acute hepatitis worldwide. Although most infections are self-limiting, mortality is particularly high in pregnant women. Chronic infections can occur in transplant and other immune-compromised patients. Successful treatment of chronic hepatitis E has been reported with ribavirin and pegylated interferon-alpha, however severe side effects were observed. We employed the cutthroat trout virus (CTV), a non-pathogenic fish virus with remarkable similarities to HEV, as a potential surrogate for HEV and established an antiviral assay against this virus using the Chinook salmon embryo (CHSE-214) cell line. Ribavirin and the respective trout interferon were found to efficiently inhibit CTV replication. Other known broad-spectrum inhibitors of RNA virus replication such as the nucleoside analog 2'-C-methylcytidine resulted only in a moderate antiviral activity. In its natural fish host, CTV levels largely fluctuate during the reproductive cycle with the virus detected mainly during spawning. We wondered whether this aspect of CTV infection may serve as a surrogate model for the peculiar pathogenesis of HEV in pregnant women. To that end the effect of three sex steroids on in vitro CTV replication was evaluated. Whereas progesterone resulted in marked inhibition of virus replication, testosterone and 17β-estradiol stimulated viral growth. Our data thus indicate that CTV may serve as a surrogate model for HEV, both for antiviral experiments and studies on the replication biology of the Hepeviridae. Copyright © 2013 Elsevier B.V. All rights reserved.

Exploiting replicative stress to treat cancer

DEFF Research Database (Denmark)

Dobbelstein, Matthias; Sørensen, Claus Storgaard

2015-01-01

DNA replication in cancer cells is accompanied by stalling and collapse of the replication fork and signalling in response to DNA damage and/or premature mitosis; these processes are collectively known as 'replicative stress'. Progress is being made to increase our understanding of the mechanisms...

Field-test of a date-rape drug detection device.

Science.gov (United States)

Quest, Dale W; Horsley, Joanne

2007-01-01

Drink Safe Technology Version 1.2 is an inexpensive color-change reagent test marketed internationally for use by consumers in settings such as a night club to detect potentially incapacitating concentrations of gamma-hydroxybutyric acid (GHB) and ketamine in beverages. The objective of this study was to compare product performance in the laboratory and performance in the hands of consumers in the field. Product performance in the laboratory adhered to the protocol defined by the manufacturer. Product performance in the hands of consumers in field settings allowed browsing participants to pipette an aliquot of their own drinks into randomly coded vials containing authentic drugs, or pure water, so as to yield the same concentrations of GHB or ketamine specified in the manufacturer-defined protocol, or blanks. Consumers were to proceed according to the directions printed on the product, and to record their results on a card with a code corresponding with the vial to which they had added an aliquot of their beverage. Diagnostic performance was calculated using two-way analysis. In the laboratory, Drink Safe Technology Version 1.2 reliably detected GHB and ketamine at concentrations specified by the manufacturer's protocol. The reactive color change denoting a positive test for GHB was rapid, but a positive test for ketamine required substantially more time to resolve. Nonetheless, test accuracy following the manufacturer's protocol in the laboratory was 100%. In the field, based on 101 paired-test results recorded by consumers, the test efficiency was 65.1%, sensitivity 50%, and specificity 91.6%. The product performed much better in the laboratory than it did in the hand of consumers in the field. There seems to be considerable potential for consumers to misinterpret a test result. The potential for consumers to record a false-negative test result for a spiked drink is cause for concern.

40 CFR 1065.935 - Emission test sequence for field testing.

Science.gov (United States)

2010-07-01

... value for the previous 2 min or until an engine thermostat controls engine temperature with coolant or... media, such as evacuated bags or tare-weighed PM sample media. (2) Operate the PEMS according to the... ambient data, and integrate measured values using a PEMS. (3) If engine starting is part of field testing...

Field assessment of soil structural quality - a development of the Peerlkamp test

DEFF Research Database (Denmark)

Ball, B C; Batey, T; Munkholm, Lars Juhl

2007-01-01

properties (bulk density, penetration resistance and porosity) and crop growth. Indicative thresholds of soil management are suggested. The assessment should be viewed as complementary to conventional laboratory assessments of soil structure. Visual soil structure assessment can indicate to the soil......Increased awareness of the role of soil structure in defining the physical fertility or quality of soil has led to the need for a simple assessment relevant to the environmental and economic sustainability of soil productivity. A test is required that is usable by farmer, consultant and researcher...... alike. Here an assessment of soil structure quality (Sq) is described which is based on a visual key linked to criteria chosen to be as objective as possible. The influences of operator, tillage and crop type on Sq value were tested. The test takes 5-15 min per location and enough replicates were...

Chromatin maturation depends on continued DNA-replication

International Nuclear Information System (INIS)

Schlaeger, E.J.; Puelm, W.; Knippers, R.

1983-01-01

The structure of [ 3 H]thymidine pulse-labeled chromatin in lymphocytes differs from that of non-replicating chromatin by several operational criteria which are related to the higher nuclease sensitivity of replicating chromatin. These structural features of replicating chromatin rapidly disappear when the [ 3 H]thymidine pulse is followed by a chase in the presence of an excess of non-radioactive thymidine. However, when the rate of DNA replication is reduced, as in cycloheximide-treated lymphocytes, chromatin maturation is retarded. No chromatin maturation is observed when nuclei from pulse-labeled lymphocytes are incubated in vitro in the absence of DNA precursors. In contrast, when these nuclei are incubated under conditions known to be optimal for DNA replication, the structure of replicating chromatin is efficiently converted to that of 'mature', non-replicating chromatin. The authors conclude that the properties of nascent DNA and/or the distance from the replication fork are important factors in chromatin maturation. (Auth.)

Failing the Future: Three Unsuccessful Attempts to Replicate Bem's ‘Retroactive Facilitation of Recall’ Effect

Science.gov (United States)

Ritchie, Stuart J.; Wiseman, Richard; French, Christopher C.

2012-01-01

Nine recently reported parapsychological experiments appear to support the existence of precognition. We describe three pre-registered independent attempts to exactly replicate one of these experiments, ‘retroactive facilitation of recall’, which examines whether performance on a memory test can be influenced by a post-test exercise. All three replication attempts failed to produce significant effects (combined n = 150; combined p = .83, one-tailed) and thus do not support the existence of psychic ability. PMID:22432019

Replicating an Intervention: The Tension between Fidelity and Adaptation

Science.gov (United States)

Morrison, Diane M.; Hoppe, Marilyn J.; Gillmore, Mary Rogers; Kluver, Carisa; Higa, Darrel; Wells, Elizabeth A.

2009-01-01

Increased awareness of the importance of tailoring interventions to participants' cultures has focused attention on the limited generalizability of a single test of an intervention to determine efficacy. Adaptation is often necessary to replicate interventions across cultures. This produces a tension between fidelity to the original intervention…

The Escherichia coli Tus-Ter replication fork barrier causes site-specific DNA replication perturbation in yeast

DEFF Research Database (Denmark)

Larsen, Nicolai B; Sass, Ehud; Suski, Catherine

2014-01-01

Replication fork (RF) pausing occurs at both 'programmed' sites and non-physiological barriers (for example, DNA adducts). Programmed RF pausing is required for site-specific DNA replication termination in Escherichia coli, and this process requires the binding of the polar terminator protein, Tus...... as a versatile, site-specific, heterologous DNA replication-perturbing system, with a variety of potential applications....

Remote field eddy current testing of ferromagnetic tubes

International Nuclear Information System (INIS)

David, B.

1990-01-01

In order to test ferromagnetic tubes using internal probes, Intercontrole and the CEA have carried out theoretical and experimental works and developed a method to adapt the Remote Field Eddy Current technique which has been known and used for 30 years now. This document briefly recalls the basic principles of the Remote Field Eddy Current technique, the various steps of the works carried out and mainly describes examples of field inspection of ferromagnetic tubes and pipes [fr

R20 Programme: Field testing of grouting materials

International Nuclear Information System (INIS)

Ranta-Korpi, R.; Karttunen, P.; Sievaenen, U.

2008-02-01

In the year 2004 Finnish nuclear waste management organisation Posiva Oy started to construct an underground rock characterisation facility in Olkiluoto, Eurajoki, Western Finland. The ONKALO was planned to be a final disposal of spent nuclear fuel in Finland. This facility is going to be finished in the end of the year 2010. In ONKALO it is important to grout the water conductive structures to minimize the leakage of the inflowing groundwater in order to keep the geohydrogical environment unchanged and the repository conditions safe. Before the construction began the development of the grouts suitable for this demanding environment was started. Especially pH, the penetration ability and rheology have been the matter of interest. One target for the grout has been that the pH is relatively low. These grouts have different properties than the normal grouts. Several laboratory and field tests have been done for low pH grouts. The suitable recipies are studied in laboratory and the properties are verified in field. This work concerns the field testing of several low pH grout recipies. The different W/DM -ratios and SPL contents were studied. Besides the customary test methods, two new measurement methods were taken into use: the sand column test for measuring the penetration ability and the stick test for determining the plastic viscosity and yield value. The relationships between different properties, the properties as a function of the time and the effect of the (orig.)

Enzymatic recognition of DNA replication origins

International Nuclear Information System (INIS)

Stayton, M.M.; Bertsch, L.; Biswas, S.

1983-01-01

In this paper we discuss the process of recognition of the complementary-strand origin with emphasis on RNA polymerase action in priming M13 DNA replication, the role of primase in G4 DNA replication, and the function of protein n, a priming protein, during primosome assembly. These phage systems do not require several of the bacterial DNA replication enzymes, particularly those involved in the regulation of chromosome copy number of the initiatiion of replication of duplex DNA. 51 references, 13 figures, 1 table

Surface Microstructure Replication in Injection Moulding

DEFF Research Database (Denmark)

Hansen, Hans Nørgaard; Arlø, Uffe Rolf

2005-01-01

topography is transcribed onto the plastic part through complex mechanisms. This replication however, is not perfect, and the replication quality depends on the plastic material properties, the topography itself, and the process conditions. This paper describes and discusses an investigation of injection...... moulding of surface microstructures. Emphasis is put on the ability to replicate surface microstructures under normal injection moulding conditions, notably with low cost materials at low mould temperatures. The replication of surface microstructures in injection moulding has been explored...... for Polypropylene at low mould temperatures. The process conditions were varied over the recommended process window for the material. The geometry of the obtained structures was analyzed. Evidence suggests that step height replication quality depends linearly on structure width in a certain range. Further...

Systematic identification of novel, essential host genes affecting bromovirus RNA replication.

Directory of Open Access Journals (Sweden)

Brandi L Gancarz

Full Text Available Positive-strand RNA virus replication involves viral proteins and cellular proteins at nearly every replication step. Brome mosaic virus (BMV is a well-established model for dissecting virus-host interactions and is one of very few viruses whose RNA replication, gene expression and encapsidation have been reproduced in the yeast Saccharomyces cerevisiae. Previously, our laboratory identified ∼100 non-essential host genes whose loss inhibited or enhanced BMV replication at least 3-fold. However, our isolation of additional BMV-modulating host genes by classical genetics and other results underscore that genes essential for cell growth also contribute to BMV RNA replication at a frequency that may be greater than that of non-essential genes. To systematically identify novel, essential host genes affecting BMV RNA replication, we tested a collection of ∼900 yeast strains, each with a single essential gene promoter replaced by a doxycycline-repressible promoter, allowing repression of gene expression by adding doxycycline to the growth medium. Using this strain array of ∼81% of essential yeast genes, we identified 24 essential host genes whose depleted expression reproducibly inhibited or enhanced BMV RNA replication. Relevant host genes are involved in ribosome biosynthesis, cell cycle regulation and protein homeostasis, among other cellular processes. BMV 2a(Pol levels were significantly increased in strains depleted for a heat shock protein (HSF1 or proteasome components (PRE1 and RPT6, suggesting these genes may affect BMV RNA replication by directly or indirectly modulating 2a(Pol localization, post-translational modification or interacting partners. Investigating the diverse functions of these newly identified essential host genes should advance our understanding of BMV-host interactions and normal cellular pathways, and suggest new modes of virus control.

Replication Protein A (RPA) Phosphorylation Prevents RPA Association with Replication Centers

OpenAIRE

Vassin, Vitaly M.; Wold, Marc S.; Borowiec, James A.

2004-01-01

Mammalian replication protein A (RPA) undergoes DNA damage-dependent phosphorylation at numerous sites on the N terminus of the RPA2 subunit. To understand the functional significance of RPA phosphorylation, we expressed RPA2 variants in which the phosphorylation sites were converted to aspartate (RPA2D) or alanine (RPA2A). Although RPA2D was incorporated into RPA heterotrimers and supported simian virus 40 DNA replication in vitro, the RPA2D mutant was selectively unable to associate with re...

IN SITU FIELD TESTING OF PROCESSES

Energy Technology Data Exchange (ETDEWEB)

J.S.Y. YANG

2004-11-08

The purpose of this scientific analysis report is to update and document the data and subsequent analyses from ambient field-testing activities performed in underground drifts and surface-based boreholes through unsaturated zone (UZ) tuff rock units. In situ testing, monitoring, and associated laboratory studies are conducted to directly assess and evaluate the waste emplacement environment and the natural barriers to radionuclide transport at Yucca Mountain. This scientific analysis report supports and provides data to UZ flow and transport model reports, which in turn contribute to the Total System Performance Assessment (TSPA) of Yucca Mountain, an important document for the license application (LA). The objectives of ambient field-testing activities are described in Section 1.1. This report is the third revision (REV 03), which supercedes REV 02. The scientific analysis of data for inputs to model calibration and validation as documented in REV 02 were developed in accordance with the Technical Work Plan (TWP) ''Technical Work Plan for: Performance Assessment Unsaturated Zone'' (BSC 2004 [DIRS 167969]). This revision was developed in accordance with the ''Technical Work Plan for: Unsaturated Zone Flow Analysis and Model Report Integration'' (BSC 2004 [DIRS 169654], Section 1.2.4) for better integrated, consistent, transparent, traceable, and more complete documentation in this scientific analysis report and associated UZ flow and transport model reports. No additional testing or analyses were performed as part of this revision. The list of relevant acceptance criteria is provided by ''Technical Work Plan for: Unsaturated Zone Flow Analysis and Model Report Integration'' (BSC 2004 [DIRS 169654]), Table 3-1. Additional deviations from the TWP regarding the features, events, and processes (FEPs) list are discussed in Section 1.3. Documentation in this report includes descriptions of how, and under what

Variance Swap Replication: Discrete or Continuous?

Directory of Open Access Journals (Sweden)

Fabien Le Floc’h

2018-02-01

Full Text Available The popular replication formula to price variance swaps assumes continuity of traded option strikes. In practice, however, there is only a discrete set of option strikes traded on the market. We present here different discrete replication strategies and explain why the continuous replication price is more relevant.

Field test of the PNNL Automated Radioxenon Sampler/Analyzer (ARSA)

International Nuclear Information System (INIS)

Lagomarsino, R.J.; Ku, E.; Latner, N.; Sanderson, C.G.

1998-07-01

As part of the requirements of the Comprehensive Test Ban Treaty (CTBT), the Automated Radioxenon/Sampler Analyzer (ARSA) was designed and engineered by the Pacific Northwest National Laboratory (PNNL). The instrument is to provide near real-time detection and measurement of the radioxenons released into the atmosphere after a nuclear test. Forty-six field tests, designed to determine the performance of the ARSA prototype under simulated field conditions, were conducted at EML from March to December 1997. This final report contains detailed results of the tests with recommendations for improvements in instrument performance

Field test of the PNNL Automated Radioxenon Sampler/Analyzer (ARSA)

Energy Technology Data Exchange (ETDEWEB)

Lagomarsino, R.J.; Ku, E.; Latner, N.; Sanderson, C.G.

1998-07-01

As part of the requirements of the Comprehensive Test Ban Treaty (CTBT), the Automated Radioxenon/Sampler Analyzer (ARSA) was designed and engineered by the Pacific Northwest National Laboratory (PNNL). The instrument is to provide near real-time detection and measurement of the radioxenons released into the atmosphere after a nuclear test. Forty-six field tests, designed to determine the performance of the ARSA prototype under simulated field conditions, were conducted at EML from March to December 1997. This final report contains detailed results of the tests with recommendations for improvements in instrument performance.

Conceptual Design and Requirements for Characterization and Field Test Boreholes: Deep Borehole Field Test

Energy Technology Data Exchange (ETDEWEB)

Kuhlman, Kristopher L. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Brady, Patrick Vane [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); MacKinnon, Robert J. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Heath, Jason E. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Herrick, Courtney G. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Jensen, Richard P. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Rigali, Mark J. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Hadgu, Teklu [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Sevougian, S. David [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Birkholzer, Jens [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Freifeld, Barry M. [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Daley, Tom [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)

2015-09-24

Deep Borehole Disposal (DBD) of high-level radioactive wastes has been considered an option for geological isolation for many years (Hess et al. 1957). Recent advances in drilling technology have decreased costs and increased reliability for large-diameter (i.e., ≥50 cm [19.7”]) boreholes to depths of several kilometers (Beswick 2008; Beswick et al. 2014). These advances have therefore also increased the feasibility of the DBD concept (Brady et al. 2009; Cornwall 2015), and the current field test, introduced herein, is a demonstration of the DBD concept and these advances.

Acceptance test report: Field test of mixer pump for 241-AN-107 caustic addition project

International Nuclear Information System (INIS)

Leshikar, G.A.

1997-01-01

The field acceptance test of a 75 HP mixer pump (Hazleton serial number N-20801) installed in Tank 241-AN-107 was conducted from October 1995 thru February 1996. The objectives defined in the acceptance test were successfully met, with two exceptions recorded. The acceptance test encompassed field verification of mixer pump turntable rotation set-up and operation, verification that the pump instrumentation functions within established limits, facilitation of baseline data collection from the mixer pump mounted ultrasonic instrumentation, verification of mixer pump water flush system operation and validation of a procedure for its operation, and several brief test runs (bump) of the mixer pump

Surface microstructure replication in injection molding

DEFF Research Database (Denmark)

Theilade, Uffe Arlø; Hansen, Hans Nørgaard

2006-01-01

topography is transcribed onto the plastic part through complex mechanisms. This replication, however, is not perfect, and the replication quality depends on the plastic material properties, the topography itself, and the process conditions. This paper describes and discusses an investigation of injection...... molding of surface microstructures. The fundamental problem of surface microstructure replication has been studied. The research is based on specific microstructures as found in lab-on-a-chip products and on rough surfaces generated from EDM (electro discharge machining) mold cavities. Emphasis is put...... on the ability to replicate surface microstructures under normal injection-molding conditions, i.e., with commodity materials within typical process windows. It was found that within typical process windows the replication quality depends significantly on several process parameters, and especially the mold...

Disaster recovery using VMware vSphere Replication and vCenter Site Recovery Manager

CERN Document Server

GB, Abhilash

2014-01-01

This is a step-by-step guide that will help you understand disaster recovery using VMware vSphere Replication 5.5 and VMware vCenter Site Recovery Manager (SRM) 5.5. The topics and configuration procedures are accompanied with relevant screenshots, flowcharts, and logical diagrams that makes grasping the concepts easier. This book is a guide for anyone who is keen on using vSphere Replication or vCenter Site Recovery Manager as a disaster recovery solution. This is an excellent handbook for solution architects, administrators, on-field engineers, and support professionals. Although the book as

Support, shape and number of replicate samples for tree foliage analysis.

Science.gov (United States)

Luyssaert, Sebastiaan; Mertens, Jan; Raitio, Hannu

2003-06-01

Many fundamental features of a sampling program are determined by the heterogeneity of the object under study and the settings for the error (alpha), the power (beta), the effect size (ES), the number of replicate samples, and sample support, which is a feature that is often overlooked. The number of replicates, alpha, beta, ES, and sample support are interconnected. The effect of the sample support and its shape on the required number of replicate samples was investigated by means of a resampling method. The method was applied to a simulated distribution of Cd in the crown of a Salix fragilis L. tree. Increasing the dimensions of the sample support results in a decrease in the variance of the element concentration under study. Analysis of the variance is often the foundation of statistical tests, therefore, valid statistical testing requires the use of a fixed sample support during the experiment. This requirement might be difficult to meet in time-series analyses and long-term monitoring programs. Sample supports have their largest dimension in the direction with the largest heterogeneity, i.e. the direction representing the crown height, and this will give more accurate results than supports with other shapes. Taking the relationships between the sample support and the variance of the element concentrations in tree crowns into account provides guidelines for sampling efficiency in terms of precision and costs. In terms of time, the optimal support to test whether the average Cd concentration of the crown exceeds a threshold value is 0.405 m3 (alpha = 0.05, beta = 0.20, ES = 1.0 mg kg(-1) dry mass). The average weight of this support is 23 g dry mass, and 11 replicate samples need to be taken. It should be noted that in this case the optimal support applies to Cd under conditions similar to those of the simulation, but not necessarily all the examinations for this tree species, element, and hypothesis test.

Activation of human herpesvirus replication by apoptosis.

Science.gov (United States)

Prasad, Alka; Remick, Jill; Zeichner, Steven L

2013-10-01

A central feature of herpesvirus biology is the ability of herpesviruses to remain latent within host cells. Classically, exposure to inducing agents, like activating cytokines or phorbol esters that stimulate host cell signal transduction events, and epigenetic agents (e.g., butyrate) was thought to end latency. We recently showed that Kaposi's sarcoma-associated herpesvirus (KSHV, or human herpesvirus-8 [HHV-8]) has another, alternative emergency escape replication pathway that is triggered when KSHV's host cell undergoes apoptosis, characterized by the lack of a requirement for the replication and transcription activator (RTA) protein, accelerated late gene kinetics, and production of virus with decreased infectivity. Caspase-3 is necessary and sufficient to initiate the alternative replication program. HSV-1 was also recently shown to initiate replication in response to host cell apoptosis. These observations suggested that an alternative apoptosis-triggered replication program might be a general feature of herpesvirus biology and that apoptosis-initiated herpesvirus replication may have clinical implications, particularly for herpesviruses that almost universally infect humans. To explore whether an alternative apoptosis-initiated replication program is a common feature of herpesvirus biology, we studied cell lines latently infected with Epstein-Barr virus/HHV-4, HHV-6A, HHV-6B, HHV-7, and KSHV. We found that apoptosis triggers replication for each HHV studied, with caspase-3 being necessary and sufficient for HHV replication. An alternative apoptosis-initiated replication program appears to be a common feature of HHV biology. We also found that commonly used cytotoxic chemotherapeutic agents activate HHV replication, which suggests that treatments that promote apoptosis may lead to activation of latent herpesviruses, with potential clinical significance.

Rock sealing - large scale field test and accessory investigations

International Nuclear Information System (INIS)

Pusch, R.

1988-03-01

The experience from the pilot field test and the basic knowledge extracted from the lab experiments have formed the basis of the planning of a Large Scale Field Test. The intention is to find out how the 'instrument of rock sealing' can be applied to a number of practical cases, where cutting-off and redirection of groundwater flow in repositories are called for. Five field subtests, which are integrated mutually or with other Stripa projects (3D), are proposed. One of them concerns 'near-field' sealing, i.e. sealing of tunnel floors hosting deposition holes, while two involve sealing of 'disturbed' rock around tunnels. The fourth concerns sealing of a natural fracture zone in the 3D area, and this latter test has the expected spin-off effect of obtaining additional information on the general flow pattern around the northeastern wing of the 3D cross. The fifth test is an option of sealing structures in the Validation Drift. The longevity of major grout types is focussed on as the most important part of the 'Accessory Investigations', and detailed plans have been worked out for that purpose. It is foreseen that the continuation of the project, as outlined in this report, will yield suitable methods and grouts for effective and long-lasting sealing of rock for use at stategic points in repositories. (author)

Field tests and commercialization of natural gas leak detectors

Energy Technology Data Exchange (ETDEWEB)

Choi, D S; Jeon, J S; Kim, K D; Cho, Y A [R and D Center, Korea Gas Corporation, Ansan (Korea)

1999-09-01

Objectives - (1) fields test of industrial gas leak detection monitoring system. (2) commericialization of residential gas leak detector. Contents - (1) five sets of gas leak detection monitoring system were installed at natural gas transmition facilities and tested long term stability and their performance. (2) improved residential gas leak detector was commercialised. Expected benefits and application fields - (1) contribution to the improvement of domestic gas sensor technology. (2) localization of fabrication technology for gas leak detectors. 23 refs., 126 figs., 37 tabs.

Evolution of Replication Machines

Science.gov (United States)

Yao, Nina Y.; O'Donnell, Mike E.

2016-01-01

The machines that decode and regulate genetic information require the translation, transcription and replication pathways essential to all living cells. Thus, it might be expected that all cells share the same basic machinery for these pathways that were inherited from the primordial ancestor cell from which they evolved. A clear example of this is found in the translation machinery that converts RNA sequence to protein. The translation process requires numerous structural and catalytic RNAs and proteins, the central factors of which are homologous in all three domains of life, bacteria, archaea and eukarya. Likewise, the central actor in transcription, RNA polymerase, shows homology among the catalytic subunits in bacteria, archaea and eukarya. In contrast, while some “gears” of the genome replication machinery are homologous in all domains of life, most components of the replication machine appear to be unrelated between bacteria and those of archaea and eukarya. This review will compare and contrast the central proteins of the “replisome” machines that duplicate DNA in bacteria, archaea and eukarya, with an eye to understanding the issues surrounding the evolution of the DNA replication apparatus. PMID:27160337

Comparison Between 30-15 Intermittent Fitness Test and Multistage Field Test on Physiological Responses in Wheelchair Basketball Players

OpenAIRE

Weissland, Thierry; Faupin, Arnaud; Borel, Benoit; Leprêtre, Pierre-Marie

2015-01-01

The intermittent nature of wheelchair court sports suggests using a similar protocol to assess repeated shuttles and recovery abilities. This study aimed to compare performances, physiological responses and perceived rating exertion obtained from the continuous multistage field test (MFT) and the 30-15 intermittent field test (30-15IFT). Eighteen trained wheelchair basketball players (WBP) (WBP: 32.0 ? 5.7 y, IWBF classification: 2.9 ? 1.1 points) performed both incremental field tests in ran...

DNA replication origins—where do we begin?

Science.gov (United States)

Prioleau, Marie-Noëlle; MacAlpine, David M.

2016-01-01

For more than three decades, investigators have sought to identify the precise locations where DNA replication initiates in mammalian genomes. The development of molecular and biochemical approaches to identify start sites of DNA replication (origins) based on the presence of defining and characteristic replication intermediates at specific loci led to the identification of only a handful of mammalian replication origins. The limited number of identified origins prevented a comprehensive and exhaustive search for conserved genomic features that were capable of specifying origins of DNA replication. More recently, the adaptation of origin-mapping assays to genome-wide approaches has led to the identification of tens of thousands of replication origins throughout mammalian genomes, providing an unprecedented opportunity to identify both genetic and epigenetic features that define and regulate their distribution and utilization. Here we summarize recent advances in our understanding of how primary sequence, chromatin environment, and nuclear architecture contribute to the dynamic selection and activation of replication origins across diverse cell types and developmental stages. PMID:27542827

1981 radon barrier field test at Grand Junction uranium mill tailings pile

International Nuclear Information System (INIS)

Hartley, J.N.; Gee, G.W.; Baker, E.G.; Freeman, H.D.

1983-04-01

Technologies to reduce radon released from uranium mill tailings are being investigated by Pacific Northwest Laboratory as part of the Department of Energy's Uranium Mill Tailings Remedial Action Project (UMTRAP) Technology development program. These technologies include: (1) earthen cover systems, (2) multilayer cover systems, and (3) asphalt emulsion radon barrier systems. During the summer of 1981, a field test was initiated at the Grand Junction, Colorado, uranium tailings pile to evaluate and compare the effectiveness of each radon barrier system. Test plots cover about 1.2 ha (3 acres). The field test has demonstrated the effectiveness of all three cover systems in reducing radon release to near background levels ( 2 s - 1 ). In conjunction with the field tests, column tests (1.8 m diameter) were initiated with cover systems similar to those in the larger field test plots. The column tests allow a direct comparison of the two test procedures and also provide detailed information on radon transport

Sample preparation composite and replicate strategy case studies for assay of solid oral drug products.

Science.gov (United States)

Nickerson, Beverly; Harrington, Brent; Li, Fasheng; Guo, Michele Xuemei

2017-11-30

Drug product assay is one of several tests required for new drug products to ensure the quality of the product at release and throughout the life cycle of the product. Drug product assay testing is typically performed by preparing a composite sample of multiple dosage units to obtain an assay value representative of the batch. In some cases replicate composite samples may be prepared and the reportable assay value is the average value of all the replicates. In previously published work by Harrington et al. (2014) [5], a sample preparation composite and replicate strategy for assay was developed to provide a systematic approach which accounts for variability due to the analytical method and dosage form with a standard error of the potency assay criteria based on compendia and regulatory requirements. In this work, this sample preparation composite and replicate strategy for assay is applied to several case studies to demonstrate the utility of this approach and its application at various stages of pharmaceutical drug product development. Copyright © 2017 Elsevier B.V. All rights reserved.

