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Sample records for single point mutants

  1. Design of thermostable rhamnogalacturonan lyase mutants from Bacillus licheniformis by combination of targeted single point mutations

    DEFF Research Database (Denmark)

    da Silva, Ines Isabel Cardoso Rodrigues; Jers, Carsten; Otten, Harm

    2014-01-01

    Rhamnogalacturonan I lyases (RGI lyases) (EC 4.2.2.-) catalyze cleavage of α-1,4 bonds between rhamnose and galacturonic acid in the backbone of pectins by β-elimination. In the present study, targeted improvement of the thermostability of a PL family 11 RGI lyase from Bacillus licheniformis (DSM...... the wild-type RGI lyase in Bacillus subtilis as opposed to in Pichia pastoris; this effect is suggested to be a negative result of glycosylation of the P. pastoris expressed enzyme. A ~ twofold improvement in thermal stability at 60 °C, accompanied by less significant increases in Tm of the enzyme mutants......, were obtained due to additive stabilizing effects of single amino acid mutations (E434L, G55V, and G326E) compared to the wild type. The crystal structure of the B. licheniformis wild-type RGI lyase was also determined; the structural analysis corroborated that especially mutation of charged amino...

  2. Single-point ACT2 gene mutation in the Arabidopsis root hair mutant der1-3 affects overall actin organization, root growth and plant development.

    Science.gov (United States)

    Vaškebová, L; Šamaj, J; Ovecka, M

    2017-12-27

    The actin cytoskeleton forms a dynamic network in plant cells. A single-point mutation in the DER1 (deformed root hairs1) locus located in the sequence of ACTIN2, a gene for major actin in vegetative tissues of Arabidopsis thaliana, leads to impaired root hair development (Ringli C, Baumberger N, Diet A, Frey B, Keller B. 2002. ACTIN2 is essential for bulge site selection and tip growth during root hair development of Arabidopsis. Plant Physiology129: 1464-1472). Only root hair phenotypes have been described so far in der1 mutants, but here we demonstrate obvious aberrations in the organization of the actin cytoskeleton and overall plant development. Organization of the actin cytoskeleton in epidermal cells of cotyledons, hypocotyls and roots was studied qualitatively and quantitatively by live-cell imaging of transgenic lines carrying the GFP-FABD2 fusion protein and in fixed cells after phalloidin labelling. Patterns of root growth were characterized by FM4-64 vital staining, light-sheet microscopy imaging and microtubule immunolabelling. Plant phenotyping included analyses of germination, root growth and plant biomass. Speed of germination, plant fresh weight and total leaf area were significantly reduced in the der1-3 mutant in comparison with the C24 wild-type. Actin filaments in root, hypocotyl and cotyledon epidermal cells of the der1-3 mutant were shorter, thinner and arranged in more random orientations, while actin bundles were shorter and had altered orientations. The wavy pattern of root growth in der1-3 mutant was connected with higher frequencies of shifted cell division planes (CDPs) in root cells, which was consistent with the shifted positioning of microtubule-based preprophase bands and phragmoplasts. The organization of cortical microtubules in the root cells of the der1-3 mutant, however, was not altered. Root growth rate of the der1-3 mutant is not reduced, but changes in the actin cytoskeleton organization can induce a wavy root growth pattern

  3. Comparative studies on the minus origin mutants of Escherichia coli spherical single-stranded DNA phages.

    Science.gov (United States)

    Kodaira, K; Godson, N G; Taketo, A

    1995-01-25

    The minus origins for complementary strand DNA synthesis (-ori) of Escherichia coli spherical single-stranded DNA (microvirid) phages G4, phi K, alpha 3, and St-1 closely resemble each other in DNA structure and contain two potential secondary hairpin loops (I and II) that have been implicated as direct recognition sites for host E. coli dnaG protein (primase). We introduced mutations (deletion or insertion) within the -ori regions of phi K and G4 by the nuclease digestion method. Mutants thus constructed produced minute plaques, showed thermosensitivity, and they remarkably reduced the phage yield and rate of viral DNA synthesis. Deletions in the phi K mutants (dTa) were ranging from 1 nucleotide (nt) to 102 nt centered at the hairpin II; a dTa8 mutant was entirely lacking in the two hairpins besides the starting point for primer RNA synthesis. On the other hand, the G4 mutants (dSa) had deletions centered at hairpin I; two mutants dSa35 and dXN completely lost the hairpin I and the primer RNA starting point. In addition, progeny phage populations of several phi K and G4 mutants contained revertant-like phages. DNA sequencing analysis revealed that these secondary phages had been generated by spontaneous DNA rearrangement with additional insertion or deletion near the parental mutation sites, via an unknown recA-independent pathway.

  4. Theory of Single Point Incremental Forming

    DEFF Research Database (Denmark)

    Martins, P.A.F.; Bay, Niels; Skjødt, Martin

    2008-01-01

    This paper presents a closed-form theoretical analysis modelling the fundamentals of single point incremental forming and explaining the experimental and numerical results available in the literature for the past couple of years. The model is based on membrane analysis with bi-directional in-plan...

  5. Molecular basis of proton uptake in single and double mutants of cytochrome c oxidase

    Energy Technology Data Exchange (ETDEWEB)

    Henry, Rowan M; Caplan, David; Pomes, Regis [Molecular Structure and Function, Hospital for Sick Children, Toronto, ON, M5G 1X8 (Canada); Fadda, Elisa, E-mail: pomes@sickkids.ca [Department of Chemistry, University of Galway (Ireland)

    2011-06-15

    Cytochrome c oxidase, the terminal enzyme of the respiratory chain, utilizes the reduction of dioxygen into water to pump protons across the mitochondrial inner membrane. The principal pathway of proton uptake into the enzyme, the D channel, is a 2.5 nm long channel-like cavity named after a conserved, negatively charged aspartic acid (D) residue thought to help recruiting protons to its entrance (D132 in the first subunit of the S. sphaeroides enzyme). The single-point mutation of D132 to asparagine (N), a neutral residue, abolishes enzyme activity. Conversely, replacing conserved N139, one-third into the D channel, by D, induces a decoupled phenotype, whereby oxygen reduction proceeds but not proton pumping. Intriguingly, the double mutant D132N/N139D, which conserves the charge of the D channel, restores the wild-type phenotype. We use molecular dynamics simulations and electrostatic calculations to examine the structural and physical basis for the coupling of proton pumping and oxygen chemistry in single and double N139D mutants. The potential of mean force for the conformational isomerization of N139 and N139D side chains reveals the presence of three rotamers, one of which faces the channel entrance. This out-facing conformer is metastable in the wild-type and in the N139D single mutant, but predominant in the double mutant thanks to the loss of electrostatic repulsion with the carboxylate group of D132. The effects of mutations and conformational isomerization on the pKa of E286, an essential proton-shuttling residue located at the top of the D channel, are shown to be consistent with the electrostatic control of proton pumping proposed recently (Fadda et al 2008 Biochim. Biophys. Acta 1777 277-84). Taken together, these results suggest that preserving the spatial distribution of charges at the entrance of the D channel is necessary to guarantee both the uptake and the relay of protons to the active site of the enzyme. These findings highlight the interplay

  6. Biochemical Analysis of Two Single Mutants that Give Rise to a Polymorphic G6PD A-Double Mutant

    Directory of Open Access Journals (Sweden)

    Edson Jiovany Ramírez-Nava

    2017-10-01

    Full Text Available Glucose-6-phosphate dehydrogenase (G6PD is a key regulatory enzyme that plays a crucial role in the regulation of cellular energy and redox balance. Mutations in the gene encoding G6PD cause the most common enzymopathy that drives hereditary nonspherocytic hemolytic anemia. To gain insights into the effects of mutations in G6PD enzyme efficiency, we have investigated the biochemical, kinetic, and structural changes of three clinical G6PD variants, the single mutations G6PD A+ (Asn126AspD and G6PD Nefza (Leu323Pro, and the double mutant G6PD A− (Asn126Asp + Leu323Pro. The mutants showed lower residual activity (≤50% of WT G6PD and displayed important kinetic changes. Although all Class III mutants were located in different regions of the three-dimensional structure of the enzyme and were not close to the active site, these mutants had a deleterious effect over catalytic activity and structural stability. The results indicated that the G6PD Nefza mutation was mainly responsible for the functional and structural alterations observed in the double mutant G6PD A−. Moreover, our study suggests that the G6PD Nefza and G6PD A− mutations affect enzyme functions in a similar fashion to those reported for Class I mutations.

  7. Biochemical Analysis of Two Single Mutants that Give Rise to a Polymorphic G6PD A-Double Mutant

    Science.gov (United States)

    Ramírez-Nava, Edson Jiovany; González-Valdez, Abigail; Vanoye-Carlo, America; Hernández-Ochoa, Beatriz; Sierra-Palacios, Edgar; Hernández-Pineda, Jessica; Rodríguez-Bustamante, Eduardo; Arreguin-Espinosa, Roberto; Oria-Hernández, Jesús; Reyes-Vivas, Horacio; Marcial-Quino, Jaime

    2017-01-01

    Glucose-6-phosphate dehydrogenase (G6PD) is a key regulatory enzyme that plays a crucial role in the regulation of cellular energy and redox balance. Mutations in the gene encoding G6PD cause the most common enzymopathy that drives hereditary nonspherocytic hemolytic anemia. To gain insights into the effects of mutations in G6PD enzyme efficiency, we have investigated the biochemical, kinetic, and structural changes of three clinical G6PD variants, the single mutations G6PD A+ (Asn126AspD) and G6PD Nefza (Leu323Pro), and the double mutant G6PD A− (Asn126Asp + Leu323Pro). The mutants showed lower residual activity (≤50% of WT G6PD) and displayed important kinetic changes. Although all Class III mutants were located in different regions of the three-dimensional structure of the enzyme and were not close to the active site, these mutants had a deleterious effect over catalytic activity and structural stability. The results indicated that the G6PD Nefza mutation was mainly responsible for the functional and structural alterations observed in the double mutant G6PD A−. Moreover, our study suggests that the G6PD Nefza and G6PD A− mutations affect enzyme functions in a similar fashion to those reported for Class I mutations. PMID:29072585

  8. Improving the Patron Experience: Sterling Memorial Library's Single Service Point

    Science.gov (United States)

    Sider, Laura Galas

    2016-01-01

    This article describes the planning process and implementation of a single service point at Yale University's Sterling Memorial Library. While much recent scholarship on single service points (SSPs) has focused on the virtues or hazards of eliminating reference desks in libraries nationwide, this essay explores the ways in which single service…

  9. Transistors - From Point Contact to Single Electron

    Indian Academy of Sciences (India)

    development of the energy band theory of solids by Wilson. ... Davydov developed bis theory independently. All these theories had shortcomings as pointed out by Sab. The correct theory of metal-semiconductor diodes based on thermionic emission of majority carriers .... watches and other portable consumer applications.

  10. Single and multispecies reference points for Baltic fish stocks

    DEFF Research Database (Denmark)

    Gislason, Henrik

    1999-01-01

    Single and multispecies models are used to examine the effect of species interaction on biological reference points for cod, herring, and sprat in the Baltic. The results demonstrate that reference points are different in single and multispecies contexts. Reference points for fishing mortality...... based on single-species yield and SSB calculations are difficult to use when natural mortality depends on the absolute abundance of the predators and their alternative prey. Reference points based on maximizing total yield from the system may lead to impractical results when species interact...... reference points. Management advice based on biomass reference points will also differ. In the single species situation the combinations of cod and pelagic fishing effort for which the equilibrium spawning- stock biomass of the three species is above the biomass reference points forms a rectangular area...

  11. Program computes single-point failures in critical system designs

    Science.gov (United States)

    Brown, W. R.

    1967-01-01

    Computer program analyzes the designs of critical systems that will either prove the design is free of single-point failures or detect each member of the population of single-point failures inherent in a system design. This program should find application in the checkout of redundant circuits and digital systems.

  12. Creating Helical Tool Paths for Single Point Incremental Forming

    DEFF Research Database (Denmark)

    Skjødt, Martin; Hancock, Michael H.; Bay, Niels

    2007-01-01

    Single point incremental forming (SPIF) is a relatively new sheet forming process. A sheet is clamped in a rig and formed incrementally using a rotating single point tool in the form of a rod with a spherical end. The process is often performed on a CNC milling machine and the tool movement...

  13. A single quantum dot-based biosensor for DNA point mutation assay.

    Science.gov (United States)

    Tang, Wei; Zhu, Guichi; Liang, Li; Zhang, Chun-Yang

    2015-09-07

    Sensitive and selective detection of point mutation is essential to molecular biology research and early clinical diagnosis. Here, we demonstrate a single quantum dot (QD)-based biosensor for DNA point mutation assay. In this assay, a mutant target (G/C) remains unchanged after the endonuclease treatment, and the polymerase chain reaction (PCR) may be initiated with the assistance of primers and polymerase, generating a large number of mutant targets. The amplified mutant targets can be captured by biotinylated probes during the process of denaturation and annealing, and Cy5-dGTP may be assembled into the biotinylated probe with the catalysis of polymerase, leading to the formation of Cy5-labeled biotinylated probes. The Cy5-labeled biotinylated probes can be further assembled onto the QD surface to obtain a Cy5-DNA-QD complex, resulting in the generation of fluorescence resonance energy transfer (FRET) between the QD donor and the Cy5 receptor. The mutant targets can be quantitatively evaluated by the measurement of Cy5 counts by total internal reflection fluorescence (TIRF) microscopy. While in the presence of wild-type targets (T/A), no Cy5-dGTP can be assembled into the biotinylated probe due to the presence of a mismatch and consequently no FRET is observed. This single QD-based biosensor exhibits high sensitivity with a detection limit of 5.3 aM (or 32 copies) and can even discriminate as low as 0.01% variant frequency from the mixture of mutant targets and wild-type ones. Importantly, this biosensor can be used for genomic analysis in human lung cancer cells, and may be further applied for an early clinical diagnosis and personalized medicine.

  14. Point defects and electric compensation in gallium arsenide single crystals

    International Nuclear Information System (INIS)

    Kretzer, Ulrich

    2007-01-01

    In the present thesis the point-defect budget of gallium arsenide single crystals with different dopings is studied. It is shown, in which way the concentration of the single point defects depende on the concentration of the dopants, the stoichiometry deviation, and the position of the Fermi level. For this serve the results of the measurement-technical characterization of a large number of samples, in the fabrication of which these parameters were directedly varied. The main topic of this thesis lies in the development of models, which allow a quantitative description of the experimentally studied electrical and optical properties of gallium arsenide single crystals starting from the point-defect concentrations. Because from point defects charge carriers can be set free, their concentration determines essentially the charge-carrier concentration in the bands. In the ionized state point defects act as scattering centers for free charge carriers and influence by this the drift mobility of the charge carriers. A thermodynamic modeling of the point-defect formation yields statements on the equilibrium concentrations of the point defects in dependence on dopant concentration and stoichiometry deviation. It is show that the electrical properties of the crystals observed at room temperature result from the kinetic suppression of processes, via which the adjustment of a thermodynamic equilibrium between the point defects is mediated [de

  15. DNA deformability changes of single base pair mutants within CDE binding sites in S. Cerevisiae centromere DNA correlate with measured chromosomal loss rates and CDE binding site symmetries

    Directory of Open Access Journals (Sweden)

    Marx Kenneth A

    2006-03-01

    Full Text Available Abstract Background The centromeres in yeast (S. cerevisiae are organized by short DNA sequences (125 bp on each chromosome consisting of 2 conserved elements: CDEI and CDEIII spaced by a CDEII region. CDEI and CDEIII are critical sequence specific protein binding sites necessary for correct centromere formation and following assembly with proteins, are positioned near each other on a specialized nucleosome. Hegemann et al. BioEssays 1993, 15: 451–460 reported single base DNA mutants within the critical CDEI and CDEIII binding sites on the centromere of chromosome 6 and quantitated centromere loss of function, which they measured as loss rates for the different chromosome 6 mutants during cell division. Olson et al. Proc Natl Acad Sci USA 1998, 95: 11163–11168 reported the use of protein-DNA crystallography data to produce a DNA dinucleotide protein deformability energetic scale (PD-scale that describes local DNA deformability by sequence specific binding proteins. We have used the PD-scale to investigate the DNA sequence dependence of the yeast chromosome 6 mutants' loss rate data. Each single base mutant changes 2 PD-scale values at that changed base position relative to the wild type. In this study, we have utilized these mutants to demonstrate a correlation between the change in DNA deformability of the CDEI and CDEIII core sites and the overall experimentally measured chromosome loss rates of the chromosome 6 mutants. Results In the CDE I and CDEIII core binding regions an increase in the magnitude of change in deformability of chromosome 6 single base mutants with respect to the wild type correlates to an increase in the measured chromosome loss rate. These correlations were found to be significant relative to 105 Monte Carlo randomizations of the dinucleotide PD-scale applied to the same calculation. A net loss of deformability also tends to increase the loss rate. Binding site position specific, 4 data-point correlations were also

  16. Single-Molecule Counting of Point Mutations by Transient DNA Binding

    Science.gov (United States)

    Su, Xin; Li, Lidan; Wang, Shanshan; Hao, Dandan; Wang, Lei; Yu, Changyuan

    2017-03-01

    High-confidence detection of point mutations is important for disease diagnosis and clinical practice. Hybridization probes are extensively used, but are hindered by their poor single-nucleotide selectivity. Shortening the length of DNA hybridization probes weakens the stability of the probe-target duplex, leading to transient binding between complementary sequences. The kinetics of probe-target binding events are highly dependent on the number of complementary base pairs. Here, we present a single-molecule assay for point mutation detection based on transient DNA binding and use of total internal reflection fluorescence microscopy. Statistical analysis of single-molecule kinetics enabled us to effectively discriminate between wild type DNA sequences and single-nucleotide variants at the single-molecule level. A higher single-nucleotide discrimination is achieved than in our previous work by optimizing the assay conditions, which is guided by statistical modeling of kinetics with a gamma distribution. The KRAS c.34 A mutation can be clearly differentiated from the wild type sequence (KRAS c.34 G) at a relative abundance as low as 0.01% mutant to WT. To demonstrate the feasibility of this method for analysis of clinically relevant biological samples, we used this technology to detect mutations in single-stranded DNA generated from asymmetric RT-PCR of mRNA from two cancer cell lines.

  17. Structure Based Thermostability Prediction Models for Protein Single Point Mutations with Machine Learning Tools.

    Directory of Open Access Journals (Sweden)

    Lei Jia

    Full Text Available Thermostability issue of protein point mutations is a common occurrence in protein engineering. An application which predicts the thermostability of mutants can be helpful for guiding decision making process in protein design via mutagenesis. An in silico point mutation scanning method is frequently used to find "hot spots" in proteins for focused mutagenesis. ProTherm (http://gibk26.bio.kyutech.ac.jp/jouhou/Protherm/protherm.html is a public database that consists of thousands of protein mutants' experimentally measured thermostability. Two data sets based on two differently measured thermostability properties of protein single point mutations, namely the unfolding free energy change (ddG and melting temperature change (dTm were obtained from this database. Folding free energy change calculation from Rosetta, structural information of the point mutations as well as amino acid physical properties were obtained for building thermostability prediction models with informatics modeling tools. Five supervised machine learning methods (support vector machine, random forests, artificial neural network, naïve Bayes classifier, K nearest neighbor and partial least squares regression are used for building the prediction models. Binary and ternary classifications as well as regression models were built and evaluated. Data set redundancy and balancing, the reverse mutations technique, feature selection, and comparison to other published methods were discussed. Rosetta calculated folding free energy change ranked as the most influential features in all prediction models. Other descriptors also made significant contributions to increasing the accuracy of the prediction models.

  18. Single-point incremental forming and formability-failure diagrams

    DEFF Research Database (Denmark)

    Silva, M.B.; Skjødt, Martin; Atkins, A.G.

    2008-01-01

    In a recent work [1], the authors constructed a closed-form analytical model that is capable of dealing with the fundamentals of single point incremental forming and explaining the experimental and numerical results published in the literature over the past couple of years. The model is based on ...

  19. Transistors-From Point Contact to Single Electron

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education; Volume 2; Issue 12. Transistors – From Point Contact to Single Electron. D N Bose. General Article Volume 2 Issue 12 December 1997 pp 39-54. Fulltext. Click here to view fulltext PDF. Permanent link: http://www.ias.ac.in/article/fulltext/reso/002/12/0039-0054 ...

  20. Revisiting the fundamentals of single point incremental forming by

    DEFF Research Database (Denmark)

    Silva, Beatriz; Skjødt, Martin; Martins, Paulo A.F.

    2008-01-01

    Knowledge of the physics behind the fracture of material at the transition between the inclined wall and the corner radius of the sheet is of great importance for understanding the fundamentals of single point incremental forming (SPIF). How the material fractures, what is the state of strain and...

  1. Common fixed points of single-valued and multivalued maps

    Directory of Open Access Journals (Sweden)

    Yicheng Liu

    2005-01-01

    Full Text Available We define a new property which contains the property (EA for a hybrid pair of single- and multivalued maps and give some new common fixed point theorems under hybrid contractive conditions. Our results extend previous ones. As an application, we give a partial answer to the problem raised by Singh and Mishra.

  2. Laser-induced single point nanowelding of silver nanowires

    Energy Technology Data Exchange (ETDEWEB)

    Dai, Shuowei; Li, Qiang, E-mail: qiangli@zju.edu.cn; Liu, Guoping; Yang, Hangbo; Yang, Yuanqing; Zhao, Ding; Wang, Wei; Qiu, Min, E-mail: minqiu@zju.edu.cn [State Key Laboratory of Modern Optical Instrumentation, College of Optical Science and Engineering, Zhejiang University, Hangzhou 310027 (China)

    2016-03-21

    Nanowelding of nanomaterials opens up an emerging set of applications in transparent conductors, thin-film solar cells, nanocatalysis, cancer therapy, and nanoscale patterning. Single point nanowelding (SPNW) is highly demanded for building complex nanostructures. In this letter, the precise control of SPNW of silver nanowires is explored in depth, where the nanowelding is laser-induced through the plasmonic resonance enhanced photothermal effect. It is shown that the illumination position is a critical factor for the nanowelding process. As an example of performance enhancement, output at wire end can be increased by 65% after welding for a plasmonic nanocoupler. Thus, single point nanowelding technique shows great potentials for high-performance electronic and photonic devices based on nanowires, such as nanoelectronic circuits and plasmonic nanodevices.

  3. Creating Helical Tool Paths for Single Point Incremental Forming

    DEFF Research Database (Denmark)

    Skjødt, Martin; Hancock, Michael H.; Bay, Niels

    2007-01-01

    Single point incremental forming (SPIF) is a relatively new sheet forming process. A sheet is clamped in a rig and formed incrementally using a rotating single point tool in the form of a rod with a spherical end. The process is often performed on a CNC milling machine and the tool movement...... is programed using CAM software intended for surface milling. Often the function called profile milling or contour milling is applied. Using this milling function the tool only has a continuous feed rate in two directions X and Y, which is the plane of the undeformed sheet. The feed in the vertical Z direction...... from the profile milling code and converts them into a helical tool path with continuous feed in all three directions. Using the helical tool path the scarring is removed, the part is otherwise unchanged and a major disadvantage of using milling software for SPIF is removed. The solution...

  4. 3D Point Cloud Reconstruction from Single Plenoptic Image

    Directory of Open Access Journals (Sweden)

    F. Murgia

    2016-06-01

    Full Text Available Novel plenoptic cameras sample the light field crossing the main camera lens. The information available in a plenoptic image must be processed, in order to create the depth map of the scene from a single camera shot. In this paper a novel algorithm, for the reconstruction of 3D point cloud of the scene from a single plenoptic image, taken with a consumer plenoptic camera, is proposed. Experimental analysis is conducted on several test images, and results are compared with state of the art methodologies. The results are very promising, as the quality of the 3D point cloud from plenoptic image, is comparable with the quality obtained with current non-plenoptic methodologies, that necessitate more than one image.

  5. Laser-induced single point nanowelding of silver nanowires

    International Nuclear Information System (INIS)

    Dai, Shuowei; Li, Qiang; Liu, Guoping; Yang, Hangbo; Yang, Yuanqing; Zhao, Ding; Wang, Wei; Qiu, Min

    2016-01-01

    Nanowelding of nanomaterials opens up an emerging set of applications in transparent conductors, thin-film solar cells, nanocatalysis, cancer therapy, and nanoscale patterning. Single point nanowelding (SPNW) is highly demanded for building complex nanostructures. In this letter, the precise control of SPNW of silver nanowires is explored in depth, where the nanowelding is laser-induced through the plasmonic resonance enhanced photothermal effect. It is shown that the illumination position is a critical factor for the nanowelding process. As an example of performance enhancement, output at wire end can be increased by 65% after welding for a plasmonic nanocoupler. Thus, single point nanowelding technique shows great potentials for high-performance electronic and photonic devices based on nanowires, such as nanoelectronic circuits and plasmonic nanodevices.

  6. Determining ERβ Binding Affinity to Singly Mutant ERE Using Dual Polarization Interferometry

    Science.gov (United States)

    Song, Hong Yan; Su, Xiaodi

    In a classic mode of estrogen action, estrogen receptors (ERs) bind to estrogen responsive element (ERE) to activate gene transcription. A perfect ERE contains a 13-base pair sequence of a palindromic repeat separated by a three-base spacer, 5‧-GGTCAnnnTGACC-3‧. In addition to the consensus or wild-type ERE (wtERE), naturally occurring EREs often have one or two base pairs’ alternation. Based on the newly constructed Thermodynamic Modeling of ChIP-seq (TherMos) model, binding energy between ERβ and a series of 34-bp mutant EREs (mutERE) was simulated to predict the binding affinity between ERs and EREs with single base pair deviation at different sites of the 13-bp inverted sequence. Experimentally, dual polarization interferometry (DPI) method was developed to measure ERβ-mutEREs binding affinity. On a biotin-NeutrAvidin (NA)-biotin treated DPI chip, wtERE is immobilized. In a direct binding assay, ERβ-wtERE binding affinity is determined. In a competition assay, ERβ was preincubated with mutant EREs before being added for competitive binding to the immobilized wtERE. This competition strategy provided a successful platform to evaluate the binding affinity variation among large number of ERE with different base mutations. The experimental result correlates well with the mathematically predicted binding energy with a Spearman correlation coefficient of 0.97.