Multiscale analysis of replication technique efficiency for 3D roughness characterization of manufactured surfaces

Science.gov (United States)

Jolivet, S.; Mezghani, S.; El Mansori, M.

2016-09-01

The replication of topography has been generally restricted to optimizing material processing technologies in terms of statistical and single-scale features such as roughness. By contrast, manufactured surface topography is highly complex, irregular, and multiscale. In this work, we have demonstrated the use of multiscale analysis on replicates of surface finish to assess the precise control of the finished replica. Five commercial resins used for surface replication were compared. The topography of five standard surfaces representative of common finishing processes were acquired both directly and by a replication technique. Then, they were characterized using the ISO 25178 standard and multiscale decomposition based on a continuous wavelet transform, to compare the roughness transfer quality at different scales. Additionally, atomic force microscope force modulation mode was used in order to compare the resins’ stiffness properties. The results showed that less stiff resins are able to replicate the surface finish along a larger wavelength band. The method was then tested for non-destructive quality control of automotive gear tooth surfaces.

Chromosomal DNA replication of Vicia faba cells

International Nuclear Information System (INIS)

Ikushima, Takaji

1976-01-01

The chromosomal DNA replication of higher plant cells has been investigated by DNA fiber autoradiography. The nuclear DNA fibers of Vicia root meristematic cells are organized into many tandem arrays of replication units or replicons which exist as clusters with respect to replication. DNA is replicated bidirectionally from the initiation points at the average rate of 0.15 μm/min at 20 0 C, and the average interinitiation interval is about 16 μm. The manner of chromosomal DNA replication in this higher plant is similar to that found in other eukaryotic cells at a subchromosomal level. (auth.)

Sequential steps in DNA replication are inhibited to ensure reduction of ploidy in meiosis

Science.gov (United States)

Hua, Hui; Namdar, Mandana; Ganier, Olivier; Gregan, Juraj; Méchali, Marcel; Kearsey, Stephen E.

2013-01-01

Meiosis involves two successive rounds of chromosome segregation without an intervening S phase. Exit from meiosis I is distinct from mitotic exit, in that replication origins are not licensed by Mcm2-7 chromatin binding, but spindle disassembly occurs during a transient interphase-like state before meiosis II. The absence of licensing is assumed to explain the block to DNA replication, but this has not been formally tested. Here we attempt to subvert this block by expressing the licensing control factors Cdc18 and Cdt1 during the interval between meiotic nuclear divisions. Surprisingly, this leads only to a partial round of DNA replication, even when these factors are overexpressed and effect clear Mcm2-7 chromatin binding. Combining Cdc18 and Cdt1 expression with modulation of cyclin-dependent kinase activity, activation of Dbf4-dependent kinase, or deletion of the Spd1 inhibitor of ribonucleotide reductase has little additional effect on the extent of DNA replication. Single-molecule analysis indicates this partial round of replication results from inefficient progression of replication forks, and thus both initiation and elongation replication steps may be inhibited in late meiosis. In addition, DNA replication or damage during the meiosis I–II interval fails to arrest meiotic progress, suggesting absence of checkpoint regulation of meiosis II entry. PMID:23303250

Surface micro topography replication in injection moulding

DEFF Research Database (Denmark)

Arlø, Uffe Rolf

Thermoplastic injection moulding is a widely used industrial process that involves surface generation by replication. The surface topography of injection moulded plastic parts can be important for aesthetical or technical reasons. With the emergence of microengineering and nanotechnology additional...... importance of surface topography follows. In general the replication is not perfect and the topography of the plastic part differs from the inverse topography of the mould cavity. It is desirable to be able to control the degree of replication perfection or replication quality. This requires an understanding...... of the physical mechanisms of replication. Such understanding can lead to improved process design and facilitate in-line process quality control with respect to surface properties. The purpose of the project is to identify critical factors that affect topography replication quality and to obtain an understanding...

DNA replication in ultraviolet light irradiated Chinese hamster cells: the nature of replicon inhibition and post-replication repair

International Nuclear Information System (INIS)

Doniger, J.

1978-01-01

DNA replication in ultraviolet light irradiated Chinese hamster cells was studied using techniques of DNA fiber autoradiography and alkaline sucrose sedimentation. Bidirectionally growing replicons were observed in the autoradiograms independent of the irradiation conditions. After a dose of 5 J/m 2 at 254 nm the rate of fork progression was the same as in unirradiated cells, while the rate of replication was reduced by 50%. After a dose of 10J/m 2 the rate of fork progression was reduced 40%, while the replication rate was only 25% of normal. Therefore, at low doses of ultraviolet light irradiation, the inhibition of DNA replication is due to reduction in the number of functioning replicons, while at higher doses the rate of fork progression is also slowed. Those replicons which no longer function after irradiation are blocked in fork movement rather than replicon initiation. After irradiation, pulse label was first incorporated into short nascent strands, the average size of which was approximately equal to the distance between pyrimidine dimers. Under conditions where post-replication repair occurs these short strands were eventually joined into larger pieces. Finally, the data show that slowing post-replication repair with caffeine does not slow fork movement. The results presented here support the post-replication repair model of 'gapped synthesis' and rule out a major role for 'replicative bypass'. (author)

Field test of coordinated ramp metering (CRM).

Science.gov (United States)

2017-03-15

This project has focused on field implementation and testing of a Coordinated Ramp Metering (CRM) algorithm at California State Route 99 : Northbound corridor in Sacramento between Calvine Road and the SR50 interchange after 12th Ave. It is a 9 mile ...

Dynamics of Escherichia coli Chromosome Segregation during Multifork Replication

DEFF Research Database (Denmark)

Nielsen, Henrik Jørck; Youngren, Brenda; Hansen, Flemming G.

2007-01-01

Slowly growing Escherichia coli cells have a simple cell cycle, with replication and progressive segregation of the chromosome completed before cell division. In rapidly growing cells, initiation of replication occurs before the previous replication rounds are complete. At cell division, the chro......Slowly growing Escherichia coli cells have a simple cell cycle, with replication and progressive segregation of the chromosome completed before cell division. In rapidly growing cells, initiation of replication occurs before the previous replication rounds are complete. At cell division......, the chromosomes contain multiple replication forks and must be segregated while this complex pattern of replication is still ongoing. Here, we show that replication and segregation continue in step, starting at the origin and progressing to the replication terminus. Thus, early-replicated markers on the multiple......-branched chromosomes continue to separate soon after replication to form separate protonucleoids, even though they are not segregated into different daughter cells until later generations. The segregation pattern follows the pattern of chromosome replication and does not follow the cell division cycle. No extensive...

Field Tests of the HYDAD-D Landmine Detector

International Nuclear Information System (INIS)

Brooks, F.D.; Drosg, M.; Smit, F.D.

2010-01-01

The HYDAD-D landmine detector has been field tested in South Africa and in Egypt, using both a dummy landmine and real (disabled) anti-personnel landmines. A hand-held version of the detector and a version that incorporates a motor-driven scanner were tested. The test results demonstrate that HYDAD-D can detect the VS 50 anti-personnel landmine (mass 185 g) when buried at a cover depth of 20 cm in dry sand. (author)

Nonequilibrium Entropic Bounds for Darwinian Replicators

Directory of Open Access Journals (Sweden)

Jordi Piñero

2018-01-01

Full Text Available Life evolved on our planet by means of a combination of Darwinian selection and innovations leading to higher levels of complexity. The emergence and selection of replicating entities is a central problem in prebiotic evolution. Theoretical models have shown how populations of different types of replicating entities exclude or coexist with other classes of replicators. Models are typically kinetic, based on standard replicator equations. On the other hand, the presence of thermodynamical constraints for these systems remain an open question. This is largely due to the lack of a general theory of statistical methods for systems far from equilibrium. Nonetheless, a first approach to this problem has been put forward in a series of novel developements falling under the rubric of the extended second law of thermodynamics. The work presented here is twofold: firstly, we review this theoretical framework and provide a brief description of the three fundamental replicator types in prebiotic evolution: parabolic, malthusian and hyperbolic. Secondly, we employ these previously mentioned techinques to explore how replicators are constrained by thermodynamics. Finally, we comment and discuss where further research should be focused on.

Rescue from replication stress during mitosis.

Science.gov (United States)

Fragkos, Michalis; Naim, Valeria

2017-04-03

Genomic instability is a hallmark of cancer and a common feature of human disorders, characterized by growth defects, neurodegeneration, cancer predisposition, and aging. Recent evidence has shown that DNA replication stress is a major driver of genomic instability and tumorigenesis. Cells can undergo mitosis with under-replicated DNA or unresolved DNA structures, and specific pathways are dedicated to resolving these structures during mitosis, suggesting that mitotic rescue from replication stress (MRRS) is a key process influencing genome stability and cellular homeostasis. Deregulation of MRRS following oncogene activation or loss-of-function of caretaker genes may be the cause of chromosomal aberrations that promote cancer initiation and progression. In this review, we discuss the causes and consequences of replication stress, focusing on its persistence in mitosis as well as the mechanisms and factors involved in its resolution, and the potential impact of incomplete replication or aberrant MRRS on tumorigenesis, aging and disease.

Personality and Academic Motivation: Replication, Extension, and Replication

Science.gov (United States)

Jones, Martin H.; McMichael, Stephanie N.

2015-01-01

Previous work examines the relationships between personality traits and intrinsic/extrinsic motivation. We replicate and extend previous work to examine how personality may relate to achievement goals, efficacious beliefs, and mindset about intelligence. Approximately 200 undergraduates responded to the survey with a 150 participants replicating…

DNA replication origins-where do we begin?

Science.gov (United States)

Prioleau, Marie-Noëlle; MacAlpine, David M

2016-08-01

For more than three decades, investigators have sought to identify the precise locations where DNA replication initiates in mammalian genomes. The development of molecular and biochemical approaches to identify start sites of DNA replication (origins) based on the presence of defining and characteristic replication intermediates at specific loci led to the identification of only a handful of mammalian replication origins. The limited number of identified origins prevented a comprehensive and exhaustive search for conserved genomic features that were capable of specifying origins of DNA replication. More recently, the adaptation of origin-mapping assays to genome-wide approaches has led to the identification of tens of thousands of replication origins throughout mammalian genomes, providing an unprecedented opportunity to identify both genetic and epigenetic features that define and regulate their distribution and utilization. Here we summarize recent advances in our understanding of how primary sequence, chromatin environment, and nuclear architecture contribute to the dynamic selection and activation of replication origins across diverse cell types and developmental stages. © 2016 Prioleau and MacAlpine; Published by Cold Spring Harbor Laboratory Press.

Pattern replication by confined dewetting

NARCIS (Netherlands)

Harkema, S.; Schäffer, E.; Morariu, M.D.; Steiner, U

2003-01-01

The dewetting of a polymer film in a confined geometry was employed in a pattern-replication process. The instability of dewetting films is pinned by a structured confining surface, thereby replicating its topographic pattern. Depending on the surface energy of the confining surface, two different

Parametrised Constants and Replication for Spatial Mobility

DEFF Research Database (Denmark)

Hüttel, Hans; Haagensen, Bjørn

2009-01-01

Parametrised replication and replication are common ways of expressing infinite computation in process calculi. While parametrised constants can be encoded using replication in the π-calculus, this changes in the presence of spatial mobility as found in e.g. the distributed π- calculus...... of the distributed π-calculus with parametrised constants and replication are incomparable. On the other hand, we shall see that there exists a simple encoding of recursion in mobile ambients....

Adenovirus sequences required for replication in vivo.

OpenAIRE

Wang, K; Pearson, G D

1985-01-01

We have studied the in vivo replication properties of plasmids carrying deletion mutations within cloned adenovirus terminal sequences. Deletion mapping located the adenovirus DNA replication origin entirely within the first 67 bp of the adenovirus inverted terminal repeat. This region could be further subdivided into two functional domains: a minimal replication origin and an adjacent auxillary region which boosted the efficiency of replication by more than 100-fold. The minimal origin occup...

USP37 deubiquitinates Cdt1 and contributes to regulate DNA replication.

Science.gov (United States)

Hernández-Pérez, Santiago; Cabrera, Elisa; Amoedo, Hugo; Rodríguez-Acebes, Sara; Koundrioukoff, Stephane; Debatisse, Michelle; Méndez, Juan; Freire, Raimundo

2016-10-01

DNA replication control is a key process in maintaining genomic integrity. Monitoring DNA replication initiation is particularly important as it needs to be coordinated with other cellular events and should occur only once per cell cycle. Crucial players in the initiation of DNA replication are the ORC protein complex, marking the origin of replication, and the Cdt1 and Cdc6 proteins, that license these origins to replicate by recruiting the MCM2-7 helicase. To accurately achieve its functions, Cdt1 is tightly regulated. Cdt1 levels are high from metaphase and during G1 and low in S/G2 phases of the cell cycle. This control is achieved, among other processes, by ubiquitination and proteasomal degradation. In an overexpression screen for Cdt1 deubiquitinating enzymes, we isolated USP37, to date the first ubiquitin hydrolase controlling Cdt1. USP37 overexpression stabilizes Cdt1, most likely a phosphorylated form of the protein. In contrast, USP37 knock down destabilizes Cdt1, predominantly during G1 and G1/S phases of the cell cycle. USP37 interacts with Cdt1 and is able to de-ubiquitinate Cdt1 in vivo and, USP37 is able to regulate the loading of MCM complexes onto the chromatin. In addition, downregulation of USP37 reduces DNA replication fork speed. Taken together, here we show that the deubiquitinase USP37 plays an important role in the regulation of DNA replication. Whether this is achieved via Cdt1, a central protein in this process, which we have shown to be stabilized by USP37, or via additional factors, remains to be tested. Copyright © 2016 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

Prototype Engineered Barrier System Field Tests (PEBSFT)

International Nuclear Information System (INIS)

Ramirez, A.L.; Wilder, D.G.

1991-02-01

This progress report presents the interpretation of data obtained (up to November 1, 1988) from the Prototype Engineered Barrier System Field Tests (PEBSFT) that are being performed for the Yucca Mountain Project (YMP) in G-Tunnel within the Nevada Test site. The PEBSFTs are being conducted to evaluate the applicability of measurement techniques, numerical models, and procedures developed for the field tests for future investigations that will be conducted in the Exploratory Shaft Facilities, at a potential high-level radioactive waste repository site in Yucca Mountain. The primary objective of the tests is to provide the basis for determining whether tests planned for Yucca Mountain have the potential to be successful. Thirteen chapters discuss the following: mapping the electromagnetic permittivity and attenuation rate of the rock mass; changes in moisture content detected by the neutron logging probe; characterization of the in-situ permeability of the fractured tuff around the heater borehole; electrical resistance heater installed in a 30-cm borehole; relative humidity measurements; the operation, design, construction, calibration, and installation of a microwave circuit that might provide partial pressure information at temperatures in excess of 200 degree C (392 degree F); pressure and temperature measurements in the G-Tunnel; the moisture collection system, which attempts to collect steam that migrates into the heater borehole; The borehole television and borescope surveys that were performed to map the location, orientation, and aperture of the fractures intersecting the boreholes; preliminary scoping calculations of the hydrothermal conditions expected for this prototype test; the Data Acquisition System; and the results of the PEBSFT, preliminary interpretations of these results, and plans for the remainder of the test. Chapters have been indexed separately for inclusion on the data base

The Center-TRACON Automation System: Simulation and field testing

Science.gov (United States)

Denery, Dallas G.; Erzberger, Heinz

1995-01-01

A new concept for air traffic management in the terminal area, implemented as the Center-TRACON Automation System, has been under development at NASA Ames in a cooperative program with the FAA since 1991. The development has been strongly influenced by concurrent simulation and field site evaluations. The role of simulation and field activities in the development process will be discussed. Results of recent simulation and field tests will be presented.

Mammalian RAD52 Functions in Break-Induced Replication Repair of Collapsed DNA Replication Forks.

Science.gov (United States)

Sotiriou, Sotirios K; Kamileri, Irene; Lugli, Natalia; Evangelou, Konstantinos; Da-Ré, Caterina; Huber, Florian; Padayachy, Laura; Tardy, Sebastien; Nicati, Noemie L; Barriot, Samia; Ochs, Fena; Lukas, Claudia; Lukas, Jiri; Gorgoulis, Vassilis G; Scapozza, Leonardo; Halazonetis, Thanos D

2016-12-15

Human cancers are characterized by the presence of oncogene-induced DNA replication stress (DRS), making them dependent on repair pathways such as break-induced replication (BIR) for damaged DNA replication forks. To better understand BIR, we performed a targeted siRNA screen for genes whose depletion inhibited G1 to S phase progression when oncogenic cyclin E was overexpressed. RAD52, a gene dispensable for normal development in mice, was among the top hits. In cells in which fork collapse was induced by oncogenes or chemicals, the Rad52 protein localized to DRS foci. Depletion of Rad52 by siRNA or knockout of the gene by CRISPR/Cas9 compromised restart of collapsed forks and led to DNA damage in cells experiencing DRS. Furthermore, in cancer-prone, heterozygous APC mutant mice, homozygous deletion of the Rad52 gene suppressed tumor growth and prolonged lifespan. We therefore propose that mammalian RAD52 facilitates repair of collapsed DNA replication forks in cancer cells. Copyright © 2016 The Author(s). Published by Elsevier Inc. All rights reserved.

Suppression of Poxvirus Replication by Resveratrol.

Science.gov (United States)

Cao, Shuai; Realegeno, Susan; Pant, Anil; Satheshkumar, Panayampalli S; Yang, Zhilong

2017-01-01

Poxviruses continue to cause serious diseases even after eradication of the historically deadly infectious human disease, smallpox. Poxviruses are currently being developed as vaccine vectors and cancer therapeutic agents. Resveratrol is a natural polyphenol stilbenoid found in plants that has been shown to inhibit or enhance replication of a number of viruses, but the effect of resveratrol on poxvirus replication is unknown. In the present study, we found that resveratrol dramatically suppressed the replication of vaccinia virus (VACV), the prototypic member of poxviruses, in various cell types. Resveratrol also significantly reduced the replication of monkeypox virus, a zoonotic virus that is endemic in Western and Central Africa and causes human mortality. The inhibitory effect of resveratrol on poxviruses is independent of VACV N1 protein, a potential resveratrol binding target. Further experiments demonstrated that resveratrol had little effect on VACV early gene expression, while it suppressed VACV DNA synthesis, and subsequently post-replicative gene expression.

Suppression of Poxvirus Replication by Resveratrol

Directory of Open Access Journals (Sweden)

Shuai Cao

2017-11-01

Full Text Available Poxviruses continue to cause serious diseases even after eradication of the historically deadly infectious human disease, smallpox. Poxviruses are currently being developed as vaccine vectors and cancer therapeutic agents. Resveratrol is a natural polyphenol stilbenoid found in plants that has been shown to inhibit or enhance replication of a number of viruses, but the effect of resveratrol on poxvirus replication is unknown. In the present study, we found that resveratrol dramatically suppressed the replication of vaccinia virus (VACV, the prototypic member of poxviruses, in various cell types. Resveratrol also significantly reduced the replication of monkeypox virus, a zoonotic virus that is endemic in Western and Central Africa and causes human mortality. The inhibitory effect of resveratrol on poxviruses is independent of VACV N1 protein, a potential resveratrol binding target. Further experiments demonstrated that resveratrol had little effect on VACV early gene expression, while it suppressed VACV DNA synthesis, and subsequently post-replicative gene expression.

Crinivirus replication and host interactions

Directory of Open Access Journals (Sweden)

Zsofia A Kiss

2013-05-01

Full Text Available Criniviruses comprise one of the genera within the family Closteroviridae. Members in this family are restricted to the phloem and rely on whitefly vectors of the genera Bemisia and/or Trialeurodes for plant-to-plant transmission. All criniviruses have bipartite, positive-sense ssRNA genomes, although there is an unconfirmed report of one having a tripartite genome. Lettuce infectious yellows virus (LIYV is the type species of the genus, the best studied so far of the criniviruses and the first for which a reverse genetics system was available. LIYV RNA 1 encodes for proteins predicted to be involved in replication, and alone is competent for replication in protoplasts. Replication results in accumulation of cytoplasmic vesiculated membranous structures which are characteristic of most studied members of the Closteroviridae. These membranous structures, often referred to as BYV-type vesicles, are likely sites of RNA replication. LIYV RNA 2 is replicated in trans when co-infecting cells with RNA 1, but is temporally delayed relative to RNA1. Efficient RNA 2 replication also is dependent on the RNA 1-encoded RNA binding protein, P34. No LIYV RNA 2-encoded proteins have been shown to affect RNA replication, but at least four, CP, CPm, Hsp70h, and p59 are virion structural components and CPm is a determinant of whitefly transmissibility. Roles of other LIYV RNA 2-encoded proteins are largely as yet unknown, but P26 is a non-virion protein that accumulates in cells as characteristic plasmalemma deposits which in plants are localized within phloem parenchyma and companion cells over plasmodesmata connections to sieve elements. The two remaining crinivirus-conserved RNA 2-encoded proteins are P5 and P9. P5 is 39 amino acid protein and is encoded at the 5’ end of RNA 2 as ORF1 and is part of the hallmark closterovirus gene array. The orthologous gene in BYV has been shown to play a role in cell-to-cell movement and indicated to be localized to the

Effector-Triggered Self-Replication in Coupled Subsystems.

Science.gov (United States)

Komáromy, Dávid; Tezcan, Meniz; Schaeffer, Gaël; Marić, Ivana; Otto, Sijbren

2017-11-13

In living systems processes like genome duplication and cell division are carefully synchronized through subsystem coupling. If we are to create life de novo, similar control over essential processes such as self-replication need to be developed. Here we report that coupling two dynamic combinatorial subsystems, featuring two separate building blocks, enables effector-mediated control over self-replication. The subsystem based on the first building block shows only self-replication, whereas that based on the second one is solely responsive toward a specific external effector molecule. Mixing the subsystems arrests replication until the effector molecule is added, resulting in the formation of a host-effector complex and the liberation of the building block that subsequently engages in self-replication. The onset, rate and extent of self-replication is controlled by the amount of effector present. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Addressing the "Replication Crisis": Using Original Studies to Design Replication Studies with Appropriate Statistical Power.

Science.gov (United States)

Anderson, Samantha F; Maxwell, Scott E

2017-01-01

Psychology is undergoing a replication crisis. The discussion surrounding this crisis has centered on mistrust of previous findings. Researchers planning replication studies often use the original study sample effect size as the basis for sample size planning. However, this strategy ignores uncertainty and publication bias in estimated effect sizes, resulting in overly optimistic calculations. A psychologist who intends to obtain power of .80 in the replication study, and performs calculations accordingly, may have an actual power lower than .80. We performed simulations to reveal the magnitude of the difference between actual and intended power based on common sample size planning strategies and assessed the performance of methods that aim to correct for effect size uncertainty and/or bias. Our results imply that even if original studies reflect actual phenomena and were conducted in the absence of questionable research practices, popular approaches to designing replication studies may result in a low success rate, especially if the original study is underpowered. Methods correcting for bias and/or uncertainty generally had higher actual power, but were not a panacea for an underpowered original study. Thus, it becomes imperative that 1) original studies are adequately powered and 2) replication studies are designed with methods that are more likely to yield the intended level of power.

TEST OF AN ANIMAL DRAWN FIELD IMPLEMENT CART

Directory of Open Access Journals (Sweden)

Paolo Spugnoli

2008-06-01

Full Text Available The field performance of a horse-drawn hitch cart equipped with a PTO system powered by the two cart ground wheels have been investigated. For this purpose field tests on clay and turf soil, with varying ballast and PTO torque, have been carried out pulling the cart by a tractor. Preliminary tests were aimed at assessing the traction capability of horse breed. These tests showed that the mean draught force given by two of these horses was 173daN, average working speed was about 1m*s-1, resulting a mean draught power developed by each horse of about 0.86kW. The PTO cart system performance has shown that the torque has not exceeded 2.4daN*m, maximum draught or PTO power was 1.15kW, rotation speed just higher than 400min-1, with mean efficiency of about 50%. These values are consistent with horse performance and small haymaking, fertilizing, seeding and chemical application machine requirements.

NanoPCR observation: different levels of DNA replication fidelity in nanoparticle-enhanced polymerase chain reactions

International Nuclear Information System (INIS)

Shen Cenchao; Yang Wenjuan; Ji Qiaoli; Zhang Zhizhou; Maki, Hisaji; Dong Anjie

2009-01-01

Nanoparticle-assisted PCR (polymerase chain reaction) technology is getting more and more attention recently. It is believed that some of the DNA recombinant technologies will be upgraded by nanotechnology in the near future, among which DNA replication is one of the core manipulation techniques. So whether or not the DNA replication fidelity is compromised in nanoparticle-assisted PCR is a question. In this study, a total of 16 different metallic and non-metallic nanoparticles (NPs) were tested for their effects on DNA replication fidelity in vitro and in vivo. Sixteen types of nanomaterials were distinctly different in enhancing the PCR efficiency, and their relative capacity to retain DNA replication fidelity was largely different from each other based on rpsL gene mutation assay. Generally speaking, metallic nanoparticles induced larger error rates in DNA replication fidelity than non-metallic nanoparticles, and non-metallic nanomaterials such as carbon nanopowder or nanotubes were still safe as PCR enhancers because they did not compromise the DNA replication fidelity in the Taq DNA polymerase-based PCR system.

Superconductors for fusion magnets tested under pulsed field in SULTAN

International Nuclear Information System (INIS)

Bruzzone, P.; Bottura, L.; Katheder, H.; Blau, B.; Rohleder, I.; Vecsey, G.

1995-01-01

The SULTAN III test facility has been upgraded with a pair of pulsed field coils to carry out AC losses and stability experiments under full operating loads on large size, fusion conductors for ITER. A fast data aquisition system records the conductor behaviour under fast field transient. The commissioning results of the pulsed coils and instrumentation are critically discussed and the test capability of the set up is assessed. (orig.)

Epoxy replication for Wolter x-ray microscope fabrication

International Nuclear Information System (INIS)

Priedhorsky, W.

1981-01-01

An epoxy replica of a test piece designed to simulate a Wolter x-ray microscope geometry showed no loss of x-ray reflectivity or resolution, compared to the original. The test piece was a diamond-turned cone with 1.5 0 half angle. A flat was fly-cut on one side, then super- and conventionally polished. The replica was separated at the 1.5 0 -draft angle, simulating a shallow angle Wolter microscope geometry. A test with 8.34 A x rays at 0.9 0 grazing angle showed a reflectivity of 67% for the replica flat surface, and 70% for the original. No spread of the reflected beam was observed with a 20-arc second wide test beam. This test verifies the epoxy replication technique for production of Wolter x-ray microscopes

40 CFR 1065.295 - PM inertial balance for field-testing analysis.

Science.gov (United States)

2010-07-01

... net PM on a sample medium for field testing. (b) Component requirements. We recommend that you use a... 40 Protection of Environment 32 2010-07-01 2010-07-01 false PM inertial balance for field-testing analysis. 1065.295 Section 1065.295 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED...

Field testing existence values for riparian ecosystems

Science.gov (United States)

John W. Duffield; Chris J. Neher; David A. Patterson; Patricia A. Champ

2007-01-01

This paper presents preliminary findings on a cash and contingent valuation (cv) experiment. The study replicates major elements of an earlier (1990) experiment, which solicited hypothetical and actual donations to benefit instream flows for Montana fisheries. Extensions of the earlier work include: repeat contacts to increase response rate, follow-up of the contingent...

Multiple determinants controlling activation of yeast replication origins late in S phase.

Science.gov (United States)

Friedman, K L; Diller, J D; Ferguson, B M; Nyland, S V; Brewer, B J; Fangman, W L

1996-07-01

Analysis of a 131-kb segment of the left arm of yeast chromosome XIV beginning 157 kb from the telomere reveals four highly active origins of replication that initiate replication late in S phase. Previous work has shown that telomeres act as determinants for late origin activation. However, at least two of the chromosome XIV origins maintain their late activation time when located on large circular plasmids, indicating that late replication is independent of telomeres. Analysis of the replication time of plasmid derivatives containing varying amounts of chromosome XIV DNA show that a minimum of three chromosomal elements, distinct from each tested origin, contribute to late activation time. These late determinants are functionally equivalent, because duplication of one set of contributing sequences can compensate for the removal of another set. Furthermore, insertion of an origin that is normally early activated into this domain results in a shift to late activation, suggesting that the chromosome XIV origins are not unique in their ability to respond to the late determinants.

Combustion Safety Simplified Test Protocol Field Study

Energy Technology Data Exchange (ETDEWEB)

Brand, L [Gas Technology Inst., Des Plaines, IL (United States); Cautley, D. [Gas Technology Inst., Des Plaines, IL (United States); Bohac, D. [Gas Technology Inst., Des Plaines, IL (United States); Francisco, P. [Gas Technology Inst., Des Plaines, IL (United States); Shen, L. [Gas Technology Inst., Des Plaines, IL (United States); Gloss, S. [Gas Technology Inst., Des Plaines, IL (United States)

2015-11-05

"9Combustions safety is an important step in the process of upgrading homes for energy efficiency. There are several approaches used by field practitioners, but researchers have indicated that the test procedures in use are complex to implement and provide too many false positives. Field failures often mean that the house is not upgraded until after remediation or not at all, if not include in the program. In this report the PARR and NorthernSTAR DOE Building America Teams provide a simplified test procedure that is easier to implement and should produce fewer false positives. A survey of state weatherization agencies on combustion safety issues, details of a field data collection instrumentation package, summary of data collected over seven months, data analysis and results are included. The project provides several key results. State weatherization agencies do not generally track combustion safety failures, the data from those that do suggest that there is little actual evidence that combustion safety failures due to spillage from non-dryer exhaust are common and that only a very small number of homes are subject to the failures. The project team collected field data on 11 houses in 2015. Of these homes, two houses that demonstrated prolonged and excessive spillage were also the only two with venting systems out of compliance with the National Fuel Gas Code. The remaining homes experienced spillage that only occasionally extended beyond the first minute of operation. Combustion zone depressurization, outdoor temperature, and operation of individual fans all provide statistically significant predictors of spillage.

Histone deacetylase inhibitors improve the replication of oncolytic herpes simplex virus in breast cancer cells.

Directory of Open Access Journals (Sweden)

James J Cody

Full Text Available New therapies are needed for metastatic breast cancer patients. Oncolytic herpes simplex virus (oHSV is an exciting therapy being developed for use against aggressive tumors and established metastases. Although oHSV have been demonstrated safe in clinical trials, a lack of sufficient potency has slowed the clinical application of this approach. We utilized histone deacetylase (HDAC inhibitors, which have been noted to impair the innate antiviral response and improve gene transcription from viral vectors, to enhance the replication of oHSV in breast cancer cells. A panel of chemically diverse HDAC inhibitors were tested at three different doses (LD50 for their ability to modulate the replication of oHSV in breast cancer cells. Several of the tested HDAC inhibitors enhanced oHSV replication at low multiplicity of infection (MOI following pre-treatment of the metastatic breast cancer cell line MDA-MB-231 and the oHSV-resistant cell line 4T1, but not in the normal breast epithelial cell line MCF10A. Inhibitors of class I HDACs, including pan-selective compounds, were more effective for increasing oHSV replication compared to inhibitors that selectively target class II HDACs. These studies demonstrate that select HDAC inhibitors increase oHSV replication in breast cancer cells and provides support for pre-clinical evaluation of this combination strategy.

Non‐Canonical Replication Initiation: You’re Fired!

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Bazilė Ravoitytė

2017-01-01

Full Text Available The division of prokaryotic and eukaryotic cells produces two cells that inherit a perfect copy of the genetic material originally derived from the mother cell. The initiation of canonical DNA replication must be coordinated to the cell cycle to ensure the accuracy of genome duplication. Controlled replication initiation depends on a complex interplay of cis‐acting DNA sequences, the so‐called origins of replication (ori, with trans‐acting factors involved in the onset of DNA synthesis. The interplay of cis‐acting elements and trans‐acting factors ensures that cells initiate replication at sequence‐specific sites only once, and in a timely order, to avoid chromosomal endoreplication. However, chromosome breakage and excessive RNA:DNA hybrid formation can cause breakinduced (BIR or transcription‐initiated replication (TIR, respectively. These non‐canonical replication events are expected to affect eukaryotic genome function and maintenance, and could be important for genome evolution and disease development. In this review, we describe the difference between canonical and non‐canonical DNA replication, and focus on mechanistic differences and common features between BIR and TIR. Finally, we discuss open issues on the factors and molecular mechanisms involved in TIR.

Measurement of replication structures at the nanometer scale using super-resolution light microscopy.

Science.gov (United States)

Baddeley, D; Chagin, V O; Schermelleh, L; Martin, S; Pombo, A; Carlton, P M; Gahl, A; Domaing, P; Birk, U; Leonhardt, H; Cremer, C; Cardoso, M C

2010-01-01

DNA replication, similar to other cellular processes, occurs within dynamic macromolecular structures. Any comprehensive understanding ultimately requires quantitative data to establish and test models of genome duplication. We used two different super-resolution light microscopy techniques to directly measure and compare the size and numbers of replication foci in mammalian cells. This analysis showed that replication foci vary in size from 210 nm down to 40 nm. Remarkably, spatially modulated illumination (SMI) and 3D-structured illumination microscopy (3D-SIM) both showed an average size of 125 nm that was conserved throughout S-phase and independent of the labeling method, suggesting a basic unit of genome duplication. Interestingly, the improved optical 3D resolution identified 3- to 5-fold more distinct replication foci than previously reported. These results show that optical nanoscopy techniques enable accurate measurements of cellular structures at a level previously achieved only by electron microscopy and highlight the possibility of high-throughput, multispectral 3D analyses.

Factors influencing microinjection molding replication quality

Science.gov (United States)

Vera, Julie; Brulez, Anne-Catherine; Contraires, Elise; Larochette, Mathieu; Trannoy-Orban, Nathalie; Pignon, Maxime; Mauclair, Cyril; Valette, Stéphane; Benayoun, Stéphane

2018-01-01

In recent years, there has been increased interest in producing and providing high-precision plastic parts that can be manufactured by microinjection molding: gears, pumps, optical grating elements, and so on. For all of these applications, the replication quality is essential. This study has two goals: (1) fabrication of high-precision parts using the conventional injection molding machine; (2) identification of robust parameters that ensure production quality. Thus, different technological solutions have been used: cavity vacuuming and the use of a mold coated with DLC or CrN deposits. AFM and SEM analyses were carried out to characterize the replication profile. The replication quality was studied in terms of the process parameters, coated and uncoated molds and crystallinity of the polymer. Specific studies were processed to quantify the replicability of injection molded parts (ABS, PC and PP). Analysis of the Taguchi experimental designs permits prioritization of the impact of each parameter on the replication quality. A discussion taking into account these new parameters and the thermal and spreading properties on the coatings is proposed. It appeared that, in general, increasing the mold temperature improves the molten polymer fill in submicron features except for the steel insert (for which the presence of a vacuum is the most important factor). Moreover, the DLC coating was the best coating to increase the quality of the replication. This result could be explained by the lower thermal diffusivity of this coating. We noted that the viscosity of the polymers is not a primordial factor of the replication quality.

Evaluation of seed vigor tests for determinig alfalfa seed quality

Directory of Open Access Journals (Sweden)

hamidreza tavakoli kakhki

2009-06-01

Full Text Available In order to determine the best seed vigor test in laboratory which may provide high correlation between alfalfa seedling stablishment in the field, two experiment were conducted in field as well as in laboratory. Four alfalfa cultivars Bami, Ghareyonjeh, Hamadani and Yazdi were used. Different seed vigor tests including standard germination, germination in low tempretaure (5c , germination rate, osmotic stress, accelerated ageing and electrical conductivity tests were laid out in a randomized completely design (RCD with four replications in laboratory experiment‌. The second experiment was carried out in a randomized completely block design (RCBD and each treatment replicated four times‌. Results revealed significant differences (p≤ 0.01 among different cultivars for establishment percentage and rate. Results also showed that there were significant correlations between electrical conductivity test and establishment percentage (r= -0.65 , p ≤ 0.01 as well as establishment rate (r= -0.80 , p ≤ 0.01. Correlation coefficients for rate and emergence percentage were significant and positive (r= 0.91, p ≤ 0.01 . Stepwise method disclosed that the regression model (y=165.23 – 0.15 x r2=0.64 , (p ≤ 0.01 for predicting rate of emergence could be recommended. In this model x (dependent variable is value of electrical conductivity and y (independent variable is rate of emergence.

Test of piezo-ceramic motor technology in ITER relevant high magnetic fields

Energy Technology Data Exchange (ETDEWEB)

Monti, Chiara, E-mail: chiara.monti@enea.it [Associazione EURATOM-ENEA sulla Fusione, via Enrico Fermi 45, 00044 Frascati, Rome (Italy); Besi Vetrella, Ugo; Mugnaini, Giampiero; Neri, Carlo; Rossi, Paolo; Viola, Rosario [Associazione EURATOM-ENEA sulla Fusione, via Enrico Fermi 45, 00044 Frascati, Rome (Italy); Dubus, Gregory; Damiani, Carlo [Fusion for Energy, c/ Josep Pla, 2 Torres Diagonal Litoral, 08019 Barcelona (Spain)

2014-10-15

In the framework of a Fusion for Energy (F4E) grant, a test campaign started in 2012 in order to assess the performance of the in-vessel viewing system (IVVS) probe concept and to verify its compatibility when exposed to ITER typical working conditions. ENEA laboratories went through with several tests simulating high magnetic fields, high temperature, high vacuum, gamma radiation and neutron radiation. A customized motor has been adopted to study the performances of ultrasonic piezo motors technology in high magnetic field conditions. This paper reports on the testing activity performed on the motor in a multi Tesla magnetic field. The job was carried out in a test facility of ENEA laboratories able to achieve 14 T. A maximum field of 10 T, fully compliant with ITER requirements (8 T), was applied. A specific mechanical assembly has been designed and manufactured to hold the motor in the region with high homogeneity of the field. Results obtained so far indicate that the motor is compatible with high magnetic fields, and are presented in the paper.

The molecular biology of Bluetongue virus replication.

Science.gov (United States)

Patel, Avnish; Roy, Polly

2014-03-01

The members of Orbivirus genus within the Reoviridae family are arthropod-borne viruses which are responsible for high morbidity and mortality in ruminants. Bluetongue virus (BTV) which causes disease in livestock (sheep, goat, cattle) has been in the forefront of molecular studies for the last three decades and now represents the best understood orbivirus at a molecular and structural level. The complex nature of the virion structure has been well characterised at high resolution along with the definition of the virus encoded enzymes required for RNA replication; the ordered assembly of the capsid shell as well as the protein and genome sequestration required for it; and the role of host proteins in virus entry and virus release. More recent developments of Reverse Genetics and Cell-Free Assembly systems have allowed integration of the accumulated structural and molecular knowledge to be tested at meticulous level, yielding higher insight into basic molecular virology, from which the rational design of safe efficacious vaccines has been possible. This article is centred on the molecular dissection of BTV with a view to understanding the role of each protein in the virus replication cycle. These areas are important in themselves for BTV replication but they also indicate the pathways that related viruses, which includes viruses that are pathogenic to man and animals, might also use providing an informed starting point for intervention or prevention. Copyright © 2014 Elsevier B.V. All rights reserved.

Investigating the conformational stability of prion strains through a kinetic replication model.

Directory of Open Access Journals (Sweden)

Mattia Zampieri

2009-07-01

Full Text Available Prion proteins are known to misfold into a range of different aggregated forms, showing different phenotypic and pathological states. Understanding strain specificities is an important problem in the field of prion disease. Little is known about which PrP(Sc structural properties and molecular mechanisms determine prion replication, disease progression and strain phenotype. The aim of this work is to investigate, through a mathematical model, how the structural stability of different aggregated forms can influence the kinetics of prion replication. The model-based results suggest that prion strains with different conformational stability undergoing in vivo replication are characterizable in primis by means of different rates of breakage. A further role seems to be played by the aggregation rate (i.e. the rate at which a prion fibril grows. The kinetic variability introduced in the model by these two parameters allows us to reproduce the different characteristic features of the various strains (e.g., fibrils' mean length and is coherent with all experimental observations concerning strain-specific behavior.

Replicating chromatin: a tale of histones

DEFF Research Database (Denmark)

Groth, Anja

2009-01-01

Chromatin serves structural and functional roles crucial for genome stability and correct gene expression. This organization must be reproduced on daughter strands during replication to maintain proper overlay of epigenetic fabric onto genetic sequence. Nucleosomes constitute the structural...... framework of chromatin and carry information to specify higher-order organization and gene expression. When replication forks traverse the chromosomes, nucleosomes are transiently disrupted, allowing the replication machinery to gain access to DNA. Histone recycling, together with new deposition, ensures...

The Savannah River environmental technology field test platform: Phase II

International Nuclear Information System (INIS)

Rossabi, J.; Riha, B.D.; May, C.P.; Pemberton, B.E.; Jarosch, T.R.; Eddy-Dilek, C.A.; Looney, B.B.; Raymond, R.

1995-01-01

The principal goal in the development of new technologies for environmental monitoring and characterization is transferring them to organizations and individuals for use in site assessment and compliance monitoring. The DOE complex has devised several strategies to facilitate this transfer including joint research projects between private industries and government laboratories or universities (CRADAs), and streamlined licensing procedures. One strategy that has been under-utilized is a planned sequence gradually moving from laboratory development and field demonstration to long term evaluation and onsite use. Industrial partnership and commercial production can be initiated at any step based on the performance, market, user needs, and costs associated with the technology. The Savannah River Technology Center (SRTC) has been developing a program to rigorously field test promising environmental technologies that have not undergone EPA equivalency testing. The infrastructure and staff expertise developed as part of the activities of the Savannah River Integrated Demonstration Program (i.e., wells, available power, conventional baseline characterization and monitoring equipment, shelter structures) allows field testing of technologies without the difficulties of providing remote field support. By providing a well-characterized site and a well-developed infrastructure, technologies can be tested for long periods of time to determine their appropriate applications in environmental characterization and monitoring activities. Situation specific evaluations of the technology following stringent test plans can be made in comparison with simultaneous baseline methods and historical data. This program is designed to help expedite regulatory approval and technology transfer to manufacturers and the user community

Varicella-zoster virus (VZV) origin of DNA replication oriS influences origin-dependent DNA replication and flanking gene transcription.

Science.gov (United States)

Khalil, Mohamed I; Sommer, Marvin H; Hay, John; Ruyechan, William T; Arvin, Ann M

2015-07-01

The VZV genome has two origins of DNA replication (oriS), each of which consists of an AT-rich sequence and three origin binding protein (OBP) sites called Box A, C and B. In these experiments, the mutation in the core sequence CGC of the Box A and C not only inhibited DNA replication but also inhibited both ORF62 and ORF63 expression in reporter gene assays. In contrast the Box B mutation did not influence DNA replication or flanking gene transcription. These results suggest that efficient DNA replication enhances ORF62 and ORF63 transcription. Recombinant viruses carrying these mutations in both sites and one with a deletion of the whole oriS were constructed. Surprisingly, the recombinant virus lacking both copies of oriS retained the capacity to replicate in melanoma and HELF cells suggesting that VZV has another origin of DNA replication. Copyright © 2015 Elsevier Inc. All rights reserved.

A study of statistical tests for near-real-time materials accountancy using field test data of Tokai reprocessing plant

International Nuclear Information System (INIS)

Ihara, Hitoshi; Nishimura, Hideo; Ikawa, Koji; Miura, Nobuyuki; Iwanaga, Masayuki; Kusano, Toshitsugu.

1988-03-01

An Near-Real-Time Materials Accountancy(NRTA) system had been developed as an advanced safeguards measure for PNC Tokai Reprocessing Plant; a minicomputer system for NRTA data processing was designed and constructed. A full scale field test was carried out as a JASPAS(Japan Support Program for Agency Safeguards) project with the Agency's participation and the NRTA data processing system was used. Using this field test data, investigation of the detection power of a statistical test under real circumstances was carried out for five statistical tests, i.e., a significance test of MUF, CUMUF test, average loss test, MUF residual test and Page's test on MUF residuals. The result shows that the CUMUF test, average loss test, MUF residual test and the Page's test on MUF residual test are useful to detect a significant loss or diversion. An unmeasured inventory estimation model for the PNC reprocessing plant was developed in this study. Using this model, the field test data from the C-1 to 85 - 2 campaigns were re-analyzed. (author)

[Mediate evaluation of replicating a Training Program in Nonverbal Communication in Gerontology].

Science.gov (United States)

Schimidt, Teresa Cristina Gioia; Duarte, Yeda Aparecida de Oliveira; Silva, Maria Julia Paes da

2015-04-01

Replicating the training program in non-verbal communication based on the theoretical framework of interpersonal communication; non-verbal coding, valuing the aging aspects in the perspective of active aging, checking its current relevance through the content assimilation index after 90 days (mediate) of its application. A descriptive and exploratory field study was conducted in three hospitals under direct administration of the state of São Paulo that caters exclusively to Unified Health System (SUS) patients. The training lasted 12 hours divided in three meetings, applied to 102 health professionals. Revealed very satisfactory and satisfactory mediate content assimilation index in 82.9%. The program replication proved to be relevant and updated the setting of hospital services, while remaining efficient for healthcare professionals.

Cdc7 is required throughout the yeast S phase to activate replication origins.

Science.gov (United States)

Donaldson, A D; Fangman, W L; Brewer, B J

1998-02-15

The long-standing conclusion that the Cdc7 kinase of Saccharomyces cerevisiae is required only to trigger S phase has been challenged by recent data that suggests it acts directly on individual replication origins. We tested the possibility that early- and late-activated origins have different requirements for Cdc7 activity. Cells carrying a cdc7(ts) allele were first arrested in G1 at the cdc7 block by incubation at 37 degrees C, and then were allowed to enter S phase by brief incubation at 23 degrees C. During the S phase, after return to 37 degrees C, early-firing replication origins were activated, but late origins failed to fire. Similarly, a plasmid with a late-activated origin was defective in replication. As a consequence of the origin activation defect, duplication of chromosomal sequences that are normally replicated from late origins was greatly delayed. Early-replicating regions of the genome duplicated at approximately their normal time. The requirements of early and late origins for Cdc7 appear to be temporally rather than quantitatively different, as reducing overall levels of Cdc7 by growth at semi-permissive temperature reduced activation at early and late origins approximately equally. Our results show that Cdc7 activates early and late origins separately, with late origins requiring the activity later in S phase to permit replication initiation.

Modes of DNA repair and replication

International Nuclear Information System (INIS)

Hanawalt, P.; Kondo, S.

1979-01-01

Modes of DNA repair and replication require close coordination as well as some overlap of enzyme functions. Some classes of recovery deficient mutants may have defects in replication rather than repair modes. Lesions such as the pyrimidine dimers produced by ultraviolet light irradiation are the blocks to normal DNA replication in vivo and in vitro. The DNA synthesis by the DNA polymerase 1 of E. coli is blocked at one nucleotide away from the dimerized pyrimidines in template strands. Thus, some DNA polymerases seem to be unable to incorporate nucleotides opposite to the non-pairing lesions in template DNA strands. The lesions in template DNA strands may block the sequential addition of nucleotides in the synthesis of daughter strands. Normal replication utilizes a constitutive ''error-free'' mode that copies DNA templates with high fidelity, but which may be totally blocked at a lesion that obscures the appropriate base pairing specificity. It might be expected that modified replication system exhibits generally high error frequency. The error rate of DNA polymerases may be controlled by the degree of phosphorylation of the enzyme. Inducible SOS system is controlled by recA genes that also control the pathways for recombination. It is possible that SOS system involves some process other than the modification of a blocked replication apparatus to permit error-prone transdimer synthesis. (Yamashita, S.)

Charter School Replication. Policy Guide

Science.gov (United States)

Rhim, Lauren Morando

2009-01-01

"Replication" is the practice of a single charter school board or management organization opening several more schools that are each based on the same school model. The most rapid strategy to increase the number of new high-quality charter schools available to children is to encourage the replication of existing quality schools. This policy guide…

Replication of kinetoplast minicircle DNA

International Nuclear Information System (INIS)

Sheline, C.T.

1989-01-01

These studies describe the isolation and characterization of early minicircle replication intermediates from Crithidia fasciculata, and Leishmania tarentolae, the mitochondrial localization of a type II topoisomerase (TIImt) in C. fasciculata, and the implication of the aforementioned TIImt in minicircle replication in L. tarentolae. Early minicircle replication intermediates from C. fasciculata were identified and characterized using isolated kinetoplasts to incorporate radiolabeled nucleotides into its DNA. The pulse-label in an apparent theta-type intermediate chase into two daughter molecules. A uniquely gapped, ribonucleotide primed, knotted molecule represents the leading strand in the model proposed, and a highly gapped molecule represents the lagging strand. This theta intermediate is repaired in vitro to a doubly nicked catenated dimer which was shown to result from the replication of a single parental molecule. Very similar intermediates were found in the heterogeneous population of minicircles of L. tarentolae. The sites of the Leishmania specific discontinuities were mapped and shown to lie within the universally conserved sequence blocks in identical positions as compared to C. fasciculata and Trypanosoma equiperdum

Manual of Cupule Replication Technology

Directory of Open Access Journals (Sweden)

Giriraj Kumar

2015-09-01

Full Text Available Throughout the world, iconic rock art is preceded by non-iconic rock art. Cupules (manmade, roughly semi-hemispherical depressions on rocks form the major bulk of the early non-iconic rock art globally. The antiquity of cupules extends back to the Lower Paleolithic in Asia and Africa, hundreds of thousand years ago. When one observes these cupules, the inquisitive mind poses so many questions with regard to understanding their technology, reasons for selecting the site, which rocks were used to make the hammer stones used, the skill and cognitive abilities employed to create the different types of cupules, the objective of their creation, their age, and so on. Replication of the cupules can provide satisfactory answers to some of these questions. Comparison of the hammer stones and cupules produced by the replication process with those obtained from excavation can provide support to observations. This paper presents a manual of cupule replication technology based on our experience of cupule replication on hard quartzite rock near Daraki-Chattan in the Chambal Basin, India.

Zika Virus RNA Replication and Persistence in Brain and Placental Tissue

Science.gov (United States)

Rabeneck, Demi B.; Martines, Roosecelis B.; Reagan-Steiner, Sarah; Ermias, Yokabed; Estetter, Lindsey B.C.; Suzuki, Tadaki; Ritter, Jana; Keating, M. Kelly; Hale, Gillian; Gary, Joy; Muehlenbachs, Atis; Lambert, Amy; Lanciotti, Robert; Oduyebo, Titilope; Meaney-Delman, Dana; Bolaños, Fernando; Saad, Edgar Alberto Parra; Shieh, Wun-Ju; Zaki, Sherif R.

2017-01-01

Zika virus is causally linked with congenital microcephaly and may be associated with pregnancy loss. However, the mechanisms of Zika virus intrauterine transmission and replication and its tropism and persistence in tissues are poorly understood. We tested tissues from 52 case-patients: 8 infants with microcephaly who died and 44 women suspected of being infected with Zika virus during pregnancy. By reverse transcription PCR, tissues from 32 (62%) case-patients (brains from 8 infants with microcephaly and placental/fetal tissues from 24 women) were positive for Zika virus. In situ hybridization localized replicative Zika virus RNA in brains of 7 infants and in placentas of 9 women who had pregnancy losses during the first or second trimester. These findings demonstrate that Zika virus replicates and persists in fetal brains and placentas, providing direct evidence of its association with microcephaly. Tissue-based reverse transcription PCR extends the time frame of Zika virus detection in congenital and pregnancy-associated infections. PMID:27959260

Targeting DNA Replication Stress for Cancer Therapy

Directory of Open Access Journals (Sweden)

Jun Zhang

2016-08-01

Full Text Available The human cellular genome is under constant stress from extrinsic and intrinsic factors, which can lead to DNA damage and defective replication. In normal cells, DNA damage response (DDR mediated by various checkpoints will either activate the DNA repair system or induce cellular apoptosis/senescence, therefore maintaining overall genomic integrity. Cancer cells, however, due to constitutive growth signaling and defective DDR, may exhibit “replication stress” —a phenomenon unique to cancer cells that is described as the perturbation of error-free DNA replication and slow-down of DNA synthesis. Although replication stress has been proven to induce genomic instability and tumorigenesis, recent studies have counterintuitively shown that enhancing replicative stress through further loosening of the remaining checkpoints in cancer cells to induce their catastrophic failure of proliferation may provide an alternative therapeutic approach. In this review, we discuss the rationale to enhance replicative stress in cancer cells, past approaches using traditional radiation and chemotherapy, and emerging approaches targeting the signaling cascades induced by DNA damage. We also summarize current clinical trials exploring these strategies and propose future research directions including the use of combination therapies, and the identification of potential new targets and biomarkers to track and predict treatment responses to targeting DNA replication stress.

Global profiling of DNA replication timing and efficiency reveals that efficient replication/firing occurs late during S-phase in S. pombe.

Directory of Open Access Journals (Sweden)

Majid Eshaghi

Full Text Available BACKGROUND: During S. pombe S-phase, initiation of DNA replication occurs at multiple sites (origins that are enriched with AT-rich sequences, at various times. Current studies of genome-wide DNA replication profiles have focused on the DNA replication timing and origin location. However, the replication and/or firing efficiency of the individual origins on the genomic scale remain unclear. METHODOLOGY/PRINCIPAL FINDINGS: Using the genome-wide ORF-specific DNA microarray analysis, we show that in S. pombe, individual origins fire with varying efficiencies and at different times during S-phase. The increase in DNA copy number plotted as a function of time is approximated to the near-sigmoidal model, when considering the replication start and end timings at individual loci in cells released from HU-arrest. Replication efficiencies differ from origin to origin, depending on the origin's firing efficiency. We have found that DNA replication is inefficient early in S-phase, due to inefficient firing at origins. Efficient replication occurs later, attributed to efficient but late-firing origins. Furthermore, profiles of replication timing in cds1Delta cells are abnormal, due to the failure in resuming replication at the collapsed forks. The majority of the inefficient origins, but not the efficient ones, are found to fire in cds1Delta cells after HU removal, owing to the firing at the remaining unused (inefficient origins during HU treatment. CONCLUSIONS/SIGNIFICANCE: Taken together, our results indicate that efficient DNA replication/firing occurs late in S-phase progression in cells after HU removal, due to efficient late-firing origins. Additionally, checkpoint kinase Cds1p is required for maintaining the efficient replication/firing late in S-phase. We further propose that efficient late-firing origins are essential for ensuring completion of DNA duplication by the end of S-phase.

Replicated Data Management for Mobile Computing

CERN Document Server

Douglas, Terry

2008-01-01

Managing data in a mobile computing environment invariably involves caching or replication. In many cases, a mobile device has access only to data that is stored locally, and much of that data arrives via replication from other devices, PCs, and services. Given portable devices with limited resources, weak or intermittent connectivity, and security vulnerabilities, data replication serves to increase availability, reduce communication costs, foster sharing, and enhance survivability of critical information. Mobile systems have employed a variety of distributed architectures from client-server

pUL34 binding near the human cytomegalovirus origin of lytic replication enhances DNA replication and viral growth.

Science.gov (United States)

Slayton, Mark; Hossain, Tanvir; Biegalke, Bonita J

2018-05-01

The human cytomegalovirus (HCMV) UL34 gene encodes sequence-specific DNA-binding proteins (pUL34) which are required for viral replication. Interactions of pUL34 with DNA binding sites represses transcription of two viral immune evasion genes, US3 and US9. 12 additional predicted pUL34-binding sites are present in the HCMV genome (strain AD169) with three binding sites concentrated near the HCMV origin of lytic replication (oriLyt). We used ChIP-seq analysis of pUL34-DNA interactions to confirm that pUL34 binds to the oriLyt region during infection. Mutagenesis of the UL34-binding sites in an oriLyt-containing plasmid significantly reduced viral-mediated oriLyt-dependent DNA replication. Mutagenesis of these sites in the HCMV genome reduced the replication efficiencies of the resulting viruses. Protein-protein interaction analyses demonstrated that pUL34 interacts with the viral proteins IE2, UL44, and UL84, that are essential for viral DNA replication, suggesting that pUL34-DNA interactions in the oriLyt region are involved in the DNA replication cascade. Copyright © 2018 Elsevier Inc. All rights reserved.

Travel Time Data Collection Field Tests - Lessons Learned

Science.gov (United States)

1999-06-28

The Los Angeles Spread Spectrum Radio (SSR) Traffic Signal Interconnect Field Operational Test (FOT) investigated the feasibility of using wireless communications as an alternative to traditional hard-wire interconnection, to extend the coverage of c...

Testing Reproducibility in Earth Sciences

Science.gov (United States)

Church, M. A.; Dudill, A. R.; Frey, P.; Venditti, J. G.