  7. ODMR of single point defects in silicon nanostructures

    Energy Technology Data Exchange (ETDEWEB)

    Bagraev, Nikolay; Danilovsky, Eduard; Gets, Dmitry; Klyachkin, Leonid; Kudryavtsev, Andrey; Kuzmin, Roman; Malyarenko, Anna [Ioffe Physical-Technical Institute, Polytekhnicheskaya st. 26, 194021 St. Petersburg (Russian Federation)

    2012-05-15

    We present the findings of the optically detected magnetic resonance technique (ODMR), which reveal single point defects in the ultra-narrow silicon quantum wells (Si-QW) confined by the superconductor {delta}-barriers. This technique allows the ODMR identification without application of an external cavity, as well as a high frequency source and recorder, and with measuring the transmission spectra within the frameworks of the excitonic normal-mode coupling caused by the microcavities embedded in the Si-QW plane. (copyright 2012 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim) (orig.)

  8. Geometry of single-point turning tools and drills

    CERN Document Server

    Astakhov, Viktor P

    2010-01-01

    Tools for metal cutting have many shapes and features, each of which is described by its angles or geometries. The selection of the right cutting tool geometry is critical because it directly affects the integrity of the machined surface, tool life, power needed for machining, and thus the overall machining efficiency. ""Geometry of Single-Point Turning Tools and Drills"" outlines clear objectives of cutting tool geometry selection and optimization, using multiple examples to provide a thorough explanation. The establishment of clear bridges between cutting theory, tool geometry, and shop prac

  9. Testing single point incremental forming molds for thermoforming operations

    Science.gov (United States)

    Afonso, Daniel; de Sousa, Ricardo Alves; Torcato, Ricardo

    2016-10-01

    Low pressure polymer processing processes as thermoforming or rotational molding use much simpler molds then high pressure processes like injection. However, despite the low forces involved with the process, molds manufacturing for this operations is still a very material, energy and time consuming operation. The goal of the research is to develop and validate a method for manufacturing plastically formed sheets metal molds by single point incremental forming (SPIF) operation for thermoforming operation. Stewart platform based SPIF machines allow the forming of thick metal sheets, granting the required structural stiffness for the mold surface, and keeping the short lead time manufacture and low thermal inertia.

  10. Point defects in electron-irradiated oxide single crystals

    Energy Technology Data Exchange (ETDEWEB)

    Caulfield, K.J.; Cooper, R. [Univ. of Melbourne, Parkville, Victoria (Australia). Dept. of Chemistry; Boas, J.F. [Australian Radiation Lab., Yallambie, Victoria (Australia)

    1995-04-01

    Point defects have been produced in CaO, MgO, and {alpha}-Al{sub 2}O{sub 3} single crystals by electron irradiation, and thresholds for atomic displacement have been measured using time-resolved luminescence spectroscopy. Oxygen displacement energies of approximately 50 eV have been determined; however, a temperature-dependent threshold observed for an emission band in MgO may arise from a magnesium displacement. A 300-nm emission in {alpha}-Al{sub 2}O{sub 3} may be due to an F-center transition. Studies of radiative recombination kinetics are consistent with an electron-detrapping model.

  11. Effective screen of CRISPR/Cas9-induced mutants in rice by single-strand conformation polymorphism.

    Science.gov (United States)

    Zheng, Xuelian; Yang, Shixin; Zhang, Dengwei; Zhong, Zhaohui; Tang, Xu; Deng, Kejun; Zhou, Jianping; Qi, Yiping; Zhang, Yong

    2016-07-01

    A method based on DNA single-strand conformation polymorphism is demonstrated for effective genotyping of CRISPR/Cas9-induced mutants in rice. Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) has been widely adopted for genome editing in many organisms. A large proportion of mutations generated by CRISPR/Cas9 are very small insertions and deletions (indels), presumably because Cas9 generates blunt-ended double-strand breaks which are subsequently repaired without extensive end-processing. CRISPR/Cas9 is highly effective for targeted mutagenesis in the important crop, rice. For example, homozygous mutant seedlings are commonly recovered from CRISPR/Cas9-treated calli. However, many current mutation detection methods are not very suitable for screening homozygous mutants that typically carry small indels. In this study, we tested a mutation detection method based on single-strand conformational polymorphism (SSCP). We found it can effectively detect small indels in pilot experiments. By applying the SSCP method for CRISRP-Cas9-mediated targeted mutagenesis in rice, we successfully identified multiple mutants of OsROC5 and OsDEP1. In conclusion, the SSCP analysis will be a useful genotyping method for rapid identification of CRISPR/Cas9-induced mutants, including the most desirable homozygous mutants. The method also has high potential for similar applications in other plant species.

  12. Failure mechanisms in single-point incremental forming of metals

    DEFF Research Database (Denmark)

    Silva, Maria B.; Nielsen, Peter Søe; Bay, Niels

    2011-01-01

    The last years saw the development of two different views on how failure develops in single-point incremental forming (SPIF). Today, researchers are split between those claiming that fracture is always preceded by necking and those considering that fracture occurs with suppression of necking. Eac...... and involves independent determination of formability limits by necking and fracture using tensile and hydraulic bulge tests in conjunction with SPIF of benchmark shapes under laboratory conditions.......The last years saw the development of two different views on how failure develops in single-point incremental forming (SPIF). Today, researchers are split between those claiming that fracture is always preceded by necking and those considering that fracture occurs with suppression of necking. Each...... on formability limits and development of fracture. The unified view conciliates the aforementioned different explanations on the role of necking in fracture and is consistent with the experimental observations that have been reported in the past years. The work is performed on aluminium AA1050-H111 sheets...

  13. Securing Single Points of Compromise (SPoC)

    Energy Technology Data Exchange (ETDEWEB)

    Belangia, David Warren [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

    2015-06-25

    Securing the Single Points of Compromise that provide central services to the institution’s environment is paramount to success when trying to protect the business. (Fisk, 2014) Time Based Security mandates protection (erecting and ensuring effective controls) that last longer than the time to detect and react to a compromise. When enterprise protections fail, providing additional layered controls for these central services provides more time to detect and react. While guidance is readily available for securing the individual critical asset, protecting these assets as a group is not often discussed. Using best business practices to protect these resources as individual assets while leveraging holistic defenses for the group increases the opportunity to maximize protection time, allowing detection and reaction time for the SPoCs that is commensurate with the inherent risk of these centralized services.

  14. The Analysis of Forming Forces in Single Point Incremental Forming

    Directory of Open Access Journals (Sweden)

    Koh Kyung Hee

    2016-01-01

    Full Text Available Incremental forming is a process to produce sheet metal parts in quick. Because there is no need for dedicated dies and molds, this process is less cost and time spent. The purpose of this study is to investigate forming forces in single point incremental forming. Producing a cone frustum of aluminum is tested for forming forces. A dynamometer is used to collect forming forces and analyze them. These forces are compared with cutting forces upon producing same geometrical shapes of experimental parts. The forming forces in Z direction are 40 times larger than the machining forces. A spindle and its axis of a forming machine should be designed enough to withstand the forming forces.

  15. Code Single Point Positioning Using Nominal GNSS Constellations (Future Perception)

    Science.gov (United States)

    Farah, A. M. A.

    Global Navigation Satellite Systems (GNSS) have an endless number of applications in industry, science, military, transportation and recreation & sports. Two systems are currently in operation, namely GPS (the USA Global Positioning System) and GLONASS (the Russian GLObal NAvigation Satellite System), and a third is planned, the European satellite navigation system GALILEO. The potential performance improvements achievable through combining these systems could be significant and expectations are high. The need is inevitable to explore the future of positioning from different nominal constellations. In this research paper, Bernese 5.0 software could be modified to simulate and process GNSS observations from three different constellations (GPS, GLONASS and Galileo) using different combinations. This study presents results of code single point positioning for five stations using the three constellations and different combinations.

  16. Single Point Incremental Forming using a Dummy Sheet

    DEFF Research Database (Denmark)

    Skjødt, Martin; Silva, Beatriz; Bay, Niels

    2007-01-01

    A new version of single point incremental forming (SPIF) is presented. This version includes a dummy sheet on top of the work piece, thus forming two sheets instead of one. The dummy sheet, which is in contact with the rotating tool pin, is discarded after forming. The new set-up influences...... the process and furthermore offers a number of new possibilities for solving some of the problems appearing in SPIF. Investigations of the influence of dummy sheet on: formability, wear, surface quality and bulging of planar sides is done by forming to test shapes: a hyperboloid and a truncated pyramid....... The possible influence of friction between the two sheets is furthermore investigated. The results show that the use of a dummy sheet reduces wear of the work piece to almost zero, but also causes a decrease in formability. Bulging of the planar sides of the pyramid is reduced and surface roughness...

  17. Single supplier single retailer inventory model controlled by the reorder and shipping points with sharing information

    Science.gov (United States)

    Ho, Wen-Tsung; Hsiao, Yu-Cheng

    2012-04-01

    This study investigated the integrated stochastic inventory problem for a two-stage supply chain consisting of a single retailer and a single supplier. By using batch shipment policy, the expected total cost can be significantly reduced. An equally sized batch shipment model, controlled by both the reorder and shipping points, with sharing information by enterprise resource planning and radio frequency identification is constructed. The problem is solved optimally by the proposed algorithm that determines the economic lot size, the optimal batch sizes and number of batches. A numerical example is included to illustrate the algorithmic procedures and to prove that the model controlled both by the reorder and shipping points is superior to the classic model controlled only by the reorder point.

  18. Surface chemical functionalization of single walled carbon nanotubes with a bacteriorhodopsin mutant.

    Science.gov (United States)

    Ingrosso, Chiara; Bianco, Giuseppe Valerio; Lopalco, Patrizia; Tamborra, Michela; Curri, Maria Lucia; Corcelli, Angela; Bruno, Giovanni; Agostiano, Angela; Siciliano, Pietro; Striccoli, Marinella

    2012-10-21

    In this work, single walled carbon nanotubes (SWNTs) have been chemically functionalized at their walls with a membrane protein, namely the mutated bacteriorhodopsin D96N, integrated in its native archaeal lipid membrane. The modification of the SWNT walls with the mutant has been carried out in different buffer solutions, at pH 5, 7.5 and 9, to investigate the anchoring process, the typical chemical and physical properties of the component materials being dependent on the pH. The SWNTs modified by interactions with bacteriorhodopsin membrane patches have been characterized by UV-vis steady state, Raman and attenuated total reflection Fourier transform infrared spectroscopy and by atomic force and transmission electron microscopy. The investigation shows that the membrane protein patches wrap the carbon walls by tight chemical interactions undergoing a conformational change; such chemical interactions increase the mechanical strength of the SWNTs and promote charge transfers which p-dope the nano-objects. The functionalization, as well as the SWNT doping, is favoured in acid and basic buffer conditions; such buffers make the nanotube walls more reactive, thus catalysing the anchoring of the membrane protein. The direct electron communication among the materials can be exploited for effectively interfacing the transport properties of carbon nanotubes with both molecular recognition capability and photoactivity of the cell membrane for sensing and photoconversion applications upon integration of the achieved hybrid materials in sensors or photovoltaic devices.

  19. Exposure of Salmonella enterica serovar Typhimurium to high level biocide challenge can select multidrug resistant mutants in a single step.

    Directory of Open Access Journals (Sweden)

    Rebekah N Whitehead

    Full Text Available Biocides are crucial to the prevention of infection by bacteria, particularly with the global emergence of multiply antibiotic resistant strains of many species. Concern has been raised regarding the potential for biocide exposure to select for antibiotic resistance due to common mechanisms of resistance, notably efflux.Salmonella enterica serovar Typhimurium was challenged with 4 biocides of differing modes of action at both low and recommended-use concentration. Flow cytometry was used to investigate the physiological state of the cells after biocide challenge. After 5 hours exposure to biocide, live cells were sorted by FACS and recovered. Cells recovered after an exposure to low concentrations of biocide had antibiotic resistance profiles similar to wild-type cells. Live cells were recovered after exposure to two of the biocides at in-use concentration for 5 hours. These cells were multi-drug resistant and accumulation assays demonstrated an efflux phenotype of these mutants. Gene expression analysis showed that the AcrEF multidrug efflux pump was de-repressed in mutants isolated from high-levels of biocide.These data show that a single exposure to the working concentration of certain biocides can select for mutant Salmonella with efflux mediated multidrug resistance and that flow cytometry is a sensitive tool for identifying biocide tolerant mutants. The propensity for biocides to select for MDR mutants varies and this should be a consideration when designing new biocidal formulations.

  20. Expression and purification of single cysteine-containing mutant variants of the mouse prion protein by oxidative refolding.

    Science.gov (United States)

    Sengupta, Ishita; Udgaonkar, Jayant B

    2017-12-01

    The folding and aggregation of proteins has been studied extensively, using multiple probes. To facilitate such experiments, introduction of spectroscopically-active moieties in to the protein of interest is often necessary. This is commonly achieved by specifically labelling cysteine residues in the protein, which are either present naturally or introduced artificially by site-directed mutagenesis. In the case of the recombinant prion protein, which is normally expressed in inclusion bodies, the presence of the native disulfide bond complicates the correct refolding of single cysteine-containing mutant variants of the protein. To overcome this major bottleneck, a simple purification strategy for single tryptophan, single cysteine-containing mutant variants of the mouse prion protein is presented, with yields comparable to that of the wild type protein. The protein(s) obtained by this method are correctly folded, with a single reduced cysteine, and the native disulfide bond between residues C178 and C213 intact. The β-sheet rich oligomers formed from these mutant variant protein(s) are identical to the wild type protein oligomer. The current strategy facilitates sample preparation for a number of high resolution spectroscopic measurements for the prion protein, which specifically require thiol labelling. Copyright © 2017 Elsevier Inc. All rights reserved.

  1. Single-frequency, single-receiver terrestrial and spaceborne point positioning

    Science.gov (United States)

    Beran, Tomas

    High-accuracy, point positioning has been an attractive research topic in the GPS community for a number of years. The overall quality of precise point positioning results is also dependent on the quality of the GPS measurements and the user's processing software. Dual-frequency, geodetic-quality GPS receivers are routinely used both in static and kinematic applications for high-accuracy point positioning. However, use of low-cost, single-frequency GPS receivers in similar applications creates a challenge because of difficulty of handling the ionosphere, multipath and other measurement error sources. Potential use of such receivers to provide horizontal positioning accuracies of a few decimetres, and vertical accuracies of less than two metres, will be examined in this dissertation. Practical applications of post-processed, high-accuracy, single-frequency point positioning include a myriad of terrestrial and space-borne applications, where the size and cost of the GPS unit is an issue. The processing technique uses pseudorange and time-differenced carrier-phase measurements in a sequential least-squares filter. In developing the approach, different techniques were investigated. Ionospheric delay grid maps are used to remove the bulk of the ionospheric error, while tropospheric error is handled by a prediction model. Pseudorange multipath errors are mitigated by means of stochastic modelling and carrier-phase cycle slips are detected and corrupted measurements are removed in a quality-control algorithm. The technique was first tested on L1 measurements extracted from datasets from static, high-quality GPS receivers. Accuracies better than two-decimetres in horizontal components (northing and easting r.m.s.), and three-decimetre accuracies in the vertical component (up-component r.m.s.), were obtained. A test dataset from a stationary low-cost GPS receiver has been processed to demonstrate the difference in data quality. Positioning results obtained are worse than

  2. Infectious mutants of cassava latent virus generated in vivo from intact recombinant DNA clones containing single copies of the genome.

    Science.gov (United States)

    Stanley, J; Townsend, R

    1986-08-11

    Intact recombinant DNAs containing single copies of either component of the cassava latent virus genome can elicit infection when mechanically inoculated to host plants in the presence of the appropriate second component. Characterisation of infectious mutant progeny viruses, by analysis of virus-specific supercoiled DNA intermediates, indicates that most if not all of the cloning vector has been deleted, achieved at least in some cases by intermolecular recombination in vivo between DNAs 1 and 2. Significant rearrangements within the intergenic region of DNA 2, predominantly external to the common region, can be tolerated without loss of infectivity suggesting a somewhat passive role in virus multiplication for the sequences in question. Although packaging constraints might impose limits on the amount of DNA within geminate particles, isolation of an infectious coat protein mutant defective in virion production suggests that packaging is not essential for systemic spread of the viral DNA.

  3. Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection.

    Science.gov (United States)

    Baba, Tomoya; Ara, Takeshi; Hasegawa, Miki; Takai, Yuki; Okumura, Yoshiko; Baba, Miki; Datsenko, Kirill A; Tomita, Masaru; Wanner, Barry L; Mori, Hirotada

    2006-01-01

    We have systematically made a set of precisely defined, single-gene deletions of all nonessential genes in Escherichia coli K-12. Open-reading frame coding regions were replaced with a kanamycin cassette flanked by FLP recognition target sites by using a one-step method for inactivation of chromosomal genes and primers designed to create in-frame deletions upon excision of the resistance cassette. Of 4288 genes targeted, mutants were obtained for 3985. To alleviate problems encountered in high-throughput studies, two independent mutants were saved for every deleted gene. These mutants-the 'Keio collection'-provide a new resource not only for systematic analyses of unknown gene functions and gene regulatory networks but also for genome-wide testing of mutational effects in a common strain background, E. coli K-12 BW25113. We were unable to disrupt 303 genes, including 37 of unknown function, which are candidates for essential genes. Distribution is being handled via GenoBase (http://ecoli.aist-nara.ac.jp/).

  4. Growth and sporulation defects in Bacillus subtilis mutants with a single rrn operon can be suppressed by amplification of the rrn operon.

    Science.gov (United States)

    Yano, Koichi; Masuda, Kenta; Akanuma, Genki; Wada, Tetsuya; Matsumoto, Takashi; Shiwa, Yuh; Ishige, Taichiro; Yoshikawa, Hirofumi; Niki, Hironori; Inaoka, Takashi; Kawamura, Fujio

    2016-01-01

    The genome of Bacillus subtilis strain 168 encodes ten rRNA (rrn) operons. We previously reported that strains with only a single rrn operon had a decreased growth and sporulation frequency. We report here the isolation and characterization of suppressor mutants from seven strains that each have a single rrn operon (rrnO, A, J, I, E, D or B). The suppressor mutants for strain RIK656 with a single rrnO operon had a higher frequency of larger colonies. These suppressor mutants had not only increased growth rates, but also increased sporulation frequencies and ribosome levels compared to the parental mutant strain RIK656. Quantitative PCR analyses showed that all these suppressor mutants had an increased number of copies of the rrnO operon. Suppressor mutants were also isolated from the six other strains with single rrn operons (rrnA, J, I, E, D or B). Next generation and capillary sequencing showed that all of the suppressor mutants had tandem repeats of the chromosomal locus containing the remaining rrn operon (amplicon). These amplicons varied in size from approximately 9 to 179 kb. The amplifications were likely to be initiated by illegitimate recombination between non- or micro-homologous sequences, followed by unequal crossing-over during DNA replication. These results are consistent with our previous report that rrn operon copy number has a major role in cellular processes such as cell growth and sporulation.

  5. Phenotypic characterization and complementation analysis of Bacillus subtilis 6S RNA single and double deletion mutants.

    Science.gov (United States)

    Hoch, Philipp G; Burenina, Olga Y; Weber, Michael H W; Elkina, Daria A; Nesterchuk, Mikhail V; Sergiev, Petr V; Hartmann, Roland K; Kubareva, Elena A

    2015-10-01

    6S RNA, a global regulator of transcription in bacteria, binds to housekeeping RNA polymerase (RNAP) holoenzymes to competitively inhibit transcription from DNA promoters. Bacillus subtilis encodes two 6S RNA homologs whose differential functions are as yet unclear. We constructed derivative strains of B. subtilis PY79 lacking 6S-1 RNA (ΔbsrA), 6S-2 RNA (ΔbsrB) or both (ΔbsrAB) to study the physiological role of the two 6S RNAs. We observed two growth phenotypes of mutant strains: (i) accelerated decrease of optical density toward extended stationary phase and (ii) faster outgrowth from stationary phase under alkaline stress conditions (pH 9.8). The first phenotype was observed for bacteria lacking bsrA, and even more pronounced for ΔbsrAB bacteria, but not for those lacking bsrB. The magnitude of the second phenotype was relatively weak for ΔbsrB, moderate for ΔbsrA and again strongest for ΔbsrAB bacteria. Whereas ΔbsrAB bacteria complemented with bsrB or bsrA (strains ΔbsrAB + B and ΔbsrAB + A) mimicked the phenotypes of the ΔbsrA and ΔbsrB strains, respectively, complementation with the gene ssrS encoding Escherichia coli 6S RNA failed to cure the "low stationary optical density" phenotype of the double mutant, despite ssrS expression, in line with previous findings. Finally, proteomics (two-dimensional differential gel electrophoresis, 2D-DIGE) of B. subtilis 6S RNA deletion strains unveiled a set of proteins that were expressed at higher levels particularly during exponential growth and preferentially in mutant strains lacking 6S-2 RNA. Several of these proteins are involved in metabolism and stress responses. Copyright © 2015 Elsevier B.V. and Société française de biochimie et biologie Moléculaire (SFBBM). All rights reserved.

  6. Comparing Single-Point and Multi-point Calibration Methods in Modulated DSC

    Energy Technology Data Exchange (ETDEWEB)

    Van Buskirk, Caleb Griffith [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

    2017-06-14

    Heat capacity measurements for High Density Polyethylene (HDPE) and Ultra-high Molecular Weight Polyethylene (UHMWPE) were performed using Modulated Differential Scanning Calorimetry (mDSC) over a wide temperature range, -70 to 115 °C, with a TA Instruments Q2000 mDSC. The default calibration method for this instrument involves measuring the heat capacity of a sapphire standard at a single temperature near the middle of the temperature range of interest. However, this method often fails for temperature ranges that exceed a 50 °C interval, likely because of drift or non-linearity in the instrument's heat capacity readings over time or over the temperature range. Therefore, in this study a method was developed to calibrate the instrument using multiple temperatures and the same sapphire standard.

  7. Nanomolar oligomerization and selective co-aggregation of α-synuclein pathogenic mutants revealed by single-molecule fluorescence

    Science.gov (United States)

    Sierecki, Emma; Giles, Nichole; Bowden, Quill; Polinkovsky, Mark E.; Steinbeck, Janina; Arrioti, Nicholas; Rahman, Diya; Bhumkar, Akshay; Nicovich, Philip R.; Ross, Ian; Parton, Robert G.; Böcking, Till; Gambin, Yann

    2016-01-01

    Protein aggregation is a hallmark of many neurodegenerative diseases, notably Alzheimer’s and Parkinson’s disease. Parkinson’s disease is characterized by the presence of Lewy bodies, abnormal aggregates mainly composed of α-synuclein. Moreover, cases of familial Parkinson’s disease have been linked to mutations in α-synuclein. In this study, we compared the behavior of wild-type (WT) α-synuclein and five of its pathological mutants (A30P, E46K, H50Q, G51D and A53T). To this end, single-molecule fluorescence detection was coupled to cell-free protein expression to measure precisely the oligomerization of proteins without purification, denaturation or labelling steps. In these conditions, we could detect the formation of oligomeric and pre-fibrillar species at very short time scale and low micromolar concentrations. The pathogenic mutants surprisingly segregated into two classes: one group forming large aggregates and fibrils while the other tending to form mostly oligomers. Strikingly, co-expression experiments reveal that members from the different groups do not generally interact with each other, both at the fibril and monomer levels. Together, this data paints a completely different picture of α-synuclein aggregation, with two possible pathways leading to the development of fibrils. PMID:27892477

  8. Nanomolar oligomerization and selective co-aggregation of α-synuclein pathogenic mutants revealed by single-molecule fluorescence.

    Science.gov (United States)

    Sierecki, Emma; Giles, Nichole; Bowden, Quill; Polinkovsky, Mark E; Steinbeck, Janina; Arrioti, Nicholas; Rahman, Diya; Bhumkar, Akshay; Nicovich, Philip R; Ross, Ian; Parton, Robert G; Böcking, Till; Gambin, Yann

    2016-11-28

    Protein aggregation is a hallmark of many neurodegenerative diseases, notably Alzheimer's and Parkinson's disease. Parkinson's disease is characterized by the presence of Lewy bodies, abnormal aggregates mainly composed of α-synuclein. Moreover, cases of familial Parkinson's disease have been linked to mutations in α-synuclein. In this study, we compared the behavior of wild-type (WT) α-synuclein and five of its pathological mutants (A30P, E46K, H50Q, G51D and A53T). To this end, single-molecule fluorescence detection was coupled to cell-free protein expression to measure precisely the oligomerization of proteins without purification, denaturation or labelling steps. In these conditions, we could detect the formation of oligomeric and pre-fibrillar species at very short time scale and low micromolar concentrations. The pathogenic mutants surprisingly segregated into two classes: one group forming large aggregates and fibrils while the other tending to form mostly oligomers. Strikingly, co-expression experiments reveal that members from the different groups do not generally interact with each other, both at the fibril and monomer levels. Together, this data paints a completely different picture of α-synuclein aggregation, with two possible pathways leading to the development of fibrils.

  9. Responsiveness of Myofascial Trigger Points to Single and Multiple Trigger Point Release Massages: A Randomized, Placebo Controlled Trial.