2017-12-01

Reproducibility represents how closely the results of independent tests agree when undertaken using the same materials but different conditions of measurement, such as operator, equipment or laboratory. The concept of reproducibility is fundamental to the scientific method as it prevents the persistence of incorrect or biased results. Yet currently the production of scientific knowledge emphasizes rapid publication of previously unreported findings, a culture that has emerged from pressures related to hiring, publication criteria and funding requirements. Awareness and critique of the disconnect between how scientific research should be undertaken, and how it actually is conducted, has been prominent in biomedicine for over a decade, with the fields of economics and psychology more recently joining the conversation. The purpose of this presentation is to stimulate the conversation in earth sciences where, despite implicit evidence in widely accepted classifications, formal testing of reproducibility is rare.As a formal test of reproducibility, two sets of experiments were undertaken with the same experimental procedure, at the same scale, but in different laboratories. Using narrow, steep flumes and spherical glass beads, grain size sorting was examined by introducing fine sediment of varying size and quantity into a mobile coarse bed. The general setup was identical, including flume width and slope; however, there were some variations in the materials, construction and lab environment. Comparison of the results includes examination of the infiltration profiles, sediment mobility and transport characteristics. The physical phenomena were qualitatively reproduced but not quantitatively replicated. Reproduction of results encourages more robust research and reporting, and facilitates exploration of possible variations in data in various specific contexts. Following the lead of other fields, testing of reproducibility can be incentivized through changes to journal

Baculovirus resistance in codling moth (Cydia pomonella L.) caused by early block of virus replication.

Science.gov (United States)

Asser-Kaiser, Sabine; Radtke, Pit; El-Salamouny, Said; Winstanley, Doreen; Jehle, Johannes A

2011-02-20

An up to 10,000-fold resistance against the biocontrol agent Cydia pomonella granulovirus (CpGV) was observed in field populations of codling moth, C. pomonella, in Europe. Following different experimental approaches, a modified peritrophic membrane, a modified midgut receptor, or a change of the innate immune response could be excluded as possible resistance mechanisms. When CpGV replication was traced by quantitative PCR in different tissues of susceptible and resistant insects after oral and intra-hemocoelic infection, no virus replication could be detected in any of the tissues of resistant insects, suggesting a systemic block prior to viral DNA replication. This conclusion was corroborated by fluorescence microscopy using a modified CpGV (bacCpGV(hsp-eGFP)) carrying enhanced green fluorescent gene (eGFP), which showed that infection in resistant insects did not spread. In conclusion, the different lines of evidence indicate that CpGV can enter but not replicate in the cells of resistant codling moth larvae. Copyright © 2010 Elsevier Inc. All rights reserved.

Replication of urban innovations - prioritization of strategies for the replication of Dhaka's community-based decentralized composting model.

Science.gov (United States)

Yedla, Sudhakar

2012-01-01

Dhaka's community-based decentralized composting (DCDC) is a successful demonstration of solid waste management by adopting low-cost technology, local resources community participation and partnerships among the various actors involved. This paper attempts to understand the model, necessary conditions, strategies and their priorities to replicate DCDC in the other developing cities of Asia. Thirteen strategies required for its replication are identified and assessed based on various criteria, namely transferability, longevity, economic viability, adaptation and also overall replication. Priority setting by multi-criteria analysis by applying analytic hierarchy process revealed that immediate transferability without long-term and economic viability consideration is not advisable as this would result in unsustainable replication of DCDC. Based on the analysis, measures to ensure the product quality control; partnership among stakeholders (public-private-community); strategies to achieve better involvement of the private sector in solid waste management (entrepreneurship in approach); simple and low-cost technology; and strategies to provide an effective interface among the complementing sectors are identified as important strategies for its replication.

Replication of bacteriophage lambda DNA

International Nuclear Information System (INIS)

Tsurimoto, T.; Matsubara, K.

1983-01-01

In this paper results of studies on the mechanism of bacteriophage lambda replication using molecular biological and biochemical approaches are reported. The purification of the initiator proteins, O and P, and the role of the O and P proteins in the initiation of lambda DNA replication through interactions with specific DNA sequences are described. 47 references, 15 figures

Direct Visualization of DNA Replication Dynamics in Zebrafish Cells.

Science.gov (United States)

Kuriya, Kenji; Higashiyama, Eriko; Avşar-Ban, Eriko; Tamaru, Yutaka; Ogata, Shin; Takebayashi, Shin-ichiro; Ogata, Masato; Okumura, Katsuzumi

2015-12-01

Spatiotemporal regulation of DNA replication in the S-phase nucleus has been extensively studied in mammalian cells because it is tightly coupled with the regulation of other nuclear processes such as transcription. However, little is known about the replication dynamics in nonmammalian cells. Here, we analyzed the DNA replication processes of zebrafish (Danio rerio) cells through the direct visualization of replicating DNA in the nucleus and on DNA fiber molecules isolated from the nucleus. We found that zebrafish chromosomal DNA at the nuclear interior was replicated first, followed by replication of DNA at the nuclear periphery, which is reminiscent of the spatiotemporal regulation of mammalian DNA replication. However, the relative duration of interior DNA replication in zebrafish cells was longer compared to mammalian cells, possibly reflecting zebrafish-specific genomic organization. The rate of replication fork progression and ori-to-ori distance measured by the DNA combing technique were ∼ 1.4 kb/min and 100 kb, respectively, which are comparable to those in mammalian cells. To our knowledge, this is a first report that measures replication dynamics in zebrafish cells.

AFSC/FMA/NPRB Halibut Bycatch Video Field Test

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — These data arise from a field test of electronic monitoring (EM) of vessels participating in the Pacific halibut fishery off Alaska. A side-by-side comparison was...

Insulated hsp70B' promoter: stringent heat-inducible activity in replication-deficient, but not replication-competent adenoviruses.

Science.gov (United States)

Rohmer, Stanimira; Mainka, Astrid; Knippertz, Ilka; Hesse, Andrea; Nettelbeck, Dirk M

2008-04-01

Key to the realization of gene therapy is the development of efficient and targeted gene transfer vectors. Therapeutic gene transfer by replication-deficient or more recently by conditionally replication-competent/oncolytic adenoviruses has shown much promise. For specific applications, however, it will be advantageous to provide vectors that allow for external control of gene expression. The efficient cellular heat shock system in combination with available technology for focused and controlled hyperthermia suggests heat-regulated transcription control as a promising tool for this purpose. We investigated the feasibility of a short fragment of the human hsp70B' promoter, with and without upstream insulator elements, for the regulation of transgene expression by replication-deficient or oncolytic adenoviruses. Two novel adenoviral vectors with an insulated hsp70B' promoter were developed and showed stringent heat-inducible gene expression with induction ratios up to 8000-fold. In contrast, regulation of gene expression from the hsp70B' promoter without insulation was suboptimal. In replication-competent/oncolytic adenoviruses regulation of the hsp70B' promoter was lost specifically during late replication in permissive cells and could not be restored by the insulators. We developed novel adenovirus gene transfer vectors that feature improved and stringent regulation of transgene expression from the hsp70B' promoter using promoter insulation. These vectors have potential for gene therapy applications that benefit from external modulation of therapeutic gene expression or for combination therapy with hyperthermia. Furthermore, our study reveals that vector replication can deregulate inserted cellular promoters, an observation which is of relevance for the development of replication-competent/oncolytic gene transfer vectors. (c) 2008 John Wiley & Sons, Ltd.

Field testing for cosmic ray soft errors in semiconductor memories

International Nuclear Information System (INIS)

O'Gorman, T.J.; Ross, J.M.; Taber, A.H.; Ziegler, J.F.; Muhlfeld, H.P.; Montrose, C.J.; Curtis, H.W.; Walsh, J.L.

1996-01-01

This paper presents a review of experiments performed by IBM to investigate the causes of soft errors in semiconductor memory chips under field test conditions. The effects of alpha-particles and cosmic rays are separated by comparing multiple measurements of the soft-error rate (SER) of samples of memory chips deep underground and at various altitudes above the earth. The results of case studies on four different memory chips show that cosmic rays are an important source of the ionizing radiation that causes soft errors. The results of field testing are used to confirm the accuracy of the modeling and the accelerated testing of chips

Replication assessment of surface texture at sub-micrometre scale

DEFF Research Database (Denmark)

Quagliotti, Danilo; Tosello, Guido; Hansen, Hans Nørgaard

2017-01-01

[2]. A replication process requires reproducing a master geometry by conveying it to a substrate material. It is typically induced by means of different energy sources (usually heat and force) and a direct physical contact between the master and the substrate. Furthermore, concepts of advanced......, because of the replication nature of molding processes, the required specifications for the manufacture of micro molded components must be ensured by means of a metrological approach to surface replication and dimensional control of both master geometry and replicated substrate [3]-[4]. Therefore...... replication was assessed by the replication fidelity, i.e., comparing the produced parts with the tool used to replicate the geometry. Furthermore, the uncertainty of the replication fidelity was achieved by propagating the uncertainties evaluated for both masters and replicas. Finally, despite the specimens...

Mechanisms and regulation of DNA replication initiation in eukaryotes.

Science.gov (United States)

Parker, Matthew W; Botchan, Michael R; Berger, James M

2017-04-01

Cellular DNA replication is initiated through the action of multiprotein complexes that recognize replication start sites in the chromosome (termed origins) and facilitate duplex DNA melting within these regions. In a typical cell cycle, initiation occurs only once per origin and each round of replication is tightly coupled to cell division. To avoid aberrant origin firing and re-replication, eukaryotes tightly regulate two events in the initiation process: loading of the replicative helicase, MCM2-7, onto chromatin by the origin recognition complex (ORC), and subsequent activation of the helicase by its incorporation into a complex known as the CMG. Recent work has begun to reveal the details of an orchestrated and sequential exchange of initiation factors on DNA that give rise to a replication-competent complex, the replisome. Here, we review the molecular mechanisms that underpin eukaryotic DNA replication initiation - from selecting replication start sites to replicative helicase loading and activation - and describe how these events are often distinctly regulated across different eukaryotic model organisms.

Reply: Birnbaum's (2012 statistical tests of independence have unknown Type-I error rates and do not replicate within participant

Directory of Open Access Journals (Sweden)

Yun-shil Cha

2013-01-01

Full Text Available Birnbaum (2011, 2012 questioned the iid (independent and identically distributed sampling assumptions used by state-of-the-art statistical tests in Regenwetter, Dana and Davis-Stober's (2010, 2011 analysis of the ``linear order model''. Birnbaum (2012 cited, but did not use, a test of iid by Smith and Batchelder (2008 with analytically known properties. Instead, he created two new test statistics with unknown sampling distributions. Our rebuttal has five components: 1 We demonstrate that the Regenwetter et al. data pass Smith and Batchelder's test of iid with flying colors. 2 We provide evidence from Monte Carlo simulations that Birnbaum's (2012 proposed tests have unknown Type-I error rates, which depend on the actual choice probabilities and on how data are coded as well as on the null hypothesis of iid sampling. 3 Birnbaum analyzed only a third of Regenwetter et al.'s data. We show that his two new tests fail to replicate on the other two-thirds of the data, within participants. 4 Birnbaum selectively picked data of one respondent to suggest that choice probabilities may have changed partway into the experiment. Such nonstationarity could potentially cause a seemingly good fit to be a Type-II error. We show that the linear order model fits equally well if we allow for warm-up effects. 5 Using hypothetical data, Birnbaum (2012 claimed to show that ``true-and-error'' models for binary pattern probabilities overcome the alleged short-comings of Regenwetter et al.'s approach. We disprove this claim on the same data.

Differential Gender Performance on the Major Field Test-Business

Science.gov (United States)

Bielinska-Kwapisz, Agnieszka; Brown, F. William

2013-01-01

The Major Field Test in Business (MFT-B), a standardized assessment test of business knowledge among undergraduate business seniors, is widely used to measure student achievement. Many previous studies analyzing scores on the MFT-B report gender differences on the exam even after controlling for student's aptitude, general intellectual ability,…

Effects of ultraviolet irradiation on the rate and sequence of DNA replication in synchronized Chinese hamster cells

International Nuclear Information System (INIS)

Meyn, R.E.; Hewitt, R.R.; Thomson, L.F.; Humphrey, R.M.

1976-01-01

The effects of ultraviolet light (uv) irradiation on the rate of DNA replication in synchronized Chinese hamster ovary (CHO) cells were investigated. A technique for measuring semiconservative DNA replication was employed that involved growing the cells in medium containing 5-bromodeoxyuridine and subsequently determining the amount of DNA that acquired hybrid buoyant density in CsCl density gradients. One of the advantages of this technique was that it allowed a characterization of the extent of DNA replication as well as rate after irradiation. It was found that while there was a dose-dependent reduction in the rate of DNA replication following uv-irradiation, doses of up to 10 J/m 2 (which produce many dimers per replicon) did not prevent the ultimate replication of the entire genome. Hence, we conclude that dimers cannot be absolute blocks to DNA replication. In order to account for the total genome replication observed, a mechanism must exist that allows genome replication between dimers. The degree of reduction in the rate of replication by uv was the same whether the cells were irradiated at the Gl-S boundary or 1 h into S-phase. Previous work had shown that cells in early S-phase are considerably more sensitive to uv than cells at the G1-S boundary. Experiments specifically designed to test for reiterative replication showed that uv does not induce a second round of DNA replication within the same S-phase

Commercial Building Partnerships Replication and Diffusion

Energy Technology Data Exchange (ETDEWEB)

Antonopoulos, Chrissi A.; Dillon, Heather E.; Baechler, Michael C.

2013-09-16

This study presents findings from survey and interview data investigating replication efforts of Commercial Building Partnership (CBP) partners that worked directly with the Pacific Northwest National Laboratory (PNNL). PNNL partnered directly with 12 organizations on new and retrofit construction projects, which represented approximately 28 percent of the entire U.S. Department of Energy (DOE) CBP program. Through a feedback survey mechanism, along with personal interviews, PNNL gathered quantitative and qualitative data relating to replication efforts by each organization. These data were analyzed to provide insight into two primary research areas: 1) CBP partners’ replication efforts of technologies and approaches used in the CBP project to the rest of the organization’s building portfolio (including replication verification), and, 2) the market potential for technology diffusion into the total U.S. commercial building stock, as a direct result of the CBP program. The first area of this research focused specifically on replication efforts underway or planned by each CBP program participant. Factors that impact replication include motivation, organizational structure and objectives firms have for implementation of energy efficient technologies. Comparing these factors between different CBP partners revealed patterns in motivation for constructing energy efficient buildings, along with better insight into market trends for green building practices. The second area of this research develops a diffusion of innovations model to analyze potential broad market impacts of the CBP program on the commercial building industry in the United States.

A reporter system for replication-competent gammaretroviruses: the inGluc-MLV-DERSE assay

Science.gov (United States)

Aloia, Amanda L.; Duffy, Lisa; Pak, Vladimir; Lee, KyeongEun; Sanchez-Martinez, Silvia; Derse, David; Heidecker, Gisela; Cornetta, Kenneth; Rein, Alan

2012-01-01

While novel retroviral vectors for use in gene-therapy products are reducing the potential for formation of replication-competent retrovirus (RCR), it remains crucial to screen products for RCR for both research and clinical purposes. For clinical grade gammaretrovirus-based vectors, RCR screening is achieved by an extended S+L− or marker rescue assay, while standard methods for replication-competent lentivirus detection are still in development. In this report, we describe a rapid and sensitive method for replication-competent gammaretrovirus detection. We used this assay to detect three members of the gammaretrovirus family and compared the sensitivity of our assay with well-established methods for retrovirus detection, including the extended S+L− assay. Results presented here demonstrate that this assay should be useful for gene-therapy product testing. PMID:22402321

Deep Borehole Field Test Requirements and Controlled Assumptions.

Energy Technology Data Exchange (ETDEWEB)

Hardin, Ernest [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

2015-07-01

This document presents design requirements and controlled assumptions intended for use in the engineering development and testing of: 1) prototype packages for radioactive waste disposal in deep boreholes; 2) a waste package surface handling system; and 3) a subsurface system for emplacing and retrieving packages in deep boreholes. Engineering development and testing is being performed as part of the Deep Borehole Field Test (DBFT; SNL 2014a). This document presents parallel sets of requirements for a waste disposal system and for the DBFT, showing the close relationship. In addition to design, it will also inform planning for drilling, construction, and scientific characterization activities for the DBFT. The information presented here follows typical preparations for engineering design. It includes functional and operating requirements for handling and emplacement/retrieval equipment, waste package design and emplacement requirements, borehole construction requirements, sealing requirements, and performance criteria. Assumptions are included where they could impact engineering design. Design solutions are avoided in the requirements discussion. Deep Borehole Field Test Requirements and Controlled Assumptions July 21, 2015 iv ACKNOWLEDGEMENTS This set of requirements and assumptions has benefited greatly from reviews by Gordon Appel, Geoff Freeze, Kris Kuhlman, Bob MacKinnon, Steve Pye, David Sassani, Dave Sevougian, and Jiann Su.

Replication Studies: Vocabulary Knowledge in Relation to Memory and Analysis--An Approximate Replication of Milton's (2007) Study on Lexical Profiles and Learning Style

Science.gov (United States)

Booth, Paul

2013-01-01

This paper presents an approximate replication of Milton's (2007) study on lexical profiles and learning style. Milton investigated the assumption that more frequent words are acquired before less frequent ones. Using a vocabulary recognition test ("X-Lex") to measure vocabulary size, Milton found that L2 English group profiles show…

The Field Lysimeter Test Facility (FLTF) at the Hanford Site: Installation and initial tests

International Nuclear Information System (INIS)

Gee, G.W.; Kirkham, R.R.; Downs, J.L.; Campbell, M.D.

1989-02-01

The objectives of this program are to test barrier design concepts and to demonstrate a barrier design that meets established performance criteria for use in isolating wastes disposed of near-surface at the Hanford Site. Specifically, the program is designed to assess how well the barriers perform in controlling biointrusion, water infiltration, and erosion, as well as evaluating interactions between environmental variables and design factors of the barriers. To assess barrier performance and design with respect to infiltration control, field lysimeters and small- and large-scale field plots are planned to test the performance of specific barrier designs under actual and modified (enhanced precipitation) climatic conditions. The Field Lysimeter Test Facility (FLTF) is located in the 600 Area of the Hanford Site just east of the 200 West Area and adjacent to the Hanford Meteorological Station. The FLTF data will be used to assess the effectiveness of selected protective barrier configurations in controlling water infiltration. The facility consists of 14 drainage lysimeters (2 m dia x 3 m deep) and four precision weighing lysimeters (1.5 m x 1.5 m x 1.7 m deep). The lysimeters are buried at grade and aligned in a parallel configuration, with nine lysimeters on each side of an underground instrument chamber. The lysimeters were filled with materials to simulate a multilayer protective barrier system. Data gathered from the FLTF will be used to compare key barrier components and to calibrate and test models for predicting long-term barrier performance

Extremal dynamics in random replicator ecosystems

Energy Technology Data Exchange (ETDEWEB)

Kärenlampi, Petri P., E-mail: petri.karenlampi@uef.fi

2015-10-02

The seminal numerical experiment by Bak and Sneppen (BS) is repeated, along with computations with replicator models, including a greater amount of features. Both types of models do self-organize, and do obey power-law scaling for the size distribution of activity cycles. However species extinction within the replicator models interferes with the BS self-organized critical (SOC) activity. Speciation–extinction dynamics ruins any stationary state which might contain a steady size distribution of activity cycles. The BS-type activity appears as a dissimilar phenomenon in comparison to speciation–extinction dynamics in the replicator system. No criticality is found from the speciation–extinction dynamics. Neither are speciations and extinctions in real biological macroevolution known to contain any diverging distributions, or self-organization towards any critical state. Consequently, biological macroevolution probably is not a self-organized critical phenomenon. - Highlights: • Extremal Dynamics organizes random replicator ecosystems to two phases in fitness space. • Replicator systems show power-law scaling of activity. • Species extinction interferes with Bak–Sneppen type mutation activity. • Speciation–extinction dynamics does not show any critical phase transition. • Biological macroevolution probably is not a self-organized critical phenomenon.

Field tests of 2- and 40-tube condensers at the East Mesa Geothermal Test Site

Energy Technology Data Exchange (ETDEWEB)

Murphy, R.W.; Domingo, N.

1982-05-01

Two water-cooled isobutane condensers, one with 2 tubes and one with 40 tubes, were subjected to field tests at the East Mesa Geothermal Test Site to assess relative heat transfer performance in both surface evaporator and direct-contact evaporator modes. The five groups of tests established that field performance was below earlier laboratory-determined levels and that direct-contact evaporator mode performance was poorer than that for the surface evaporator mode. In all test situations, fluted condenser tubes performed better than smooth condenser tubes. Cooling water quality had no significant effect on performance, but brine preflash in the direct-contact mode did promote some relative performance improvement. Important implications of these results for binary geothermal power plants are that (1) working-fluid-side impurities can significantly degrade heat transfer performance of the power plant condensers and (2) provisions for minimizing such impurities may be required.

Experimental investigation on shrinkage and surface replication of injection moulded ceramic parts

DEFF Research Database (Denmark)

Islam, Aminul; Giannekas, Nikolaos; Marhöfer, David Maximilian

2014-01-01

Ceramic moulded parts are increasingly being used in advanced components and devices due to their unprecedented material and performance attributes. The surface finish, replication quality and material shrinkage are of immense importance for moulded ceramic parts intended for precision applications....... The current paper presents a thorough investigation on the process of ceramic moulding where it systematically characterizes the surface replication and shrinkage behaviours of precision moulded ceramic components. The test parts are moulded from Catamold TZP-A which is Y2O3-stabilised ZrO2 having widespread...... distribution for the moulded ceramic parts is presented....

A quick behavioral dichotic word test is prognostic for clinical response to cognitive therapy for depression: A replication study.

Science.gov (United States)

Bruder, Gerard E; Haggerty, Agnes; Siegle, Greg J

2017-02-01

There are no commonly used clinical indicators of whether an individual will benefit from cognitive therapy (CT) for depression. A prior study found right ear (left hemisphere) advantage for perceiving dichotic words predicted CT response. This study replicates this finding at a different research center in clinical trials that included clinically representative samples and community therapists. Right-handed individuals with unipolar major depressive disorder who subsequently received 12-14 weeks of CT at the University of Pittsburgh were tested on dichotic fused words and complex tones tests. Responders to CT showed twice the mean right ear advantage in dichotic fused words performance than non-responders. Patients with a right ear advantage greater than the mean for healthy controls had an 81% response rate to CT, whereas those with performance lower than the mean for controls had a 46% response rate. Individuals with a right ear advantage, indicative of strong left hemisphere language dominance, may be better at utilizing cognitive processes and left frontotemporal cortical regions critical for success of CT for depression. Findings at two clinical research centers suggest that verbal dichotic listening may be a clinically disseminative brief, inexpensive and easily automated test prognostic for response to CT across diverse clinical settings. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Conducting field studies for testing pesticide leaching models

Science.gov (United States)

Smith, Charles N.; Parrish, Rudolph S.; Brown, David S.

1990-01-01

A variety of predictive models are being applied to evaluate the transport and transformation of pesticides in the environment. These include well known models such as the Pesticide Root Zone Model (PRZM), the Risk of Unsaturated-Saturated Transport and Transformation Interactions for Chemical Concentrations Model (RUSTIC) and the Groundwater Loading Effects of Agricultural Management Systems Model (GLEAMS). The potentially large impacts of using these models as tools for developing pesticide management strategies and regulatory decisions necessitates development of sound model validation protocols. This paper offers guidance on many of the theoretical and practical problems encountered in the design and implementation of field-scale model validation studies. Recommendations are provided for site selection and characterization, test compound selection, data needs, measurement techniques, statistical design considerations and sampling techniques. A strategy is provided for quantitatively testing models using field measurements.

FUELS IN SOIL TEST KIT: FIELD USE OF DIESEL DOG SOIL TEST KITS

Energy Technology Data Exchange (ETDEWEB)

Unknown

2001-05-31

Western Research Institute (WRI) is commercializing Diesel Dog Portable Soil Test Kits for performing analysis of fuel-contaminated soils in the field. The technology consists of a method developed by WRI (U.S. Patents 5,561,065 and 5,976,883) and hardware developed by WRI that allows the method to be performed in the field (patent pending). The method is very simple and does not require the use of highly toxic reagents. The aromatic components in a soil extract are measured by absorption at 254 nm with a field-portable photometer. WRI added significant value to the technology by taking the method through the American Society for Testing and Materials (ASTM) approval and validation processes. The method is designated ASTM Method D-5831-96, Standard Test Method for Screening Fuels in Soils. This ASTM designation allows the method to be used for federal compliance activities. In FY 99, twenty-five preproduction kits were successfully constructed in cooperation with CF Electronics, Inc., of Laramie, Wyoming. The kit components work well and the kits are fully operational. In the calendar year 2000, kits were provided to the following entities who agreed to participate as FY 99 and FY 00 JSR (Jointly Sponsored Research) cosponsors and use the kits as opportunities arose for field site work: Wyoming Department of Environmental Quality (DEQ) (3 units), F.E. Warren Air Force Base, Gradient Corporation, The Johnson Company (2 units), IT Corporation (2 units), TRC Environmental Corporation, Stone Environmental, ENSR, Action Environmental, Laco Associates, Barenco, Brown and Caldwell, Dames and Moore Lebron LLP, Phillips Petroleum, GeoSyntek, and the State of New Mexico. By early 2001, ten kits had been returned to WRI following the six-month evaluation period. On return, the components of all ten kits were fully functional. The kits were upgraded with circuit modifications, new polyethylene foam inserts, and updated instruction manuals.

Field Testing Research at the NWTC (Fact Sheet)

Energy Technology Data Exchange (ETDEWEB)

2015-02-01

The National Wind Technology Center (NWTC) at the National Renewable Laboratory (NREL) has extensive field testing capabilities that have been used in collaboration with the wind industry to accelerate wind technology development and deployment for more than 30 years.

The Genomic Replication of the Crenarchaeal Virus SIRV2

DEFF Research Database (Denmark)

Martinez Alvarez, Laura

reinitiation events may partially explain the branched topology of the viral replication intermediates. We also analyzed the intracellular location of viral replication, showing the formation of viral peripheral replication centers in SIRV2-infected cells, where viral DNA synthesis and replication...

Self-Replication of Localized Vegetation Patches in Scarce Environments

Science.gov (United States)

Bordeu, Ignacio; Clerc, Marcel G.; Couteron, Piere; Lefever, René; Tlidi, Mustapha

2016-09-01

Desertification due to climate change and increasing drought periods is a worldwide problem for both ecology and economy. Our ability to understand how vegetation manages to survive and propagate through arid and semiarid ecosystems may be useful in the development of future strategies to prevent desertification, preserve flora—and fauna within—or even make use of scarce resources soils. In this paper, we study a robust phenomena observed in semi-arid ecosystems, by which localized vegetation patches split in a process called self-replication. Localized patches of vegetation are visible in nature at various spatial scales. Even though they have been described in literature, their growth mechanisms remain largely unexplored. Here, we develop an innovative statistical analysis based on real field observations to show that patches may exhibit deformation and splitting. This growth mechanism is opposite to the desertification since it allows to repopulate territories devoid of vegetation. We investigate these aspects by characterizing quantitatively, with a simple mathematical model, a new class of instabilities that lead to the self-replication phenomenon observed.

The Role of the Transcriptional Response to DNA Replication Stress.

Science.gov (United States)

Herlihy, Anna E; de Bruin, Robertus A M

2017-03-02

During DNA replication many factors can result in DNA replication stress. The DNA replication stress checkpoint prevents the accumulation of replication stress-induced DNA damage and the potential ensuing genome instability. A critical role for post-translational modifications, such as phosphorylation, in the replication stress checkpoint response has been well established. However, recent work has revealed an important role for transcription in the cellular response to DNA replication stress. In this review, we will provide an overview of current knowledge of the cellular response to DNA replication stress with a specific focus on the DNA replication stress checkpoint transcriptional response and its role in the prevention of replication stress-induced DNA damage.

The Role of the Transcriptional Response to DNA Replication Stress

Science.gov (United States)

Herlihy, Anna E.; de Bruin, Robertus A.M.

2017-01-01

During DNA replication many factors can result in DNA replication stress. The DNA replication stress checkpoint prevents the accumulation of replication stress-induced DNA damage and the potential ensuing genome instability. A critical role for post-translational modifications, such as phosphorylation, in the replication stress checkpoint response has been well established. However, recent work has revealed an important role for transcription in the cellular response to DNA replication stress. In this review, we will provide an overview of current knowledge of the cellular response to DNA replication stress with a specific focus on the DNA replication stress checkpoint transcriptional response and its role in the prevention of replication stress-induced DNA damage. PMID:28257104

Replication of swine and human influenza viruses in juvenile and layer turkey hens.

Science.gov (United States)

Ali, Ahmed; Yassine, Hadi; Awe, Olusegun O; Ibrahim, Mahmoud; Saif, Yehia M; Lee, Chang-Won

2013-04-12

Since the first reported isolation of swine influenza viruses (SIVs) in turkeys in the 1980s, transmission of SIVs to turkeys was frequently documented. Recently, the 2009 pandemic H1N1 virus, that was thought to be of swine origin, was detected in turkeys with a severe drop in egg production. In this study, we assessed the infectivity of different mammalian influenza viruses including swine, pandemic H1N1 and seasonal human influenza viruses in both juvenile and layer turkeys. In addition, we investigated the potential influenza virus dissemination in the semen of experimentally infected turkey toms. Results showed that all mammalian origin influenza viruses tested can infect turkeys. SIVs were detected in respiratory and digestive tracts of both juvenile and layer turkeys. Variations in replication efficiencies among SIVs were observed especially in the reproductive tract of layer turkeys. Compared to SIVs, limited replication of seasonal human H1N1 and no detectable replication of recent human-like swine H1N2, pandemic H1N1 and seasonal human H3N2 viruses was noticed. All birds seroconverted to all tested viruses regardless of their replication level. In turkey toms, we were able to detect swine H3N2 virus in semen and reproductive tract of infected toms by real-time RT-PCR although virus isolation was not successful. These data suggest that turkey hens could be affected by diverse influenza strains especially SIVs. Moreover, the differences in the replication efficiency we demonstrated among SIVs and between SIV and human influenza viruses in layer turkeys suggest a possible use of turkeys as an animal model to study host tropism and pathogenesis of influenza viruses. Our results also indicate a potential risk of venereal transmission of influenza viruses in turkeys. Copyright © 2012 Elsevier B.V. All rights reserved.

Macrophage replication screen identifies a novel Francisella hydroperoxide resistance protein involved in virulence.

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Anna C Llewellyn

Full Text Available Francisella tularensis is a gram-negative facultative intracellular pathogen and the causative agent of tularemia. Recently, genome-wide screens have identified Francisella genes required for virulence in mice. However, the mechanisms by which most of the corresponding proteins contribute to pathogenesis are still largely unknown. To further elucidate the roles of these virulence determinants in Francisella pathogenesis, we tested whether each gene was required for replication of the model pathogen F. novicida within macrophages, an important virulence trait. Fifty-three of the 224 genes tested were involved in intracellular replication, including many of those within the Francisella pathogenicity island (FPI, validating our results. Interestingly, over one third of the genes identified are annotated as hypothetical, indicating that F. novicida likely utilizes novel virulence factors for intracellular replication. To further characterize these virulence determinants, we selected two hypothetical genes to study in more detail. As predicted by our screen, deletion mutants of FTN_0096 and FTN_1133 were attenuated for replication in macrophages. The mutants displayed differing levels of attenuation in vivo, with the FTN_1133 mutant being the most attenuated. FTN_1133 has sequence similarity to the organic hydroperoxide resistance protein Ohr, an enzyme involved in the bacterial response to oxidative stress. We show that FTN_1133 is required for F. novicida resistance to, and degradation of, organic hydroperoxides as well as resistance to the action of the NADPH oxidase both in macrophages and mice. Furthermore, we demonstrate that F. holarctica LVS, a strain derived from a highly virulent human pathogenic species of Francisella, also requires this protein for organic hydroperoxide resistance as well as replication in macrophages and mice. This study expands our knowledge of Francisella's largely uncharacterized intracellular lifecycle and

Mediate evaluation of replicating a Training Program in Nonverbal Communication in Gerontology

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Teresa Cristina Gioia Schimidt

2015-04-01

Full Text Available OBJECTIVE Replicating the training program in non-verbal communication based on the theoretical framework of interpersonal communication; non-verbal coding, valuing the aging aspects in the perspective of active aging, checking its current relevance through the content assimilation index after 90 days (mediate of its application. METHOD A descriptive and exploratory field study was conducted in three hospitals under direct administration of the state of São Paulo that caters exclusively to Unified Health System (SUS patients. The training lasted 12 hours divided in three meetings, applied to 102 health professionals. RESULTS Revealed very satisfactory and satisfactory mediate content assimilation index in 82.9%. CONCLUSION The program replication proved to be relevant and updated the setting of hospital services, while remaining efficient for healthcare professionals.

DNA Copy-Number Control through Inhibition of Replication Fork Progression

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Jared T. Nordman

2014-11-01

Full Text Available Proper control of DNA replication is essential to ensure faithful transmission of genetic material and prevent chromosomal aberrations that can drive cancer progression and developmental disorders. DNA replication is regulated primarily at the level of initiation and is under strict cell-cycle regulation. Importantly, DNA replication is highly influenced by developmental cues. In Drosophila, specific regions of the genome are repressed for DNA replication during differentiation by the SNF2 domain-containing protein SUUR through an unknown mechanism. We demonstrate that SUUR is recruited to active replication forks and mediates the repression of DNA replication by directly inhibiting replication fork progression instead of functioning as a replication fork barrier. Mass spectrometry identification of SUUR-associated proteins identified the replicative helicase member CDC45 as a SUUR-associated protein, supporting a role for SUUR directly at replication forks. Our results reveal that control of eukaryotic DNA copy number can occur through the inhibition of replication fork progression.

Initiation of Replication in Escherichia coli

DEFF Research Database (Denmark)

Frimodt-Møller, Jakob

The circular chromosome of Escherichia coli is replicated by two replisomes assembled at the unique origin and moving in the opposite direction until they meet in the less well defined terminus. The key protein in initiation of replication, DnaA, facilitates the unwinding of double-stranded DNA...... to single-stranded DNA in oriC. Although DnaA is able to bind both ADP and ATP, DnaA is only active in initiation when bound to ATP. Although initiation of replication, and the regulation of this, is thoroughly investigated it is still not fully understood. The overall aim of the thesis was to investigate...... the regulation of initiation, the effect on the cell when regulation fails, and if regulation was interlinked to chromosomal organization. This thesis uncovers that there exists a subtle balance between chromosome replication and reactive oxygen species (ROS) inflicted DNA damage. Thus, failure in regulation...

Sampling hydrometeors in clouds in-situ - the replicator technique

Science.gov (United States)

Wex, Heike; Löffler, Mareike; Griesche, Hannes; Bühl, Johannes; Stratmann, Frank; Schmitt, Carl; Dirksen, Ruud; Reichardt, Jens; Wolf, Veronika; Kuhn, Thomas; Prager, Lutz; Seifert, Patric

2017-04-01

For the examination of ice crystals in clouds, concerning their number concentrations, sizes and shapes, often instruments mounted on fast flying aircraft are used. One related disadvantage is possible shattering of the ice crystals on inlets, which has been improved with the introduction of the "Korolev-tip" and by accounting for inter-arrival times (Korolev et al., 2013, 2015), but additionally, the typically fast flying aircraft allow only for a low spatial resolution. Alternative sampling methods have been introduced as e.g., a replicator by Miloshevich & Heymsfield (1997) and an in-situ imager by by Kuhn & Heymsfield (2016). They both sample ice crystals onto an advancing stripe while ascending on a balloon, conserving the ice crystals either in formvar for later off-line analysis under a microscope (Miloshevich & Heymsfield, 1997) or imaging them upon their impaction on silicone oil (Kuhn & Heymsfield, 2016), both yielding vertical profiles for different ice crystal properties. A measurement campaign was performed at the Lindenberg Meteorological Observatory of the German Meteorological Service (DWD) in Germany in October 2016, during which both types of instruments were used during balloon ascents, while ground-based Lidar and cloud-radar measurements were performed simultaneously. The two ice particle sondes were operated by people from the Lulea University of Technology and from TROPOS, where the latter one was made operational only recently. Here, we will show first results of the TROPOS replicator on ice crystals sampled during one ascent, for which the collected ice crystals were analyzed off-line using a microscope. Literature: Korolev, A., E. Emery, and K. Creelman (2013), Modification and tests of particle probe tips to mitigate effects of ice shattering, J. Atmos. Ocean. Tech., 30, 690-708, 2013. Korolev, A., and P. R. Field (2015), Assessment of the performance of the inter-arrival time algorithm to identify ice shattering artifacts in cloud

Ultrastructural Characterization of Zika Virus Replication Factories

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Mirko Cortese

2017-02-01

Full Text Available Summary: A global concern has emerged with the pandemic spread of Zika virus (ZIKV infections that can cause severe neurological symptoms in adults and newborns. ZIKV is a positive-strand RNA virus replicating in virus-induced membranous replication factories (RFs. Here we used various imaging techniques to investigate the ultrastructural details of ZIKV RFs and their relationship with host cell organelles. Analyses of human hepatic cells and neural progenitor cells infected with ZIKV revealed endoplasmic reticulum (ER membrane invaginations containing pore-like openings toward the cytosol, reminiscent to RFs in Dengue virus-infected cells. Both the MR766 African strain and the H/PF/2013 Asian strain, the latter linked to neurological diseases, induce RFs of similar architecture. Importantly, ZIKV infection causes a drastic reorganization of microtubules and intermediate filaments forming cage-like structures surrounding the viral RF. Consistently, ZIKV replication is suppressed by cytoskeleton-targeting drugs. Thus, ZIKV RFs are tightly linked to rearrangements of the host cell cytoskeleton. : Cortese et al. show that ZIKV infection in both human hepatoma and neuronal progenitor cells induces drastic structural modification of the cellular architecture. Microtubules and intermediate filaments surround the viral replication factory composed of vesicles corresponding to ER membrane invagination toward the ER lumen. Importantly, alteration of microtubule flexibility impairs ZIKV replication. Keywords: Zika virus, flavivirus, human neural progenitor cells, replication factories, replication organelles, microtubules, intermediate filaments, electron microscopy, electron tomography, live-cell imaging

Tombusviruses upregulate phospholipid biosynthesis via interaction between p33 replication protein and yeast lipid sensor proteins during virus replication in yeast

International Nuclear Information System (INIS)

Barajas, Daniel; Xu, Kai; Sharma, Monika; Wu, Cheng-Yu; Nagy, Peter D.

2014-01-01

Positive-stranded RNA viruses induce new membranous structures and promote membrane proliferation in infected cells to facilitate viral replication. In this paper, the authors show that a plant-infecting tombusvirus upregulates transcription of phospholipid biosynthesis genes, such as INO1, OPI3 and CHO1, and increases phospholipid levels in yeast model host. This is accomplished by the viral p33 replication protein, which interacts with Opi1p FFAT domain protein and Scs2p VAP protein. Opi1p and Scs2p are phospholipid sensor proteins and they repress the expression of phospholipid genes. Accordingly, deletion of OPI1 transcription repressor in yeast has a stimulatory effect on TBSV RNA accumulation and enhanced tombusvirus replicase activity in an in vitro assay. Altogether, the presented data convincingly demonstrate that de novo lipid biosynthesis is required for optimal TBSV replication. Overall, this work reveals that a (+)RNA virus reprograms the phospholipid biosynthesis pathway in a unique way to facilitate its replication in yeast cells. - Highlights: • Tombusvirus p33 replication protein interacts with FFAT-domain host protein. • Tombusvirus replication leads to upregulation of phospholipids. • Tombusvirus replication depends on de novo lipid synthesis. • Deletion of FFAT-domain host protein enhances TBSV replication. • TBSV rewires host phospholipid synthesis

A replicated field intervention study evaluating the impact of a highly adjustable chair and office ergonomics training on visual symptoms☆

Science.gov (United States)

Menéndez, Cammie Chaumont; Amick, Benjamin C.; Robertson, Michelle; Bazzani, Lianna; DeRango, Kelly; Rooney, Ted; Moore, Anne

2016-01-01

Objective Examine the effects of two office ergonomics interventions in reducing visual symptoms at a private sector worksite. Methods A quasi-experimental study design evaluated the effects of a highly adjustable chair with office ergonomics training intervention (CWT group) and the training only (TO group) compared with no intervention (CO group). Data collection occurred 2 and 1 month(s) pre-intervention and 2, 6 and 12 months post-intervention. During each data collection period, a work environment and health questionnaire (covariates) and daily health diary (outcomes) were completed. Multilevel statistical models tested hypotheses. Results Both the training only intervention (p office ergonomics training alone and coupled with a highly adjustable chair reduced visual symptoms. In replicating results from a public sector worksite at a private sector worksite the external validity of the interventions is strengthened, thus broadening its generalizability. PMID:22030069

A replicated field intervention study evaluating the impact of a highly adjustable chair and office ergonomics training on visual symptoms.

Science.gov (United States)

Menéndez, Cammie Chaumont; Amick, Benjamin C; Robertson, Michelle; Bazzani, Lianna; DeRango, Kelly; Rooney, Ted; Moore, Anne

2012-07-01

Examine the effects of two office ergonomics interventions in reducing visual symptoms at a private sector worksite. A quasi-experimental study design evaluated the effects of a highly adjustable chair with office ergonomics training intervention (CWT group) and the training only (TO group) compared with no intervention (CO group). Data collection occurred 2 and 1 month(s) pre-intervention and 2, 6 and 12 months post-intervention. During each data collection period, a work environment and health questionnaire (covariates) and daily health diary (outcomes) were completed. Multilevel statistical models tested hypotheses. Both the training only intervention (poffice ergonomics training alone and coupled with a highly adjustable chair reduced visual symptoms. In replicating results from a public sector worksite at a private sector worksite the external validity of the interventions is strengthened, thus broadening its generalizability. Published by Elsevier Ltd.

Regulation of beta cell replication

DEFF Research Database (Denmark)

Lee, Ying C; Nielsen, Jens Høiriis

2008-01-01

Beta cell mass, at any given time, is governed by cell differentiation, neogenesis, increased or decreased cell size (cell hypertrophy or atrophy), cell death (apoptosis), and beta cell proliferation. Nutrients, hormones and growth factors coupled with their signalling intermediates have been...... suggested to play a role in beta cell mass regulation. In addition, genetic mouse model studies have indicated that cyclins and cyclin-dependent kinases that determine cell cycle progression are involved in beta cell replication, and more recently, menin in association with cyclin-dependent kinase...... inhibitors has been demonstrated to be important in beta cell growth. In this review, we consider and highlight some aspects of cell cycle regulation in relation to beta cell replication. The role of cell cycle regulation in beta cell replication is mostly from studies in rodent models, but whether...

Layout of the objects of underground nuclear tests at the Balapan test field of the former Semipalatinsk test site

International Nuclear Information System (INIS)

Konovalov, V.E.; Gryaznov, O.V.

2000-01-01

Integrated research of practical and scientific interest is conducted at the Balapan test field of the Semipalatinsk test site. The lack of the reliable locations for features associated with nuclear testing causes considerable difficulties while carrying out the research. To fill this gap the authors present data available at the Institute of Geophysical Research of the National Nuclear Center of the Republic of Kazakhstan. (author)

Phosphorylation of NS5A Serine-235 is essential to hepatitis C virus RNA replication and normal replication compartment formation

Energy Technology Data Exchange (ETDEWEB)

Eyre, Nicholas S., E-mail: nicholas.eyre@adelaide.edu.au [School of Biological Sciences and Research Centre for Infectious Diseases, University of Adelaide, Adelaide (Australia); Centre for Cancer Biology, SA Pathology, Adelaide (Australia); Hampton-Smith, Rachel J.; Aloia, Amanda L. [School of Biological Sciences and Research Centre for Infectious Diseases, University of Adelaide, Adelaide (Australia); Centre for Cancer Biology, SA Pathology, Adelaide (Australia); Eddes, James S. [Adelaide Proteomics Centre, School of Biological Sciences, University of Adelaide, Adelaide (Australia); Simpson, Kaylene J. [Victorian Centre for Functional Genomics, Peter MacCallum Cancer Centre, East Melbourne (Australia); The Sir Peter MacCallum Department of Oncology, University of Melbourne, Parkville (Australia); Hoffmann, Peter [Adelaide Proteomics Centre, School of Biological Sciences, University of Adelaide, Adelaide (Australia); Institute for Photonics and Advanced Sensing (IPAS), University of Adelaide, Adelaide (Australia); Beard, Michael R. [School of Biological Sciences and Research Centre for Infectious Diseases, University of Adelaide, Adelaide (Australia); Centre for Cancer Biology, SA Pathology, Adelaide (Australia)

2016-04-15

Hepatitis C virus (HCV) NS5A protein is essential for HCV RNA replication and virus assembly. Here we report the identification of NS5A phosphorylation sites Ser-222, Ser-235 and Thr-348 during an infectious HCV replication cycle and demonstrate that Ser-235 phosphorylation is essential for HCV RNA replication. Confocal microscopy revealed that both phosphoablatant (S235A) and phosphomimetic (S235D) mutants redistribute NS5A to large juxta-nuclear foci that display altered colocalization with known replication complex components. Using electron microscopy (EM) we found that S235D alters virus-induced membrane rearrangements while EM using ‘APEX2’-tagged viruses demonstrated S235D-mediated enrichment of NS5A in irregular membranous foci. Finally, using a customized siRNA screen of candidate NS5A kinases and subsequent analysis using a phospho-specific antibody, we show that phosphatidylinositol-4 kinase III alpha (PI4KIIIα) is important for Ser-235 phosphorylation. We conclude that Ser-235 phosphorylation of NS5A is essential for HCV RNA replication and normal replication complex formation and is regulated by PI4KIIIα. - Highlights: • NS5A residues Ser-222, Ser-235 and Thr-348 are phosphorylated during HCV infection. • Phosphorylation of Ser-235 is essential to HCV RNA replication. • Mutation of Ser-235 alters replication compartment localization and morphology. • Phosphatidylinositol-4 kinase III alpha is important for Ser-235 phosphorylation.

Novel Field Test Equipment for Lithium-Ion Batteries in Hybrid Electrical Vehicle Applications

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Goran Lindbergh

2011-04-01

Full Text Available Lifetime testing of batteries for hybrid-electrical vehicles (HEV is usually performed in the lab, either at the cell, module or battery pack level. Complementary field tests of battery packs in vehicles are also often performed. There are, however, difficulties related to field testing of battery-packs. Some examples are cost issues and the complexity of continuously collecting battery performance data, such as capacity fade and impedance increase. In this paper, a novel field test equipment designed primarily for lithium-ion battery cell testing is presented. This equipment is intended to be used on conventional vehicles, not hybrid vehicles, as a cheaper and faster field testing method for batteries, compared to full scale HEV testing. The equipment emulates an HEV environment for the tested battery cell by using real time vehicle sensor information and the existing starter battery as load and source. In addition to the emulated battery cycling, periodical capacity and pulse testing capability are implemented as well. This paper begins with presenting some background information about hybrid electrical vehicles and describing the limitations with today’s HEV battery testing. Furthermore, the functionality of the test equipment is described in detail and, finally, results from verification of the equipment are presented and discussed.

The theory and method of two-well field test for in-situ leaching uranium

International Nuclear Information System (INIS)

Yao Yixuan; Huo Jiandang; Xiang Qiulin; Tang Baobin

2007-01-01

Because leaching area in field test for in-situ leaching uranium is not accounted exactly, the reliability of obtaining parameters by calculating can not be ensured, and the whole test needs a long time and great investment. In two-well field test, lixiviant is injected from one well, pregnant solution is pumped out from the other, flow rate of the production well is more than that of the injection well, and uranium is not recoveried. In the case of keeping invariable ratio of pumping capacity to injecting capacity during the testing process, leaching area is not variable, can be exactly calculated. The full field test needs six months to one year. Two-well test is a scientific, rapid, minimal spending field test method, and is widely used in Commonwealth of Independent States. (authors)

Uncoupling of Sister Replisomes during Eukaryotic DNA Replication

NARCIS (Netherlands)

Yardimci, Hasan; Loveland, Anna B.; Habuchi, Satoshi; van Oijen, Antoine M.; Walter, Johannes C.

2010-01-01

The duplication of eukaryotic genomes involves the replication of DNA from multiple origins of replication. In S phase, two sister replisomes assemble at each active origin, and they replicate DNA in opposite directions. Little is known about the functional relationship between sister replisomes.

Visualizing Single-molecule DNA Replication with Fluorescence Microscopy

NARCIS (Netherlands)

Tanner, Nathan A.; Loparo, Joseph J.; Oijen, Antoine M. van

2009-01-01

We describe a simple fluorescence microscopy-based real-time method for observing DNA replication at the single-molecule level. A circular, forked DNA template is attached to a functionalized glass coverslip and replicated extensively after introduction of replication proteins and nucleotides. The

Replication of somatic micronuclei in bovine enucleated oocytes

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Canel Natalia

2012-11-01

Full Text Available Abstract Background Microcell-mediated chromosome transfer (MMCT was developed to introduce a low number of chromosomes into a host cell. We have designed a novel technique combining part of MMCT with somatic cell nuclear transfer, which consists of injecting a somatic micronucleus into an enucleated oocyte, and inducing its cellular machinery to replicate such micronucleus. It would allow the isolation and manipulation of a single or a low number of somatic chromosomes. Methods Micronuclei from adult bovine fibroblasts were produced by incubation in 0.05 μg/ml demecolcine for 46 h followed by 2 mg/ml mitomycin for 2 h. Cells were finally treated with 10 μg/ml cytochalasin B for 1 h. In vitro matured bovine oocytes were mechanically enucleated and intracytoplasmatically injected with one somatic micronucleus, which had been previously exposed [Micronucleus- injected (+] or not [Micronucleus- injected (−] to a transgene (50 ng/μl pCX-EGFP during 5 min. Enucleated oocytes [Enucleated (+] and parthenogenetic [Parthenogenetic (+] controls were injected into the cytoplasm with less than 10 pl of PVP containing 50 ng/μl pCX-EGFP. A non-injected parthenogenetic control [Parthenogenetic (−] was also included. Two hours after injection, oocytes and reconstituted embryos were activated by incubation in 5 μM ionomycin for 4 min + 1.9 mM 6-DMAP for 3 h. Cleavage stage and egfp expression were evaluated. DNA replication was confirmed by DAPI staining. On day 2, Micronucleus- injected (−, Parthenogenetic (− and in vitro fertilized (IVF embryos were karyotyped. Differences among treatments were determined by Fisher′s exact test (p≤0.05. Results All the experimental groups underwent the first cell divisions. Interestingly, a low number of Micronucleus-injected embryos showed egfp expression. DAPI staining confirmed replication of micronuclei in most of the evaluated embryos. Karyotype analysis revealed that all Micronucleus-injected embryos had

Dynamic behavior of DNA replication domains

NARCIS (Netherlands)

Manders, E. M.; Stap, J.; Strackee, J.; van Driel, R.; Aten, J. A.

1996-01-01

Like many nuclear processes, DNA replication takes place in distinct domains that are scattered throughout the S-phase nucleus. Recently we have developed a fluorescent double-labeling procedure that allows us to visualize nascent DNA simultaneously with "newborn" DNA that had replicated earlier in

The cis-acting replication signal at the 3' end of Flock House virus RNA2 is RNA3-dependent

International Nuclear Information System (INIS)

Albarino, Cesar G.; Eckerle, Lance D.; Ball, L. Andrew

2003-01-01

The nodavirus Flock House virus has a bipartite positive-sense RNA genome consisting of RNAs 1 and 2, which encode the viral RNA-dependent RNA polymerase (RdRp) and capsid protein precursor, respectively. The RdRp catalyzes replication of both genome segments and produces from RNA1 a subgenomic RNA (RNA3) that transactivates RNA2 replication. Here, we replaced internal sequences of RNAs 1 and 2 with a common heterologous core and were thereby able to test the RNA termini for compatibility in supporting the replication of chimeric RNAs. The results showed that the 3' 50 nt of RNA2 contained an RNA3-dependent cis-acting replication signal. Since covalent RNA dimers can direct the synthesis of monomeric replication products, the RdRp can evidently respond to cis-acting replication signals located internally. Accordingly, RNA templates containing the 3' termini of both RNAs 1 and 2 in tandem generated different replication products depending on the presence or absence of RNA3

Intrapulmonary Human Cytomegalovirus Replication in Lung Transplant Recipients Is Associated With a Rise of CCL-18 and CCL-20 Chemokine Levels.

Science.gov (United States)

Weseslindtner, Lukas; Görzer, Irene; Roedl, Kevin; Küng, Erik; Jaksch, Peter; Klepetko, Walter; Puchhammer-Stöckl, Elisabeth

2017-01-01

In lung transplant recipients (LTRs), human cytomegalovirus (HCMV) DNA detection in the bronchoalveolar lavage fluid (BALF) indicates HCMV replication in the pulmonary compartment. Such local HCMV replication episodes may remain asymptomatic or may lead to symptomatic HCMV disease. Here, we investigated LTRs with intrapulmonary HCMV replication for the chemokines CCL-18 and CCL-20. In particular, we analyzed whether these chemokines rise in the allograft and/or the blood and are associated with HCMV disease. CCL-18 and CCL-20 levels were quantitated by ELISA in BALF and serum samples from 60 LTRs. During the posttransplant follow-up, these LTRs displayed HCMV DNA detection in the BALF by PCR, whereas other infectious agents were undetectable. Furthermore, we investigated samples from 10 controls who did not display any HCMV replication episode during the follow-up. HCMV replication in the allograft was associated with a significant increase of CCL-18 and CCL-20 BALF levels (P Wilcoxon signed-rank test) and a significant rise of CCL-20 (P Wilcoxon signed-rank test) but not of CCL-18 in the blood. In controls, no such chemokine increase was observed. Furthermore, CCL-18 BALF levels were significantly higher in 8 LTRs who additionally developed HCMV disease, as compared with the other 52 patients in whom HCMV replication remained asymptomatic (P test). HCMV replication in the allograft causes an intrapulmonary increase of CCL-18 and CCL-20 and a systemic rise of CCL-20 serum levels. Strong intrapulmonary CCL-18 responses are associated with symptomatic HCMV disease, proposing that CCL-18 BALF levels could serve as a marker.

Test-particle motion in Einstein's unified field theory. III. Magnetic monopoles and charged particles

International Nuclear Information System (INIS)

Johnson, C.R.

1986-01-01

In a previous paper (paper I), we developed a method for finding the exact equations of structure and motion of multipole test particles in Einstein's unified field theory: the theory of the nonsymmetric field. In that paper we also applied the method and found in Einstein's unified field theory the equations of structure and motion of neutral pole-dipole test particles possessing no electromagnetic multipole moments. In a second paper (paper II), we applied the method and found in Einstein's unified field theory the exact equations of structure and motion of charged test particles possessing no magnetic monopole moments. In the present paper (paper III), we apply the method and find in Einstein's unified field theory the exact equations of structure and motion of charged test particles possessing magnetic monopole moments. It follows from the form of these equations of structure and motion that in general in Einstein's unified field theory a test particle possessing a magnetic monopole moment in a background electromagnetic field must also possess spin

A Fuzzy Modeling Approach for Replicated Response Measures Based on Fuzzification of Replications with Descriptive Statistics and Golden Ratio

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Özlem TÜRKŞEN

2018-03-01

Full Text Available Some of the experimental designs can be composed of replicated response measures in which the replications cannot be identified exactly and may have uncertainty different than randomness. Then, the classical regression analysis may not be proper to model the designed data because of the violation of probabilistic modeling assumptions. In this case, fuzzy regression analysis can be used as a modeling tool. In this study, the replicated response values are newly formed to fuzzy numbers by using descriptive statistics of replications and golden ratio. The main aim of the study is obtaining the most suitable fuzzy model for replicated response measures through fuzzification of the replicated values by taking into account the data structure of the replications in statistical framework. Here, the response and unknown model coefficients are considered as triangular type-1 fuzzy numbers (TT1FNs whereas the inputs are crisp. Predicted fuzzy models are obtained according to the proposed fuzzification rules by using Fuzzy Least Squares (FLS approach. The performances of the predicted fuzzy models are compared by using Root Mean Squared Error (RMSE criteria. A data set from the literature, called wheel cover component data set, is used to illustrate the performance of the proposed approach and the obtained results are discussed. The calculation results show that the combined formulation of the descriptive statistics and the golden ratio is the most preferable fuzzification rule according to the well-known decision making method, called TOPSIS, for the data set.

Rapid transient production in plants by replicating and non-replicating vectors yields high quality functional anti-HIV antibody.