    Science.gov (United States)

    Moraska, Albert F; Schmiege, Sarah J; Mann, John D; Butryn, Nathan; Krutsch, Jason P

    2017-09-01

    This study aimed to assess the effects of single and multiple massage treatments on pressure-pain threshold (PPT) at myofascial trigger points (MTrPs) in people with myofascial pain syndrome expressed as tension-type headache. Individuals (n = 62) with episodic or chronic tension-type headache were randomized to receive 12 twice-weekly 45-min massage or sham ultrasound sessions or wait-list control. Massage focused on trigger point release (ischemic compression) of MTrPs in the bilateral upper trapezius and suboccipital muscles. PPT was measured at MTrPs with a pressure algometer pre and post the first and final (12th) treatments. PPT increased across the study timeframe in all four muscle sites tested for massage, but not sham ultrasound or wait-list groups (P trigger points to myofascial pain; (2) Describe an effective treatment for decreasing tenderness of a myofascial trigger point; and (3) Discuss the relative values of single vs. multiple massage sessions on increasing pressure-pain thresholds at myofascial trigger points. Advanced ACCREDITATION: The Association of Academic Physiatrists is accredited by the Accreditation Council for Continuing Medical Education to provide continuing medical education for physicians.The Association of Academic Physiatrists designates this Journal-based CME activity for a maximum of 0.5 AMA PRA Category 1 Credit(s)™. Physicians should only claim credit commensurate with the extent of their participation in the activity.

  10. Comparison of Single-Point and Continuous Sampling Methods for Estimating Residential Indoor Temperature and Humidity.

    Science.gov (United States)

    Johnston, James D; Magnusson, Brianna M; Eggett, Dennis; Collingwood, Scott C; Bernhardt, Scott A

    2015-01-01

    Residential temperature and humidity are associated with multiple health effects. Studies commonly use single-point measures to estimate indoor temperature and humidity exposures, but there is little evidence to support this sampling strategy. This study evaluated the relationship between single-point and continuous monitoring of air temperature, apparent temperature, relative humidity, and absolute humidity over four exposure intervals (5-min, 30-min, 24-hr, and 12-days) in 9 northern Utah homes, from March-June 2012. Three homes were sampled twice, for a total of 12 observation periods. Continuous data-logged sampling was conducted in homes for 2-3 wks, and simultaneous single-point measures (n = 114) were collected using handheld thermo-hygrometers. Time-centered single-point measures were moderately correlated with short-term (30-min) data logger mean air temperature (r = 0.76, β = 0.74), apparent temperature (r = 0.79, β = 0.79), relative humidity (r = 0.70, β = 0.63), and absolute humidity (r = 0.80, β = 0.80). Data logger 12-day means were also moderately correlated with single-point air temperature (r = 0.64, β = 0.43) and apparent temperature (r = 0.64, β = 0.44), but were weakly correlated with single-point relative humidity (r = 0.53, β = 0.35) and absolute humidity (r = 0.52, β = 0.39). Of the single-point RH measures, 59 (51.8%) deviated more than ±5%, 21 (18.4%) deviated more than ±10%, and 6 (5.3%) deviated more than ±15% from data logger 12-day means. Where continuous indoor monitoring is not feasible, single-point sampling strategies should include multiple measures collected at prescribed time points based on local conditions.

  11. Single Point Adjustments: A New Definition with Examples. Acquisition Review Quarterly, Fall 2001

    National Research Council Canada - National Science Library

    Bachman, David

    2002-01-01

    .... A single point adjustment (SPA) is made when a contract's existing cost and/or schedule variances are set to zero and all the remaining work is replanned with the goal of completing the project on schedule and on budget...

  12. Glutamate Excitotoxicity Is Involved in the Induction of Paralysis in Mice after Infection by a Human Coronavirus with a Single Point Mutation in Its Spike Protein▿

    Science.gov (United States)

    Brison, Elodie; Jacomy, Hélène; Desforges, Marc; Talbot, Pierre J.

    2011-01-01

    Human coronaviruses (HCoV) are recognized respiratory pathogens, and some strains, including HCoV-OC43, can infect human neuronal and glial cells of the central nervous system (CNS) and activate neuroinflammatory mechanisms. Moreover, HCoV-OC43 is neuroinvasive, neurotropic, and neurovirulent in susceptible mice, where it induces chronic encephalitis. Herein, we show that a single point mutation in the viral spike (S) glycoprotein (Y241H), acquired during viral persistence in human neural cells, led to a hind-limb paralytic disease in infected mice. Inhibition of glutamate excitotoxicity using a 2-amino-3-(5-methyl-3-oxo-1,2-oxazol-4-yl)propranoic acid (AMPA) receptor antagonist (GYKI-52466) improved clinical scores related to the paralysis and motor disabilities in S mutant virus-infected mice, as well as protected the CNS from neuronal dysfunctions, as illustrated by restoration of the phosphorylation state of neurofilaments. Expression of the glial glutamate transporter GLT-1, responsible for glutamate homeostasis, was downregulated following infection, and GYKI-52466 also significantly restored its steady-state expression level. Finally, GYKI-52466 treatment of S mutant virus-infected mice led to reduced microglial activation, which may lead to improvement in the regulation of CNS glutamate homeostasis. Taken together, our results strongly suggest an involvement of excitotoxicity in the paralysis-associated neuropathology induced by an HCoV-OC43 mutant which harbors a single point mutation in its spike protein that is acquired upon persistent virus infection. PMID:21957311

  13. SEE Transient Response of Crane Interpoint Single Output Point of Load DC-DC Converters

    Science.gov (United States)

    Sanders, Anthony B.; Chen, Dakai; Kim, Hak S.; Phan, Anthony M.

    2011-01-01

    This study was undertaken to determine the single event effect and transient susceptibility of the Crane Interpoint Maximum Flexible Power (MFP) Single Output Point of Load DC/DC Converters for transient interruptions in the output signal and for destructive and non destructive events induced by exposing it to a heavy ion beam..

  14. Detection of kinetic change points in piece-wise linear single molecule motion

    Science.gov (United States)

    Hill, Flynn R.; van Oijen, Antoine M.; Duderstadt, Karl E.

    2018-03-01

    Single-molecule approaches present a powerful way to obtain detailed kinetic information at the molecular level. However, the identification of small rate changes is often hindered by the considerable noise present in such single-molecule kinetic data. We present a general method to detect such kinetic change points in trajectories of motion of processive single molecules having Gaussian noise, with a minimum number of parameters and without the need of an assumed kinetic model beyond piece-wise linearity of motion. Kinetic change points are detected using a likelihood ratio test in which the probability of no change is compared to the probability of a change occurring, given the experimental noise. A predetermined confidence interval minimizes the occurrence of false detections. Applying the method recursively to all sub-regions of a single molecule trajectory ensures that all kinetic change points are located. The algorithm presented allows rigorous and quantitative determination of kinetic change points in noisy single molecule observations without the need for filtering or binning, which reduce temporal resolution and obscure dynamics. The statistical framework for the approach and implementation details are discussed. The detection power of the algorithm is assessed using simulations with both single kinetic changes and multiple kinetic changes that typically arise in observations of single-molecule DNA-replication reactions. Implementations of the algorithm are provided in ImageJ plugin format written in Java and in the Julia language for numeric computing, with accompanying Jupyter Notebooks to allow reproduction of the analysis presented here.

  15. Single purse string with four-point traction for better haemorrhoid retraction.

    Science.gov (United States)

    Chen, Shao-Quan; Cai, Ai-Zhen; Wang, Ning; Chen, Lin

    2012-10-01

    Traction method is important to stapled haemorrhoidopexy (SH) for its impact on haemorrhoidal prolapse reduction. Single purse string with one-point traction is most commonly used in SH. This traction method often results in an irregular mucosal doughnut with inadequate height, which leads to insignificant and uneven haemorrhoidal prolapse reduction. Single purse string with two-point traction is a modified traction method. According to some authors, it has significantly improved the height and regularity of the mucosal doughnut in SH. However, the reduction of haemorrhoidal prolapse is not always satisfactory, especially in patients with large prolapse. Our assumption is that single purse string with four-point traction, a more balanced traction method, will provide better control to increase the doughnut height using the same amount of traction. This study was designed to evaluate the outcomes of single purse string with four-point traction as compared with two-point traction in SH. Consecutive patients with symptomatic grade III haemorrhoids were randomized to two groups: Group 1, G(1) ; Group 2, G(2) . Each group underwent SH by colorectal surgeons. The patients in G(1) underwent single purse string with two-point traction. The patients in G(2) underwent SH using single purse string with four-point traction. Preoperative, intraoperative and post-operative patient characteristics were evaluated. Sixty patients with a median age of 44 (range, 21-78) years were identified. Patient demographic and clinical characteristics were similar in the two groups. The height of the mucosectomy specimens significantly increased in patients of G(2) (P string with four-point traction ensures a taller and more regular mucosal doughnut during SH, thus it brings a more satisfactory haemorrhoid retraction. © 2012 The Authors. ANZ Journal of Surgery © 2012 Royal Australasian College of Surgeons.

  16. Dynamic fluorescence spectroscopy on single tryptophan mutants of EIImtl in detergent micelles : Effects of substrate binding and phosphorylation on the fluorescence and anisotropy decay

    NARCIS (Netherlands)

    Swaving Dijkstra, Dolf; Broos, J.; Visser, Antonie J.W.G.; van Hoek, A.; Robillard, George

    1997-01-01

    The effects of substrate and substrate analogue binding and phosphorylation on the conformational dynamics of the mannitol permease of Escherichia coli were investigated, using time-resolved fluorescence spectroscopy on mutants containing five single tryptophans situated in the membrane-embedded C

  17. Single chain variable fragment antibodies block aggregation and toxicity induced by familial ALS-linked mutant forms of SOD1.

    Science.gov (United States)

    Ghadge, Ghanashyam D; Pavlovic, John D; Koduvayur, Sujatha P; Kay, Brian K; Roos, Raymond P

    2013-08-01

    Approximately 10% of amyotrophic lateral sclerosis (ALS) cases are familial (known as FALS) with an autosomal dominant inheritance pattern, and ~25% of FALS cases are caused by mutations in Cu/Zn superoxide dismutase (SOD1). There is convincing evidence that mutant SOD1 (mtSOD1) kills motor neurons (MNs) because of a gain-of-function toxicity, most likely related to aggregation of mtSOD1. A number of recent reports have suggested that antibodies can be used to treat mtSOD1-induced FALS. To follow up on the use of antibodies as potential therapeutics, we generated single chain fragments of variable region antibodies (scFvs) against SOD1, and then expressed them as 'intrabodies' within a motor neuron cell line. In the present study, we describe isolation of human scFvs that interfere with mtSOD1 in vitro aggregation and toxicity. These scFvs may have therapeutic potential in sporadic ALS, as well as FALS, given that sporadic ALS may also involve abnormalities in the SOD1 protein or activity. Copyright © 2013 Elsevier Inc. All rights reserved.

  18. Conversion of a beta-strand to an alpha-helix induced by a single-site mutation observed in the crystal structure of Fis mutant Pro26Ala.

    OpenAIRE

    Yang, W. Z.; Ko, T. P.; Corselli, L.; Johnson, R. C.; Yuan, H. S.

    1998-01-01

    The conversion from an alpha-helix to a beta-strand has received extensive attention since this structural change may induce many amyloidogenic proteins to self-assemble into fibrils and cause fatal diseases. Here we report the conversion of a peptide segment from a beta-strand to an alpha-helix by a single-site mutation as observed in the crystal structure of Fis mutant Pro26Ala determined at 2.0 A resolution. Pro26 in Fis occurs at the point where a flexible extended beta-hairpin arm leaves...

  19. A broad phenotypic screen identifies novel phenotypes driven by a single mutant allele in Huntington's disease CAG knock-in mice.

    Directory of Open Access Journals (Sweden)

    Sabine M Hölter

    Full Text Available Huntington's disease (HD is an autosomal dominant neurodegenerative disorder caused by the expansion of a CAG trinucleotide repeat in the HTT gene encoding huntingtin. The disease has an insidious course, typically progressing over 10-15 years until death. Currently there is no effective disease-modifying therapy. To better understand the HD pathogenic process we have developed genetic HTT CAG knock-in mouse models that accurately recapitulate the HD mutation in man. Here, we describe results of a broad, standardized phenotypic screen in 10-46 week old heterozygous HdhQ111 knock-in mice, probing a wide range of physiological systems. The results of this screen revealed a number of behavioral abnormalities in HdhQ111/+ mice that include hypoactivity, decreased anxiety, motor learning and coordination deficits, and impaired olfactory discrimination. The screen also provided evidence supporting subtle cardiovascular, lung, and plasma metabolite alterations. Importantly, our results reveal that a single mutant HTT allele in the mouse is sufficient to elicit multiple phenotypic abnormalities, consistent with a dominant disease process in patients. These data provide a starting point for further investigation of several organ systems in HD, for the dissection of underlying pathogenic mechanisms and for the identification of reliable phenotypic endpoints for therapeutic testing.

  20. Estimating Total Heliospheric Magnetic Flux from Single-Point in Situ Measurements

    Science.gov (United States)

    Owens, M. J.; Arge, C. N.; Crooker, N. U.; Schwardron, N. A.; Horbury, T. S.

    2008-01-01

    A fraction of the total photospheric magnetic flux opens to the heliosphere to form the interplanetary magnetic field carried by the solar wind. While this open flux is critical to our understanding of the generation and evolution of the solar magnetic field, direct measurements are generally limited to single-point measurements taken in situ by heliospheric spacecraft. An observed latitude invariance in the radial component of the magnetic field suggests that extrapolation from such single-point measurements to total heliospheric magnetic flux is possible. In this study we test this assumption using estimates of total heliospheric flux from well-separated heliospheric spacecraft and conclude that single-point measurements are indeed adequate proxies for the total heliospheric magnetic flux, though care must be taken when comparing flux estimates from data collected at different heliocentric distances.

  1. The impact of a single-nucleotide mutation of bgl2 on cellulase induction in a Trichoderma reesei mutant.

    Science.gov (United States)

    Shida, Yosuke; Yamaguchi, Kaori; Nitta, Mikiko; Nakamura, Ayana; Takahashi, Machiko; Kidokoro, Shun-Ichi; Mori, Kazuki; Tashiro, Kosuke; Kuhara, Satoru; Matsuzawa, Tomohiko; Yaoi, Katsuro; Sakamoto, Yasumitsu; Tanaka, Nobutada; Morikawa, Yasushi; Ogasawara, Wataru

    2015-01-01

    The filamentous fungus Trichoderma reesei (anamorph of Hypocrea jecorina) produces increased cellulase expression when grown on cellulose or its derivatives as a sole carbon source. It has been believed that β-glucosidases of T. reesei not only metabolize cellobiose but also contribute in the production of inducers of cellulase gene expression by their transglycosylation activity. The cellulase hyper-producing mutant PC-3-7 developed in Japan has enhanced cellulase production ability when cellobiose is used as the inducer. The comparative genomics analysis of PC-3-7 and its parent revealed a single-nucleotide mutation within the bgl2 gene encoding intracellular β-glucosidase II (BGLII/Cel1a), giving rise to an amino acid substitution in PC-3-7, which could potentially account for the enhanced cellulase expression when these strains are cultivated on cellulose and cellobiose. To analyze the effects of the BGLII mutation in cellulase induction, we constructed both a bgl2 revertant and a disruptant. Enzymatic analysis of the transformant lysates showed that the strain expressing mutant BGLII exhibited weakened cellobiose hydrolytic activity, but produced some transglycosylation products, suggesting that the SNP in bgl2 strongly diminished cellobiase activity, but did not result in complete loss of function of BGLII. The analysis of the recombinant BGLII revealed that transglycosylation products might be oligosaccharides, composed probably of glucose linked β-1,4, β-1,3, or a mixture of both. PC-3-7 revertants of bgl2 exhibited reduced expression and inducibility of cellulase during growth on cellulose and cellobiose substrates. Furthermore, the effect of this bgl2 mutation was reproduced in the common strain QM9414 in which the transformants showed cellulase production comparable to that of PC-3-7. We conclude that BGLII plays an important role in cellulase induction in T. reesei and that the bgl2 mutation in PC-3-7 brought about enhanced cellulase expression on

  2. Selective Imaging of VEGFR-1 and VEGFR-2 Using 89Zr-Labeled Single-Chain VEGF Mutants.

    Science.gov (United States)

    Meyer, Jan-Philip; Edwards, Kimberly J; Kozlowski, Paul; Backer, Marina V; Backer, Joseph M; Lewis, Jason S

    2016-11-01

    Vascular endothelial growth factor-A (VEGF-A) acts via 2 vascular endothelial growth factor receptors, VEGFR-1 and VEGFR-2, that play important and distinct roles in tumor biology. We reasoned that selective imaging of these receptors could provide unique information for diagnostics and for monitoring and optimizing responses to anticancer therapy, including antiangiogenic therapy. Herein, we report the development of 2 first-in-class 89 Zr-labeled PET tracers that enable the selective imaging of VEGFR-1 and VEGFR-2. Functionally active mutants of scVEGF (an engineered single-chain version of pan-receptor VEGF-A with an N-terminal cysteine-containing tag for site-specific conjugation), named scVR1 and scVR2 with enhanced affinity to, respectively, VEGFR-1 and VEGFR-2, were constructed. Parental scVEGF and its receptor-specific mutants were site-specifically derivatized with the 89 Zr chelator desferroxamine B via a 3.4-kDa PEG linker. 89 Zr labeling of the desferroxamine B conjugates furnished scV/Zr, scVR1/Zr, and scVR2/Zr tracers with high radiochemical yield (>87%), high specific activity (≥9.8 MBq/nmol), and purity (>99%). Tracers were tested in an orthotopic breast cancer model using 4T1luc-bearing syngeneic BALB/c mice. For testing tracer specificity, tracers were coinjected with an excess of cold proteins of the same or opposite receptor specificity or pan-receptor scVEGF. PET imaging, biodistribution, and dosimetry studies in mice, as well as immunohistochemical analysis of harvested tumors, were performed. All tracers rapidly accumulated in orthotopic 4T1luc tumors, allowing for the successful PET imaging of the tumors as early as 2 h after injection. Blocking experiments with an excess of pan-receptor or receptor-specific cold proteins indicated that more than 80% of tracer tumor uptake is VEGFR-mediated, whereas uptake in all major organs is not affected by blocking within the margin of error. Critically, blocking experiments indicated that VEGFR

  3. Soft-Switched Neutral-Point-Clamped Single-Phase Boost Rectifier

    Science.gov (United States)

    Itoh, Ryozo; Ishizaka, Kouichi

    A soft-switched neutral-point-clamped single-phase boost rectifier capable of compensating the imbalance load voltage is studied. This is based on a single-phase rectifier, in which an inductor is placed in series with the AC supply to resonate with a capacitor connected across the DC output of a full-bridge rectifier and the switching transition is mainly governed by a series resonance. The experimental prototype using insulated-gate bipolar transistors is implemented to investigate the operation under the charge control. The experimental results confirm that the rectifier has a neutral-point-clamp feature providing a good quality AC current.

  4. A Study of Impulsive Multiterm Fractional Differential Equations with Single and Multiple Base Points and Applications

    Directory of Open Access Journals (Sweden)

    Yuji Liu

    2014-01-01

    Full Text Available We discuss the existence and uniqueness of solutions for initial value problems of nonlinear singular multiterm impulsive Caputo type fractional differential equations on the half line. Our study includes the cases for a single base point fractional differential equation as well as multiple base points fractional differential equation. The asymptotic behavior of solutions for the problems is also investigated. We demonstrate the utility of our work by applying the main results to fractional-order logistic models.

  5. SCAP-82, Single Scattering, Albedo Scattering, Point-Kernel Analysis in Complex Geometry

    International Nuclear Information System (INIS)

    Disney, R.K.; Vogtman, S.E.

    1987-01-01

    1 - Description of problem or function: SCAP solves for radiation transport in complex geometries using the single or albedo scatter point kernel method. The program is designed to calculate the neutron or gamma ray radiation level at detector points located within or outside a complex radiation scatter source geometry or a user specified discrete scattering volume. Geometry is describable by zones bounded by intersecting quadratic surfaces within an arbitrary maximum number of boundary surfaces per zone. Anisotropic point sources are describable as pointwise energy dependent distributions of polar angles on a meridian; isotropic point sources may also be specified. The attenuation function for gamma rays is an exponential function on the primary source leg and the scatter leg with a build- up factor approximation to account for multiple scatter on the scat- ter leg. The neutron attenuation function is an exponential function using neutron removal cross sections on the primary source leg and scatter leg. Line or volumetric sources can be represented as a distribution of isotropic point sources, with un-collided line-of-sight attenuation and buildup calculated between each source point and the detector point. 2 - Method of solution: A point kernel method using an anisotropic or isotropic point source representation is used, line-of-sight material attenuation and inverse square spatial attenuation between the source point and scatter points and the scatter points and detector point is employed. A direct summation of individual point source results is obtained. 3 - Restrictions on the complexity of the problem: - The SCAP program is written in complete flexible dimensioning so that no restrictions are imposed on the number of energy groups or geometric zones. The geometric zone description is restricted to zones defined by boundary surfaces defined by the general quadratic equation or one of its degenerate forms. The only restriction in the program is that the total

  6. Bayesian unknown change-point models to investigate immediacy in single case designs.

    Science.gov (United States)

    Natesan, Prathiba; Hedges, Larry V

    2017-12-01

    Although immediacy is one of the necessary criteria to show strong evidence of a causal relation in single case designs (SCDs), no inferential statistical tool is currently used to demonstrate it. We propose a Bayesian unknown change-point model to investigate and quantify immediacy in SCD analysis. Unlike visual analysis that considers only 3-5 observations in consecutive phases to investigate immediacy, this model considers all data points. Immediacy is indicated when the posterior distribution of the unknown change-point is narrow around the true value of the change-point. This model can accommodate delayed effects. Monte Carlo simulation for a 2-phase design shows that the posterior standard deviations of the change-points decrease with increase in standardized mean difference between phases and decrease in test length. This method is illustrated with real data. (PsycINFO Database Record (c) 2017 APA, all rights reserved).

  7. Evaluating Change in Behavioral Preferences: Multidimensional Scaling Single-Ideal Point Model

    Science.gov (United States)

    Ding, Cody

    2016-01-01

    The purpose of the article is to propose a multidimensional scaling single-ideal point model as a method to evaluate changes in individuals' preferences under the explicit methodological framework of behavioral preference assessment. One example is used to illustrate the approach for a clear idea of what this approach can accomplish.

  8. Mechanism of DNA–binding loss upon single-point mutation in p53

    Indian Academy of Sciences (India)

    PRAKASH KUMAR

    provides an excellent prototype system to elucidate the different mechanisms underlying the loss in DNA binding affinity and specificity upon single point mutation because over 50% of all human cancers involve p53 mutations, which occur mostly in the sequence–specific DNA–binding central domain (residues 102–292, ...

  9. Single stage buck-boost DC-AC neutral point clamped inverter

    DEFF Research Database (Denmark)

    Mo, Wei; Loh, Poh Chiang; Andrew, A.

    2012-01-01

    This paper proposes a new single stage buck-boost DC-AC neutral point clamped inverter topology which integrates the cascaded configurations of recently introduced inductor-capacitor-capacitor-transformer impedance source network (by Adamowicz) and classic NPC configuration. As a consequence...

  10. Real-time multi-GNSS single-frequency precise point positioning

    NARCIS (Netherlands)

    de Bakker, P.F.; Tiberius, C.C.J.M.

    2017-01-01

    Precise Point Positioning (PPP) is a popular Global Positioning System (GPS) processing strategy, thanks to its high precision without requiring additional GPS infrastructure. Single-Frequency PPP (SF-PPP) takes this one step further by no longer relying on expensive dual-frequency GPS receivers,

  11. SINGLE TREE DETECTION FROM AIRBORNE LASER SCANNING DATA USING A MARKED POINT PROCESS BASED METHOD

    Directory of Open Access Journals (Sweden)

    J. Zhang

    2013-05-01

    Full Text Available Tree detection and reconstruction is of great interest in large-scale city modelling. In this paper, we present a marked point process model to detect single trees from airborne laser scanning (ALS data. We consider single trees in ALS recovered canopy height model (CHM as a realization of point process of circles. Unlike traditional marked point process, we sample the model in a constraint configuration space by making use of image process techniques. A Gibbs energy is defined on the model, containing a data term which judge the fitness of the model with respect to the data, and prior term which incorporate the prior knowledge of object layouts. We search the optimal configuration through a steepest gradient descent algorithm. The presented hybrid framework was test on three forest plots and experiments show the effectiveness of the proposed method.

  12. Generating and executing programs for a floating point single instruction multiple data instruction set architecture

    Science.gov (United States)

    Gschwind, Michael K

    2013-04-16

    Mechanisms for generating and executing programs for a floating point (FP) only single instruction multiple data (SIMD) instruction set architecture (ISA) are provided. A computer program product comprising a computer recordable medium having a computer readable program recorded thereon is provided. The computer readable program, when executed on a computing device, causes the computing device to receive one or more instructions and execute the one or more instructions using logic in an execution unit of the computing device. The logic implements a floating point (FP) only single instruction multiple data (SIMD) instruction set architecture (ISA), based on data stored in a vector register file of the computing device. The vector register file is configured to store both scalar and floating point values as vectors having a plurality of vector elements.

  13. A single point mutation in Tomato spotted wilt virus NSs protein is sufficient to overcome Tsw-gene-mediated resistance in pepper.