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Frank Sainsbury

2010-11-01

Full Text Available The capacity of plants and plant cells to produce large amounts of recombinant protein has been well established. Due to advantages in terms of speed and yield, attention has recently turned towards the use of transient expression systems, including viral vectors, to produce proteins of pharmaceutical interest in plants. However, the effects of such high level expression from viral vectors and concomitant effects on host cells may affect the quality of the recombinant product.To assess the quality of antibodies transiently expressed to high levels in plants, we have expressed and characterised the human anti-HIV monoclonal antibody, 2G12, using both replicating and non-replicating systems based on deleted versions of Cowpea mosaic virus (CPMV RNA-2. The highest yield (approximately 100 mg/kg wet weight leaf tissue of affinity purified 2G12 was obtained when the non-replicating CPMV-HT system was used and the antibody was retained in the endoplasmic reticulum (ER. Glycan analysis by mass-spectrometry showed that the glycosylation pattern was determined exclusively by whether the antibody was retained in the ER and did not depend on whether a replicating or non-replicating system was used. Characterisation of the binding and neutralisation properties of all the purified 2G12 variants from plants showed that these were generally similar to those of the Chinese hamster ovary (CHO cell-produced 2G12.Overall, the results demonstrate that replicating and non-replicating CPMV-based vectors are able to direct the production of a recombinant IgG similar in activity to the CHO-produced control. Thus, a complex recombinant protein was produced with no apparent effect on its biochemical properties using either high-level expression or viral replication. The speed with which a recombinant pharmaceutical with excellent biochemical characteristics can be produced transiently in plants makes CPMV-based expression vectors an attractive option for

Dynamics of picornavirus RNA replication within infected cells

DEFF Research Database (Denmark)

Belsham, Graham; Normann, Preben

2008-01-01

Replication of many picornaviruses is inhibited by low concentrations of guanidine. Guanidine-resistant mutants are readily isolated and the mutations map to the coding region for the 2C protein. Using in vitro replication assays it has been determined previously that guanidine blocks the initiat......Replication of many picornaviruses is inhibited by low concentrations of guanidine. Guanidine-resistant mutants are readily isolated and the mutations map to the coding region for the 2C protein. Using in vitro replication assays it has been determined previously that guanidine blocks...... the initiation of negative-strand synthesis. We have now examined the dynamics of RNA replication, measured by quantitative RT-PCR, within cells infected with either swine vesicular disease virus (an enterovirus) or foot-and-mouth disease virus as regulated by the presence or absence of guanidine. Following...... the removal of guanidine from the infected cells, RNA replication occurs after a significant lag phase. This restoration of RNA synthesis requires de novo protein synthesis. Viral RNA can be maintained for at least 72 h within cells in the absence of apparent replication but guanidine-resistant virus can...

Pyrimidine dimers block simian virus 40 replication forks

International Nuclear Information System (INIS)

Berger, C.A.; Edenberg, H.J.

1986-01-01

UV light produces lesions, predominantly pyrimidine dimers, which inhibit DNA replication in mammalian cells. The mechanism of inhibition is controversial: is synthesis of a daughter strand halted at a lesion while the replication fork moves on and reinitiates downstream, or is fork progression itself blocked for some time at the site of a lesion? We directly addressed this question by using electron microscopy to examine the distances of replication forks from the origin in unirradiated and UV-irradiated simian virus 40 chromosomes. If UV lesions block replication fork progression, the forks should be asymmetrically located in a large fraction of the irradiated molecules; if replication forks move rapidly past lesions, the forks should be symmetrically located. A large fraction of the simian virus 40 replication forks in irradiated molecules were asymmetrically located, demonstrating that UV lesions present at the frequency of pyrimidine dimers block replication forks. As a mechanism for this fork blockage, we propose that polymerization of the leading strand makes a significant contribution to the energetics of fork movement, so any lesion in the template for the leading strand which blocks polymerization should also block fork movement

Autonomous model protocell division driven by molecular replication.

Science.gov (United States)

Taylor, J W; Eghtesadi, S A; Points, L J; Liu, T; Cronin, L

2017-08-10

The coupling of compartmentalisation with molecular replication is thought to be crucial for the emergence of the first evolvable chemical systems. Minimal artificial replicators have been designed based on molecular recognition, inspired by the template copying of DNA, but none yet have been coupled to compartmentalisation. Here, we present an oil-in-water droplet system comprising an amphiphilic imine dissolved in chloroform that catalyses its own formation by bringing together a hydrophilic and a hydrophobic precursor, which leads to repeated droplet division. We demonstrate that the presence of the amphiphilic replicator, by lowering the interfacial tension between droplets of the reaction mixture and the aqueous phase, causes them to divide. Periodic sampling by a droplet-robot demonstrates that the extent of fission is increased as the reaction progresses, producing more compartments with increased self-replication. This bridges a divide, showing how replication at the molecular level can be used to drive macroscale droplet fission.Coupling compartmentalisation and molecular replication is essential for the development of evolving chemical systems. Here the authors show an oil-in-water droplet containing a self-replicating amphiphilic imine that can undergo repeated droplet division.

Initiation preference at a yeast origin of replication.

Science.gov (United States)

Brewer, B J; Fangman, W L

1994-04-12

Replication origins in the yeast Saccharomyces cerevisiae are identified as autonomous replication sequence (ARS) elements. To examine the effect of origin density on replication initiation, we have analyzed the replication of a plasmid that contains two copies of the same origin, ARS1. The activation of origins and the direction that replication forks move through flanking sequences can be physically determined by analyzing replication intermediates on two-dimensional agarose gels. We find that only one of the two identical ARSs on the plasmid initiates replication on any given plasmid molecule; that is, this close spacing of ARSs results in an apparent interference between the potential origins. Moreover, in the particular plasmid that we constructed, one of the two identical copies of ARS1 is used four times more frequently than the other one. These results show that the plasmid context is critical for determining the preferred origin. This origin preference is also exhibited when the tandem copies of ARS1 are introduced into a yeast chromosome. The sequences responsible for establishing the origin preference have been identified by deletion analysis and are found to reside in a portion of the yeast URA3 gene.

Gene organization inside replication domains in mammalian genomes

Science.gov (United States)

Zaghloul, Lamia; Baker, Antoine; Audit, Benjamin; Arneodo, Alain

2012-11-01

We investigate the large-scale organization of human genes with respect to "master" replication origins that were previously identified as bordering nucleotide compositional skew domains. We separate genes in two categories depending on their CpG enrichment at the promoter which can be considered as a marker of germline DNA methylation. Using expression data in mouse, we confirm that CpG-rich genes are highly expressed in germline whereas CpG-poor genes are in a silent state. We further show that, whether tissue-specific or broadly expressed (housekeeping genes), the CpG-rich genes are over-represented close to the replication skew domain borders suggesting some coordination of replication and transcription. We also reveal that the transcription of the longest CpG-rich genes is co-oriented with replication fork progression so that the promoter of these transcriptionally active genes be located into the accessible open chromatin environment surrounding the master replication origins that border the replication skew domains. The observation of a similar gene organization in the mouse genome confirms the interplay of replication, transcription and chromatin structure as the cornerstone of mammalian genome architecture.

Mapping replication origins in yeast chromosomes.

Science.gov (United States)

Brewer, B J; Fangman, W L

1991-07-01

The replicon hypothesis, first proposed in 1963 by Jacob and Brenner, states that DNA replication is controlled at sites called origins. Replication origins have been well studied in prokaryotes. However, the study of eukaryotic chromosomal origins has lagged behind, because until recently there has been no method for reliably determining the identity and location of origins from eukaryotic chromosomes. Here, we review a technique we developed with the yeast Saccharomyces cerevisiae that allows both the mapping of replication origins and an assessment of their activity. Two-dimensional agarose gel electrophoresis and Southern hybridization with total genomic DNA are used to determine whether a particular restriction fragment acquires the branched structure diagnostic of replication initiation. The technique has been used to localize origins in yeast chromosomes and assess their initiation efficiency. In some cases, origin activation is dependent upon the surrounding context. The technique is also being applied to a variety of eukaryotic organisms.

Assembly of Slx4 signaling complexes behind DNA replication forks.

Science.gov (United States)

Balint, Attila; Kim, TaeHyung; Gallo, David; Cussiol, Jose Renato; Bastos de Oliveira, Francisco M; Yimit, Askar; Ou, Jiongwen; Nakato, Ryuichiro; Gurevich, Alexey; Shirahige, Katsuhiko; Smolka, Marcus B; Zhang, Zhaolei; Brown, Grant W

2015-08-13

Obstructions to replication fork progression, referred to collectively as DNA replication stress, challenge genome stability. In Saccharomyces cerevisiae, cells lacking RTT107 or SLX4 show genome instability and sensitivity to DNA replication stress and are defective in the completion of DNA replication during recovery from replication stress. We demonstrate that Slx4 is recruited to chromatin behind stressed replication forks, in a region that is spatially distinct from that occupied by the replication machinery. Slx4 complex formation is nucleated by Mec1 phosphorylation of histone H2A, which is recognized by the constitutive Slx4 binding partner Rtt107. Slx4 is essential for recruiting the Mec1 activator Dpb11 behind stressed replication forks, and Slx4 complexes are important for full activity of Mec1. We propose that Slx4 complexes promote robust checkpoint signaling by Mec1 by stably recruiting Dpb11 within a discrete domain behind the replication fork, during DNA replication stress. © 2015 The Authors.

Checkpoint responses to replication stalling: inducing tolerance and preventing mutagenesis

Energy Technology Data Exchange (ETDEWEB)

Kai, Mihoko; Wang, Teresa S.-F

2003-11-27

Replication mutants often exhibit a mutator phenotype characterized by point mutations, single base frameshifts, and the deletion or duplication of sequences flanked by homologous repeats. Mutation in genes encoding checkpoint proteins can significantly affect the mutator phenotype. Here, we use fission yeast (Schizosaccharomyces pombe) as a model system to discuss the checkpoint responses to replication perturbations induced by replication mutants. Checkpoint activation induced by a DNA polymerase mutant, aside from delay of mitotic entry, up-regulates the translesion polymerase DinB (Pol{kappa}). Checkpoint Rad9-Rad1-Hus1 (9-1-1) complex, which is loaded onto chromatin by the Rad17-Rfc2-5 checkpoint complex in response to replication perturbation, recruits DinB onto chromatin to generate the point mutations and single nucleotide frameshifts in the replication mutator. This chain of events reveals a novel checkpoint-induced tolerance mechanism that allows cells to cope with replication perturbation, presumably to make possible restarting stalled replication forks. Fission yeast Cds1 kinase plays an essential role in maintaining DNA replication fork stability in the face of DNA damage and replication fork stalling. Cds1 kinase is known to regulate three proteins that are implicated in maintaining replication fork stability: Mus81-Eme1, a hetero-dimeric structure-specific endonuclease complex; Rqh1, a RecQ-family helicase involved in suppressing inappropriate recombination during replication; and Rad60, a protein required for recombinational repair during replication. These Cds1-regulated proteins are thought to cooperatively prevent mutagenesis and maintain replication fork stability in cells under replication stress. These checkpoint-regulated processes allow cells to survive replication perturbation by preventing stalled replication forks from degenerating into deleterious DNA structures resulting in genomic instability and cancer development.

Checkpoint responses to replication stalling: inducing tolerance and preventing mutagenesis

International Nuclear Information System (INIS)

Kai, Mihoko; Wang, Teresa S.-F.

2003-01-01

Replication mutants often exhibit a mutator phenotype characterized by point mutations, single base frameshifts, and the deletion or duplication of sequences flanked by homologous repeats. Mutation in genes encoding checkpoint proteins can significantly affect the mutator phenotype. Here, we use fission yeast (Schizosaccharomyces pombe) as a model system to discuss the checkpoint responses to replication perturbations induced by replication mutants. Checkpoint activation induced by a DNA polymerase mutant, aside from delay of mitotic entry, up-regulates the translesion polymerase DinB (Polκ). Checkpoint Rad9-Rad1-Hus1 (9-1-1) complex, which is loaded onto chromatin by the Rad17-Rfc2-5 checkpoint complex in response to replication perturbation, recruits DinB onto chromatin to generate the point mutations and single nucleotide frameshifts in the replication mutator. This chain of events reveals a novel checkpoint-induced tolerance mechanism that allows cells to cope with replication perturbation, presumably to make possible restarting stalled replication forks. Fission yeast Cds1 kinase plays an essential role in maintaining DNA replication fork stability in the face of DNA damage and replication fork stalling. Cds1 kinase is known to regulate three proteins that are implicated in maintaining replication fork stability: Mus81-Eme1, a hetero-dimeric structure-specific endonuclease complex; Rqh1, a RecQ-family helicase involved in suppressing inappropriate recombination during replication; and Rad60, a protein required for recombinational repair during replication. These Cds1-regulated proteins are thought to cooperatively prevent mutagenesis and maintain replication fork stability in cells under replication stress. These checkpoint-regulated processes allow cells to survive replication perturbation by preventing stalled replication forks from degenerating into deleterious DNA structures resulting in genomic instability and cancer development

Analytical strategies for discovery and replication of genetic effects in pharmacogenomic studies

Directory of Open Access Journals (Sweden)

Kohler JR

2014-08-01

Full Text Available Jared R Kohler, Tobias Guennel, Scott L MarshallBioStat Solutions, Inc., Frederick, MD, USAAbstract: In the past decade, the pharmaceutical industry and biomedical research sector have devoted considerable resources to pharmacogenomics (PGx with the hope that understanding genetic variation in patients would deliver on the promise of personalized medicine. With the advent of new technologies and the improved collection of DNA samples, the roadblock to advancements in PGx discovery is no longer the lack of high-density genetic information captured on patient populations, but rather the development, adaptation, and tailoring of analytical strategies to effectively harness this wealth of information. The current analytical paradigm in PGx considers the single-nucleotide polymorphism (SNP as the genomic feature of interest and performs single SNP association tests to discover PGx effects – ie, genetic effects impacting drug response. While it can be straightforward to process single SNP results and to consider how this information may be extended for use in downstream patient stratification, the rate of replication for single SNP associations has been low and the desired success of producing clinically and commercially viable biomarkers has not been realized. This may be due to the fact that single SNP association testing is suboptimal given the complexities of PGx discovery in the clinical trial setting, including: 1 relatively small sample sizes; 2 diverse clinical cohorts within and across trials due to genetic ancestry (potentially impacting the ability to replicate findings; and 3 the potential polygenic nature of a drug response. Subsequently, a shift in the current paradigm is proposed: to consider the gene as the genomic feature of interest in PGx discovery. The proof-of-concept study presented in this manuscript demonstrates that genomic region-based association testing has the potential to improve the power of detecting single SNP or

Suppression of Zika Virus Infection and Replication in Endothelial Cells and Astrocytes by PKA Inhibitor PKI 14-22.

Science.gov (United States)

Cheng, Fan; Ramos da Silva, Suzane; Huang, I-Chueh; Jung, Jae U; Gao, Shou-Jiang

2018-02-15

The recent outbreak of Zika virus (ZIKV), a reemerging flavivirus, and its associated neurological disorders, such as Guillain-Barré (GB) syndrome and microcephaly, have generated an urgent need to develop effective ZIKV vaccines and therapeutic agents. Here, we used human endothelial cells and astrocytes, both of which represent key cell types for ZIKV infection, to identify potential inhibitors of ZIKV replication. Because several pathways, including the AMP-activated protein kinase (AMPK), protein kinase A (PKA), and mitogen-activated protein kinase (MAPK) signaling pathways, have been reported to play important roles in flavivirus replication, we tested inhibitors and agonists of these pathways for their effects on ZIKV replication. We identified the PKA inhibitor PKI 14-22 (PKI) to be a potent inhibitor of ZIKV replication. PKI effectively suppressed the replication of ZIKV from both the African and Asian/American lineages with a high efficiency and minimal cytotoxicity. While ZIKV infection does not induce PKA activation, endogenous PKA activity is essential for supporting ZIKV replication. Interestingly, in addition to PKA, PKI also inhibited another unknown target(s) to block ZIKV replication. PKI inhibited ZIKV replication at the postentry stage by preferentially affecting negative-sense RNA synthesis as well as viral protein translation. Together, these results have identified a potential inhibitor of ZIKV replication which could be further explored for future therapeutic application. IMPORTANCE There is an urgent need to develop effective vaccines and therapeutic agents against Zika virus (ZIKV) infection, a reemerging flavivirus associated with neurological disorders, including Guillain-Barré (GB) syndrome and microcephaly. By screening for inhibitors of several cellular pathways, we have identified the PKA inhibitor PKI 14-22 (PKI) to be a potent inhibitor of ZIKV replication. We show that PKI effectively suppresses the replication of all ZIKV

Hyperthermia stimulates HIV-1 replication.

Directory of Open Access Journals (Sweden)

Ferdinand Roesch

Full Text Available HIV-infected individuals may experience fever episodes. Fever is an elevation of the body temperature accompanied by inflammation. It is usually beneficial for the host through enhancement of immunological defenses. In cultures, transient non-physiological heat shock (42-45°C and Heat Shock Proteins (HSPs modulate HIV-1 replication, through poorly defined mechanisms. The effect of physiological hyperthermia (38-40°C on HIV-1 infection has not been extensively investigated. Here, we show that culturing primary CD4+ T lymphocytes and cell lines at a fever-like temperature (39.5°C increased the efficiency of HIV-1 replication by 2 to 7 fold. Hyperthermia did not facilitate viral entry nor reverse transcription, but increased Tat transactivation of the LTR viral promoter. Hyperthermia also boosted HIV-1 reactivation in a model of latently-infected cells. By imaging HIV-1 transcription, we further show that Hsp90 co-localized with actively transcribing provirus, and this phenomenon was enhanced at 39.5°C. The Hsp90 inhibitor 17-AAG abrogated the increase of HIV-1 replication in hyperthermic cells. Altogether, our results indicate that fever may directly stimulate HIV-1 replication, in a process involving Hsp90 and facilitation of Tat-mediated LTR activity.

Replication and Robustness in Developmental Research

Science.gov (United States)

Duncan, Greg J.; Engel, Mimi; Claessens, Amy; Dowsett, Chantelle J.

2014-01-01

Replications and robustness checks are key elements of the scientific method and a staple in many disciplines. However, leading journals in developmental psychology rarely include explicit replications of prior research conducted by different investigators, and few require authors to establish in their articles or online appendices that their key…

Seed vigour tests for predicting field emergence of maize under severe conditions

OpenAIRE

García de Yzaguirre, Álvaro; Lasa Dolhagaray, José Manuel

1989-01-01

[EN] With 40 to 50 different seed vigour tests available, appropiate procedures for choosing the best single test or combination the best predictors of seedling emergence of maize (Zea Mays L.) under severe conditions. Thirteen vigour tests and various field emergence trials were performed on six inbred lines and two commercial hybrids. The best single predictors of field emergence were identified by calculating simple correlation coefficients. The calculation of the geometric mean of the res...

Biomarkers of replicative senescence revisited

DEFF Research Database (Denmark)

Nehlin, Jan

2016-01-01

Biomarkers of replicative senescence can be defined as those ultrastructural and physiological variations as well as molecules whose changes in expression, activity or function correlate with aging, as a result of the gradual exhaustion of replicative potential and a state of permanent cell cycle...... arrest. The biomarkers that characterize the path to an irreversible state of cell cycle arrest due to proliferative exhaustion may also be shared by other forms of senescence-inducing mechanisms. Validation of senescence markers is crucial in circumstances where quiescence or temporary growth arrest may...... be triggered or is thought to be induced. Pre-senescence biomarkers are also important to consider as their presence indicate that induction of aging processes is taking place. The bona fide pathway leading to replicative senescence that has been extensively characterized is a consequence of gradual reduction...

3X-100 blade field test.

Energy Technology Data Exchange (ETDEWEB)

Zayas, Jose R.; Johnson, Wesley D.

2008-03-01

In support of a Work-For-Other (WFO) agreement between the Wind Energy Technology Department at Sandia National Laboratories and 3TEX, one of the three Micon 65/13M wind turbines at the USDA Agriculture Research Service (ARS) center in Bushland, Texas, has been used to test a set of 9 meter wind turbine blades, manufactured by TPI composites using the 3TEX carbon material for the spar cap. Data collected from the test has been analyzed to evaluate both the aerodynamic performance and the structural response from the blades. The blades aerodynamic and structural performance, the meteorological inflow and the wind turbine structural response has been monitored with an array of 57 instruments: 15 to characterize the blades, 13 to characterize inflow, and 15 to characterize the time-varying state of the turbine. For the test, data was sampled at a rate of 40 Hz using the ATLAS II (Accurate GPS Time-Linked Data Acquisition System) data acquisition system. The system features a time-synchronized continuous data stream and telemetered data from the turbine rotor. This paper documents the instruments and infrastructure that have been developed to monitor these blades, turbines and inflow, as well as both modeling and field testing results.

Enzymes involved in organellar DNA replication in photosynthetic eukaryotes.

Science.gov (United States)

Moriyama, Takashi; Sato, Naoki

2014-01-01

Plastids and mitochondria possess their own genomes. Although the replication mechanisms of these organellar genomes remain unclear in photosynthetic eukaryotes, several organelle-localized enzymes related to genome replication, including DNA polymerase, DNA primase, DNA helicase, DNA topoisomerase, single-stranded DNA maintenance protein, DNA ligase, primer removal enzyme, and several DNA recombination-related enzymes, have been identified. In the reference Eudicot plant Arabidopsis thaliana, the replication-related enzymes of plastids and mitochondria are similar because many of them are dual targeted to both organelles, whereas in the red alga Cyanidioschyzon merolae, plastids and mitochondria contain different replication machinery components. The enzymes involved in organellar genome replication in green plants and red algae were derived from different origins, including proteobacterial, cyanobacterial, and eukaryotic lineages. In the present review, we summarize the available data for enzymes related to organellar genome replication in green plants and red algae. In addition, based on the type and distribution of replication enzymes in photosynthetic eukaryotes, we discuss the transitional history of replication enzymes in the organelles of plants.

Mutational analysis of the hypervariable region of hepatitis e virus reveals its involvement in the efficiency of viral RNA replication.

Science.gov (United States)

Pudupakam, R S; Kenney, Scott P; Córdoba, Laura; Huang, Yao-Wei; Dryman, Barbara A; Leroith, Tanya; Pierson, F William; Meng, Xiang-Jin

2011-10-01

The RNA genome of the hepatitis E virus (HEV) contains a hypervariable region (HVR) in ORF1 that tolerates small deletions with respect to infectivity. To further investigate the role of the HVR in HEV replication, we constructed a panel of mutants with overlapping deletions in the N-terminal, central, and C-terminal regions of the HVR by using a genotype 1 human HEV luciferase replicon and analyzed the effects of deletions on viral RNA replication in Huh7 cells. We found that the replication levels of the HVR deletion mutants were markedly reduced in Huh7 cells, suggesting a role of the HVR in viral replication efficiency. To further verify the results, we constructed HVR deletion mutants by using a genetically divergent, nonmammalian avian HEV, and similar effects on viral replication efficiency were observed when the avian HEV mutants were tested in LMH cells. Furthermore, the impact of complete HVR deletion on virus infectivity was tested in chickens, using an avian HEV mutant with a complete HVR deletion. Although the deletion mutant was still replication competent in LMH cells, the complete HVR deletion resulted in a loss of avian HEV infectivity in chickens. Since the HVR exhibits extensive variations in sequence and length among different HEV genotypes, we further examined the interchangeability of HVRs and demonstrated that HVR sequences are functionally exchangeable between HEV genotypes with regard to viral replication and infectivity in vitro, although genotype-specific HVR differences in replication efficiency were observed. The results showed that although the HVR tolerates small deletions with regard to infectivity, it may interact with viral and host factors to modulate the efficiency of HEV replication.

A New Facility for Testing Superconducting Solenoid Magnets with Large Fringe Fields at Fermilab

Energy Technology Data Exchange (ETDEWEB)

Orris, D. [Fermilab; Carcagno, R. [Fermilab; Nogiec, J. [Fermilab; Rabehl, R. [Fermilab; Sylvester, C. [Fermilab; Tartaglia, M. [Fermilab

2013-09-01

Testing superconducting solenoid with no iron flux return can be problematic for a magnet test facility due to the large magnetic fringe fields generated. These large external fields can interfere with the operation of equipment while precautions must be taken for personnel supporting the test. The magnetic forces between the solenoid under test and the external infrastructure must also be taken under consideration. A new test facility has been designed and built at Fermilab specifically for testing superconducting magnets with large external fringe fields. This paper discusses the test stand design, capabilities, and details of the instrumentation and controls with data from the first solenoid tested in this facility: the Muon Ionization Cooling Experiment (MICE) coupling coil.

The virtual fields method applied to spalling tests on concrete

Directory of Open Access Journals (Sweden)

Forquin P.

2012-08-01

Full Text Available For one decade spalling techniques based on the use of a metallic Hopkinson bar put in contact with a concrete sample have been widely employed to characterize the dynamic tensile strength of concrete at strain-rates ranging from a few tens to two hundreds of s−1. However, the processing method mainly based on the use of the velocity profile measured on the rear free surface of the sample (Novikov formula remains quite basic and an identification of the whole softening behaviour of the concrete is out of reach. In the present paper a new processing method is proposed based on the use of the Virtual Fields Method (VFM. First, a digital high speed camera is used to record the pictures of a grid glued on the specimen. Next, full-field measurements are used to obtain the axial displacement field at the surface of the specimen. Finally, a specific virtual field has been defined in the VFM equation to use the acceleration map as an alternative ‘load cell’. This method applied to three spalling tests allowed to identify Young’s modulus during the test. It was shown that this modulus is constant during the initial compressive part of the test and decreases in the tensile part when micro-damage exists. It was also shown that in such a simple inertial test, it was possible to reconstruct average axial stress profiles using only the acceleration data. Then, it was possible to construct local stress-strain curves and derive a tensile strength value.

Viral hijacking of a replicative helicase loader and its implications for helicase loading control and phage replication

Energy Technology Data Exchange (ETDEWEB)

Hood, Iris V.; Berger, James M.

2016-05-31

Replisome assembly requires the loading of replicative hexameric helicases onto origins by AAA+ ATPases. How loader activity is appropriately controlled remains unclear. Here, we use structural and biochemical analyses to establish how an antimicrobial phage protein interferes with the function of theStaphylococcus aureusreplicative helicase loader, DnaI. The viral protein binds to the loader’s AAA+ ATPase domain, allowing binding of the host replicative helicase but impeding loader self-assembly and ATPase activity. Close inspection of the complex highlights an unexpected locus for the binding of an interdomain linker element in DnaI/DnaC-family proteins. We find that the inhibitor protein is genetically coupled to a phage-encoded homolog of the bacterial helicase loader, which we show binds to the host helicase but not to the inhibitor itself. These findings establish a new approach by which viruses can hijack host replication processes and explain how loader activity is internally regulated to prevent aberrant auto-association.

Diagonalization of replicated transfer matrices for disordered Ising spin systems

International Nuclear Information System (INIS)

Nikoletopoulos, T; Coolen, A C C

2004-01-01

We present an alternative procedure for solving the eigenvalue problem of replicated transfer matrices describing disordered spin systems with (random) 1D nearest neighbour bonds and/or random fields, possibly in combination with (random) long range bonds. Our method is based on transforming the original eigenvalue problem for a 2 n x 2 n matrix (where n → 0) into an eigenvalue problem for integral operators. We first develop our formalism for the Ising chain with random bonds and fields, where we recover known results. We then apply our methods to models of spins which interact simultaneously via a one-dimensional ring and via more complex long-range connectivity structures, e.g., (1 + ∞)-dimensional neural networks and 'small-world' magnets. Numerical simulations confirm our predictions satisfactorily

The transcription elongation factor Bur1-Bur2 interacts with replication protein A and maintains genome stability during replication stress

DEFF Research Database (Denmark)

Clausing, Emanuel; Mayer, Andreas; Chanarat, Sittinan

2010-01-01

Multiple DNA-associated processes such as DNA repair, replication, and recombination are crucial for the maintenance of genome integrity. Here, we show a novel interaction between the transcription elongation factor Bur1-Bur2 and replication protein A (RPA), the eukaryotic single-stranded DNA......-binding protein with functions in DNA repair, recombination, and replication. Bur1 interacted via its C-terminal domain with RPA, and bur1-¿C mutants showed a deregulated DNA damage response accompanied by increased sensitivity to DNA damage and replication stress as well as increased levels of persisting Rad52...... foci. Interestingly, the DNA damage sensitivity of an rfa1 mutant was suppressed by bur1 mutation, further underscoring a functional link between these two protein complexes. The transcription elongation factor Bur1-Bur2 interacts with RPA and maintains genome integrity during DNA replication stress....

Field Test Kit for Gun Residue Detection; TOPICAL

International Nuclear Information System (INIS)

WALKER, PAMELA K.; RODACY, PHILIP J.