    Science.gov (United States)

    Almási, Asztéria; Nemes, Katalin; Csömör, Zsófia; Tóbiás, István; Palkovics, László; Salánki, Katalin

    2017-06-01

    The nonstructural protein (NSs) of Tomato spotted wilt virus (TSWV) was previously identified as an avirulence determinant for Tsw-based resistance on pepper. The NSs of wild-type (WT) and resistance-breaking (RB) TSWV strains isolated in Hungary had only two amino acid substitutions (104, 461). We have analysed the ability of the NSs and their point mutant variants to trigger Tsw-mediated hypersensitive responses and RNA silencing suppressor (RSS) activity in patch assays. We identified a single amino acid change at position 104 (T-A) that was responsible for the necrosis induction or loss, while a significant difference was not detected in the RSS activity of the two parental strains. We have successfully complemented the infection of the WT strain on resistant pepper cultivar with the infectious S RNA transcript of the RB strain and the WT-T104A point mutant. Our work provides direct evidence that a single amino acid change can induce an RB phenotype.

  14. Point defects in lines in single crystalline phosphorene: directional migration and tunable band gaps.

    Science.gov (United States)

    Li, Xiuling; Ma, Liang; Wang, Dayong; Zeng, Xiao Cheng; Wu, Xiaojun; Yang, Jinlong

    2016-10-20

    Extended line defects in two-dimensional (2D) materials can play an important role in modulating their electronic properties. During the experimental synthesis of 2D materials, line defects are commonly generated at grain boundaries between domains of different orientations. In this work, twelve types of line-defect structures in single crystalline phosphorene are examined by using first-principles calculations. These line defects are typically formed via migration and aggregation of intrinsic point defects, including the Stone-Wales (SW), single or double vacancy (SV or DV) defects. Our calculated results demonstrate that the migration of point defects in phosphorene is anisotropic, for instance, the lowest migration energy barriers are 1.39 (or 0.40) and 2.58 (or 0.49) eV for SW (or SV) defects in zigzag and armchair directions, respectively. The aggregation of point defects into lines is energetically favorable compared with the separated point defects in phosphorene. In particular, the axis of line defects in phosphorene is direction-selective, depending on the composed point defects. The presence of line defects effectively modulates the electronic properties of phosphorene, rendering the defect-containing phosphorene either metallic or semiconducting with a tunable band gap. Of particular interest is the fact that the SV-based line defect can behave as a metallic wire, suggesting a possibility to fabricate a circuit with subnanometer widths in the semiconducting phosphorene for nanoscale electronic application.

  15. Design of an omnidirectional single-point photodetector for large-scale spatial coordinate measurement

    Science.gov (United States)

    Xie, Hongbo; Mao, Chensheng; Ren, Yongjie; Zhu, Jigui; Wang, Chao; Yang, Lei

    2017-10-01

    In high precision and large-scale coordinate measurement, one commonly used approach to determine the coordinate of a target point is utilizing the spatial trigonometric relationships between multiple laser transmitter stations and the target point. A light receiving device at the target point is the key element in large-scale coordinate measurement systems. To ensure high-resolution and highly sensitive spatial coordinate measurement, a high-performance and miniaturized omnidirectional single-point photodetector (OSPD) is greatly desired. We report one design of OSPD using an aspheric lens, which achieves an enhanced reception angle of -5 deg to 45 deg in vertical and 360 deg in horizontal. As the heart of our OSPD, the aspheric lens is designed in a geometric model and optimized by LightTools Software, which enables the reflection of a wide-angle incident light beam into the single-point photodiode. The performance of home-made OSPD is characterized with working distances from 1 to 13 m and further analyzed utilizing developed a geometric model. The experimental and analytic results verify that our device is highly suitable for large-scale coordinate metrology. The developed device also holds great potential in various applications such as omnidirectional vision sensor, indoor global positioning system, and optical wireless communication systems.

  16. Identification of Intensity Ratio Break Points from Photon Arrival Trajectories in Ratiometric Single Molecule Spectroscopy

    Science.gov (United States)

    Bingemann, Dieter; Allen, Rachel M.

    2012-01-01

    We describe a statistical method to analyze dual-channel photon arrival trajectories from single molecule spectroscopy model-free to identify break points in the intensity ratio. Photons are binned with a short bin size to calculate the logarithm of the intensity ratio for each bin. Stochastic photon counting noise leads to a near-normal distribution of this logarithm and the standard student t-test is used to find statistically significant changes in this quantity. In stochastic simulations we determine the significance threshold for the t-test’s p-value at a given level of confidence. We test the method’s sensitivity and accuracy indicating that the analysis reliably locates break points with significant changes in the intensity ratio with little or no error in realistic trajectories with large numbers of small change points, while still identifying a large fraction of the frequent break points with small intensity changes. Based on these results we present an approach to estimate confidence intervals for the identified break point locations and recommend a bin size to choose for the analysis. The method proves powerful and reliable in the analysis of simulated and actual data of single molecule reorientation in a glassy matrix. PMID:22837704

  17. Single point incremental forming of tailored blanks produced by friction stir welding

    DEFF Research Database (Denmark)

    Silva, M.B.; Skjødt, Martin; Vilaca, P.

    2009-01-01

    This paper is focused on the single point incremental forming (SPIF) of tailored welded blanks produced by friction stirwelding (FSW). Special emphasis is placed on the know-how for producing the tailored blanks and on the utilization of innovative forming strategies to protect thewelding joint...... fromthe rotating single point-forming tool. Formability of the tailor welded blanks (TWB) is evaluated by means of benchmark tests carried out on truncated conical and pyramidal shapes and results are compared with similar tests performed on conventional reference blanks of the same material. Results show...... that the combination of SPIF with tailored welded blanks produced by FSW seems promising in the manufacture of complex sheet metal parts with high depths....

  18. Point defects and magnetic properties of neutron irradiated MgO single crystal

    Directory of Open Access Journals (Sweden)

    Mengxiong Cao

    2017-05-01

    Full Text Available (100-oriented MgO single crystals were irradiated to introduce point defects with different neutron doses ranging from 1.0×1016 to 1.0×1020 cm-2. The point defect configurations were studied with X-ray diffuse scattering and UV-Vis absorption spectra. The isointensity profiles of X-ray diffuse scattering caused by the cubic and double-force point defects in MgO were theoretically calculated based on the Huang scattering theory. The magnetic properties at different temperature were measured with superconducting quantum interference device (SQUID. The reciprocal space mappings (RSMs of irradiated MgO revealed notable diffuse scattering. The UV-Vis spectra indicated the presence of O Frenkel defects in irradiated MgO. Neutron-irradiated MgO was diamagnetic at room temperature and became ferromagnetic at low temperature due to O Frenkel defects induced by neutron-irradiation.

  19. Exposure buildup factors for a cobalt-60 point isotropic source for single and two layer slabs

    International Nuclear Information System (INIS)

    Chakarova, R.

    1992-01-01

    Exposure buildup factors for point isotropic cobalt-60 sources are calculated by the Monte Carlo method with statistical errors ranging from 1.5 to 7% for 1-5 mean free paths (mfp) thick water and iron single slabs and for 1 and 2 mfp iron layers followed by water layers 1-5 mfp thick. The computations take into account Compton scattering. The Monte Carlo data for single slab geometries are approximated by Geometric Progression formula. Kalos's formula using the calculated single slab buildup factors may be applied to reproduce the data for two-layered slabs. The presented results and discussion may help when choosing the manner in which the radiation field gamma irradiation units will be described. (author)

  20. The consistency condition for the three-point function in dissipative single-clock inflation

    International Nuclear Information System (INIS)

    Nacir, Diana López; Porto, Rafael A.; Zaldarriaga, Matias

    2012-01-01

    We generalize the consistency condition for the three-point function in single field inflation to the case of dissipative, multi-field, single-clock models. We use the recently introduced extension of the effective field theory of inflation that accounts for dissipative effects, to provide an explicit proof to leading (non-trivial) order in the generalized slow roll parameters and mixing with gravity scales. Our results illustrate the conditions necessary for the validity of the consistency relation in situations with many degrees of freedom relevant during inflation, namely that there is a preferred clock. Departures from this condition in forthcoming experiments would rule out not only single field but also a large class of multi-field models

  1. The consistency condition for the three-point function in dissipative single-clock inflation

    Energy Technology Data Exchange (ETDEWEB)

    Nacir, Diana López [Departamento de Física, Facultad de Ciencias Exactas y Naturales, UBA and IFIBA, CONICET, Ciudad Universitaria, Pabellón 1, 1428, Buenos Aires (Argentina); Porto, Rafael A.; Zaldarriaga, Matias, E-mail: dnacir@df.uba.ar, E-mail: rporto@ias.edu, E-mail: matiasz@ias.edu [School of Natural Sciences, Institute for Advanced Study, Einstein Drive, Princeton, NJ 08540 (United States)

    2012-09-01

    We generalize the consistency condition for the three-point function in single field inflation to the case of dissipative, multi-field, single-clock models. We use the recently introduced extension of the effective field theory of inflation that accounts for dissipative effects, to provide an explicit proof to leading (non-trivial) order in the generalized slow roll parameters and mixing with gravity scales. Our results illustrate the conditions necessary for the validity of the consistency relation in situations with many degrees of freedom relevant during inflation, namely that there is a preferred clock. Departures from this condition in forthcoming experiments would rule out not only single field but also a large class of multi-field models.

  2. Identification of enhanced hydrogen and ethanol Escherichia coli producer strains in a glycerol-based medium by screening in single-knock out mutant collections.

    Science.gov (United States)

    Valle, Antonio; Cabrera, Gema; Cantero, Domingo; Bolivar, Jorge

    2015-06-28

    Earth's climate is warming as a result of anthropogenic emissions of greenhouse gases from fossil fuel combustion. Bioenergy, which includes biodiesel, biohydrogen and bioethanol, has emerged as a sustainable alternative fuel source. For this reason, in recent years biodiesel production has become widespread but this industry currently generates a huge amount of glycerol as a by-product, which has become an environmental problem in its own right. A feasible possibility to solve this problem is the use of waste glycerol as a carbon source for microbial transformation into biofuels such as hydrogen and ethanol. For instance, Escherichia coli is a microorganism that can synthesize these compounds under anaerobic conditions. In this work an experimental procedure was established for screening E. coli single mutants to identify strains with enhanced ethanol and/or H2 productions compared to the wild type strain. In an initial screening of 150 single mutants, 12 novel strains (gnd, tdcE, rpiA nanE, tdcB, deoB, sucB, cpsG, frmA, glgC, fumA and gadB) were found to provide enhanced yields for at least one of the target products. The mutations, that improve most significantly the parameters evaluated (gnd and tdcE genes), were combined with other mutations in three engineered E. coli mutant strains in order to further redirect carbon flux towards the desired products. This methodology can be a useful tool to disclose the metabolic pathways that are more susceptible to manipulation in order to obtain higher molar yields of hydrogen and ethanol using glycerol as main carbon source in multiple E. coli mutants.

  3. Mms Sensitivity of All Amino Acid-Requiring Mutants in Aspergillus and Its Suppression by Mutations in a Single Gene

    OpenAIRE

    Käfer, Etta

    1987-01-01

    All available amino acid-requiring mutants of Aspergillus nidulans were found to be hypersensitive to MMS (methyl methanesulfonate) to various degrees. On MMS media, secondary mutations could be selected which suppress this MMS sensitivity but do not affect the requirement. Many such mutations were analyzed and found to be alleles of one gene, smsA (= suppressor of MMS sensitivity), which mapped distal on the right arm of chromosome V. This gene is more likely to be involved in general regula...

  4. Introduction of a point mutation into an HLA class I single-chain trimer induces enhancement of CTL priming and antitumor immunity

    Directory of Open Access Journals (Sweden)

    Masanori Matsui

    2014-01-01

    Full Text Available We previously discovered one particular HLA-A*02:01 mutant that enhanced peptide-specific cytotoxic T lymphocyte (CTL recognition in vitro compared to wild-type HLA-A*02:01. This mutant contains a single amino acid substitution from histidine to leucine at position 74 (H74L that is located in the peptide-binding groove. To investigate the effect of the H74L mutation on the in vivo CTL priming, we took advantage of the technology of the HLA class I single-chain trimer (SCT in which three components involving a peptide, β2 microglobulin and the HLA class I heavy chain are joined together via flexible linkers. We generated recombinant adenovirus expressing SCT comprised influenza A matrix protein (FMP-derived peptide, β2 microglobulin and the H74L heavy chain. HLA-A*02:01 transgenic mice were immunized with the adenovirus, and the induction of peptide-specific CTLs and antitumor immunity was investigated. It was clearly shown that the H74L mutation enabled the HLA-A*02:01 SCT molecule to dramatically enhance both in vivo priming of FMP-specific CTLs and protection against a lethal challenge of tumor cells expressing FMP. These data present the first evidence that a simple point mutation in the HLA class I heavy chain of SCT is beneficial for improving CTL-based immunotherapy and prophylaxis to control tumors.

  5. A single point mutation within the coding sequence of cholera toxin B subunit will increase its expression yield.

    Science.gov (United States)

    Bakhshi, Bita; Boustanshenas, Mina; Ghorbani, Masoud

    2014-07-01

    Cholera toxin B subunit (CTB) has been extensively considered as an immunogenic and adjuvant protein, but its yield of expression is not satisfactory in many studies. The aim of this study was to compare the expression of native and mutant recombinant CTB (rCTB) in pQE vector. ctxB fragment from Vibrio cholerae O1 ATCC14035 containing the substitution of mutant ctxB for amino acid S128T was amplified by PCR and cloned in pGETM-T easy vector. It was then transformed to E. coli Top 10F' and cultured on LB agar plate containing ampicillin. Sequence analysis confirmed the mature ctxB gene sequence and the mutant one in both constructs which were further subcloned to pQE-30 vector. Both constructs were subsequently transformed to E. coli M15 (pREP4) for expression of mature and mutant rCTB. SDS-PAGE analysis showed the maximum expression of rCTB in both systems at 5 hours after induction and Western-blot analysis confirmed the presence of rCTB in blotting membranes. The expression of mutant rCTB was much higher than mature rCTB, which may be the result of serine-to-threonine substitution at position 128 of mature rCTB amino acid sequence created by PCR mutagenesis. The mutant rCTB retained pentameric stability and its ability to bind to anti- cholera toxin IgG antibodies. Point mutation in ctxB sequence resulted in over-expression of rCTB, probably due to the increase of solubility of produced rCTB. Consequently, this expression system can be used to produce rCTB in high yield.

  6. Ultrasonography for suspected deep vein thrombosis: how useful is single-point augmentation?

    Energy Technology Data Exchange (ETDEWEB)

    McQueen, A.S. [Department of Radiology, Freeman Hospital, Newcastle upon Tyne (United Kingdom)], E-mail: andrewmcqueen7@hotmail.com; Elliott, S.T. [Department of Radiology, Freeman Hospital, Newcastle upon Tyne (United Kingdom); Keir, M.J. [Department of Medical Physics, Royal Victoria Infirmary, Newcastle upon Tyne (United Kingdom)

    2009-02-15

    Aims: To assess the role of single-point augmentation of spectral Doppler flow in the diagnosis of acute deep vein thrombosis (DVT). Secondary objectives included identifying the augmentation response in non-DVT diagnoses. Methods: Patients attending the ultrasound departments of two hospitals for investigation of suspected acute DVT during an 8-month period were recruited to the study group. Spectral Doppler assessment of the superficial femoral vein was recorded during Valsalva and calf compression manoeuvres in the asymptomatic and symptomatic legs. The Doppler waveforms from the symptomatic limb were characterized as 'normal' or 'abnormal' by the operator. Standard compression ultrasonography of the symptomatic leg was then performed with the presence of DVT or an alternative diagnosis documented. Results: One hundred and sixty-seven patients underwent ultrasound examinations using the study methodology. Nine patients were subsequently excluded due to bilateral DVT or inability to tolerate calf compression. The mean age of the remaining 158 patients was 65.4 years with 28 DVTs identified (18% of patients). Calf compression elicited a normal response in 118/130 of non-DVT examinations (specificity 91%) and an abnormal response in 18/28 DVT examinations (sensitivity 64%). Diminished or absent augmentation was identified in alternative diagnoses that included haematoma and Baker's cyst. Conclusions: This study demonstrates that single-point augmentation has a low sensitivity in suspected lower-limb DVT, and that the majority of undetected DVTs are isolated to the calf veins. An abnormal augmentation response is a poor predictor of lower-limb DVT as alternative diagnoses can produce diminished or reduced augmentation. Therefore, single-point augmentation does not add to the standard compression ultrasound examination for suspected DVT and should not be routinely performed.

  7. Ultrasonography for suspected deep vein thrombosis: how useful is single-point augmentation?

    International Nuclear Information System (INIS)

    McQueen, A.S.; Elliott, S.T.; Keir, M.J.

    2009-01-01

    Aims: To assess the role of single-point augmentation of spectral Doppler flow in the diagnosis of acute deep vein thrombosis (DVT). Secondary objectives included identifying the augmentation response in non-DVT diagnoses. Methods: Patients attending the ultrasound departments of two hospitals for investigation of suspected acute DVT during an 8-month period were recruited to the study group. Spectral Doppler assessment of the superficial femoral vein was recorded during Valsalva and calf compression manoeuvres in the asymptomatic and symptomatic legs. The Doppler waveforms from the symptomatic limb were characterized as 'normal' or 'abnormal' by the operator. Standard compression ultrasonography of the symptomatic leg was then performed with the presence of DVT or an alternative diagnosis documented. Results: One hundred and sixty-seven patients underwent ultrasound examinations using the study methodology. Nine patients were subsequently excluded due to bilateral DVT or inability to tolerate calf compression. The mean age of the remaining 158 patients was 65.4 years with 28 DVTs identified (18% of patients). Calf compression elicited a normal response in 118/130 of non-DVT examinations (specificity 91%) and an abnormal response in 18/28 DVT examinations (sensitivity 64%). Diminished or absent augmentation was identified in alternative diagnoses that included haematoma and Baker's cyst. Conclusions: This study demonstrates that single-point augmentation has a low sensitivity in suspected lower-limb DVT, and that the majority of undetected DVTs are isolated to the calf veins. An abnormal augmentation response is a poor predictor of lower-limb DVT as alternative diagnoses can produce diminished or reduced augmentation. Therefore, single-point augmentation does not add to the standard compression ultrasound examination for suspected DVT and should not be routinely performed

  8. Assessment of Grade-Level Differences in Coping Behavior among Adolescents Using Multidimensional Scaling Single-Ideal-Point Model

    Science.gov (United States)

    Ding, Cody; Yang, Dong

    2013-01-01

    The purpose of the study was to examine grade-level differences in coping behaviors among adolescents using a probabilistic multidimensional scaling (MDS) single-ideal-point model. Using data from students in middle school and at college, this article illustrated the MDS single-ideal-point model as an alternative to examine students' typical…

  9. Probing Critical Point Energies of Transition Metal Dichalcogenides: Surprising Indirect Gap of Single Layer WSe 2

    KAUST Repository

    Zhang, Chendong

    2015-09-21

    By using a comprehensive form of scanning tunneling spectroscopy, we have revealed detailed quasi-particle electronic structures in transition metal dichalcogenides, including the quasi-particle gaps, critical point energy locations, and their origins in the Brillouin zones. We show that single layer WSe surprisingly has an indirect quasi-particle gap with the conduction band minimum located at the Q-point (instead of K), albeit the two states are nearly degenerate. We have further observed rich quasi-particle electronic structures of transition metal dichalcogenides as a function of atomic structures and spin-orbit couplings. Such a local probe for detailed electronic structures in conduction and valence bands will be ideal to investigate how electronic structures of transition metal dichalcogenides are influenced by variations of local environment.

  10. Efficient Background Segmentation and Seed Point Generation for a Single-Shot Stereo System

    Science.gov (United States)

    Chen, Xiaobo; Xi, Juntong

    2017-01-01

    Single-shot stereo 3D shape measurement is becoming more popular due to its advantages of noise robustness and short acquisition period. One of the key problems is stereo matching, which is related to the efficiency of background segmentation and seed point generation, etc. In this paper, a more efficient and automated matching algorithm based on digital image correlation (DIC) is proposed. The standard deviation of image gradients and an adaptive threshold are employed to segment the background. Scale-invariant feature transform (SIFT)-based feature matching and two-dimensional triangulation are combined to estimate accurate initial parameters for seed point generation. The efficiency of background segmentation and seed point generation, as well as the measuring precision, are evaluated by experimental simulation and real tests. Experimental results show that the average segmentation time for an image with a resolution of 1280 × 960 pixels is 240 milliseconds. The efficiency of seed point generation is verified to be high with different convergence criteria. PMID:29194415

  11. Optical Properties And Thermal Stability Of Single-Point Diamond-Machined Aluminum Alloys

    Science.gov (United States)

    Ogloza, A. A.; Decker, D. L.; Archibald, P. C.; O'Connor, D. A.; Bueltmann, E. R.

    1989-01-01

    This paper presents the results of diamond-turned surfaces of a wide range of aluminum alloys. The alloys machined included a sand-cast A201 alloy manufactured by Specialty Aluminum Inc., conventionally extruded plate alloys 2024, 3003, 5052, 6061, 7075, and for comparison as a best and worst case possible a high-purity aluminum single crystal, and tooling plate. The surfaces were obtained by diamond single-point machining using an interferometrically controlled two-axis, air-bearing lathe. The effect of tool-rake angle and machining fluid on surface quality is examined. Surface characterization was performed by Nomarski microscopy and noncontact optical surface profilometry. The optical properties measured included absolute reflectance at 3.8 μm, total integrated scatter at 752.5 nm, and bidirectional reflection distribution function measurements at 632.8 nm. The dimensional stability of the aluminum alloys subject to thermal cycling is examined.

  12. Synthesis of a wild-type and three mutant Cucurbita maxima trypsin inhibitor-encoding genes by a single-strand approach.

    Science.gov (United States)

    Botes, D P; Qobose, M D; Corfield, V A

    1991-09-15

    A single-strand approach to gene assembly, based on a modification of an in vitro complementary oligodeoxyribonucleotide template-directed ligation of the desired sequence to a linearized vector [Chen et al., Nucleic Acids Res. 18 (1990) 871-878], is described. The gene coding for the wild-type Cucurbita maxima trypsin inhibitor of 29 amino acid residues [Bode et al., FEBS Lett. 242 (1989) 285-292], as well as three mutant forms of the gene, in which two of the three disulfide bonds have been replaced singly or as a pair, have been synthesized in a single synthesis run with minimal manual intervention. Subsequent to ligation to pUC9 and in vivo gapped duplex repair by Escherichia coli, their sequences have been verified.

  13. Expression, purification and characterization of a novel double-sites mutant of the single-chain sweet-tasting protein monellin (MNEI) with both improved sweetness and stability.

    Science.gov (United States)

    Zheng, Weiwei; Yang, Liu; Cai, Chenggu; Ni, Jinfeng; Liu, Bo

    2018-03-01

    The sweet protein monellin has high sweet potency with limited stability. In this study, 3 double-sites mutants (E2N/E23A, E2N/Y65R and E23A/Y65R) of the single-chain monellin (MNEI) were constructed. The proteins were expressed in E. coli BL21 and purified to homogeneity by nickel affinity chromatography with yields above 10 mg/L cell culture. Introduction of a sweeter mutant E2N into E23A or Y65R (E2N/E23A and E2N/Y65R) led to about 3-fold increase of sweetness, while addition of a more stable mutant E23A into E2N or Y65R (E2N/E23A and E23A/Y65R) resulted in improved thermal stability (about 10 °C). The results indicate that residues E2 and E23 mediate the sweetness and thermal stability of the protein, respectively. Multiple mutations of different residues (E2N/E23A) led to an additive performance with both improved sweetness and stability, suggesting that the sweetness and stability could be modulated by the independent molecular mechanism. The sweeter and thermal stable variant has a potential in further industrial applications. Copyright © 2017 Elsevier Inc. All rights reserved.

  14. Toxicity of CuO nanoparticles to yeast Saccharomyces cerevisiae BY4741 wild-type and its nine isogenic single-gene deletion mutants.

    Science.gov (United States)

    Kasemets, Kaja; Suppi, Sandra; Künnis-Beres, Kai; Kahru, Anne

    2013-03-18

    A suite of eight tentatively oxidative stress response-deficient Saccharomyces cerevisiae BY4741 single-gene mutants (sod1Δ, sod2Δ, yap1Δ, cta1Δ, ctt1Δ, gsh1Δ, glr1Δ, and ccs1Δ) and one copper-vulnerable mutant (cup2Δ) was used to elucidate weather the toxicity of CuO nanoparticles to S. cerevisiae is mediated by oxidative stress (OS). Specifically, sensitivity profiles of mutants' phenotypes and wild-type (wt) upon exposure to nano-CuO were compared. As controls, CuSO4 (solubility), bulk-CuO (size), H2O2, and menadione (OS) were used. Growth inhibition of wt and mutant strains was studied in rich YPD medium and cell viability in deionized water (DI). Dissolved Cu-ions were quantified by recombinant metal-sensing bacteria and chemical analysis. To wt strain nano-CuO was 32-fold more toxic than bulk-CuO: 24-h IC50 4.8 and 155 mg/L in DI and 643 and >20000 mg/L in YPD, respectively. In toxicant-free YPD medium, all mutants had practically similar growth patterns as wt. However, the mutant strains sod1Δ, sod2Δ, ccs1Δ, and yap1Δ showed up to 12-fold elevated sensitivity toward OS standard chemicals menadione and H2O2 but not to nano-CuO, indicating that CuO nanoparticles exerted toxicity to yeast cells via different mechanisms. The most vulnerable strain to all studied Cu compounds was the copper stress response-deficient strain cup2Δ (∼16-fold difference with wt), indicating that the toxic effect of CuO (nano)particles proceeds via dissolved Cu-ions. The dissolved copper solely explained the toxicity of nano-CuO in DI but not in YPD. Assumingly, in YPD nano-CuO acquired a coating of peptides/proteins and sorbed onto the yeast's outer surface, resulting in their increased solubility in the close vicinity of yeast cells and increased uptake of Cu-ions that was not registered by the assays used for the analysis of dissolved Cu-ions in the test medium. Lastly, as yeast retained its viability in DI even by 24th hour of incubation, the profiling of the acute

  15. Potent and selective antisense oligonucleotides targeting single-nucleotide polymorphisms in the Huntington disease gene / allele-specific silencing of mutant huntingtin

    DEFF Research Database (Denmark)

    Carroll, Jeffrey B; Warby, Simon C; Southwell, Amber L

    2011-01-01

    Huntington disease (HD) is an autosomal dominant neurodegenerative disorder caused by CAG-expansion in the huntingtin gene (HTT) that results in a toxic gain of function in the mutant huntingtin protein (mHTT). Reducing the expression of mHTT is therefore an attractive therapy for HD. However, wild......-type HTT protein is essential for development and has critical roles in maintaining neuronal health. Therapies for HD that reduce wild-type HTT may therefore generate unintended negative consequences. We have identified single-nucleotide polymorphism (SNP) targets in the human HD population for the disease...