2002-01-01

One of the major needs of the law enforcement field is a product that quickly, accurately, and inexpensively identifies whether a person has recently fired a gun--even if the suspect has attempted to wash the traces of gunpowder off. The Field Test Kit for Gunshot Residue Identification based on Sandia National Laboratories technology works with a wide variety of handguns and other weaponry using gunpowder. There are several organic chemicals in small arms propellants such as nitrocellulose, nitroglycerine, dinitrotoluene, and nitrites left behind after the firing of a gun that result from the incomplete combustion of the gunpowder. Sandia has developed a colorimetric shooter identification kit for in situ detection of gunshot residue (GSR) from a suspect. The test kit is the first of its kind and is small, inexpensive, and easily transported by individual law enforcement personnel requiring minimal training for effective use. It will provide immediate information identifying gunshot residue

Modeling DNA Replication.

Science.gov (United States)

Bennett, Joan

1998-01-01

Recommends the use of a model of DNA made out of Velcro to help students visualize the steps of DNA replication. Includes a materials list, construction directions, and details of the demonstration using the model parts. (DDR)

Distinct functions of human RecQ helicases during DNA replication.

Science.gov (United States)

Urban, Vaclav; Dobrovolna, Jana; Janscak, Pavel

2017-06-01

DNA replication is the most vulnerable process of DNA metabolism in proliferating cells and therefore it is tightly controlled and coordinated with processes that maintain genomic stability. Human RecQ helicases are among the most important factors involved in the maintenance of replication fork integrity, especially under conditions of replication stress. RecQ helicases promote recovery of replication forks being stalled due to different replication roadblocks of either exogenous or endogenous source. They prevent generation of aberrant replication fork structures and replication fork collapse, and are involved in proper checkpoint signaling. The essential role of human RecQ helicases in the genome maintenance during DNA replication is underlined by association of defects in their function with cancer predisposition. Copyright © 2016 Elsevier B.V. All rights reserved.

Evolution of complexity in RNA-like replicator systems

Directory of Open Access Journals (Sweden)

Hogeweg Paulien

2008-03-01

Full Text Available Abstract Background The evolution of complexity is among the most important questions in biology. The evolution of complexity is often observed as the increase of genetic information or that of the organizational complexity of a system. It is well recognized that the formation of biological organization – be it of molecules or ecosystems – is ultimately instructed by the genetic information, whereas it is also true that the genetic information is functional only in the context of the organization. Therefore, to obtain a more complete picture of the evolution of complexity, we must study the evolution of both information and organization. Results Here we investigate the evolution of complexity in a simulated RNA-like replicator system. The simplicity of the system allows us to explicitly model the genotype-phenotype-interaction mapping of individual replicators, whereby we avoid preconceiving the functionality of genotypes (information or the ecological organization of replicators in the model. In particular, the model assumes that interactions among replicators – to replicate or to be replicated – depend on their secondary structures and base-pair matching. The results showed that a population of replicators, originally consisting of one genotype, evolves to form a complex ecosystem of up to four species. During this diversification, the species evolve through acquiring unique genotypes with distinct ecological functionality. The analysis of this diversification reveals that parasitic replicators, which have been thought to destabilize the replicator's diversity, actually promote the evolution of diversity through generating a novel "niche" for catalytic replicators. This also makes the current replicator system extremely stable upon the evolution of parasites. The results also show that the stability of the system crucially depends on the spatial pattern formation of replicators. Finally, the evolutionary dynamics is shown to

Molecular Mechanisms of DNA Replication Checkpoint Activation

Directory of Open Access Journals (Sweden)

Bénédicte Recolin

2014-03-01

Full Text Available The major challenge of the cell cycle is to deliver an intact, and fully duplicated, genetic material to the daughter cells. To this end, progression of DNA synthesis is monitored by a feedback mechanism known as replication checkpoint that is untimely linked to DNA replication. This signaling pathway ensures coordination of DNA synthesis with cell cycle progression. Failure to activate this checkpoint in response to perturbation of DNA synthesis (replication stress results in forced cell division leading to chromosome fragmentation, aneuploidy, and genomic instability. In this review, we will describe current knowledge of the molecular determinants of the DNA replication checkpoint in eukaryotic cells and discuss a model of activation of this signaling pathway crucial for maintenance of genomic stability.

A new MCM modification cycle regulates DNA replication initiation.

Science.gov (United States)

Wei, Lei; Zhao, Xiaolan

2016-03-01

The MCM DNA helicase is a central regulatory target during genome replication. MCM is kept inactive during G1, and it initiates replication after being activated in S phase. During this transition, the only known chemical change to MCM is the gain of multisite phosphorylation that promotes cofactor recruitment. Because replication initiation is intimately linked to multiple biological cues, additional changes to MCM can provide further regulatory points. Here, we describe a yeast MCM SUMOylation cycle that regulates replication. MCM subunits undergo SUMOylation upon loading at origins in G1 before MCM phosphorylation. MCM SUMOylation levels then decline as MCM phosphorylation levels rise, thus suggesting an inhibitory role of MCM SUMOylation during replication. Indeed, increasing MCM SUMOylation impairs replication initiation, partly through promoting the recruitment of a phosphatase that decreases MCM phosphorylation and activation. We propose that MCM SUMOylation counterbalances kinase-based regulation, thus ensuring accurate control of replication initiation.

A CI-Independent Form of Replicative Inhibition: Turn Off of Early Replication of Bacteriophage Lambda

Science.gov (United States)

Hayes, Sidney; Horbay, Monique A.; Hayes, Connie

2012-01-01

Several earlier studies have described an unusual exclusion phenotype exhibited by cells with plasmids carrying a portion of the replication region of phage lambda. Cells exhibiting this inhibition phenotype (IP) prevent the plating of homo-immune and hybrid hetero-immune lambdoid phages. We have attempted to define aspects of IP, and show that it is directed to repλ phages. IP was observed in cells with plasmids containing a λ DNA fragment including oop, encoding a short OOP micro RNA, and part of the lambda origin of replication, oriλ, defined by iteron sequences ITN1-4 and an adjacent high AT-rich sequence. Transcription of the intact oop sequence from its promoter, pO is required for IP, as are iterons ITN3–4, but not the high AT-rich portion of oriλ. The results suggest that IP silencing is directed to theta mode replication initiation from an infecting repλ genome, or an induced repλ prophage. Phage mutations suppressing IP, i.e., Sip, map within, or adjacent to cro or in O, or both. Our results for plasmid based IP suggest the hypothesis that there is a natural mechanism for silencing early theta-mode replication initiation, i.e. the buildup of λ genomes with oop + oriλ+ sequence. PMID:22590552

Field Test of Driven Pile Group under Lateral Loading

Science.gov (United States)

Gorska, Karolina; Rybak, Jaroslaw; Wyjadlowski, Marek

2017-12-01

All the geotechnical works need to be tested because the diversity of soil parameters is much higher than in other fields of construction. Horizontal load tests are necessary to determine the lateral capacity of driven piles subject to lateral load. Various load tests were carried out altogether on the test field in Kutno (Poland). While selecting the piles for load tests, different load combinations were taken into account. The piles with diverse length were chosen, on the basis of the previous tests of their length and integrity. The subsoil around the piles consisted of mineral soils: clays and medium compacted sands with the density index ID>0.50. The pile heads were free. The points of support of the “base” to which the dial gauges (displacement sensors) were fastened were located at the distance of 0.7 m from the side surface of the pile loaded laterally. In order to assure the independence of measurement, additional control (verifying) geodetic survey of the displacement of the piles subject to the load tests was carried out (by means of the alignment method). The trial load was imposed in stages by means of a hydraulic jack. The oil pressure in the actuator was corrected by means of a manual pump in order to ensure the constant value of the load in the on-going process of the displacement of the pile under test. On the basis of the obtained results it is possible to verify the numerical simulations of the behaviour of piles loaded by a lateral force.

Replication of cultured lung epithelial cells

International Nuclear Information System (INIS)

Guzowski, D.; Bienkowski, R.

1986-01-01

The authors have investigated the conditions necessary to support replication of lung type 2 epithelial cells in culture. Cells were isolated from mature fetal rabbit lungs (29d gestation) and cultured on feeder layers of mitotically inactivated 3T3 fibroblasts. The epithelial nature of the cells was demonstrated by indirect immunofluorescent staining for keratin and by polyacid dichrome stain. Ultrastructural examination during the first week showed that the cells contained myofilaments, microvilli and lamellar bodies (markers for type 2 cells). The following changes were observed after the first week: increase in cell size; loss of lamellar bodies and appearance of multivesicular bodies; increase in rough endoplasmic reticulum and golgi; increase in tonafilaments and well-defined junctions. General cell morphology was good for up to 10 wk. Cells cultured on plastic surface degenerated after 1 wk. Cell replication was assayed by autoradiography of cultures exposed to ( 3 H)-thymidine and by direct cell counts. The cells did not replicate during the first week; however, between 2-10 wk the cells incorporated the label and went through approximately 6 population doublings. They have demonstrated that lung alveolar epithelial cells can replicate in culture if they are maintained on an appropriate substrate. The coincidence of ability to replicate and loss of markers for differentiation may reflect the dichotomy between growth and differentiation commonly observed in developing systems

In Situ Field Testing of Processes

International Nuclear Information System (INIS)

Wang, J.

2001-01-01

The purpose of this Analysis/Model Report (AMR) is to update and document the data and subsequent analyses from ambient field-testing activities performed in underground drifts of the Yucca Mountain Site Characterization Project (YMP). This revision updates data and analyses presented in the initial issue of this AMR. This AMR was developed in accordance with the ''Technical Work Plan for Unsaturated Zone (UZ) Flow and Transport Process Model Report'' and ''Technical Work Plan for UZ Flow, Transport, and Coupled Processes Process Model Report. These activities were performed to investigate in situ flow and transport processes. The evaluations provide the necessary framework to: (1) refine and confirm the conceptual model of matrix and fracture processes in the unsaturated zone (UZ) and (2) analyze the impact of excavation (including use of construction water and effect of ventilation) on the UZ flow and transport processes. This AMR is intended to support revisions to ''Conceptual and Numerical Models for UZ Flow and Transport'' and ''Unsaturated Zone Flow and Transport Model Process Model Report''. In general, the results discussed in this AMR are from studies conducted using a combination or a subset of the following three approaches: (1) air-injection tests, (2) liquid-release tests, and (3) moisture monitoring using in-drift sensors or in-borehole sensors, to evaluate the impact of excavation, ventilation, and construction-water usage on the surrounding rocks. The liquid-release tests and air-injection tests provide an evaluation of in situ fracture flow and the competing processes of matrix imbibition. Only the findings from testing and data not covered in the ''Seepage Calibration Model and Seepage Testing Data'' are analyzed in detail in the AMR

Correlation between two parameters of mice behaviour in the open field test

OpenAIRE

Stojanović, Nikola M.; Ranđelović, Pavle J.; Radulović, Niko S.

2017-01-01

The open field test is being used extensively for the determination of different aspects of animal behaviour for over seventy years. The correlation between different behavioural parameters obtained in this test, although previously studied, is still debatable. Thus, we aimed to analyze and correlate behaviour scores to estimate the importance of individual parameters in this type of experiment. The open field test was performed on male BALB/c mice treated with either saline (10 ml/kg) or dia...

Repurposing Lesogaberan to Promote Human Islet Cell Survival and β-Cell Replication

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Jide Tian

2017-01-01

Full Text Available The activation of β-cell’s A- and B-type gamma-aminobutyric acid receptors (GABAA-Rs and GABAB-Rs can promote their survival and replication, and the activation of α-cell GABAA-Rs promotes their conversion into β-cells. However, GABA and the most clinically applicable GABA-R ligands may be suboptimal for the long-term treatment of diabetes due to their pharmacological properties or potential side-effects on the central nervous system (CNS. Lesogaberan (AZD3355 is a peripherally restricted high-affinity GABAB-R-specific agonist, originally developed for the treatment of gastroesophageal reflux disease (GERD that appears to be safe for human use. This study tested the hypothesis that lesogaberan could be repurposed to promote human islet cell survival and β-cell replication. Treatment with lesogaberan significantly enhanced replication of human islet cells in vitro, which was abrogated by a GABAB-R antagonist. Immunohistochemical analysis of human islets that were grafted into immune-deficient mice revealed that oral treatment with lesogaberan promoted human β-cell replication and islet cell survival in vivo as effectively as GABA (which activates both GABAA-Rs and GABAB-Rs, perhaps because of its more favorable pharmacokinetics. Lesogaberan may be a promising drug candidate for clinical studies of diabetes intervention and islet transplantation.

Spacetime replication of continuous variable quantum information

International Nuclear Information System (INIS)

Hayden, Patrick; Nezami, Sepehr; Salton, Grant; Sanders, Barry C

2016-01-01

The theory of relativity requires that no information travel faster than light, whereas the unitarity of quantum mechanics ensures that quantum information cannot be cloned. These conditions provide the basic constraints that appear in information replication tasks, which formalize aspects of the behavior of information in relativistic quantum mechanics. In this article, we provide continuous variable (CV) strategies for spacetime quantum information replication that are directly amenable to optical or mechanical implementation. We use a new class of homologically constructed CV quantum error correcting codes to provide efficient solutions for the general case of information replication. As compared to schemes encoding qubits, our CV solution requires half as many shares per encoded system. We also provide an optimized five-mode strategy for replicating quantum information in a particular configuration of four spacetime regions designed not to be reducible to previously performed experiments. For this optimized strategy, we provide detailed encoding and decoding procedures using standard optical apparatus and calculate the recovery fidelity when finite squeezing is used. As such we provide a scheme for experimentally realizing quantum information replication using quantum optics. (paper)

COPI is required for enterovirus 71 replication.

Directory of Open Access Journals (Sweden)

Jianmin Wang

Full Text Available Enterovirus 71 (EV71, a member of the Picornaviridae family, is found in Asian countries where it causes a wide range of human diseases. No effective therapy is available for the treatment of these infections. Picornaviruses undergo RNA replication in association with membranes of infected cells. COPI and COPII have been shown to be involved in the formation of picornavirus-induced vesicles. Replication of several picornaviruses, including poliovirus and Echovirus 11 (EV11, is dependent on COPI or COPII. Here, we report that COPI, but not COPII, is required for EV71 replication. Replication of EV71 was inhibited by brefeldin A and golgicide A, inhibitors of COPI activity. Furthermore, we found EV71 2C protein interacted with COPI subunits by co-immunoprecipitation and GST pull-down assay, indicating that COPI coatomer might be directed to the viral replication complex through viral 2C protein. Additionally, because the pathway is conserved among different species of enteroviruses, it may represent a novel target for antiviral therapies.

DNA replication after mutagenic treatment in Hordeum vulgare.

Science.gov (United States)

Kwasniewska, Jolanta; Kus, Arita; Swoboda, Monika; Braszewska-Zalewska, Agnieszka

2016-12-01

The temporal and spatial properties of DNA replication in plants related to DNA damage and mutagenesis is poorly understood. Experiments were carried out to explore the relationships between DNA replication, chromatin structure and DNA damage in nuclei from barley root tips. We quantitavely analysed the topological organisation of replication foci using pulse EdU labelling during the S phase and its relationship with the DNA damage induced by mutagenic treatment with maleic hydrazide (MH), nitroso-N-methyl-urea (MNU) and gamma ray. Treatment with mutagens did not change the characteristic S-phase patterns in the nuclei; however, the frequencies of the S-phase-labelled cells after treatment differed from those observed in the control cells. The analyses of DNA replication in barley nuclei were extended to the micronuclei induced by mutagens. Replication in the chromatin of the micronuclei was rare. The results of simultanous TUNEL reaction to identify cells with DNA strand breaks and the labelling of the S-phase cells with EdU revealed the possibility of DNA replication occurring in damaged nuclei. For the first time, the intensity of EdU fluorescence to study the rate of DNA replication was analysed. Copyright © 2016 Elsevier B.V. All rights reserved.

Dispersant field testing : a review of procedures and considerations

International Nuclear Information System (INIS)

Fingas, M.F.

2004-01-01

The effectiveness of a dispersant is defined by the amount of oil that the dispersant puts into the water column compared to the amount of oil that was initially spilled. Effectiveness is generally determined visually in plumes of dispersed oil that are visible from ships and aircraft. This paper describes 25 specific issues and technical concerns regarding field testing of dispersant effectiveness. Recent field tests were reviewed and literature that relates to testing procedures was sited. The 25 factors that are important for the appropriate outcome of dispersant field experiments include: mass balance; proper controls; analytical method; differential plume movement; time lag and length of time followed; mathematics of calculation and integration; lower and upper limits of analytical methods; use of remote sensing; thickness measurement; behaviour of oil with surfactant content; surfactant stripping; tracking surface oil and dispersed oil; recovering surface oil; visibility of oil from the surface; background levels of hydrocarbons; fluorescence of dispersant; herding; emulsion breaking; application success; heterogeneity of slick and plume; deposition measurements; true analytical standards; effect of wind on dispersant and slick; dispersant run-off; and weathering of the oil. It was concluded that the most important factors are the ability to determine mass balance, use proper controls, analytical methods and to avoid procedures that give incorrect results. 34 refs., 4 tabs., 1 fig

Test plan for the field evaluation and demonstration of the Contamination Control Unit

International Nuclear Information System (INIS)

Winberg, M.R.; Thompson, D.N.

1993-06-01

This report describes test details of a full demonstration of the Contamination Control Unit (CCU). The CCU is a mobile trailer capable of employing the use of soil fixatives, dust suppression agents, misting, and vacuum systems. These systems can perform a large number of contamination control functions to support the Office of Waste Technology Development (OTD) Buried Waste Integrated Demonstration (BWID) projects, transuranic (TRU) waste retrieval operations, and emergency response for hazardous and radioactive materials incidents. The demonstration will include both performance testing at the North Holmes Laboratory Facility (NHLF) and field testing in conjunction with the Remote Excavation System Demonstration at the Cold Test Pit. The NHLF will test operational parameters using water only, and the field demonstration at the Cold Test Pit involves full scale operation of vacuum, fixant, misting, and dust suppression systems. Test objectives, detailed experimental procedures, and data quality objectives necessary to perform the field demonstration are included in this test plan

Emotionality in growing pigs: is the open field a valid test?

Science.gov (United States)

Donald, Ramona D; Healy, Susan D; Lawrence, Alistair B; Rutherford, Kenneth M D

2011-10-24

The ability to assess emotionality is important within animal welfare research. Yet, for farm animals, few tests of emotionality have been well validated. Here we investigated the construct validity of behavioural measures of pig emotionality in an open-field test by manipulating the experiences of pigs in three ways. In Experiment One (pharmacological manipulation), pigs pre-treated with Azaperone, a drug used to reduce stress in commercial pigs, were more active, spent more time exploring and vocalised less than control pigs. In Experiment Two (social manipulation), pigs that experienced the open-field arena with a familiar companion were also more exploratory, spent less time behaviourally idle, and were less vocal than controls although to a lesser degree than in Experiment One. In Experiment Three (novelty manipulation), pigs experiencing the open field for a second time were less active, explored less and vocalised less than they had done in the first exposure to the arena. A principal component analysis was conducted on data from all three trials. The first two components could be interpreted as relating to the form (cautious to exploratory) and magnitude (low to high arousal) of the emotional response to open-field testing. Based on these dimensions, in Experiment One, Azaperone pigs appeared to be less fearful than saline-treated controls. However, in Experiment Two, exposure to the arena with a conspecific did not affect the first two dimensions but did affect a third behavioural dimension, relating to oro-nasal exploration of the arena floor. In Experiment Three, repeat exposure altered the form but not the magnitude of emotional response: pigs were less exploratory in the second test. In conclusion, behavioural measures taken from pigs in an open-field test are sensitive to manipulations of their prior experience in a manner that suggests they reflect underlying emotionality. Behavioural measures taken during open-field exposure can be useful for making

rMATS: robust and flexible detection of differential alternative splicing from replicate RNA-Seq data.

Science.gov (United States)

Shen, Shihao; Park, Juw Won; Lu, Zhi-xiang; Lin, Lan; Henry, Michael D; Wu, Ying Nian; Zhou, Qing; Xing, Yi

2014-12-23

Ultra-deep RNA sequencing (RNA-Seq) has become a powerful approach for genome-wide analysis of pre-mRNA alternative splicing. We previously developed multivariate analysis of transcript splicing (MATS), a statistical method for detecting differential alternative splicing between two RNA-Seq samples. Here we describe a new statistical model and computer program, replicate MATS (rMATS), designed for detection of differential alternative splicing from replicate RNA-Seq data. rMATS uses a hierarchical model to simultaneously account for sampling uncertainty in individual replicates and variability among replicates. In addition to the analysis of unpaired replicates, rMATS also includes a model specifically designed for paired replicates between sample groups. The hypothesis-testing framework of rMATS is flexible and can assess the statistical significance over any user-defined magnitude of splicing change. The performance of rMATS is evaluated by the analysis of simulated and real RNA-Seq data. rMATS outperformed two existing methods for replicate RNA-Seq data in all simulation settings, and RT-PCR yielded a high validation rate (94%) in an RNA-Seq dataset of prostate cancer cell lines. Our data also provide guiding principles for designing RNA-Seq studies of alternative splicing. We demonstrate that it is essential to incorporate biological replicates in the study design. Of note, pooling RNAs or merging RNA-Seq data from multiple replicates is not an effective approach to account for variability, and the result is particularly sensitive to outliers. The rMATS source code is freely available at rnaseq-mats.sourceforge.net/. As the popularity of RNA-Seq continues to grow, we expect rMATS will be useful for studies of alternative splicing in diverse RNA-Seq projects.

The Design of Finite State Machine for Asynchronous Replication Protocol

Science.gov (United States)

Wang, Yanlong; Li, Zhanhuai; Lin, Wei; Hei, Minglei; Hao, Jianhua

Data replication is a key way to design a disaster tolerance system and to achieve reliability and availability. It is difficult for a replication protocol to deal with the diverse and complex environment. This means that data is less well replicated than it ought to be. To reduce data loss and to optimize replication protocols, we (1) present a finite state machine, (2) run it to manage an asynchronous replication protocol and (3) report a simple evaluation of the asynchronous replication protocol based on our state machine. It's proved that our state machine is applicable to guarantee the asynchronous replication protocol running in the proper state to the largest extent in the event of various possible events. It also can helpful to build up replication-based disaster tolerance systems to ensure the business continuity.

Cytomegalovirus-specific T-cell responses and viral replication in kidney transplant recipients

Directory of Open Access Journals (Sweden)

Sester Urban

2008-06-01

Full Text Available Abstract Background Cytomegalovirus (CMV seronegative recipients (R- of kidney transplants (KT from seropositive donors (D+ are at higher risk for CMV replication and ganciclovir(GCV-resistance than CMV R(+. We hypothesized that low CMV-specific T-cell responses are associated with increased risk of CMV replication in R(+-patients with D(+ or D(- donors. Methods We prospectively evaluated 73 consecutive KT-patients [48 R(+, 25 D(+R(-] undergoing routine testing for CMV replication as part of a preemptive strategy. We compared CMV-specific interferon-γ (IFN-γ responses of CD4+CD3+ lymphocytes in peripheral blood mononuclear cells (PBMC using three different antigen preparation (CMV-lysate, pp72- and pp65-overlapping peptide pools using intracellular cytokine staining and flow cytometry. Results Median CD4+ and CD8+T-cell responses to CMV-lysate, pp72- and pp65-overlapping peptide pools were lower in D(+R(- than in R(+patients or in non-immunosuppressed donors. Comparing subpopulations we found that CMV-lysate favored CD4+- over CD8+-responses, whereas the reverse was observed for pp72, while pp65-CD4+- and -CD8+-responses were similar. Concurrent CMV replication in R(+-patients was associated with significantly lower T-cell responses (pp65 median CD4+ 0.00% vs. 0.03%, p = 0.001; CD8+ 0.01% vs. 0.03%; p = 0.033. Receiver operated curve analysis associated CMV-pp65 CD4+ responses of > 0.03% in R(+-patients with absence of concurrent (p = 0.003 and future CMV replication in the following 8 weeks (p = 0.036. GCV-resistant CMV replication occurred in 3 R(+-patients (6.3% with pp65- CD4+ frequencies Conclusion The data suggest that pp65-specific CD4+ T-cells might be useful to identify R(+-patients at increased risk of CMV replication. Provided further corroborating evidence, CMV-pp65 CD4+ responses above 0.03% in PBMCs of KT patients under stable immunosuppression are associated with lower risk of concurrent and future CMV replication during the

DNA replication stress restricts ribosomal DNA copy number.

Science.gov (United States)

Salim, Devika; Bradford, William D; Freeland, Amy; Cady, Gillian; Wang, Jianmin; Pruitt, Steven C; Gerton, Jennifer L

2017-09-01

Ribosomal RNAs (rRNAs) in budding yeast are encoded by ~100-200 repeats of a 9.1kb sequence arranged in tandem on chromosome XII, the ribosomal DNA (rDNA) locus. Copy number of rDNA repeat units in eukaryotic cells is maintained far in excess of the requirement for ribosome biogenesis. Despite the importance of the repeats for both ribosomal and non-ribosomal functions, it is currently not known how "normal" copy number is determined or maintained. To identify essential genes involved in the maintenance of rDNA copy number, we developed a droplet digital PCR based assay to measure rDNA copy number in yeast and used it to screen a yeast conditional temperature-sensitive mutant collection of essential genes. Our screen revealed that low rDNA copy number is associated with compromised DNA replication. Further, subculturing yeast under two separate conditions of DNA replication stress selected for a contraction of the rDNA array independent of the replication fork blocking protein, Fob1. Interestingly, cells with a contracted array grew better than their counterparts with normal copy number under conditions of DNA replication stress. Our data indicate that DNA replication stresses select for a smaller rDNA array. We speculate that this liberates scarce replication factors for use by the rest of the genome, which in turn helps cells complete DNA replication and continue to propagate. Interestingly, tumors from mini chromosome maintenance 2 (MCM2)-deficient mice also show a loss of rDNA repeats. Our data suggest that a reduction in rDNA copy number may indicate a history of DNA replication stress, and that rDNA array size could serve as a diagnostic marker for replication stress. Taken together, these data begin to suggest the selective pressures that combine to yield a "normal" rDNA copy number.

DNA replication stress restricts ribosomal DNA copy number

Science.gov (United States)

Salim, Devika; Bradford, William D.; Freeland, Amy; Cady, Gillian; Wang, Jianmin

2017-01-01

Ribosomal RNAs (rRNAs) in budding yeast are encoded by ~100–200 repeats of a 9.1kb sequence arranged in tandem on chromosome XII, the ribosomal DNA (rDNA) locus. Copy number of rDNA repeat units in eukaryotic cells is maintained far in excess of the requirement for ribosome biogenesis. Despite the importance of the repeats for both ribosomal and non-ribosomal functions, it is currently not known how “normal” copy number is determined or maintained. To identify essential genes involved in the maintenance of rDNA copy number, we developed a droplet digital PCR based assay to measure rDNA copy number in yeast and used it to screen a yeast conditional temperature-sensitive mutant collection of essential genes. Our screen revealed that low rDNA copy number is associated with compromised DNA replication. Further, subculturing yeast under two separate conditions of DNA replication stress selected for a contraction of the rDNA array independent of the replication fork blocking protein, Fob1. Interestingly, cells with a contracted array grew better than their counterparts with normal copy number under conditions of DNA replication stress. Our data indicate that DNA replication stresses select for a smaller rDNA array. We speculate that this liberates scarce replication factors for use by the rest of the genome, which in turn helps cells complete DNA replication and continue to propagate. Interestingly, tumors from mini chromosome maintenance 2 (MCM2)-deficient mice also show a loss of rDNA repeats. Our data suggest that a reduction in rDNA copy number may indicate a history of DNA replication stress, and that rDNA array size could serve as a diagnostic marker for replication stress. Taken together, these data begin to suggest the selective pressures that combine to yield a “normal” rDNA copy number. PMID:28915237

DNA replication stress restricts ribosomal DNA copy number.

Directory of Open Access Journals (Sweden)

Devika Salim

2017-09-01

Full Text Available Ribosomal RNAs (rRNAs in budding yeast are encoded by ~100-200 repeats of a 9.1kb sequence arranged in tandem on chromosome XII, the ribosomal DNA (rDNA locus. Copy number of rDNA repeat units in eukaryotic cells is maintained far in excess of the requirement for ribosome biogenesis. Despite the importance of the repeats for both ribosomal and non-ribosomal functions, it is currently not known how "normal" copy number is determined or maintained. To identify essential genes involved in the maintenance of rDNA copy number, we developed a droplet digital PCR based assay to measure rDNA copy number in yeast and used it to screen a yeast conditional temperature-sensitive mutant collection of essential genes. Our screen revealed that low rDNA copy number is associated with compromised DNA replication. Further, subculturing yeast under two separate conditions of DNA replication stress selected for a contraction of the rDNA array independent of the replication fork blocking protein, Fob1. Interestingly, cells with a contracted array grew better than their counterparts with normal copy number under conditions of DNA replication stress. Our data indicate that DNA replication stresses select for a smaller rDNA array. We speculate that this liberates scarce replication factors for use by the rest of the genome, which in turn helps cells complete DNA replication and continue to propagate. Interestingly, tumors from mini chromosome maintenance 2 (MCM2-deficient mice also show a loss of rDNA repeats. Our data suggest that a reduction in rDNA copy number may indicate a history of DNA replication stress, and that rDNA array size could serve as a diagnostic marker for replication stress. Taken together, these data begin to suggest the selective pressures that combine to yield a "normal" rDNA copy number.