  16. MMS sensitivity of all amino acid-requiring mutants in aspergillus and its suppression by mutations in a single gene.

    Science.gov (United States)

    Käfer, E

    1987-04-01

    All available amino acid-requiring mutants of Aspergillus nidulans were found to be hypersensitive to MMS (methyl methanesulfonate) to various degrees. On MMS media, secondary mutations could be selected which suppress this MMS sensitivity but do not affect the requirement. Many such mutations were analyzed and found to be alleles of one gene, smsA (= suppressor of MMS sensitivity), which mapped distal on the right arm of chromosome V. This gene is more likely to be involved in general regulation of amino acid biosynthesis than MMS uptake, since a variety of pathway interactions were clearly modified by smsA suppressors in the absence of MMS.

  17. Detection of Janus Kinase 2 gene single point mutation in real samples with electrochemical DNA biosensor.

    Science.gov (United States)

    Topkaya, Seda Nur; Kosova, Buket; Ozsoz, Mehmet

    2014-02-15

    Janus Kinase 2 (JAK2) gene single point mutations, which have been reported to be associated with myeloproliferative disorders, are usually detected through conventional methods such as melting curve assays, allele-specific and quantitative Polymerase Chain Reactions (PCRs). Herein, an electrochemical biosensor for the detection of a Guanine (G) to Thymine (T) transversion at nucleotide position 1849 of the JAK2 gene was reported. Due to clinical importance of this mutation, easy and sensitive tests are needed to be developed. Our aim was to design a biosensor system that is capable of detecting the mutation within less than 1h with high sensitivity. For these purposes, an electrochemical sensing system was developed based on detecting hybridization. Hybridization between probe and its target and discrimination of single point mutation was investigated by monitoring guanine oxidation signals observed at +1.0 V with Differential Pulse Voltammetry (DPV) by using synthetic oligonucleotides and Polymerase Chain Reaction (PCR) amplicons. Hybridization between probe and PCR amplicons was also determined with Electrochemical Impedance Spectroscopy (EIS). We successfully detect hybridization first in synthetic samples, and ultimately in real samples involving blood samples from patients as well as additional healthy controls. The limit of detection (S/N=3) was calculated as 44 pmol of target sequence in a 40-μl reaction volume in real samples. Copyright © 2013 Elsevier B.V. All rights reserved.

  18. A single mutation results in diploid gamete formation and parthenogenesis in a Drosophila yemanuclein-alpha meiosis I defective mutant.

    Science.gov (United States)

    Meyer, Régis E; Delaage, Michèle; Rosset, Roland; Capri, Michèle; Aït-Ahmed, Ounissa

    2010-11-16

    Sexual reproduction relies on two key events: formation of cells with a haploid genome (the gametes) and restoration of diploidy after fertilization. Therefore the underlying mechanisms must have been evolutionary linked and there is a need for evidence that could support such a model. We describe the identification and the characterization of yem1, the first yem-alpha mutant allele (V478E), which to some extent affects diploidy reduction and its restoration. Yem-alpha is a member of the Ubinuclein/HPC2 family of proteins that have recently been implicated in playing roles in chromatin remodeling in concert with HIRA histone chaperone. The yem1 mutant females exhibited disrupted chromosome behavior in the first meiotic division and produced very low numbers of viable progeny. Unexpectedly these progeny did not display paternal chromosome markers, suggesting that they developed from diploid gametes that underwent gynogenesis, a form of parthenogenesis that requires fertilization. We focus here on the analysis of the meiotic defects exhibited by yem1 oocytes that could account for the formation of diploid gametes. Our results suggest that yem1 affects chromosome segregation presumably by affecting kinetochores function in the first meiotic division. This work paves the way to further investigations on the evolution of the mechanisms that support sexual reproduction.

  19. A single mutation results in diploid gamete formation and parthenogenesis in a Drosophila yemanuclein-alpha meiosis I defective mutant

    Science.gov (United States)

    2010-01-01

    Background Sexual reproduction relies on two key events: formation of cells with a haploid genome (the gametes) and restoration of diploidy after fertilization. Therefore the underlying mechanisms must have been evolutionary linked and there is a need for evidence that could support such a model. Results We describe the identification and the characterization of yem1, the first yem-alpha mutant allele (V478E), which to some extent affects diploidy reduction and its restoration. Yem-alpha is a member of the Ubinuclein/HPC2 family of proteins that have recently been implicated in playing roles in chromatin remodeling in concert with HIRA histone chaperone. The yem1 mutant females exhibited disrupted chromosome behavior in the first meiotic division and produced very low numbers of viable progeny. Unexpectedly these progeny did not display paternal chromosome markers, suggesting that they developed from diploid gametes that underwent gynogenesis, a form of parthenogenesis that requires fertilization. Conclusions We focus here on the analysis of the meiotic defects exhibited by yem1 oocytes that could account for the formation of diploid gametes. Our results suggest that yem1 affects chromosome segregation presumably by affecting kinetochores function in the first meiotic division. This work paves the way to further investigations on the evolution of the mechanisms that support sexual reproduction. PMID:21080953

  20. A single mutation results in diploid gamete formation and parthenogenesis in a Drosophila yemanuclein-alpha meiosis I defective mutant

    Directory of Open Access Journals (Sweden)

    Capri Michèle

    2010-11-01

    Full Text Available Abstract Background Sexual reproduction relies on two key events: formation of cells with a haploid genome (the gametes and restoration of diploidy after fertilization. Therefore the underlying mechanisms must have been evolutionary linked and there is a need for evidence that could support such a model. Results We describe the identification and the characterization of yem1, the first yem-alpha mutant allele (V478E, which to some extent affects diploidy reduction and its restoration. Yem-alpha is a member of the Ubinuclein/HPC2 family of proteins that have recently been implicated in playing roles in chromatin remodeling in concert with HIRA histone chaperone. The yem1 mutant females exhibited disrupted chromosome behavior in the first meiotic division and produced very low numbers of viable progeny. Unexpectedly these progeny did not display paternal chromosome markers, suggesting that they developed from diploid gametes that underwent gynogenesis, a form of parthenogenesis that requires fertilization. Conclusions We focus here on the analysis of the meiotic defects exhibited by yem1 oocytes that could account for the formation of diploid gametes. Our results suggest that yem1 affects chromosome segregation presumably by affecting kinetochores function in the first meiotic division. This work paves the way to further investigations on the evolution of the mechanisms that support sexual reproduction.

  1. Automatic tracking of vessel-like structures from a single starting point.

    Science.gov (United States)

    Oliveira, Dário Augusto Borges; Leal-Taixé, Laura; Feitosa, Raul Queiroz; Rosenhahn, Bodo

    2016-01-01

    The identification of vascular networks is an important topic in the medical image analysis community. While most methods focus on single vessel tracking, the few solutions that exist for tracking complete vascular networks are usually computationally intensive and require a lot of user interaction. In this paper we present a method to track full vascular networks iteratively using a single starting point. Our approach is based on a cloud of sampling points distributed over concentric spherical layers. We also proposed a vessel model and a metric of how well a sample point fits this model. Then, we implement the network tracking as a min-cost flow problem, and propose a novel optimization scheme to iteratively track the vessel structure by inherently handling bifurcations and paths. The method was tested using both synthetic and real images. On the 9 different data-sets of synthetic blood vessels, we achieved maximum accuracies of more than 98%. We further use the synthetic data-set to analyze the sensibility of our method to parameter setting, showing the robustness of the proposed algorithm. For real images, we used coronary, carotid and pulmonary data to segment vascular structures and present the visual results. Still for real images, we present numerical and visual results for networks of nerve fibers in the olfactory system. Further visual results also show the potential of our approach for identifying vascular networks topologies. The presented method delivers good results for the several different datasets tested and have potential for segmenting vessel-like structures. Also, the topology information, inherently extracted, can be used for further analysis to computed aided diagnosis and surgical planning. Finally, the method's modular aspect holds potential for problem-oriented adjustments and improvements. Copyright © 2015 Elsevier Ltd. All rights reserved.

  2. Real-Time Single Frequency Precise Point Positioning Using SBAS Corrections

    Directory of Open Access Journals (Sweden)

    Liang Li

    2016-08-01

    Full Text Available Real-time single frequency precise point positioning (PPP is a promising technique for high-precision navigation with sub-meter or even centimeter-level accuracy because of its convenience and low cost. The navigation performance of single frequency PPP heavily depends on the real-time availability and quality of correction products for satellite orbits and satellite clocks. Satellite-based augmentation system (SBAS provides the correction products in real-time, but they are intended to be used for wide area differential positioning at 1 meter level precision. By imposing the constraints for ionosphere error, we have developed a real-time single frequency PPP method by sufficiently utilizing SBAS correction products. The proposed PPP method are tested with static and kinematic data, respectively. The static experimental results show that the position accuracy of the proposed PPP method can reach decimeter level, and achieve an improvement of at least 30% when compared with the traditional SBAS method. The positioning convergence of the proposed PPP method can be achieved in 636 epochs at most in static mode. In the kinematic experiment, the position accuracy of the proposed PPP method can be improved by at least 20 cm relative to the SBAS method. Furthermore, it has revealed that the proposed PPP method can achieve decimeter level convergence within 500 s in the kinematic mode.

  3. Simple and efficient site-directed mutagenesis using two single-primer reactions in parallel to generate mutants for protein structure-function studies

    Directory of Open Access Journals (Sweden)

    Edelheit Oded

    2009-06-01

    Full Text Available Abstract Background In protein engineering, site-directed mutagenesis methods are used to generate DNA sequences with mutated codons, insertions or deletions. In a widely used method, mutations are generated by PCR using a pair of oligonucleotide primers designed with mismatching nucleotides at the center of the primers. In this method, primer-primer annealing may prevent cloning of mutant cDNAs. To circumvent this problem we developed an alternative procedure that does not use forward-reverse primer pair in the same reaction. Results In initial studies we used a double-primer PCR mutagenesis protocol, but sequencing of products showed tandem repeats of primer in cloned DNA. We developed an alternative method that starts with two Single-Primer Reactions IN Parallel using high-fidelity Pwo DNA polymerase. Thus, we call the method with the acronym SPRINP. The SPRINP reactions are then combined, denatured at 95°C, and slowly cooled, promoting random annealing of the parental DNA and the newly synthesized strands. The products are digested with DpnI that digests methylated parental strands, and then transformed into E. coli. Using this method we generated >40 mutants in cDNAs coding for human Epithelial Na+ Channel (ENaC subunits. The method has been tested for 1–3 bp codon mutation and insertion of a 27 bp epitope tag into cDNAs. Conclusion The SPRINP mutagenesis protocol yields mutants reliably and with high fidelity. The use of a single primer in each amplification reaction increases the probability of success of primers relative to previous methods employing a forward and reverse primer pair in the same reaction.

  4. Simple and efficient site-directed mutagenesis using two single-primer reactions in parallel to generate mutants for protein structure-function studies.

    Science.gov (United States)

    Edelheit, Oded; Hanukoglu, Aaron; Hanukoglu, Israel

    2009-06-30

    In protein engineering, site-directed mutagenesis methods are used to generate DNA sequences with mutated codons, insertions or deletions. In a widely used method, mutations are generated by PCR using a pair of oligonucleotide primers designed with mismatching nucleotides at the center of the primers. In this method, primer-primer annealing may prevent cloning of mutant cDNAs. To circumvent this problem we developed an alternative procedure that does not use forward-reverse primer pair in the same reaction. In initial studies we used a double-primer PCR mutagenesis protocol, but sequencing of products showed tandem repeats of primer in cloned DNA. We developed an alternative method that starts with two Single-Primer Reactions IN Parallel using high-fidelity Pwo DNA polymerase. Thus, we call the method with the acronym SPRINP. The SPRINP reactions are then combined, denatured at 95 degrees C, and slowly cooled, promoting random annealing of the parental DNA and the newly synthesized strands. The products are digested with DpnI that digests methylated parental strands, and then transformed into E. coli. Using this method we generated >40 mutants in cDNAs coding for human Epithelial Na+ Channel (ENaC) subunits. The method has been tested for 1-3 bp codon mutation and insertion of a 27 bp epitope tag into cDNAs. The SPRINP mutagenesis protocol yields mutants reliably and with high fidelity. The use of a single primer in each amplification reaction increases the probability of success of primers relative to previous methods employing a forward and reverse primer pair in the same reaction.

  5. A single point mutation changes the crystallization behavior of Mycoplasma arthritidis-derived mitogen

    International Nuclear Information System (INIS)

    Guo, Yi; Li, Zhong; Van Vranken, Sandra J.; Li, Hongmin

    2006-01-01

    The mutagenesis, crystallization and preliminary crystallographic analysis of M. arthritidis-derived mitogen is described. Mycoplasma arthritidis-derived mitogen (MAM) functions as a conventional superantigen (SAg). Although recombinant MAM has been crystallized by the hanging-drop vapour-diffusion method, the crystals diffracted poorly to only 5.0 Å resolution, with large unit-cell parameters a = 163.8, b = 93.0, c = 210.9 Å, β = 93.7° in the monoclinic space group P2 1 . Unit-cell content analysis revealed that as many as 24 molecules could be present in the asymmetric unit. Systematic alanine mutagenesis was applied in order to search for mutants that give crystals of better quality. Two mutants, L50A and K201A, were crystallized under the same conditions as wild-type MAM (MAM wt ). Crystals of the L50A mutant are isomorphous with those of MAM wt , while a new crystal form was obtained for the K201 mutant, belonging to the cubic space group P4 1 32 with unit-cell parameters a = b = c = 181.9 Å. Diffraction data were collected to 3.6 and 2.8 Å resolution from crystals of the MAM L50A and K201A mutants, respectively. Molecular-replacement calculations suggest the presence of two molecules in the asymmetric unit for the MAM K201A mutant crystal, resulting in a V M of 5.0 Å Da −1 and a solvent content of 75%. An interpretable electron-density map for the MAM K201A mutant crystal was produced using the molecular-replacement method

  6. From single point of measurement to distributed sensing in long-term glacier monitoring

    International Nuclear Information System (INIS)

    Cesarini, Daniel; Avvenuti, Marco; Jelicic, Vana; Bilas, Vedran

    2013-01-01

    Glacial environment monitoring is a key task in understanding natural phenomena related to global warming. For the last 30 years, Automatic Weather Stations (AWSs) have been spreading among the meteorological and geophysical community, and are on the way to become a de facto standard to perform long-lasting unattended data acquisitions in single localized points of interest. Sensor Networks (SNs), on the other hand, promise the possibility to perform measurements with a higher spatial density and lower cost. Designing and developing a SN for glacial environment face particular challenges for embedded electronics and sensor systems, which is why SNs are still under research and development in this field. This paper surveys the AWSs and SNs for glacial monitoring applications and compares their characteristics.

  7. Simulation study based on the single-point temperature monitoring system of LabVIEW

    Science.gov (United States)

    Wu, Yongling; Yang, Na; Liu, Shuping; Pan, Xiaohui; Wang, Wenjiang

    2014-12-01

    This paper takes LabVIEW2012 as a development platform, creating a J-type thermocouple sensor and the NI USB-6229 data acquisition card and other hardware emulation circuitry which combined with the PC designed a single-point temperature monitoring system. Through simulation experiments, the system has a collection interval, the sampling rate per channel sampling on the temperature limit set by the user function and it also has the function of real-time display the current temperature, the temperature limit alarm, maximum temperature, minimum temperature display and a temperature history data query. This system can be used for temperature monitoring of life, research, industrial control, environmental monitoring, biomedical, tobacco processing, greenhouse cultivation, livestock breeding and other fields, which has important significance and practical value.

  8. Plastic flow and failure in single point incremental forming of PVC sheets

    Directory of Open Access Journals (Sweden)

    P. A. F. Martins

    2014-05-01

    Full Text Available This paper presents an innovative and effective methodology to characterize plastic flow and failure in single point incremental forming (SPIF of polymers that allows determining the stresses and the accumulated values of ductile damage directly from the experimental values of strain at various positions over the deformed polymer sheets. The approach traces the deformation path of material elements in conical and pyramidal SPIF parts, undergoing linear strain loading paths from beginning until failure, and is built upon the generalization of the analytical framework conditions assumed by Glover et al. [1] to the pressure-sensitive yield surfaces of polymers under incompressible, non-associated, plastic flow. Experimentation in conventional and multi-stage SPIF of Polyvinylchloride (PVC sheets confirms the effectiveness of the proposed methodology and demonstrates that standard non-coupled damage models currently utilized in sheet metal forming are inapplicable to describe failure in polymers. Instead fracture forming limit lines (FFL’s should be employed.

  9. Monitoring single event upsets in SRAM-based FPGAs at the SuperKEKB interaction point

    Science.gov (United States)

    Giordano, R.; Tortone, G.; Perrella, S.; Izzo, V.; Aloisio, A.

    2017-07-01

    In February 2016, the SuperKEKB positron-electron high-luminosity collider of the KEK laboratory (Tsukuba, Japan) started being commissioned. A dedicated commissioning detector, named BEAST2, has been used to characterize beam backgrounds before the Belle2 detector was rolled into the beams and to provide tuning parameters for Monte Carlo simulations. BEAST2 consists of a fiberglass support structure and several sub-detectors mounted onto it, including time projection chambers (TPCs) and He-3 tubes. In this work, we present direct measurements of radiation-induced single event upsets in a SRAM-based FPGA device installed in BEAST2 at a distance of ~ 1 m from the beam interaction point. Our goal was to provide experimental results of the expected radiation-induced configuration upset rate and power consumption variation at Belle2 and at other experiments operating in similar radiation environments. Beam currents for both electron and positron rings spanned a range between 50 and 500 mA, therefore providing data about the FPGA operation in different radiation conditions. Even if the machine has not been providing collisions yet, as the beams were not focused to the interaction point, our results show a rate of 0.15 upsets/day averaged over the whole commissioning time frame. We had neither evidence of total dose effects on the FPGA power consumption nor of permanent damage to the device.

  10. Spherical Projection Based Straight Line Segment Extraction for Single Station Terrestrial Laser Point Cloud

    Directory of Open Access Journals (Sweden)

    ZHANG Fan

    2015-06-01

    Full Text Available Due to the discrete distribution computing errors and lack of adaptability are ubiquitous in the current straight line extraction for TLS data methods. A 3D straight line segment extraction method is proposed based on spherical projection for single station terrestrial laser point clouds. Firstly, horizontal and vertical angles of each laser point are calculated by means of spherical coordinates, intensity panoramic image according to the two angles is generated. Secondly, edges which include straight line features are detected from intensity panoramic image by using of edge detection algorithm. Thirdly, great circles are detected from edges of panoramic image using spherical Hough transform. According to the axiom that a straight line segment in 3D space is a spherical great circle after spherical projection, detecting great circles from spherical projected data sets is essentially detecting straight line segments from 3D data sets without spherical projection. Finally, a robust 3D straight line fitting method is employed to fitting the straight lines and calculating parameters of the straight line segments. Experiments using different data sets and comparison with other methods show the accuracy and applicability of the proposed method.

  11. Sensor system for multi-point monitoring using bending loss of single mode optical fiber

    International Nuclear Information System (INIS)

    Kim, Heon Young; Kim, Dae Hyun

    2015-01-01

    Applications of smart sensors have been extended to safety systems in the aerospace, transportation and civil engineering fields. In particular, structural health monitoring techniques using smart sensors have gradually become necessary and have been developed to prevent dangers to human life and damage to assets. Generally, smart sensors are based on electro-magnets and have several weaknesses, including electro-magnetic interference and distortion. Therefore, fiber optic sensors are an outstanding alternative to overcome the weaknesses of electro-magnetic sensors. However, they require expensive devices and complex systems. This paper proposes a new, affordable and simple sensor system that uses a single fiber to monitor pressures at multiple-points. Moreover, a prototype of the sensor system was manufactured and tested for a feasibility study. Based on the results of this experimental test, a relationship was carefully observed between the bend loss conditions and light-intensity. As a result, it was shown that impacts at multiple-points could be monitored.

  12. ATM mutants

    Indian Academy of Sciences (India)

    First page Back Continue Last page Graphics. ATM mutants. ATM (Ataxia Telangiectasia Mutated). AT2BE and AT5B1 cells – fibroblast cell lines from Ataxia telangiectasia patients. Deletion mutants expressing truncated ATM protein which is inactive. Have been used in studies looking at the role of ATM in DNA damage ...

  13. Criteria for evaluating protection from single points of failure for partially expanded fault trees

    International Nuclear Information System (INIS)

    Aswani, D.; Badreddine, B.; Malone, M.; Gauthier, G.; Proietty, J.

    2008-01-01

    Fault tree analysis (FTA) is a technique that describes the combinations of events in a system which result in an undesirable outcome. FTA is used as a tool to quantitatively assess a system's probability for an undesirable outcome. Time constraints from concept to production in modern engineering often limit the opportunity for a thorough statistical analysis of a system. Furthermore, when undesirable outcomes are considered such as hazard to human(s), it becomes difficult to identify strict statistical targets for what is acceptable. Consequently, when hazard to human(s) is concerned a common design target is to protect the system from single points of failure (SPOF) which means that no failure mode caused by a single event, concern, or error has a critical consequence on the system. Such a design target is common with 'by-wire' systems. FTA can be used to verify if a system is protected from SPOF. In this paper, sufficient criteria for evaluating protection from SPOF for partially expanded fault trees are proposed along with proof. The proposed criteria consider potential interactions between the lowest drawn events of a partial fault tree expansion which otherwise easily leads to an overly optimistic analysis of protection from SPOF. The analysis is limited to fault trees that are coherent and static

  14. Analysis of Point Based Image Registration Errors With Applications in Single Molecule Microscopy.

    Science.gov (United States)

    Cohen, E A K; Ober, R J

    2013-12-15

    We present an asymptotic treatment of errors involved in point-based image registration where control point (CP) localization is subject to heteroscedastic noise; a suitable model for image registration in fluorescence microscopy. Assuming an affine transform, CPs are used to solve a multivariate regression problem. With measurement errors existing for both sets of CPs this is an errors-in-variable problem and linear least squares is inappropriate; the correct method being generalized least squares. To allow for point dependent errors the equivalence of a generalized maximum likelihood and heteroscedastic generalized least squares model is achieved allowing previously published asymptotic results to be extended to image registration. For a particularly useful model of heteroscedastic noise where covariance matrices are scalar multiples of a known matrix (including the case where covariance matrices are multiples of the identity) we provide closed form solutions to estimators and derive their distribution. We consider the target registration error (TRE) and define a new measure called the localization registration error (LRE) believed to be useful, especially in microscopy registration experiments. Assuming Gaussianity of the CP localization errors, it is shown that the asymptotic distribution for the TRE and LRE are themselves Gaussian and the parameterized distributions are derived. Results are successfully applied to registration in single molecule microscopy to derive the key dependence of the TRE and LRE variance on the number of CPs and their associated photon counts. Simulations show asymptotic results are robust for low CP numbers and non-Gaussianity. The method presented here is shown to outperform GLS on real imaging data.

  15. Simian virus 40 large T-antigen point mutants that are defective in viral DNA replication but competent in oncogenic transformation.

    OpenAIRE

    Manos, M M; Gluzman, Y

    1984-01-01

    The large T antigen of simian virus 40 (SV40) is a multifunctional protein that is essential in both the virus lytic cycle and the oncogenic transformation of cells by SV40. To investigate the role of the numerous biochemical and physiological activities of T antigen in the lytic and transformation processes, we have studied DNA replication-deficient, transformation-competent large T-antigen mutants. Here we describe the genetic and biochemical analyses of two such mutants, C2/SV40 and C11/SV...

  16. Evaluations of Three-Dimensional Building Model Reconstruction from LiDAR Point Clouds and Single-View Perspective Imagery

    Directory of Open Access Journals (Sweden)

    F. Tsai

    2014-06-01

    Full Text Available This paper briefly presents two approaches for effective three-dimensional (3D building model reconstruction from terrestrial laser scanning (TLS data and single perspective view imagery and assesses their applicability to the reconstruction of 3D models of landmark or historical buildings. The collected LiDAR point clouds are registered based on conjugate points identified using a seven-parameter transformation system. Three dimensional models are generated using plan and surface fitting algorithms. The proposed single-view reconstruction (SVR method is based on vanishing points and single-view metrology. More detailed models can also be generated according to semantic analysis of the façade images. Experimental results presented in this paper demonstrate that both TLS and SVR approaches can successfully produce accurate and detailed 3D building models from LiDAR point clouds or different types of single-view perspective images.

  17. Mechanism of DNA–binding loss upon single-point mutation in p53

    Indian Academy of Sciences (India)

    PRAKASH KUMAR

    Despite our current understanding of protein−DNA recognition, the mechanism(s) underlying the loss in protein−DNA binding affinity/ ... Keywords. p53 mutants; protein-DNA interactions; molecular dynamics simulations; free energy decomposition. Abbreviations ... of the salt–bridge with Asp 281, resulting in the loss of DNA.

  18. Towards single-valued polylogarithms in two variables for the seven-point remainder function in multi-Regge kinematics

    Directory of Open Access Journals (Sweden)

    Johannes Broedel

    2017-02-01

    Full Text Available We investigate single-valued polylogarithms in two complex variables, which are relevant for the seven-point remainder function in N=4 super-Yang–Mills theory in the multi-Regge regime. After constructing these two-dimensional polylogarithms, we determine the leading logarithmic approximation of the seven-point remainder function up to and including five loops.

  19. Femtosecond few- to single-electron point-projection microscopy for nanoscale dynamic imaging

    Science.gov (United States)

    Bainbridge, A. R.; Barlow Myers, C. W.; Bryan, W. A.

    2016-01-01

    Femtosecond electron microscopy produces real-space images of matter in a series of ultrafast snapshots. Pulses of electrons self-disperse under space-charge broadening, so without compression, the ideal operation mode is a single electron per pulse. Here, we demonstrate femtosecond single-electron point projection microscopy (fs-ePPM) in a laser-pump fs-e-probe configuration. The electrons have an energy of only 150 eV and take tens of picoseconds to propagate to the object under study. Nonetheless, we achieve a temporal resolution with a standard deviation of 114 fs (equivalent to a full-width at half-maximum of 269 ± 40 fs) combined with a spatial resolution of 100 nm, applied to a localized region of charge at the apex of a nanoscale metal tip induced by 30 fs 800 nm laser pulses at 50 kHz. These observations demonstrate real-space imaging of reversible processes, such as tracking charge distributions, is feasible whilst maintaining femtosecond resolution. Our findings could find application as a characterization method, which, depending on geometry, could resolve tens of femtoseconds and tens of nanometres. Dynamically imaging electric and magnetic fields and charge distributions on sub-micron length scales opens new avenues of ultrafast dynamics. Furthermore, through the use of active compression, such pulses are an ideal seed for few-femtosecond to attosecond imaging applications which will access sub-optical cycle processes in nanoplasmonics. PMID:27158637

  20. Femtosecond few- to single-electron point-projection microscopy for nanoscale dynamic imaging

    Directory of Open Access Journals (Sweden)

    A. R. Bainbridge

    2016-03-01

    Full Text Available Femtosecond electron microscopy produces real-space images of matter in a series of ultrafast snapshots. Pulses of electrons self-disperse under space-charge broadening, so without compression, the ideal operation mode is a single electron per pulse. Here, we demonstrate femtosecond single-electron point projection microscopy (fs-ePPM in a laser-pump fs-e-probe configuration. The electrons have an energy of only 150 eV and take tens of picoseconds to propagate to the object under study. Nonetheless, we achieve a temporal resolution with a standard deviation of 114 fs (equivalent to a full-width at half-maximum of 269 ± 40 fs combined with a spatial resolution of 100 nm, applied to a localized region of charge at the apex of a nanoscale metal tip induced by 30 fs 800 nm laser pulses at 50 kHz. These observations demonstrate real-space imaging of reversible processes, such as tracking charge distributions, is feasible whilst maintaining femtosecond resolution. Our findings could find application as a characterization method, which, depending on geometry, could resolve tens of femtoseconds and tens of nanometres. Dynamically imaging electric and magnetic fields and charge distributions on sub-micron length scales opens new avenues of ultrafast dynamics. Furthermore, through the use of active compression, such pulses are an ideal seed for few-femtosecond to attosecond imaging applications which will access sub-optical cycle processes in nanoplasmonics.

  1. Obtaining Global Picture From Single Point Observations by Combining Data Assimilation and Machine Learning Tools

    Science.gov (United States)

    Shprits, Y.; Zhelavskaya, I. S.; Kellerman, A. C.; Spasojevic, M.; Kondrashov, D. A.; Ghil, M.; Aseev, N.; Castillo Tibocha, A. M.; Cervantes Villa, J. S.; Kletzing, C.; Kurth, W. S.

    2017-12-01

    Increasing volume of satellite measurements requires deployment of new tools that can utilize such vast amount of data. Satellite measurements are usually limited to a single location in space, which complicates the data analysis geared towards reproducing the global state of the space environment. In this study we show how measurements can be combined by means of data assimilation and how machine learning can help analyze large amounts of data and can help develop global models that are trained on single point measurement. Data Assimilation: Manual analysis of the satellite measurements is a challenging task, while automated analysis is complicated by the fact that measurements are given at various locations in space, have different instrumental errors, and often vary by orders of magnitude. We show results of the long term reanalysis of radiation belt measurements along with fully operational real-time predictions using data assimilative VERB code. Machine Learning: We present application of the machine learning tools for the analysis of NASA Van Allen Probes upper-hybrid frequency measurements. Using the obtained data set we train a new global predictive neural network. The results for the Van Allen Probes based neural network are compared with historical IMAGE satellite observations. We also show examples of predictions of geomagnetic indices using neural networks. Combination of machine learning and data assimilation: We discuss how data assimilation tools and machine learning tools can be combine so that physics-based insight into the dynamics of the particular system can be combined with empirical knowledge of it's non-linear behavior.

  2. Is equilibrium point control feasible for fast goal-directed single-joint movements?

    Science.gov (United States)

    Kistemaker, Dinant A; Van Soest, Arthur J; Bobbert, Maarten F

    2006-05-01

    Several types of equilibrium point (EP) controllers have been proposed for the control of posture and movement. EP controllers are appealing from a computational perspective because they do not require solving the "inverse dynamic problem" (i.e., computation of the torques required to move a system along a desired trajectory). It has been argued that EP controllers are not capable of controlling fast single-joint movements. To refute this statement, several extensions have been proposed, although these have been tested using models in which only the tendon compliance, force-length-velocity relation, and mechanical interaction between tendon and contractile element were not adequately represented. In the present study, fast elbow-joint movements were measured and an attempt was made to reproduce these using a realistic musculoskeletal model of the human arm. Three types of EP controllers were evaluated: an open-loop alpha-controller, a closed-loop lambda-controller, and a hybrid open- and closed-loop controller. For each controller we considered a continuous version and a version in which the control signals were sent out intermittently. Only the intermittent hybrid EP controller was capable of generating movements that were as fast as those of the subjects. As a result of the nonlinear muscle properties, the hybrid EP controller requires a more detailed representation of static muscle properties than generally assumed in the context of EP control. In sum, this study shows that fast single-joint movements can be realized without explicitly solving the inverse dynamics problem, but in a less straightforward manner than implied by proponents of conventional EP controllers.

  3. Prediction and control of pillow defect in single point incremental forming using numerical simulations

    Energy Technology Data Exchange (ETDEWEB)

    Isidore, B. B. Lemopi [Eastern Mediterranean University, Gazimagusa (Turkmenistan); Hussain, G.; Khan, Wasim A. [GIK Institute of Engineering, Swabi (Pakistan); Shamachi, S. Pourhassan [University of Minho, Guimaraes (Portugal)

    2016-05-15

    Pillows formed at the center of sheets in Single point incremental forming (SPIF) are fabrication defects which adversely affect the geometrical accuracy and formability of manufactured parts. This study is focused on using FEA as a tool to predict and control pillowing in SPIF by varying tool size and shape. 3D Finite element analysis (FEA) and experiments are carried out using annealed Aluminum 1050. From FEA, it is found out that the stress/strain state in the immediate vicinity of the forming tool in the transverse direction plays a determinant role on sheet pillowing. Furthermore, pillow height increases as compression in the sheet-plane increases. The nature of in-plane stresses in the transverse direction varies from compressive to tensile as the tool-end geometry is changed from spherical to flat. Additionally, the magnitude of corresponding in-plane stresses decreases as the tool radius increases. According to measurements from the FEA model, flat end tools and large radii both retard pillow formation. However, the influence of changing tool end shape from hemispherical to flat is observed to be more important than the effect of varying tool radius, because the deformation zone remains in tension in the transverse direction while forming with flat end tools. These findings are verified by conducting a set of experiments. A fair agreement between the FEM and empirical results show that FEM can be employed as a tool to predict and control the pillow defect in SPIF.

  4. Pan European Phenological database (PEP725): a single point of access for European data

    Science.gov (United States)

    Templ, Barbara; Koch, Elisabeth; Bolmgren, Kjell; Ungersböck, Markus; Paul, Anita; Scheifinger, Helfried; Rutishauser, This; Busto, Montserrat; Chmielewski, Frank-M.; Hájková, Lenka; Hodzić, Sabina; Kaspar, Frank; Pietragalla, Barbara; Romero-Fresneda, Ramiro; Tolvanen, Anne; Vučetič, Višnja; Zimmermann, Kirsten; Zust, Ana

    2018-02-01

    The Pan European Phenology (PEP) project is a European infrastructure to promote and facilitate phenological research, education, and environmental monitoring. The main objective is to maintain and develop a Pan European Phenological database (PEP725) with an open, unrestricted data access for science and education. PEP725 is the successor of the database developed through the COST action 725 "Establishing a European phenological data platform for climatological applications" working as a single access point for European-wide plant phenological data. So far, 32 European meteorological services and project partners from across Europe have joined and supplied data collected by volunteers from 1868 to the present for the PEP725 database. Most of the partners actively provide data on a regular basis. The database presently holds almost 12 million records, about 46 growing stages and 265 plant species (including cultivars), and can be accessed via http://www.pep725.eu/. Users of the PEP725 database have studied a diversity of topics ranging from climate change impact, plant physiological question, phenological modeling, and remote sensing of vegetation to ecosystem productivity.

  5. Pan European Phenological database (PEP725): a single point of access for European data.

    Science.gov (United States)

    Templ, Barbara; Koch, Elisabeth; Bolmgren, Kjell; Ungersböck, Markus; Paul, Anita; Scheifinger, Helfried; Rutishauser, This; Busto, Montserrat; Chmielewski, Frank-M; Hájková, Lenka; Hodzić, Sabina; Kaspar, Frank; Pietragalla, Barbara; Romero-Fresneda, Ramiro; Tolvanen, Anne; Vučetič, Višnja; Zimmermann, Kirsten; Zust, Ana

    2018-02-18

    The Pan European Phenology (PEP) project is a European infrastructure to promote and facilitate phenological research, education, and environmental monitoring. The main objective is to maintain and develop a Pan European Phenological database (PEP725) with an open, unrestricted data access for science and education. PEP725 is the successor of the database developed through the COST action 725 "Establishing a European phenological data platform for climatological applications" working as a single access point for European-wide plant phenological data. So far, 32 European meteorological services and project partners from across Europe have joined and supplied data collected by volunteers from 1868 to the present for the PEP725 database. Most of the partners actively provide data on a regular basis. The database presently holds almost 12 million records, about 46 growing stages and 265 plant species (including cultivars), and can be accessed via http://www.pep725.eu/ . Users of the PEP725 database have studied a diversity of topics ranging from climate change impact, plant physiological question, phenological modeling, and remote sensing of vegetation to ecosystem productivity.

  6. An application of eddy current damping effect on single point diamond turning of titanium alloys

    Science.gov (United States)

    Yip, W. S.; To, S.

    2017-11-01

    Titanium alloys Ti6Al4V (TC4) have been popularly applied in many industries. They have superior material properties including an excellent strength-to-weight ratio and corrosion resistance. However, they are regarded as difficult to cut materials; serious tool wear, a high level of cutting vibration and low surface integrity are always involved in machining processes especially in ultra-precision machining (UPM). In this paper, a novel hybrid machining technology using an eddy current damping effect is firstly introduced in UPM to suppress machining vibration and improve the machining performance of titanium alloys. A magnetic field was superimposed on samples during single point diamond turning (SPDT) by exposing the samples in between two permanent magnets. When the titanium alloys were rotated within a magnetic field in the SPDT, an eddy current was generated through a stationary magnetic field inside the titanium alloys. An eddy current generated its own magnetic field with the opposite direction of the external magnetic field leading a repulsive force, compensating for the machining vibration induced by the turning process. The experimental results showed a remarkable improvement in cutting force variation, a significant reduction in adhesive tool wear and an extreme long chip formation in comparison to normal SPDT of titanium alloys, suggesting the enhancement of the machinability of titanium alloys using an eddy current damping effect. An eddy current damping effect was firstly introduced in the area of UPM to deliver the results of outstanding machining performance.

  7. An application of eddy current damping effect on single point diamond turning of titanium alloys

    International Nuclear Information System (INIS)

    Yip, W S; To, S

    2017-01-01

    Titanium alloys Ti6Al4V (TC4) have been popularly applied in many industries. They have superior material properties including an excellent strength-to-weight ratio and corrosion resistance. However, they are regarded as difficult to cut materials; serious tool wear, a high level of cutting vibration and low surface integrity are always involved in machining processes especially in ultra-precision machining (UPM). In this paper, a novel hybrid machining technology using an eddy current damping effect is firstly introduced in UPM to suppress machining vibration and improve the machining performance of titanium alloys. A magnetic field was superimposed on samples during single point diamond turning (SPDT) by exposing the samples in between two permanent magnets. When the titanium alloys were rotated within a magnetic field in the SPDT, an eddy current was generated through a stationary magnetic field inside the titanium alloys. An eddy current generated its own magnetic field with the opposite direction of the external magnetic field leading a repulsive force, compensating for the machining vibration induced by the turning process. The experimental results showed a remarkable improvement in cutting force variation, a significant reduction in adhesive tool wear and an extreme long chip formation in comparison to normal SPDT of titanium alloys, suggesting the enhancement of the machinability of titanium alloys using an eddy current damping effect. An eddy current damping effect was firstly introduced in the area of UPM to deliver the results of outstanding machining performance. (paper)

  8. Mental Health Conditions and Symptoms in Pediatric Hospitalizations: A Single-Center Point Prevalence Study.

    Science.gov (United States)

    Doupnik, Stephanie K; Henry, M Katherine; Bae, Hanah; Litman, Jessica; Turner, Shanarra; Scharko, Alexander M; Feudtner, Chris

    2017-03-01

    Children and adolescents necessitating hospitalization for physical health conditions are at high risk for mental health conditions; however, the prevalence of mental health conditions and symptoms among hospitalized children and adolescents is uncertain. The objective of this study was to determine the proportion of hospitalized children and adolescents who have diagnosed mental health disorders or undiagnosed mental health problems. In this single-center point prevalence study of hospitalized children between the ages of 4 and 21 years, patients or their parents reported known mental health diagnoses and use of services using the Services Assessment for Children and Adolescent, and they reported patient mental health symptoms using the Pediatric Symptom Checklist, 17-item form (PSC-17). Of 229 eligible patients, 119 agreed to participate. Demographic characteristics of patients who enrolled were not statistically significantly different from those of patients who declined to participate. Among participants, 26% (95% confidence interval [CI], 18%-35%) reported a known mental health diagnosis. On the PSC-17, 29% (95% CI, 21%-38%) of participants had a positive screen for mental health symptoms. Of those with a positive screen, 38% (95% CI, 21%-55%) had no known mental health diagnosis, and 26% (95% CI, 12%-43%) had not received ambulatory mental health services in the 12 months before hospitalization. Mental health conditions and symptoms are common among patients hospitalized in a tertiary children's hospital, and many affected patients are not receiving ambulatory mental health services. Copyright © 2016 Academic Pediatric Association. Published by Elsevier Inc. All rights reserved.

  9. Estimating Uncertainty of Point-Cloud Based Single-Tree Segmentation with Ensemble Based Filtering

    Directory of Open Access Journals (Sweden)

    Matthew Parkan

    2018-02-01

    Full Text Available Individual tree crown segmentation from Airborne Laser Scanning data is a nodal problem in forest remote sensing. Focusing on single layered spruce and fir dominated coniferous forests, this article addresses the problem of directly estimating 3D segment shape uncertainty (i.e., without field/reference surveys, using a probabilistic approach. First, a coarse segmentation (marker controlled watershed is applied. Then, the 3D alpha hull and several descriptors are computed for each segment. Based on these descriptors, the alpha hulls are grouped to form ensembles (i.e., groups of similar tree shapes. By examining how frequently regions of a shape occur within an ensemble, it is possible to assign a shape probability to each point within a segment. The shape probability can subsequently be thresholded to obtain improved (filtered tree segments. Results indicate this approach can be used to produce segmentation reliability maps. A comparison to manually segmented tree crowns also indicates that the approach is able to produce more reliable tree shapes than the initial (unfiltered segmentation.

  10. Single point incremental sheet forming investigated by in-process 3D digital image correlation

    Directory of Open Access Journals (Sweden)

    Bernhart G.

    2010-06-01

    Full Text Available Single Point Incremental Forming (SPIF is a promising sheet metal forming process for prototyping and small batches, in which the blank is formed in a stepwise fashion by a displacement-controlled small-sized tool. Due to specific strain paths induced by the process and limited plastic zones in the contact region between the tool and the workpiece, forming diagrams and forming strategies are different from the classical stamping processes. One major limitation of SPIF is the lack of accuracy of the obtained final parts because of the poor knowledge of the state of stress during the process that requires a good description of the material models and a right choice of the process parameters. In this paper, the SPIF process is experimentally investigated by the mean of surface 3D digital image correlation during the forming of a AW-5086-H111 grade aluminium alloy. Development of strain fields encountered in incremental forming is reported and material formability is evaluated on several formed shapes, taking into account a wide range of straining conditions of this process.

  11. Single point incremental sheet forming investigated by in-process 3D digital image correlation

    Science.gov (United States)

    Decultot, N.; Robert, L.; Velay, V.; Bernhart, G.

    2010-06-01

    Single Point Incremental Forming (SPIF) is a promising sheet metal forming process for prototyping and small batches, in which the blank is formed in a stepwise fashion by a displacement-controlled small-sized tool. Due to specific strain paths induced by the process and limited plastic zones in the contact region between the tool and the workpiece, forming diagrams and forming strategies are different from the classical stamping processes. One major limitation of SPIF is the lack of accuracy of the obtained final parts because of the poor knowledge of the state of stress during the process that requires a good description of the material models and a right choice of the process parameters. In this paper, the SPIF process is experimentally investigated by the mean of surface 3D digital image correlation during the forming of a AW-5086-H111 grade aluminium alloy. Development of strain fields encountered in incremental forming is reported and material formability is evaluated on several formed shapes, taking into account a wide range of straining conditions of this process.

  12. Prediction and control of pillow defect in single point incremental forming using numerical simulations

    International Nuclear Information System (INIS)

    Isidore, B. B. Lemopi; Hussain, G.; Khan, Wasim A.; Shamachi, S. Pourhassan

    2016-01-01

    Pillows formed at the center of sheets in Single point incremental forming (SPIF) are fabrication defects which adversely affect the geometrical accuracy and formability of manufactured parts. This study is focused on using FEA as a tool to predict and control pillowing in SPIF by varying tool size and shape. 3D Finite element analysis (FEA) and experiments are carried out using annealed Aluminum 1050. From FEA, it is found out that the stress/strain state in the immediate vicinity of the forming tool in the transverse direction plays a determinant role on sheet pillowing. Furthermore, pillow height increases as compression in the sheet-plane increases. The nature of in-plane stresses in the transverse direction varies from compressive to tensile as the tool-end geometry is changed from spherical to flat. Additionally, the magnitude of corresponding in-plane stresses decreases as the tool radius increases. According to measurements from the FEA model, flat end tools and large radii both retard pillow formation. However, the influence of changing tool end shape from hemispherical to flat is observed to be more important than the effect of varying tool radius, because the deformation zone remains in tension in the transverse direction while forming with flat end tools. These findings are verified by conducting a set of experiments. A fair agreement between the FEM and empirical results show that FEM can be employed as a tool to predict and control the pillow defect in SPIF.

  13. The use of single point incremental forming for customized implants of unicondylar knee arthroplasty: a review

    Directory of Open Access Journals (Sweden)

    Pankaj Kailasrao Bhoyar

    Full Text Available Abstract Introduction The implantable devices are having enormous market. These products are basically made by traditional manufacturing process, but for the custom-made implants Incremental Sheet Forming is a paramount alternative. Single Point Incremental Forming (SPIF is a manufacturing process to form intricate, asymmetrical components. It forms the component using stretching and bending by maintaining materials crystal structure. SPIF process can be performed using conventional Computer Numerical Control (CNC milling machine. Review This review paper elaborates the various manufacturing processes carried on various biocompatible metallic and nonmetallic customised implantable devices. Conclusion Ti-6Al-4V alloy is broadly used for biomedical implants, but in this alloy, Vanadium is toxic so this alloy is not compatible for implants. The attention of researchers is towards the non toxic and suitable biocompatible materials. For this reason, a novel approach was developed in order to enhance the mechanical properties of this material. . The development of incremental forming technique can improve the formability of existing alloys and may meet the current strict requirements for performance of dies and punches.

  14. Modeling and optimization of surface roughness in single point incremental forming process

    Directory of Open Access Journals (Sweden)

    Suresh Kurra

    2015-07-01

    Full Text Available Single point incremental forming (SPIF is a novel and potential process for sheet metal prototyping and low volume production applications. This article is focuses on the development of predictive models for surface roughness estimation in SPIF process. Surface roughness in SPIF has been modeled using three different techniques namely, Artificial Neural Networks (ANN, Support Vector Regression (SVR and Genetic Programming (GP. In the development of these predictive models, tool diameter, step depth, wall angle, feed rate and lubricant type have been considered as model variables. Arithmetic mean surface roughness (Ra and maximum peak to valley height (Rz are used as response variables to assess the surface roughness of incrementally formed parts. The data required to generate, compare and evaluate the proposed models have been obtained from SPIF experiments performed on Computer Numerical Control (CNC milling machine using Box–Behnken design. The developed models are having satisfactory goodness of fit in predicting the surface roughness. Further, the GP model has been used for optimization of Ra and Rz using genetic algorithm. The optimum process parameters for minimum surface roughness in SPIF have been obtained and validated with the experiments and found highly satisfactory results within 10% error.

  15. Readout of a single electron spin in a double quantum dot using a quantum point contact

    International Nuclear Information System (INIS)

    Zhang Jianping; Ouyang Shihua; You, J Q; Lam, C.-H.

    2008-01-01

    We study the dynamics of a single electron spin in a double quantum dot (DQD) and its readout via a quantum point contact (QPC). We model the system microscopically and derive rate equations for the reduced electron density matrix of the DQD. Two cases with one and two electrons in the DQD are studied. In the one-electron case, with different Zeeman splittings in the two dots, the electron spin states are distinctly characterized by a constant and an oscillatory current through the QPC. In the two-electron case, the readout of the spin state of the electron in one of the dots called the qubit dot is essentially similar after considering hyperfine interactions between the electrons and the nuclear spins of the host materials and a uniform magnetic field applied to the DQD. Moreover, to ensure that an electron is properly injected into the qubit dot, we propose to determine the success of the electron injection from the variations of the QPC current after applying an oscillating magnetic field to the qubit dot

  16. State-of-health monitoring of 18650 4S packs with a single-point impedance diagnostic

    Science.gov (United States)

    Love, Corey T.; Virji, Maheboob B. V.; Rocheleau, Richard E.; Swider-Lyons, Karen E.

    2014-11-01

    The state-of-health (SOH) of Li-ion batteries and battery packs must be monitored effectively for abuse to prevent failure and accidents. In a previous publication, we described a single-point impedance diagnostic method for detecting damage in single prismatic lithium polymer rechargeable cells subjected to overcharge abuse. We now determine whether the single-point impedance diagnostic method is applicable to 4S battery packs. At 316 Hz, commercial 18650 LiCoO2 cells are determined to have the least change in impedance response when cycled between 3.0 and 4.2 V, for states-of-charge (SOC) of 0-100%. The impedance response of single cells at 316 Hz changes dramatically during overcharge (SOC = 125%), presumably due to change in their solid electrolyte interface (SEI) layers at the electrodes. When a single cell is purposely subjected to such overcharge abuse and then integrated into a 4S pack with 3 other healthy cells, the impedance response of the 4S pack at 316 Hz also changes, despite variances in the impedance response of each of the 3 healthy cells. The results suggest that this single-point impedance method could serve as a diagnostic in an all-inclusive battery management system to identify overcharge abuse of single cells without individual cell monitoring.

  17. Absence of linkage of apparently single gene mediated ADHD with the human syntenic region of the mouse mutant coloboma

    Energy Technology Data Exchange (ETDEWEB)

    Hess, E.J.; Rogan, P.K.; Domoto, M. [Pennsylvania State Univ. College of Medicine, Hershey, PA (United States)] [and others

    1995-12-18

    Attention deficit disorder (ADHD) is a complex biobehavioral phenotype which affects up to 8% of the general population and often impairs social, academic, and job performance. Its origins are heterogeneous, but a significant genetic component is suggested by family and twin studies. The murine strain, coloboma, displays a spontaneously hyperactive phenotype that is responsive to dextroamphetamine and has been proposed as a genetic model for ADHD. Coloboma is a semi-dominant mutation that is caused by a hemizygous deletion of the SNAP-25 and other genes on mouse chromosome 2q. To test the possibility that the human homolog of the mouse coloboma gene(s) could be responsible for ADHD, we have carried out linkage studies with polymorphic markers in the region syntenic to coloboma (20p11-p12). Five families in which the pattern of inheritance of ADHD appears to be autosomal dominant were studied. Segregation analysis of the traits studied suggested that the best fitting model was a sex-influenced, single gene, Mendelian pattern. Several genetic models were evaluated based on estimates of penetrance, phenocopy rate, and allele frequency derived from our patient population and those of other investigators. No significant linkage was detected between the disease locus and markers spanning this chromosome 20 interval. 39 refs., 2 figs., 1 tab.

  18. Use of a Cellulase-Derepressed Mutant of Cellulomonas in the Production of a Single-Cell Protein Product from Cellulose †

    Science.gov (United States)

    Hitchner, E. V.; Leatherwood, J. M.