On-site cell field test support program

Science.gov (United States)

Staniunas, J. W.; Merten, G. P.

1982-09-01

Utility sites for data monitoring were reviewed and selected. Each of these sites will be instrumented and its energy requirements monitored and analyzed for one year prior to the selection of 40 Kilowatt fuel cell field test sites. Analyses in support of the selection of sites for instrumentation shows that many building sectors offered considerable market potential. These sectors include nursing home, health club, restaurant, industrial, hotel/motel and apartment.

Modeling of a Parabolic Trough Solar Field for Acceptance Testing: A Case Study

Energy Technology Data Exchange (ETDEWEB)

Wagner, M. J.; Mehos, M. S.; Kearney, D. W.; McMahan, A. C.

2011-01-01

As deployment of parabolic trough concentrating solar power (CSP) systems ramps up, the need for reliable and robust performance acceptance test guidelines for the solar field is also amplified. Project owners and/or EPC contractors often require extensive solar field performance testing as part of the plant commissioning process in order to ensure that actual solar field performance satisfies both technical specifications and performance guaranties between the involved parties. Performance test code work is currently underway at the National Renewable Energy Laboratory (NREL) in collaboration with the SolarPACES Task-I activity, and within the ASME PTC-52 committee. One important aspect of acceptance testing is the selection of a robust technology performance model. NREL1 has developed a detailed parabolic trough performance model within the SAM software tool. This model is capable of predicting solar field, sub-system, and component performance. It has further been modified for this work to support calculation at subhourly time steps. This paper presents the methodology and results of a case study comparing actual performance data for a parabolic trough solar field to the predicted results using the modified SAM trough model. Due to data limitations, the methodology is applied to a single collector loop, though it applies to larger subfields and entire solar fields. Special consideration is provided for the model formulation, improvements to the model formulation based on comparison with the collected data, and uncertainty associated with the measured data. Additionally, this paper identifies modeling considerations that are of particular importance in the solar field acceptance testing process and uses the model to provide preliminary recommendations regarding acceptable steady-state testing conditions at the single-loop level.

Large Field Photogrammetry Techniques in Aircraft and Spacecraft Impact Testing

Science.gov (United States)

Littell, Justin D.

2010-01-01

The Landing and Impact Research Facility (LandIR) at NASA Langley Research Center is a 240 ft. high A-frame structure which is used for full-scale crash testing of aircraft and rotorcraft vehicles. Because the LandIR provides a unique capability to introduce impact velocities in the forward and vertical directions, it is also serving as the facility for landing tests on full-scale and sub-scale Orion spacecraft mass simulators. Recently, a three-dimensional photogrammetry system was acquired to assist with the gathering of vehicle flight data before, throughout and after the impact. This data provides the basis for the post-test analysis and data reduction. Experimental setups for pendulum swing tests on vehicles having both forward and vertical velocities can extend to 50 x 50 x 50 foot cubes, while weather, vehicle geometry, and other constraints make each experimental setup unique to each test. This paper will discuss the specific calibration techniques for large fields of views, camera and lens selection, data processing, as well as best practice techniques learned from using the large field of view photogrammetry on a multitude of crash and landing test scenarios unique to the LandIR.

Fast automatic quantitative cell replication with fluorescent live cell imaging

Directory of Open Access Journals (Sweden)

Wang Ching-Wei

2012-01-01

Full Text Available Abstract Background live cell imaging is a useful tool to monitor cellular activities in living systems. It is often necessary in cancer research or experimental research to quantify the dividing capabilities of cells or the cell proliferation level when investigating manipulations of the cells or their environment. Manual quantification of fluorescence microscopic image is difficult because human is neither sensitive to fine differences in color intensity nor effective to count and average fluorescence level among cells. However, auto-quantification is not a straightforward problem to solve. As the sampling location of the microscopy changes, the amount of cells in individual microscopic images varies, which makes simple measurement methods such as the sum of stain intensity values or the total number of positive stain within each image inapplicable. Thus, automated quantification with robust cell segmentation techniques is required. Results An automated quantification system with robust cell segmentation technique are presented. The experimental results in application to monitor cellular replication activities show that the quantitative score is promising to represent the cell replication level, and scores for images from different cell replication groups are demonstrated to be statistically significantly different using ANOVA, LSD and Tukey HSD tests (p-value Conclusion A robust automated quantification method of live cell imaging is built to measure the cell replication level, providing a robust quantitative analysis system in fluorescent live cell imaging. In addition, the presented unsupervised entropy based cell segmentation for live cell images is demonstrated to be also applicable for nuclear segmentation of IHC tissue images.

Relationships between field performance tests in high-level soccer players

DEFF Research Database (Denmark)

Ingebrigtsen, Jørgen; Brochmann, Marit; Castagna, Carlo

2014-01-01

after two and four minutes of the Yo-Yo IR tests by testing 57 high-level soccer players. All players played regularly in one of the three highest levels of Norwegian soccer and were tested during three sessions on three consecutive days. Large correlations were observed between Yo-Yo IR1 and IR2 test...... using only one of the Yo-Yo tests and a RSA test, in a general soccer-specific field test protocol. The sub-maximal heart rate measures during Yo-Yo tests are reproducible and may be utilized for frequent, time-efficient and non-exhaustive testing of intermittent exercise capacity of high-level soccer...

Chaotic interactions of self-replicating RNA.

Science.gov (United States)

Forst, C V

1996-03-01

A general system of high-order differential equations describing complex dynamics of replicating biomolecules is given. Symmetry relations and coordinate transformations of general replication systems leading to topologically equivalent systems are derived. Three chaotic attractors observed in Lotka-Volterra equations of dimension n = 3 are shown to represent three cross-sections of one and the same chaotic regime. Also a fractal torus in a generalized three-dimensional Lotka-Volterra Model has been linked to one of the chaotic attractors. The strange attractors are studied in the equivalent four-dimensional catalytic replicator network. The fractal torus has been examined in adapted Lotka-Volterra equations. Analytic expressions are derived for the Lyapunov exponents of the flow in the replicator system. Lyapunov spectra for different pathways into chaos has been calculated. In the generalized Lotka-Volterra system a second inner rest point--coexisting with (quasi)-periodic orbits--can be observed; with an abundance of different bifurcations. Pathways from chaotic tori, via quasi-periodic tori, via limit cycles, via multi-periodic orbits--emerging out of periodic doubling bifurcations--to "simple" chaotic attractors can be found.

A Case for Developing a Ground Based Replication of the Earth, Moon and Mars Spaceflight Infrastructure

Science.gov (United States)

Bradford, Robert N.; Best, Susan L.

2006-01-01

When the systems are developed and in place to provide the services needed to operate en route and on the Lunar and Martian surfaces, an Earth based replication will need to be in place for the safety and protection of mission success. The replication will entail all aspects of the flight configuration end to end but will not include any closed loop systems. This would replicate the infrastructure from Lunar and Martian robots, manned surface excursions, through man and unmanned terrestrial bases, through the various types of communication systems and technologies, manned and un-manned space vehicles (large and small), to Earth based systems and control centers. An Earth based replicated infrastructure will enable checkout and test of new technologies, hardware, software updates and upgrades and procedures without putting humans and missions at risk. Analysis of events, what ifs and trouble resolution could be played out on the ground to remove as much risk as possible from any type of proposed change to flight operational systems. With adequate detail, it is possible that failures could be predicted with a high probability and action taken to eliminate failures. A major factor in any mission to the Moon and to Mars is the complexity of systems, interfaces, processes, their limitations, associated risks and the factor of the unknown including the development by many contractors and NASA centers. The need to be able to introduce new technologies over the life of the program requires an end to end test bed to analyze and evaluate these technologies and what will happen when they are introduced into the flight system. The ability to analyze system behaviors end to end under varying conditions would enhance safety e.g. fault tolerances. This analysis along with the ability to mine data from the development environment (e.g. test data), flight ops and modeling/simulations data would provide a level of information not currently available to operations and astronauts. In

Preliminary investigation on determination of radionuclide distribution in field tracing test site

International Nuclear Information System (INIS)

Tanaka, Tadao; Mukai, Masayuki; Takebe, Shinichi; Guo Zede; Li Shushen; Kamiyama, Hideo.

1993-12-01

Field tracing tests for radionuclide migration have been conducted by using 3 H, 60 Co, 85 Sr and 134 Cs, in the natural unsaturated loess zone at field test site of China Institute for Radiation Protection. It is necessary to obtain confidable distribution data of the radionuclides in the test site, in order to evaluate exactly the migration behavior of the radionuclides in situ. An available method to determine the distribution was proposed on the basis of preliminary discussing results on sampling method of soils from the test site and analytical method of radioactivity in the soils. (author)

Insights into the Initiation of Eukaryotic DNA Replication.

Science.gov (United States)

Bruck, Irina; Perez-Arnaiz, Patricia; Colbert, Max K; Kaplan, Daniel L

2015-01-01

The initiation of DNA replication is a highly regulated event in eukaryotic cells to ensure that the entire genome is copied once and only once during S phase. The primary target of cellular regulation of eukaryotic DNA replication initiation is the assembly and activation of the replication fork helicase, the 11-subunit assembly that unwinds DNA at a replication fork. The replication fork helicase, called CMG for Cdc45-Mcm2-7, and GINS, assembles in S phase from the constituent Cdc45, Mcm2-7, and GINS proteins. The assembly and activation of the CMG replication fork helicase during S phase is governed by 2 S-phase specific kinases, CDK and DDK. CDK stimulates the interaction between Sld2, Sld3, and Dpb11, 3 initiation factors that are each required for the initiation of DNA replication. DDK, on the other hand, phosphorylates the Mcm2, Mcm4, and Mcm6 subunits of the Mcm2-7 complex. Sld3 recruits Cdc45 to Mcm2-7 in a manner that depends on DDK, and recent work suggests that Sld3 binds directly to Mcm2-7 and also to single-stranded DNA. Furthermore, recent work demonstrates that Sld3 and its human homolog Treslin substantially stimulate DDK phosphorylation of Mcm2. These data suggest that the initiation factor Sld3/Treslin coordinates the assembly and activation of the eukaryotic replication fork helicase by recruiting Cdc45 to Mcm2-7, stimulating DDK phosphorylation of Mcm2, and binding directly to single-stranded DNA as the origin is melted.

Antioxidant effects of nerolidol in mice hippocampus after open field test.

Science.gov (United States)

Nogueira Neto, José Damasceno; de Almeida, Antonia Amanda Cardoso; da Silva Oliveira, Johanssy; Dos Santos, Pauline Sousa; de Sousa, Damião Pergentino; de Freitas, Rivelilson Mendes

2013-09-01

The aim of this study was to evaluate the neuroprotective effects of nerolidol in mice hippocampus against oxidative stress in neuronal cells compared to ascorbic acid (positive control) as well as evaluated the nerolidol sedative effects by open field test compared to diazepam (positive control). Thirty minutes prior to behavioral observation on open field test, mice were intraperitoneally treated with vehicle, nerolidol (25, 50 and 75 mg/kg), diazepam (1 mg/kg) or ascorbic acid (250 mg/kg). To clarify the action mechanism of of nerolidol on oxidative stress in animals subjected to the open field test, Western blot analysis of Mn-superoxide dismutase and catalase in mice hippocampus were performed. In nerolidol group, there was a significant decrease in lipid peroxidation and nitrite levels when compared to negative control (vehicle). However, a significant increase was observed in superoxide dismutase and catalase activities in this group when compared to the other groups. Vehicle, diazepam, ascorbic acid and nerolidol groups did not affected Mn-superoxide dismutase, catalase mRNA or protein levels. Our findings strongly support the hypothesis that oxidative stress occurs in hippocampus. Nerolidol showed sedative effects in animals subjected to the open field test. Oxidative process plays a crucial role on neuronal pathological consequence, and implies that antioxidant effects could be achieved using this sesquiterpene.

Yeast as a model host to study replication and recombination of defective interfering RNA of Tomato bushy stunt virus

International Nuclear Information System (INIS)

Panavas, Tadas; Nagy, Peter D.

2003-01-01

Defective interfering (DI) RNA associated with Tomato bushy stunt virus (TBSV), which is a plus-strand RNA virus, requires p33 and p92 proteins of TBSV or the related Cucumber necrosis virus (CNV), for replication in plants. To test if DI RNA can replicate in a model host, we coexpressed TBSV DI RNA and p33/p92 of CNV in yeast. We show evidence for replication of DI RNA in yeast, including (i) dependence on p33 and p92 for DI replication; (ii) presence of active CNV RNA-dependent RNA polymerase in isolated membrane-containing preparations; (iii) increasing amount of DI RNA(+) over time; (iv) accumulation of (-)stranded DI RNA; (v) presence of correct 5' and 3' ends in DI RNA; (vi) inhibition of replication by mutations in the replication enhancer; and (vii) evolution of DI RNA over time, as shown by sequence heterogeneity. We also produced evidence supporting the occurrence of DI RNA recombinants in yeast. In summary, development of yeast as a host for replication of TBSV DI RNA will facilitate studies on the roles of viral and host proteins in replication/recombination

Autonomous replication of plasmids bearing monkey DNA origin-enriched sequences

International Nuclear Information System (INIS)

Frappier, L.; Zannis-Hadjopoulos, M.

1987-01-01

Twelve clones of origin-enriched sequences (ORS) isolated from early replicating monkey (CV-1) DNA were examined for transient episomal replication in transfected CV-1, COS-7, and HeLa cells. Plasmid DNA was isolated at time intervals after transfection and screened by the Dpn I resistance assay or by the bromodeoxyuridine substitution assay to differentiate between input and replicated DNA. The authors have identified four monkey ORS (ORS3, -8, -9, and -12) that can support plasmid replication in mammalian cells. This replication is carried out in a controlled and semiconservative manner characteristic of mammalian replicons. ORS replication was most efficient in HeLa cells. Electron microscopy showed ORS8 and ORS12 plasmids of the correct size with replication bubbles. Using a unique restriction site in ORS12, we have mapped the replication bubble within the monkey DNA sequence

A semi-field approach to testing effects of fresh pesticide residues on bees in multiple-rate test

NARCIS (Netherlands)

Bakker, F.; Calis, J.N.M.

2003-01-01

We describe a semi-field cage test specifically designed to test effects of delayed exposure to plant protection products. The trial involved the use of standardised mini-beehives. The principle of the trial was to prepare two groups of potted test plants per treatment. The first group of plants

Research Note The reliability of a field test kit for the detection and ...

African Journals Online (AJOL)

Research Note The reliability of a field test kit for the detection and the persistence of ... Open Access DOWNLOAD FULL TEXT ... The objectives were to test a field kit for practicality and reliability, to assess the spread of the bacteria among ...

In situ respiration testing: A field treatability test for bioventing

International Nuclear Information System (INIS)

Kittel, J.A.; Hinchee, R.E.; Miller, R.; Vogel, C.; Hoeppel, R.

1993-01-01

Bioventing is the process of aerating subsurface soils to stimulate in situ biological activity and promote bioremediation. Bioventing differs from soil venting in remedial approach. Soil venting is designed and operated to maximize the volatilization of low-molecular-weight compounds, with some biodegradation occurring. In contrast, bioventing is designed to maximize biodegradation of aerobically biodegradable compounds, regardless of their molecular weight, with some volatilization occurring. Bioventing is gaining wide acceptance as a remediation alternative at petroleum-contaminated sites. However, site variability usually requires that a short term treatability test be conducted in situ at potential sites to determine the applicability of bioventing. Battelle has worked with the US Air Force and the US Navy to develop a simple and inexpensive field test to evaluate bioventing potential-contaminated sites. This test has been used to evaluate the applicability of bioventing at over 50 sites. The in situ respiration test consists of injecting air and an inert tracer gas (helium) over a 24-hour period to aerate soils at an oxygen-deficient, petroleum-contaminated site. Soil vapor samples are collected to determine oxygen utilization rates and carbon dioxide production rates. The stoichiometric relationship for the oxidation of hexane is used to calculate the biodegradation rate. The tracer gas is monitored to estimate the effect of diffusion on changes in soil-gas concentrations

Beta Testing StraboSpot: Perspectives on mobile field mapping and data collection

Science.gov (United States)

Bunse, E.; Graham, K. A.; Rufledt, C.; Walker, J. D.; Müller, A.; Tikoff, B.

2017-12-01

Geologic field mapping has recently transitioned away from traditional techniques (e.g. field notebooks, paper mapping, Brunton compasses) and towards mobile `app' mapping technology. The StraboSpot system (Strabo) is an open-source solution for collection and storage for geologic field, microstructural, and lab-based data. Strabo's mission is to "enable recording and sharing data within the geoscience community, encourage interdisciplinary research, and facilitate the investigation of scientific questions that cannot currently be addressed" (Walker et al., 2015). Several mobile application beta tests of the system, on both Android and Apple iOS platforms using smartphones and tablets, began in Summer 2016. Students at the 2016 and 2017 University of Kansas Field Camps used Strabo in place of ArcGIS for Desktop on Panasonic Toughbooks, to field map two study areas. Strabo was also field tested by students of graduate and undergraduate level for both geo/thermochronologic sample collection and reconnaissance mapping associated with regional tectonic analysis in California. Throughout this period of testing, the app was geared toward structural and tectonic geologic data collection, but is versatile enough for other communities to currently use and is expanding to accommodate the sedimentology and petrology communities. Overall, users in each of the beta tests acclimated quickly to using Strabo for field data collection. Some key advantages to using Strabo over traditional mapping methods are: (1) Strabo allows for consolidation of materials in the field; (2) helps students track their position in the field with integrated GPS; and (3) Strabo data is in a uniform format making it simple for geologists to collaborate. While traditional field methods are not likely to go out of style in the near future, Strabo acts as a bridge between professional and novice geologists by providing a tool that is intuitive on all levels of geological and technological experience and

Reproducibility and replicability of rodent phenotyping in preclinical studies.

Science.gov (United States)

Kafkafi, Neri; Agassi, Joseph; Chesler, Elissa J; Crabbe, John C; Crusio, Wim E; Eilam, David; Gerlai, Robert; Golani, Ilan; Gomez-Marin, Alex; Heller, Ruth; Iraqi, Fuad; Jaljuli, Iman; Karp, Natasha A; Morgan, Hugh; Nicholson, George; Pfaff, Donald W; Richter, S Helene; Stark, Philip B; Stiedl, Oliver; Stodden, Victoria; Tarantino, Lisa M; Tucci, Valter; Valdar, William; Williams, Robert W; Würbel, Hanno; Benjamini, Yoav

2018-04-01

The scientific community is increasingly concerned with the proportion of published "discoveries" that are not replicated in subsequent studies. The field of rodent behavioral phenotyping was one of the first to raise this concern, and to relate it to other methodological issues: the complex interaction between genotype and environment; the definitions of behavioral constructs; and the use of laboratory mice and rats as model species for investigating human health and disease mechanisms. In January 2015, researchers from various disciplines gathered at Tel Aviv University to discuss these issues. The general consensus was that the issue is prevalent and of concern, and should be addressed at the statistical, methodological and policy levels, but is not so severe as to call into question the validity and the usefulness of model organisms as a whole. Well-organized community efforts, coupled with improved data and metadata sharing, have a key role in identifying specific problems and promoting effective solutions. Replicability is closely related to validity, may affect generalizability and translation of findings, and has important ethical implications. Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.

Fiber optic sensor applications in field testing

International Nuclear Information System (INIS)

Perea, J.A.

1984-01-01

Fiber optic sensors (F.O.S.) are defined, and the application of this technology to measuring various phenomonon in diverse and hostile environments are discussed. F.O.S. advantages and disavantages both technically and operationally are summarized. Three sensor techniques - intensity, interferometric, and polarization - are then discussed in some detail. General environmental instrumentation and controls that support the Nuclear Weapons Test Program at the Nevada Test Site are discussed next to provide the reader with a basic understanding of the programmatic task. This will aid in recognizing the various difficulties of the traditional measurement techniques at the NTS and the potential advantages that fiber optic measurement systems can provide. An F.O.S. development program is then outlined, depicting a plan to design and fabricate a prototype sensor to be available for field testing by the end of FY84. We conclude with future plans for further development of F.O.S. to measure more of the desired physical parameters for the Test Program, and to eventually become an integral part of an overall measurement and control system

Spatio-temporal re-organization of replication foci accompanies replication domain consolidation during human pluripotent stem cell lineage specification

Science.gov (United States)

Wilson, Korey A.; Elefanty, Andrew G.; Stanley, Edouard G.; Gilbert, David M.

2016-01-01

ABSTRACT Lineage specification of both mouse and human pluripotent stem cells (PSCs) is accompanied by spatial consolidation of chromosome domains and temporal consolidation of their replication timing. Replication timing and chromatin organization are both established during G1 phase at the timing decision point (TDP). Here, we have developed live cell imaging tools to track spatio-temporal replication domain consolidation during differentiation. First, we demonstrate that the fluorescence ubiquitination cell cycle indicator (Fucci) system is incapable of demarcating G1/S or G2/M cell cycle transitions. Instead, we employ a combination of fluorescent PCNA to monitor S phase progression, cytokinesis to demarcate mitosis, and fluorescent nucleotides to label early and late replication foci and track their 3D organization into sub-nuclear chromatin compartments throughout all cell cycle transitions. We find that, as human PSCs differentiate, the length of S phase devoted to replication of spatially clustered replication foci increases, coincident with global compartmentalization of domains into temporally clustered blocks of chromatin. Importantly, re-localization and anchorage of domains was completed prior to the onset of S phase, even in the context of an abbreviated PSC G1 phase. This approach can also be employed to investigate cell fate transitions in single PSCs, which could be seen to differentiate preferentially from G1 phase. Together, our results establish real-time, live-cell imaging methods for tracking cell cycle transitions during human PSC differentiation that can be applied to study chromosome domain consolidation and other aspects of lineage specification. PMID:27433885

Normal Threshold Size of Stimuli in Children Using a Game-Based Visual Field Test.

Science.gov (United States)

Wang, Yanfang; Ali, Zaria; Subramani, Siddharth; Biswas, Susmito; Fenerty, Cecilia; Henson, David B; Aslam, Tariq

2017-06-01

The aim of this study was to demonstrate and explore the ability of novel game-based perimetry to establish normal visual field thresholds in children. One hundred and eighteen children (aged 8.0 ± 2.8 years old) with no history of visual field loss or significant medical history were recruited. Each child had one eye tested using a game-based visual field test 'Caspar's Castle' at four retinal locations 12.7° (N = 118) from fixation. Thresholds were established repeatedly using up/down staircase algorithms with stimuli of varying diameter (luminance 20 cd/m 2 , duration 200 ms, background luminance 10 cd/m 2 ). Relationships between threshold and age were determined along with measures of intra- and intersubject variability. The Game-based visual field test was able to establish threshold estimates in the full range of children tested. Threshold size reduced with increasing age in children. Intrasubject variability and intersubject variability were inversely related to age in children. Normal visual field thresholds were established for specific locations in children using a novel game-based visual field test. These could be used as a foundation for developing a game-based perimetry screening test for children.

In Situ Field Testing of Processes

Energy Technology Data Exchange (ETDEWEB)

J. Wang

2001-12-14

The purpose of this Analysis/Model Report (AMR) is to update and document the data and subsequent analyses from ambient field-testing activities performed in underground drifts of the Yucca Mountain Site Characterization Project (YMP). This revision updates data and analyses presented in the initial issue of this AMR. This AMR was developed in accordance with the ''Technical Work Plan for Unsaturated Zone (UZ) Flow and Transport Process Model Report'' and ''Technical Work Plan for UZ Flow, Transport, and Coupled Processes Process Model Report. These activities were performed to investigate in situ flow and transport processes. The evaluations provide the necessary framework to: (1) refine and confirm the conceptual model of matrix and fracture processes in the unsaturated zone (UZ) and (2) analyze the impact of excavation (including use of construction water and effect of ventilation) on the UZ flow and transport processes. This AMR is intended to support revisions to ''Conceptual and Numerical Models for UZ Flow and Transport'' and ''Unsaturated Zone Flow and Transport Model Process Model Report''. In general, the results discussed in this AMR are from studies conducted using a combination or a subset of the following three approaches: (1) air-injection tests, (2) liquid-release tests, and (3) moisture monitoring using in-drift sensors or in-borehole sensors, to evaluate the impact of excavation, ventilation, and construction-water usage on the surrounding rocks. The liquid-release tests and air-injection tests provide an evaluation of in situ fracture flow and the competing processes of matrix imbibition. Only the findings from testing and data not covered in the ''Seepage Calibration Model and Seepage Testing Data'' are analyzed in detail in the AMR.

[Test Reviews in Child Psychology: Test Users Wish to Obtain Practical Information Relevant to their Respective Field of Work].

Science.gov (United States)

Renner, Gerolf; Irblich, Dieter

2016-11-01

Test Reviews in Child Psychology: Test Users Wish to Obtain Practical Information Relevant to their Respective Field of Work This study investigated to what extent diagnosticians use reviews of psychometric tests for children and adolescents, how they evaluate their quality, and what they expect concerning content. Test users (n = 323) from different areas of work (notably social pediatrics, early intervention, special education, speech and language therapy) rated test reviews as one of the most important sources of information. Readers of test reviews value practically oriented descriptions and evaluations of tests that are relevant to their respective field of work. They expect independent reviews that critically discuss opportunities and limits of the tests under scrutiny. The results show that authors of test reviews should not only have a background in test theory but should also be familiar with the practical application of tests in various settings.

Field Test Data for Detecting Vibrations of a Building Using High-Speed Video Cameras

Science.gov (United States)

2017-10-01

ARL-TR-8185 ● OCT 2017 US Army Research Laboratory Field Test Data for Detecting Vibrations of a Building Using High-Speed Video...Field Test Data for Detecting Vibrations of a Building Using High-Speed Video Cameras by Caitlin P Conn and Geoffrey H Goldman Sensors and...June 2016 – October 2017 4. TITLE AND SUBTITLE Field Test Data for Detecting Vibrations of a Building Using High-Speed Video Cameras 5a. CONTRACT

Mutational Analysis of the Hypervariable Region of Hepatitis E Virus Reveals Its Involvement in the Efficiency of Viral RNA Replication ▿

Science.gov (United States)

Pudupakam, R. S.; Kenney, Scott P.; Córdoba, Laura; Huang, Yao-Wei; Dryman, Barbara A.; LeRoith, Tanya; Pierson, F. William; Meng, Xiang-Jin

2011-01-01

The RNA genome of the hepatitis E virus (HEV) contains a hypervariable region (HVR) in ORF1 that tolerates small deletions with respect to infectivity. To further investigate the role of the HVR in HEV replication, we constructed a panel of mutants with overlapping deletions in the N-terminal, central, and C-terminal regions of the HVR by using a genotype 1 human HEV luciferase replicon and analyzed the effects of deletions on viral RNA replication in Huh7 cells. We found that the replication levels of the HVR deletion mutants were markedly reduced in Huh7 cells, suggesting a role of the HVR in viral replication efficiency. To further verify the results, we constructed HVR deletion mutants by using a genetically divergent, nonmammalian avian HEV, and similar effects on viral replication efficiency were observed when the avian HEV mutants were tested in LMH cells. Furthermore, the impact of complete HVR deletion on virus infectivity was tested in chickens, using an avian HEV mutant with a complete HVR deletion. Although the deletion mutant was still replication competent in LMH cells, the complete HVR deletion resulted in a loss of avian HEV infectivity in chickens. Since the HVR exhibits extensive variations in sequence and length among different HEV genotypes, we further examined the interchangeability of HVRs and demonstrated that HVR sequences are functionally exchangeable between HEV genotypes with regard to viral replication and infectivity in vitro, although genotype-specific HVR differences in replication efficiency were observed. The results showed that although the HVR tolerates small deletions with regard to infectivity, it may interact with viral and host factors to modulate the efficiency of HEV replication. PMID:21775444

How Often Is p[subscript rep] Close to the True Replication Probability?

Science.gov (United States)

Trafimow, David; MacDonald, Justin A.; Rice, Stephen; Clason, Dennis L.

2010-01-01

Largely due to dissatisfaction with the standard null hypothesis significance testing procedure, researchers have begun to consider alternatives. For example, Killeen (2005a) has argued that researchers should calculate p[subscript rep] that is purported to indicate the probability that, if the experiment in question were replicated, the obtained…