    1980-01-01

    A cellulase-derepressed mutant of a Cellulomonas species was used to produce single-cell protein from crystalline cellulose. In preliminary tests, maximum yield of single-cell protein was obtained at 30°C (pH 7.0) with urea as the nitrogen source. A continuous-flow foam flotation procedure was developed for rapid and efficient separation of bacteria from the culture liquid and cellulose residue. A pH of 4.5 was optimum for foam flotation of this organism. In preliminary trials, recovery was 85% of the cells with the flotation procedure. Cellulomonas was 68% true protein and had an essential amino acid profile featuring a high lysine content (6.5% of protein). The Cellulomonas product was evaluated nutritionally with weanling rats. The net protein utilization value for the protein supplemented with methionine was 50.4% Weight gain of rats on the Cellulomonas diet was similar to that of rats fed a casein diet. PMID:16345511

  19. Detection of Single Tree Stems in Forested Areas from High Density ALS Point Clouds Using 3d Shape Descriptors

    Science.gov (United States)

    Amiri, N.; Polewski, P.; Yao, W.; Krzystek, P.; Skidmore, A. K.

    2017-09-01

    Airborne Laser Scanning (ALS) is a widespread method for forest mapping and management purposes. While common ALS techniques provide valuable information about the forest canopy and intermediate layers, the point density near the ground may be poor due to dense overstory conditions. The current study highlights a new method for detecting stems of single trees in 3D point clouds obtained from high density ALS with a density of 300 points/m2. Compared to standard ALS data, due to lower flight height (150-200 m) this elevated point density leads to more laser reflections from tree stems. In this work, we propose a three-tiered method which works on the point, segment and object levels. First, for each point we calculate the likelihood that it belongs to a tree stem, derived from the radiometric and geometric features of its neighboring points. In the next step, we construct short stem segments based on high-probability stem points, and classify the segments by considering the distribution of points around them as well as their spatial orientation, which encodes the prior knowledge that trees are mainly vertically aligned due to gravity. Finally, we apply hierarchical clustering on the positively classified segments to obtain point sets corresponding to single stems, and perform ℓ1-based orthogonal distance regression to robustly fit lines through each stem point set. The ℓ1-based method is less sensitive to outliers compared to the least square approaches. From the fitted lines, the planimetric tree positions can then be derived. Experiments were performed on two plots from the Hochficht forest in Oberösterreich region located in Austria.We marked a total of 196 reference stems in the point clouds of both plots by visual interpretation. The evaluation of the automatically detected stems showed a classification precision of 0.86 and 0.85, respectively for Plot 1 and 2, with recall values of 0.7 and 0.67.

  20. Herpes Simplex Virus 1 Mutant with Point Mutations in UL39 Is Impaired for Acute Viral Replication in Mice, Establishment of Latency, and Explant-Induced Reactivation.

    Science.gov (United States)

    Mostafa, Heba H; Thompson, Thornton W; Konen, Adam J; Haenchen, Steve D; Hilliard, Joshua G; Macdonald, Stuart J; Morrison, Lynda A; Davido, David J

    2018-04-01

    In the process of generating herpes simplex virus 1 (HSV-1) mutations in the viral regulatory gene encoding infected cell protein 0 (ICP0), we isolated a viral mutant, termed KOS-NA, that was severely impaired for acute replication in the eyes and trigeminal ganglia (TG) of mice, defective in establishing a latent infection, and reactivated poorly from explanted TG. To identify the secondary mutation(s) responsible for the impaired phenotypes of this mutant, we sequenced the KOS-NA genome and noted that it contained two nonsynonymous mutations in UL39 , which encodes the large subunit of ribonucleotide reductase, ICP6. These mutations resulted in lysine-to-proline (residue 393) and arginine-to-histidine (residue 950) substitutions in ICP6. To determine whether alteration of these amino acids was responsible for the KOS-NA phenotypes in vivo , we recombined the wild-type UL39 gene into the KOS-NA genome and rescued its acute replication phenotypes in mice. To further establish the role of UL39 in KOS-NA's decreased pathogenicity, the UL39 mutations were recombined into HSV-1 (generating UL39 mut ), and this mutant virus showed reduced ocular and TG replication in mice comparable to that of KOS-NA. Interestingly, ICP6 protein levels were reduced in KOS-NA-infected cells relative to the wild-type protein. Moreover, we observed that KOS-NA does not counteract caspase 8-induced apoptosis, unlike wild-type strain KOS. Based on alignment studies with other HSV-1 ICP6 homologs, our data suggest that amino acid 950 of ICP6 likely plays an important role in ICP6 accumulation and inhibition of apoptosis, consequently impairing HSV-1 pathogenesis in a mouse model of HSV-1 infection. IMPORTANCE HSV-1 is a major human pathogen that infects ∼80% of the human population and can be life threatening to infected neonates or immunocompromised individuals. Effective therapies for treatment of recurrent HSV-1 infections are limited, which emphasizes a critical need to understand in

  1. Single-cell genetic expression of mutant GABAA receptors causing Human genetic epilepsy alters dendritic spine and GABAergic bouton formation in a mutation-specific manner

    Directory of Open Access Journals (Sweden)

    Pamela eLachance-Touchette

    2014-10-01

    Full Text Available Mutations in genes encoding for GABAA receptor subunits is a well-established cause of genetic generalized epilepsy. GABA neurotransmission is implicated in several developmental processes including neurite outgrowth and synapse formation. Alteration in excitatory/inhibitory synaptic activities plays a critical role in epilepsy, thus here we investigated whether mutations in α1 subunit of GABAA receptor may affect dendritic spine and GABAergic bouton formation. In particular, we examined the effects of three mutations of the GABRA1 gene (D219N, A322D and K353delins18X that were found in a cohort of families with genetic generalized epilepsy. We used a novel single-cell genetic approach, by preparing cortical organotypic cultures from GABRA1flox/flox mice and simultaneously inactivating endogenous GABRA1 and transfecting mutant α1 subunits in single glutamatergic pyramidal cells and basket GABAergic interneurons by biolistic transfection. We found that GABRA1-/- GABAergic cells showed reduced innervation field, which was rescued by co-expressing α1-A322D and α1-WT but not α1-D219N. We further found that the expression of the most severe GABRA1 missense mutation (α1-A322D induced a striking increase of spine density in pyramidal cells along with an increase in the number of mushroom-like spines. In addition, α1-A322D expression in GABAergic cells slightly increased perisomatic bouton density, whereas other mutations did not alter bouton formation. All together, these results suggest that the effects of different GABAAR mutations on GABAergic bouton and dendritic spine formation are specific to the mutation and cannot be always explained by a simple loss-of-function gene model. The use of single cell genetic manipulation in organotypic cultures may provide a better understanding of the specific and distinct neural circuit alterations caused by different GABAA receptor subunit mutations and will help define the pathophysiology of genetic

  2. Analysis of ATP-citrate lyase and malic enzyme mutants of Yarrowia lipolytica points out the importance of mannitol metabolism in fatty acid synthesis.

    Science.gov (United States)

    Dulermo, Thierry; Lazar, Zbigniew; Dulermo, Rémi; Rakicka, Magdalena; Haddouche, Ramedane; Nicaud, Jean-Marc

    2015-09-01

    The role of the two key enzymes of fatty acid (FA) synthesis, ATP-citrate lyase (Acl) and malic enzyme (Mae), was analyzed in the oleaginous yeast Yarrowia lipolytica. In most oleaginous yeasts, Acl and Mae are proposed to provide, respectively, acetyl-CoA and NADPH for FA synthesis. Acl was mainly studied at the biochemical level but no strain depleted for this enzyme was analyzed in oleaginous microorganisms. On the other hand the role of Mae in FA synthesis in Y. lipolytica remains unclear since it was proposed to be a mitochondrial NAD(H)-dependent enzyme and not a cytosolic NADP(H)-dependent enzyme. In this study, we analyzed for the first time strains inactivated for corresponding genes. Inactivation of ACL1 decreases FA synthesis by 60 to 80%, confirming its essential role in FA synthesis in Y. lipolytica. Conversely, inactivation of MAE1 has no effects on FA synthesis, except in a FA overaccumulating strain where it improves FA synthesis by 35%. This result definitively excludes Mae as a major key enzyme for FA synthesis in Y. lipolytica. During the analysis of both mutants, we observed a negative correlation between FA and mannitol level. As mannitol and FA pathways may compete for carbon storage, we inactivated YlSDR, encoding a mannitol dehydrogenase converting fructose and NADPH into mannitol and NADP+. The FA content of the resulting mutant was improved by 60% during growth on fructose, demonstrating that mannitol metabolism may modulate FA synthesis in Y. lipolytica. Copyright © 2015. Published by Elsevier B.V.

  3. A Direct Maximum Power Point Tracking Method for Single-Phase Grid Connected PV Inverters

    DEFF Research Database (Denmark)

    EL Aamri, Faicel; Maker, Hattab; Sera, Dezso

    2018-01-01

    in dynamic conditions, especially in low irradiance when the measurement of signals becomes more sensitive to noise. The proposed MPPT is designed for single-phase single-stage grid-connected PV inverters, and is based on estimating the instantaneous PV power and voltage ripples, using second...

  4. Single-Carrier Modulation for Neutral-Point-Clamped Inverters in Three-Phase Transformerless Photovoltaic Systems

    DEFF Research Database (Denmark)

    Guo, Xiaoqiang; Cavalcanti, Marcelo C.; Farias, Alexandre M.

    2013-01-01

    Modulation strategy is one of the most important issues for three-level neutral-point-clamped inverters in three-phase transformerless photovoltaic systems. A challenge for modulation is how to keep the common-mode voltages constant to reduce the leakage currents. A single-carrier modulation...

  5. Electrically continuous graphene from single crystal copper verified by terahertz conductance spectroscopy and micro four-point probe

    DEFF Research Database (Denmark)

    Buron, Jonas Christian Due; Pizzocchero, Filippo; Jessen, Bjarke Sørensen

    2014-01-01

    . Micro four-point probe resistance values measured on graphene grown on single crystalline copper in two different voltage-current configurations show close agreement with the expected distributions for a continuous 2D conductor, in contrast with previous observations on graphene grown on commercial...... noninvasive conductance characterization methods: ultrabroadband terahertz time-domain spectroscopy and micro four-point probe, which probe the electrical properties of the graphene film on different length scales, 100 nm and 10 μm, respectively. Ultrabroadband terahertz time-domain spectroscopy allows...... copper foil. The terahertz and micro four-point probe conductance values of the graphene grown on single crystalline copper shows a close to unity correlation, in contrast with those of the graphene grown on commercial copper foil, which we explain by the absence of extended defects on the microscale...

  6. A Starting Point for Fluorescence-Based Single-Molecule Measurements in Biomolecular Research

    Directory of Open Access Journals (Sweden)

    Alexander Gust

    2014-09-01

    Full Text Available Single-molecule fluorescence techniques are ideally suited to provide information about the structure-function-dynamics relationship of a biomolecule as static and dynamic heterogeneity can be easily detected. However, what type of single-molecule fluorescence technique is suited for which kind of biological question and what are the obstacles on the way to a successful single-molecule microscopy experiment? In this review, we provide practical insights into fluorescence-based single-molecule experiments aiming for scientists who wish to take their experiments to the single-molecule level. We especially focus on fluorescence resonance energy transfer (FRET experiments as these are a widely employed tool for the investigation of biomolecular mechanisms. We will guide the reader through the most critical steps that determine the success and quality of diffusion-based confocal and immobilization-based total internal reflection fluorescence microscopy. We discuss the specific chemical and photophysical requirements that make fluorescent dyes suitable for single-molecule fluorescence experiments. Most importantly, we review recently emerged photoprotection systems as well as passivation and immobilization strategies that enable the observation of fluorescently labeled molecules under biocompatible conditions. Moreover, we discuss how the optical single-molecule toolkit has been extended in recent years to capture the physiological complexity of a cell making it even more relevant for biological research.

  7. Analysis of single point and continuous wave of condensation root filling techniques by micro-computed tomography

    Directory of Open Access Journals (Sweden)

    Daniele Angerame

    2012-01-01

    Full Text Available The aim of the present microtomographic study was to investigate the quality of root canal filling and the voids formation in canals of extracted teeth instrumented with a simultaneous technique and filled with two different methods. Twenty-four single-rooted teeth were assigned to two experimental groups (no. = 12; canals were shaped with NiTi rotary files, irrigated with NaOCl and filled either with the single point (group 1 or the continuous wave of condensation technique (group 2. Specimens underwent microtomographic scanning. Collected data were statistically analyzed by nonparametric methods. Void mean percentages were found to be limited and similar between the two groups; the single point technique led to greater sealer thickness in partially oval canals.

  8. Contribution of single-strand breaks and alkali-labile bonds to the loss of infectivity of γ-irradiated phiX174 RF-DNA in E. coli cells mutant in various repair functions

    International Nuclear Information System (INIS)

    McKee, R.H.

    1975-01-01

    Twenty-one radiation sensitive mutants have been examined for their capacity to support gamma-irradiated phiX174 RF-DNA. The survival of phiX174 RF-DNA was reduced in essentially all of the sensitive mutants. The irradiated phiX174 RF-DNA was then separated into populations containing either single-strand breaks or alkali-labile bonds to examine the capacity of the mutants to repair each of the classes of lesions. It was found that all E. coli strains are unable to repair 22 percent of the single-strand breaks and all sensitive mutants are unable to repair an additional 10 percent of the breaks. All the repair functions examined are involved in single-strand break repair and none are more or less necessary than any of the others. PhiX174 RF-DNA is also inactivated by alkali-labile bonds. In the normal strains the inactivation efficiency is 0.16 lethal events per lesion with a threshold dose of 15 to 20 krads. The mutants are divided into two classes by their sensitivity to alkali-labile bonds. Both classes of mutants are also inactivated by alkali-labile bonds with efficiencies of about 0.17 and 0.29 lethal events per lesion, respectively. It is proposed that the differences seen in survival curves of phiX174 measured in the sensitive mutants is due to this difference. Although in normal cells the efficiency of inactivation of phiX174 by single-strand breaks is 50 percent greater than by alkali-labile bonds, alkali-labile bonds are produced at approximately twice the rate of single-strand breaks so alkali-labile bonds account for about 61 percent of the overall inactivation. In the mutants of least sensitivity alkali-labile bonds account for about 54 percent of the inactivating events and in the most sensitive about 67 percent

  9. Single Point Incremental Forming to increase material knowledge and production flexibility

    Science.gov (United States)

    Habraken, A. M.

    2016-08-01

    Nowadays, manufactured pieces can be divided into two groups: mass production and production of low volume number of parts. Within the second group (prototyping or small batch production), an emerging solution relies on Incremental Sheet Forming or ISF. ISF refers to processes where the plastic deformation occurs by repeated contact with a relatively small tool. More specifically, many publications over the past decade investigate Single Point Incremental Forming (SPIF) where the final shape is determined only by the tool movement. This manufacturing process is characterized by the forming of sheets by means of a CNC controlled generic tool stylus, with the sheets clamped by means of a non-workpiece-specific clamping system and in absence of a partial or a full die. The advantage is no tooling requirements and often enhanced formability, however it poses a challenge in term of process control and accuracy assurance. Note that the most commonly used materials in incremental forming are aluminum and steel alloys however other alloys are also used especially for medical industry applications, such as cobalt and chromium alloys, stainless steel and titanium alloys. Some scientists have applied incremental forming on PVC plates and other on sandwich panels composed of propylene with mild steel and aluminum metallic foams with aluminum sheet metal. Micro incremental forming of thin foils has also been developed. Starting from the scattering of the results of Finite Element (FE) simulations, when one tries to predict the tool force (see SPIF benchmark of 2014 Numisheet conference), we will see how SPIF and even micro SPIF (process applied on thin metallic sheet with a few grains within the thickness) allow investigating the material behavior. This lecture will focus on the identification of constitutive laws, on the SPIF forming mechanisms and formability as well as the failure mechanism. Different hypotheses have been proposed to explain SPIF formability, they will be

  10. Single species biofilm-forming ability of root canal isolates on gutta-percha points.

    Science.gov (United States)

    Takemura, Naoki; Noiri, Yuichiro; Ehara, Atsushi; Kawahara, Takashi; Noguchi, Nobuo; Ebisu, Shigeyuki

    2004-12-01

    The participation of bacterial biofilms in the over-filled gutta-percha points associated with refractory periapical periodontitis has recently been reported. This study investigated the initial biofilm-forming ability of root canal isolates (Enterococcus faecalis, Streptococcus sanguis, Strep. intermedius, Strep. pyogenes, Staphylococcus aureus, Fusobacterium nucleatum, Propionibacterium acnes, Porphyromonas gingivalis and Prevotella intermedia) on gutta-percha points in vitro. Each bacterial strain was suspended in 100% cell culture medium or in culture medium containing 4.5, 45 or 90% (vol/vol) serum. The bacterial suspensions were then co-incubated anaerobically with gutta-percha points for 7 d. The gutta-percha points were processed for scanning electron microscopic observation and examined for biofilm presence and thickness. E. faecalis, Strep. sanguis, Strep. intermedius, Strep. pyogenes and Staph. aureus biofilms were generated on the surfaces of the specimens incubated in culture medium supplemented with 45 or 90% (vol/vol) serum. The E. faecalis and Strep. sanguis biofilms were significantly thicker than those of Strep. intermedius, Strep. pyogenes and Staph. aureus. No biofilms were detected on the specimens incubated with F. nucleatum, Prop. acnes, Porph. gingivalis and Prev. intermedia. These findings suggest that Gram-positive facultative anaerobes have the ability to colonize and form extracellular matrices on gutta-percha points, while serum plays a crucial role in biofilm formation. Copyright Eur J Oral Sci, 2004.

  11. Insight into a Novel p53 Single Point Mutation (G389E by Molecular Dynamics Simulations

    Directory of Open Access Journals (Sweden)

    Maria Cristina De Rosa

    2010-12-01

    Full Text Available The majority of inactivating mutations of p53 reside in the central core DNA binding domain of the protein. In this computational study, we investigated the structural effects of a novel p53 mutation (G389E, identified in a patient with congenital adrenal hyperplasia, which is located within the extreme C-terminal domain (CTD of p53, an unstructured, flexible region (residues 367–393 of major importance for the regulation of the protein. Based on the three-dimensional structure of a carboxyl-terminal peptide of p53 in complex with the S100B protein, which is involved in regulation of the tumor suppressor activity, a model of wild type (WT and mutant extreme CTD was developed by molecular modeling and molecular dynamics simulation. It was found that the G389E amino acid replacement has negligible effects on free p53 in solution whereas it significantly affects the interactions of p53 with the S100B protein. The results suggest that the observed mutation may interfere with p53 transcription activation and provide useful information for site-directed mutagenesis experiments.

  12. X-Ray Radiography of Three-Point Bending of Single Human Trabecula

    Czech Academy of Sciences Publication Activity Database

    Jiroušek, Ondřej; Doktor, Tomáš; Kytýř, Daniel; Zlámal, Petr

    2012-01-01

    Roč. 45, S1 (2012), s. 261-261 ISSN 0021-9290 R&D Projects: GA ČR(CZ) GAP105/10/2305 Institutional support: RVO:68378297 Keywords : microspheres * strain measurement * three-point bending * trabecular bone * X-ray radiography Subject RIV: FI - Traumatology, Orthopedics Impact factor: 2.716, year: 2012

  13. Shape measurement system for single point incremental forming (SPIF) manufacts by using trinocular vision and random pattern

    International Nuclear Information System (INIS)

    Setti, Francesco; Bini, Ruggero; Lunardelli, Massimo; Bosetti, Paolo; Bruschi, Stefania; De Cecco, Mariolino

    2012-01-01

    Many contemporary works show the interest of the scientific community in measuring the shape of artefacts made by single point incremental forming. In this paper, we will present an algorithm able to detect feature points with a random pattern, check the compatibility of associations exploiting multi-stereo constraints and reject outliers and perform a 3D reconstruction by dense random patterns. The algorithm is suitable for a real-time application, in fact it needs just three images and a synchronous relatively fast processing. The proposed method has been tested on a simple geometry and results have been compared with a coordinate measurement machine acquisition. (paper)

  14. Problem-Solving Test: Tryptophan Operon Mutants

    Science.gov (United States)

    Szeberenyi, Jozsef

    2010-01-01

    This paper presents a problem-solving test that deals with the regulation of the "trp" operon of "Escherichia coli." Two mutants of this operon are described: in mutant A, the operator region of the operon carries a point mutation so that it is unable to carry out its function; mutant B expresses a "trp" repressor protein unable to bind…

  15. The single N-glycan deletion mutant of soluble ErbB3 protein attenuates heregulin β1-induced tumor progression by blocking of the HIF-1 and Nrf2 pathway

    Energy Technology Data Exchange (ETDEWEB)

    Takamiya, Rina, E-mail: rinataka0429@gmail.com; Takahashi, Motoko; Uehara, Yasuaki; Ariki, Shigeru; Hashimoto, Jiro; Hasegawa, Yoshihiro; Kuroki, Yoshio

    2014-11-21

    Highlights: • The sErbB3 N418Q mutant blocks heregulin β1 induced nuclear accumulation of HIF-1α. • The sErbB3 N418Q mutant attenuates cancer cell migration induced by heregulin β1. • The sErbB3 N418Q mutant blocks heregulin β1 induced nuclear accumulation of Nrf2. • The sErbB3 N418Q mutant may be a potential therapeutic application for tumor. - Abstract: It has been well documented that activation of the ErbB3–PI3K–Akt pathway is implicated in tumor survival and progression. We previously demonstrated that the single N-glycan deletion mutant of soluble ErbB3 protein (sErbB3 N418Q) attenuates heregulin β1-induced ErbB3 signaling. The active PI3K–Akt pathway augments the nuclear accumulation of hypoxia inducible factor (HIF)-1α, which activates the transcription of many target genes and drives cancer progression. In this study, we focused on the effects of sErbB3 N418Q mutant on nuclear accumulation of HIF-1α. Pretreatment with the sErbB3 N418Q mutant suppressed heregulin β1-induced HIF-1α activation in MCF7 cells. Similar results were also obtained in other breast cancer cell lines, T47D and BT474. Interestingly, these suppressive effects were not observed with the sErbB3 wild type. In addition, pretreatment with the sErbB3 N418Q mutant suppressed the cell migration of MCF7 cells induced by heregulin β1. Furthermore, incubation with heregulin β1 also induced the nuclear accumulation of Nrf2, and this effect was also reduced by the sErbB3 N418Q mutant, but not the sErbB3 wild type. These findings indicated that the sErbB3 N418Q mutant suppressed malignant formation of cancer cells by blocking of the HIF-1α and Nrf2 pathways.

  16. A High-Power-Density Single-Phase Rectifier Based on Three-Level Neutral-Point Clamped Circuits

    Directory of Open Access Journals (Sweden)

    Tao Zhou

    2017-05-01

    Full Text Available A single-phase three-level converter is suitable for medium-power applications, with an interface voltage that is higher than that of a traditional two-level configuration. The three-level neutral-point clamped converter is adopted using four switches in each bridge arm, which, compared to a two-level rectifier, leads to less voltage stress, a lower switching frequency, and switching loss on switches. The transient current control strategy is designed to control the active power. The single-phase space vector pulse width modulation (SVPWM with a voltage balance strategy is designed to solve the neutral point voltage fluctuation problem and keep the dc-link voltage stable. A 1.3 kW high-power-density prototype based on SiC MOSFET was built and tested. The experimental results verified the high performance of steady-state and dynamic responses.

  17. Highly macroscopically degenerated single-point ground states as source of specific heat capacity anomalies in magnetic frustrated systems

    Science.gov (United States)

    Jurčišinová, E.; Jurčišin, M.

    2018-04-01

    Anomalies of the specific heat capacity are investigated in the framework of the exactly solvable antiferromagnetic spin- 1 / 2 Ising model in the external magnetic field on the geometrically frustrated tetrahedron recursive lattice. It is shown that the Schottky-type anomaly in the behavior of the specific heat capacity is related to the existence of unique highly macroscopically degenerated single-point ground states which are formed on the borders between neighboring plateau-like ground states. It is also shown that the very existence of these single-point ground states with large residual entropies predicts the appearance of another anomaly in the behavior of the specific heat capacity for low temperatures, namely, the field-induced double-peak structure, which exists, and should be observed experimentally, along with the Schottky-type anomaly in various frustrated magnetic system.

  18. Characteristics of X-point lobe structures in single-null discharges on MAST

    Czech Academy of Sciences Publication Activity Database

    Harrison, J.R.; Kirk, A.; Chapman, I.T.; Cahyna, Pavel; Liu, Y.; Nardon, E.; Thornton, A.J.

    2014-01-01

    Roč. 54, č. 6 (2014), 064015-064015 ISSN 0029-5515 R&D Projects: GA ČR GAP205/11/2341 Institutional support: RVO:61389021 Keywords : tokamak * MAST * X-point * resonant magnetic perturbation Subject RIV: BL - Plasma and Gas Discharge Physics Impact factor: 3.062, year: 2014 http://iopscience.iop.org/0029-5515/54/6/064015/pdf/0029-5515_54_6_064015.pdf

  19. Characterization of Point Defects in Lithium Aluminate (LiAlO2) Single Crystals

    Science.gov (United States)

    2015-09-17

    insulating wide-band gap material currently under development for tritium breeding and radiation detection and dosimetry appli- cations. Point defects...110 xi List of Tables Table Page 1.1. Anion - Cation Bond Lengths...the valence band from the bottom of the conduction band is known as the band gap energy, and is normally represented by Eg. If a photon with an energy

  20. A QRS Detection and R Point Recognition Method for Wearable Single-Lead ECG Devices.

    Science.gov (United States)

    Chen, Chieh-Li; Chuang, Chun-Te

    2017-08-26

    In the new-generation wearable Electrocardiogram (ECG) system, signal processing with low power consumption is required to transmit data when detecting dangerous rhythms and to record signals when detecting abnormal rhythms. The QRS complex is a combination of three of the graphic deflection seen on a typical ECG. This study proposes a real-time QRS detection and R point recognition method with low computational complexity while maintaining a high accuracy. The enhancement of QRS segments and restraining of P and T waves are carried out by the proposed ECG signal transformation, which also leads to the elimination of baseline wandering. In this study, the QRS fiducial point is determined based on the detected crests and troughs of the transformed signal. Subsequently, the R point can be recognized based on four QRS waveform templates and preliminary heart rhythm classification can be also achieved at the same time. The performance of the proposed approach is demonstrated using the benchmark of the MIT-BIH Arrhythmia Database, where the QRS detected sensitivity (Se) and positive prediction (+P) are 99.82% and 99.81%, respectively. The result reveals the approach's advantage of low computational complexity, as well as the feasibility of the real-time application on a mobile phone and an embedded system.

  1. Optimal fits of diffusion constants from single-time data points of Brownian trajectories.

    Science.gov (United States)

    Boyer, Denis; Dean, David S; Mejía-Monasterio, Carlos; Oshanin, Gleb

    2012-12-01

    Experimental methods based on single particle tracking (SPT) are being increasingly employed in the physical and biological sciences, where nanoscale objects are visualized with high temporal and spatial resolution. SPT can probe interactions between a particle and its environment but the price to be paid is the absence of ensemble averaging and a consequent lack of statistics. Here we address the benchmark question of how to accurately extract the diffusion constant of one single Brownian trajectory. We analyze a class of estimators based on weighted functionals of the square displacement. For a certain choice of the weight function these functionals provide the true ensemble averaged diffusion coefficient, with a precision that increases with the trajectory resolution.

  2. Point defects and electric compensation in gallium arsenide single crystals; Punktdefekte und elektrische Kompensation in Galliumarsenid-Einkristallen

    Energy Technology Data Exchange (ETDEWEB)

    Kretzer, Ulrich

    2007-12-10

    In the present thesis the point-defect budget of gallium arsenide single crystals with different dopings is studied. It is shown, in which way the concentration of the single point defects depende on the concentration of the dopants, the stoichiometry deviation, and the position of the Fermi level. For this serve the results of the measurement-technical characterization of a large number of samples, in the fabrication of which these parameters were directedly varied. The main topic of this thesis lies in the development of models, which allow a quantitative description of the experimentally studied electrical and optical properties of gallium arsenide single crystals starting from the point-defect concentrations. Because from point defects charge carriers can be set free, their concentration determines essentially the charge-carrier concentration in the bands. In the ionized state point defects act as scattering centers for free charge carriers and influence by this the drift mobility of the charge carriers. A thermodynamic modeling of the point-defect formation yields statements on the equilibrium concentrations of the point defects in dependence on dopant concentration and stoichiometry deviation. It is show that the electrical properties of the crystals observed at room temperature result from the kinetic suppression of processes, via which the adjustment of a thermodynamic equilibrium between the point defects is mediated. [German] In der vorliegenden Arbeit wird der Punktdefekthaushalt von Galliumarsenid-Einkristallen mit unterschiedlichen Dotierungen untersucht. Es wird gezeigt, in welcher Weise die Konzentration der einzelnen Punktdefekte von der Konzentration der Dotierstoffe, der Stoechiometrieabweichung und der Lage des Ferminiveaus abhaengen. Dazu dienen die Ergebnisse der messtechnischen Charakterisierung einer grossen Anzahl von Proben, bei deren Herstellung diese Parameter gezielt variiert wurden. Der Schwerpunkt der Arbeit liegt in der Entwicklung

  3. Single-point reactive power control method on voltage rise mitigation in residential networks with high PV penetration

    DEFF Research Database (Denmark)

    Hasheminamin, Maryam; Agelidis, Vassilios; Ahmadi, Abdollah

    2018-01-01

    Voltage rise (VR) due to reverse power flow is an important obstacle for high integration of Photovoltaic (PV) into residential networks. This paper introduces and elaborates a novel methodology of an index-based single-point-reactive power-control (SPRPC) methodology to mitigate voltage rise by ...... system with high r/x ratio. Efficacy, effectiveness and cost study of SPRPC is compared to droop control to evaluate its advantages....... by absorbing adequate reactive power from one selected point. The proposed index utilizes short circuit analysis to select the best point to apply this Volt/Var control method. SPRPC is supported technically and financially by distribution network operator that makes it cost effective, simple and efficient...

  4. New, multi-dimensional reconstructions for the 12-detector, scanned focal point, single-photon tomograph

    International Nuclear Information System (INIS)

    Stoddart, H.A.; Stoddart, H.F.

    1990-01-01

    The heuristic reconstruction process used for the highly-focused, scanned focal-point tomograph described by us in 1979 has remained unchanged until recently. This paper describes an entirely new and completely rigorous reconstruction for this unique geometry which produces dramatically improved images that are least square estimates of the objects. The handling of attenuation, scattering, and object power spectra are discussed. Reconstructions are described for 2D, 3D from three slices, time varying 3D from three slices, and 3D from a large set of slices. Reconstructions for simulated and actual clinical data are shown and the accuracy for quantitation is estimated

  5. Ultrasonically Assisted Single Point Diamond Turning of Optical Mold of Tungsten Carbide

    Directory of Open Access Journals (Sweden)

    Zhanjie Li

    2018-02-01

    Full Text Available To realize high efficiency, low/no damage and high precision machining of tungsten carbide used for lens mold, a high frequency ultrasonic vibration cutting system was developed at first. Then, tungsten carbide was precisely machined with a polycrystalline diamond (PCD tool assisted by the self-developed high frequency ultrasonic vibration cutting system. Tool wear mechanism was investigated in ductile regime machining of tungsten carbide. The cutter back-off phenomenon in the process was analyzed. The subsequent experimental results of ultra-precision machining with a single crystal diamond tool showed that: under the condition of high frequency ultrasonic vibration cutting, nano-scale surface roughness can be obtained by the diamond tool with smaller tip radius and no defects like those of ground surface were found on the machined surface. Tool wear mechanisms of the single crystal diamond tool are mainly abrasive wear and micro-chipping. To solve the problem, a method of inclined ultrasonic vibration cutting with negative rake angle was put forward according to force analysis, which can further reduce tool wear and roughness of the machined surface. The investigation was important to high efficiency and quality ultra-precision machining of tungsten carbide.

  6. CScape: a tool for predicting oncogenic single-point mutations in the cancer genome.

    Science.gov (United States)

    Rogers, Mark F; Shihab, Hashem A; Gaunt, Tom R; Campbell, Colin

    2017-09-14

    For somatic point mutations in coding and non-coding regions of the genome, we propose CScape, an integrative classifier for predicting the likelihood that mutations are cancer drivers. Tested on somatic mutations, CScape tends to outperform alternative methods, reaching 91% balanced accuracy in coding regions and 70% in non-coding regions, while even higher accuracy may be achieved using thresholds to isolate high-confidence predictions. Positive predictions tend to cluster in genomic regions, so we apply a statistical approach to isolate coding and non-coding regions of the cancer genome that appear enriched for high-confidence predicted disease-drivers. Predictions and software are available at http://CScape.biocompute.org.uk/ .

  7. Potential use of point shear wave elastography for the pancreas: A single center prospective study

    Energy Technology Data Exchange (ETDEWEB)

    Kawada, Natsuko, E-mail: kawada-na@mc.pref.osaka.jp [Department of Pathology, Osaka Medical Center for Cancer and Cardiovascular Diseases, 1-3-3 Nakamichi, Higashinari, Osaka 537-8511 (Japan); Department of Gastroenterology, Kanazawa Medical University, 1-1 Daigaku, Uchinada, Ishikawa 920-0293 (Japan); Tanaka, Sachiko, E-mail: sachi686@cocoa.plala.or.jp [Osaka Center for Cancer and Cardiovascular Disease Prevention, 1-6-107, Morinomiya, Johtoh, Osaka 536-8588 (Japan); Uehara, Hiroyuki, E-mail: uehara-hi@mc.pref.osaka.jp [Department of Gastroenterology, Osaka Medical Center for Cancer and Cardiovascular Diseases, 1-3-3 Nakamichi, Higashinari, Osaka 537-8511 (Japan); Ohkawa, Kazuyoshi, E-mail: okawa-ka@mc.pref.osaka.jp [Department of Gastroenterology, Osaka Medical Center for Cancer and Cardiovascular Diseases, 1-3-3 Nakamichi, Higashinari, Osaka 537-8511 (Japan); Yamai, Takuo, E-mail: yamai-ta@mc.pref.osaka.jp [Department of Gastroenterology, Osaka Medical Center for Cancer and Cardiovascular Diseases, 1-3-3 Nakamichi, Higashinari, Osaka 537-8511 (Japan); Takada, Ryoji, E-mail: takada-ry@mc.pref.osaka.jp [Department of Gastroenterology, Osaka Medical Center for Cancer and Cardiovascular Diseases, 1-3-3 Nakamichi, Higashinari, Osaka 537-8511 (Japan); Shiroeda, Hisakazu, E-mail: shiroeda@kanazawa-med.ac.jp [Department of Gastroenterology, Kanazawa Medical University, 1-1 Daigaku, Uchinada, Ishikawa 920-0293 (Japan); Arisawa, Tomiyasu, E-mail: tarisawa@kanazawa-med.ac.jp [Department of Gastroenterology, Kanazawa Medical University, 1-1 Daigaku, Uchinada, Ishikawa 920-0293 (Japan); Tomita, Yasuhiko, E-mail: tomota-ya@mc.pref.osaka.jp [Department of Pathology, Osaka Medical Center for Cancer and Cardiovascular Diseases, 1-3-3 Nakamichi, Higashinari, Osaka 537-8511 (Japan)

    2014-04-15

    Aim: Clinical use of point shear wave elastography for the liver has been established, however, few studies demonstrated its usefulness for the pancreas. A prospective study was conducted to clarify its feasibility for the pancreas and its usefulness for the identification of high risk group for pancreatic cancer. Patients and methods: Consecutive eighty-five patients underwent point shear wave elastography for the pancreas. The success rate of shear wave velocity (SWV) measurement, that is the number of successful measurements over total 10 measurements, was recorded. The SWV of the pancreas measured at non-tumorous area was compared between patients with and without pancreatic cancer. Factors associated with high SWV were determined by logistic regression model. Results: Sixty patients were included, of these 18 had pancreatic cancer. The success rate of 100% was achieved at the head, the body and the tail of the pancreas in 80%, 83%, and 68% of the patients, respectively. The success rate of ≥80% was achieved in 100%, 100%, and 96% of the patients, respectively. Although mean SWV of the pancreas harboring pancreatic cancer tended to be higher compared with that of the pancreas without cancer (1.51 ± 0.45 m/s vs 1.43 ± 0.28 m/s), they did not reach statistical significance. Multivariate analysis showed that increased amount of alcohol intake was associated with high SWV. Conclusion: The SWV of the pancreas was measured with excellent success rate. However, tendency of higher SWV obtained from the pancreas harboring pancreatic cancer needed to be further investigated.

  8. A study of point defect aggregates in #betta#-irradiated LiF single crystals

    International Nuclear Information System (INIS)

    Frugoli, P.A.; Pimentel, C.A.F.

    1982-11-01

    Diffuse X-ray scattering near the Bragg Reflection and Bragg profile analaysis have been made in #betta#-irradiated LiF single crystal X-ray diffractometer. An estimate of the half-width of the diffraction patterns was done and preferential alteration in the profile parameters was observed. Clusters with mean parameter sizes from hundreds to thousands of angstroms were observed but each sample has presented a set of average size values. The nature of clusters was found to be dependent on the #betta#-dose: vacancy at low dose (approximately 10 MRad) and interstitial at high dose (approximately 50 MRad). Some process of coalescence at 50 MRad seems to occur. (Author) [pt

  9. Additivity of the Stabilization Effect of Single Amino Acid Substitutions in Triple Mutants of Recombinant Formate Dehydrogenase from the Soybean Glycine max.

    Science.gov (United States)

    Alekseeva, A A; Kargov, I S; Kleimenov, S Yu; Savin, S S; Tishkov, V I

    2015-01-01

    Recently, we demonstrated that the amino acid substitutions Ala267Met and Ala267Met/Ile272Val (Alekseeva et al., Biochemistry, 2012), Phe290Asp, Phe290Asn and Phe290Ser (Alekseeva et al., Prot. Eng. Des. Select, 2012) in recombinant formate dehydrogenase from soya Glycine max (SoyFDH) lead to a significant (up to 30-100 times) increase in the thermal stability of the enzyme. The substitutions Phe290Asp, Phe290Asn and Phe290Ser were introduced into double mutant SoyFDH Ala267Met/Ile272Val by site-directed mutagenesis. Combinations of three substitutions did not lead to a noticeable change in the catalytic properties of the mutant enzymes. The stability of the resultant triple mutants was studied through thermal inactivation kinetics and differential scanning calorimetry. The thermal stability of the new mutant SoyFDHs was shown to be much higher than that of their precursors. The stability of the best mutant SoyFDH Ala267Met/Ile272Val/Phe290Asp turned out to be comparable to that of the most stable wild-type formate dehydrogenases from other sources. The results obtained with both methods indicate a great synergistic contribution of individual amino acid substitutions to the common stabilization effect.

  10. High-performance analysis of single interphase cells with custom DNA probes spanning translocation break points

    Science.gov (United States)

    Weier, Heinz-Ulli G.; Munne, S.; Lersch, Robert A.; Marquez, C.; Wu, J.; Pedersen, Roger A.; Fung, Jingly

    1999-06-01

    The chromatin organization of interphase cell nuclei, albeit an object of intense investigation, is only poorly understood. In the past, this has hampered the cytogenetic analysis of tissues derived from specimens where only few cells were actively proliferating or a significant number of metaphase cells could be obtained by induction of growth. Typical examples of such hard to analyze cell systems are solid tumors, germ cells and, to a certain extent, fetal cells such as amniocytes, blastomeres or cytotrophoblasts. Balanced reciprocal translocations that do not disrupt essential genes and thus do not led to disease symptoms exit in less than one percent of the general population. Since the presence of translocations interferes with homologue pairing in meiosis, many of these individuals experience problems in their reproduction, such as reduced fertility, infertility or a history of spontaneous abortions. The majority of translocation carriers enrolled in our in vitro fertilization (IVF) programs carry simple translocations involving only two autosomes. While most translocations are relatively easy to spot in metaphase cells, the majority of cells biopsied from embryos produced by IVF are in interphase and thus unsuitable for analysis by chromosome banding or FISH-painting. We therefore set out to analyze single interphase cells for presence or absence of specific translocations. Our assay, based on fluorescence in situ hybridization (FISH) of breakpoint-spanning DNA probes, detects translocations in interphase by visual microscopic inspection of hybridization domains. Probes are prepared so that they span a breakpoint and cover several hundred kb of DNA adjacent to the breakpoint. On normal chromosomes, such probes label a contiguous stretch of DNA and produce a single hybridization domain per chromosome in interphase cells. The translocation disrupts the hybridization domain and the resulting two fragments appear as physically separated hybridization domains in

  11. A Single Phase Doubly Grounded Semi-Z-Source Inverter for Photovoltaic (PV Systems with Maximum Power Point Tracking (MPPT

    Directory of Open Access Journals (Sweden)

    Tofael Ahmed

    2014-06-01

    Full Text Available In this paper, a single phase doubly grounded semi-Z-source inverter with maximum power point tracking (MPPT is proposed for photovoltaic (PV systems. This proposed system utilizes a single-ended primary inductor (SEPIC converter as DC-DC converter to implement the MPPT algorithm for tracking the maximum power from a PV array and a single phase semi-Z-source inverter for integrating the PV with AC power utilities. The MPPT controller utilizes a fast-converging algorithm to track the maximum power point (MPP and the semi-Z-source inverter utilizes a nonlinear SPWM to produce sinusoidal voltage at the output. The proposed system is able to track the MPP of PV arrays and produce an AC voltage at its output by utilizing only three switches. Experimental results show that the fast-converging MPPT algorithm has fast tracking response with appreciable MPP efficiency. In addition, the inverter shows the minimization of common mode leakage current with its ground sharing feature and reduction of the THD as well as DC current components at the output during DC-AC conversion.

  12. Fast computation of quadrupole and hexadecapole approximations in microlensing with a single point-source evaluation

    Science.gov (United States)

    Cassan, Arnaud

    2017-07-01

    The exoplanet detection rate from gravitational microlensing has grown significantly in recent years thanks to a great enhancement of resources and improved observational strategy. Current observatories include ground-based wide-field and/or robotic world-wide networks of telescopes, as well as space-based observatories such as satellites Spitzer or Kepler/K2. This results in a large quantity of data to be processed and analysed, which is a challenge for modelling codes because of the complexity of the parameter space to be explored and the intensive computations required to evaluate the models. In this work, I present a method that allows to compute the quadrupole and hexadecapole approximations of the finite-source magnification with more efficiency than previously available codes, with routines about six times and four times faster, respectively. The quadrupole takes just about twice the time of a point-source evaluation, which advocates for generalizing its use to large portions of the light curves. The corresponding routines are available as open-source python codes.

  13. High-resolution mapping of epitopes on the C2 domain of factor VIII by analysis of point mutants using surface plasmon resonance

    Science.gov (United States)

    Nguyen, Phuong-Cac T.; Lewis, Kenneth B.; Ettinger, Ruth A.; Schuman, Jason T.; Lin, Jasper C.; Healey, John F.; Meeks, Shannon L.; Lollar, Pete

    2014-01-01

    Neutralizing anti-factor VIII (FVIII) antibodies that develop in patients with hemophilia A and in murine hemophilia A models, clinically termed “inhibitors,” bind to several distinct surfaces on the FVIII-C2 domain. To map these epitopes at high resolution, 60 recombinant FVIII-C2 proteins were generated, each having a single surface-exposed residue mutated to alanine or a conservative substitution. The binding kinetics of these muteins to 11 monoclonal, inhibitory anti-FVIII-C2 antibodies were evaluated by surface plasmon resonance and the results compared with those obtained for wild-type FVIII-C2. Clusters of residues with significantly altered binding kinetics identified “functional” B-cell epitopes, defined as those residues contributing appreciable antigen–antibody avidity. These antibodies were previously shown to neutralize FVIII activity by interfering with proteolytic activation of FVIII by thrombin or factor Xa, or with its binding to phospholipid surfaces, von Willebrand factor, or other components of the intrinsic tenase complex. Fine mapping of epitopes by surface plasmon resonance also indicated surfaces through which FVIII interacts with proteins and phospholipids as it participates in coagulation. Mutations that significantly altered the dissociation times/half-lives identified functionally important interactions within antigen–antibody interfaces and suggested specific sequence modifications to generate novel, less antigenic FVIII proteins with possible therapeutic potential for treatment of inhibitor patients. PMID:24591205

  14. On the estimation of the current density in space plasmas: Multi- versus single-point techniques

    Science.gov (United States)

    Perri, Silvia; Valentini, Francesco; Sorriso-Valvo, Luca; Reda, Antonio; Malara, Francesco

    2017-06-01

    Thanks to multi-spacecraft mission, it has recently been possible to directly estimate the current density in space plasmas, by using magnetic field time series from four satellites flying in a quasi perfect tetrahedron configuration. The technique developed, commonly called ;curlometer; permits a good estimation of the current density when the magnetic field time series vary linearly in space. This approximation is generally valid for small spacecraft separation. The recent space missions Cluster and Magnetospheric Multiscale (MMS) have provided high resolution measurements with inter-spacecraft separation up to 100 km and 10 km, respectively. The former scale corresponds to the proton gyroradius/ion skin depth in ;typical; solar wind conditions, while the latter to sub-proton scale. However, some works have highlighted an underestimation of the current density via the curlometer technique with respect to the current computed directly from the velocity distribution functions, measured at sub-proton scales resolution with MMS. In this paper we explore the limit of the curlometer technique studying synthetic data sets associated to a cluster of four artificial satellites allowed to fly in a static turbulent field, spanning a wide range of relative separation. This study tries to address the relative importance of measuring plasma moments at very high resolution from a single spacecraft with respect to the multi-spacecraft missions in the current density evaluation.

  15. Ambiguity resolution in precise point positioning with hourly data for global single receiver

    Science.gov (United States)

    Zhang, Xiaohong; Li, Pan; Guo, Fei

    2013-01-01

    Integer ambiguity resolution (IAR) can improve precise point positioning (PPP) performance significantly. IAR for PPP became a highlight topic in global positioning system (GPS) community in recent years. More and more researchers focus on this issue. Progress has been made in the latest years. In this paper, we aim at investigating and demonstrating the performance of a global zero-differenced (ZD) PPP IAR service for GPS users by providing routine ZD uncalibrated fractional offsets (UFOs) for wide-lane and narrow-lane. Data sets from all IGS stations collected on DOY 1, 100, 200 and 300 of 2010 are used to validate and demonstrate this global service. Static experiment results show that an accuracy better than 1 cm in horizontal and 1-2 cm in vertical could be achieved in ambiguity-fixed PPP solution with only hourly data. Compared with PPP float solution, an average improvement reaches 58.2% in east, 28.3% in north and 23.8% in vertical for all tested stations. Results of kinematic experiments show that the RMS of kinematic PPP solutions can be improved from 21.6, 16.6 and 37.7 mm to 12.2, 13.3 and 34.3 mm for the fixed solutions in the east, north and vertical components, respectively. Both static and kinematic experiments show that wide-lane and narrow-lane UFO products of all satellites can be generated and provided in a routine way accompanying satellite orbit and clock products for the PPP user anywhere around the world, to obtain accurate and reliable ambiguity-fixed PPP solutions.

  16. Channel capacity of TDD-OFDM-MIMO for multiple access points in a wireless single-frequency-network

    DEFF Research Database (Denmark)

    Takatori, Y.; Fitzek, Frank; Tsunekawa, K.

    2005-01-01

    MIMO data transmission scheme, which combines Single-Frequency-Network (SFN) with TDD-OFDM-MIMO applied for wireless LAN networks. In our proposal, we advocate to use SFN for multiple access points (MAP) MIMO data transmission. The goal of this approach is to achieve very high channel capacity in both...... LOS and non line of sight (NLOS) environments. The channel capacity of the proposed method is derived for the direct path environments and it confirms the effectiveness of the proposed scheme in the LOS scenario. Moreover, solid computer simulation results confirm the effectiveness of the proposed...

  17. Theoretical Basis for Stochastic Optimization Starting from a Single Point in the Search Space Formed by Real DNA Molecules

    Science.gov (United States)

    Someya, Hiroshi; Yamamura, Masayuki; Sakamoto, Kensaku

    This paper discusses DNA-based stochastic optimizations under the constraint that the search starts from a given point in a search space. Generally speaking, a stochastic optimization method explores a search space and finds out the optimum or a sub-optimum after many cycles of trials and errors. This search process could be implemented efficiently by ``molecular computing'', which processes DNA molecules by the techniques of molecular biology to generate and evaluate a vast number of solution candidates at a time. We assume the exploration starting from a single point, and propose a method to embody DNA-based optimization under this constraint, because this method has a promising application in the research field of protein engineering. In this application, a string of nucleotide bases (a base sequence) encodes a protein possessing a specific activity, which could be given as a value of an objective function. Thus, a problem of obtaining a protein with the optimum or a sub-optimum about the desired activity corresponds to a combinatorial problem of obtaining a base sequence giving the optimum or a sub-optimum in the sequence space. Biologists usually modify a base sequence corresponding to a naturally occurring protein into another sequence giving a desired activity. In other words, they explore the space in the proximity of a natural protein as a start point. We first examined if the optimization methods that involve a single start point, such as simulated annealing, Gibbs sampler, and MH algorithms, can be implemented by DNA-based operations. Then, we proposed an application of genetic algorithm, and examined the performance of this application on a model fitness landscape by computer experiments. These experiments gave helpful guidelines in the embodiments of DNA-based stochastic optimization, including a better design of crossover operator.

  18. Detecting shifts in gene regulatory networks during time-course experiments at single-time-point temporal resolution.

    Science.gov (United States)

    Takenaka, Yoichi; Seno, Shigeto; Matsuda, Hideo

    2015-10-01

    Comprehensively understanding the dynamics of biological systems is one of the greatest challenges in biology. Vastly improved biological technologies have provided vast amounts of information that must be understood by bioinformatics and systems biology researchers. Gene regulations have been frequently modeled by ordinary differential equations or graphical models based on time-course gene expression profiles. The state-of-the-art computational approaches for analyzing gene regulations assume that their models are same throughout time-course experiments. However, these approaches cannot easily analyze transient changes at a time point, such as diauxic shift. We propose a score that analyzes the gene regulations at each time point. The score is based on the information gains of information criterion values. The method detects the shifts in gene regulatory networks (GRNs) during time-course experiments with single-time-point resolution. The effectiveness of the method is evaluated on the diauxic shift from glucose to lactose in Escherichia coli. Gene regulation shifts were detected at two time points: the first corresponding to the time at which the growth of E. coli ceased and the second corresponding to the end of the experiment, when the nutrient sources (glucose and lactose) had become exhausted. According to these results, the proposed score and method can appropriately detect the time of gene regulation shifts. The method based on the proposed score provides a new tool for analyzing dynamic biological systems. Because the score value indicates the strength of gene regulation at each time point in a gene expression profile, it can potentially infer hidden GRNs from time-course experiments.

  19. Evaluation of the Expression of Amyloid Precursor Protein and the Ratio of Secreted Amyloid Beta 42 to Amyloid Beta 40 in SH-SY5Y Cells Stably Transfected with Wild-Type, Single-Mutant and Double-Mutant Forms of the APP Gene for the Study of Alzheimer's Disease Pathology.

    Science.gov (United States)

    Pahrudin Arrozi, Aslina; Shukri, Siti Nur Syazwani; Wan Ngah, Wan Zurinah; Mohd Yusof, Yasmin Anum; Ahmad Damanhuri, Mohd Hanafi; Makpol, Suzana

    2017-11-01

    Neuroblastoma cell lines such as SH-SY5Y are the most frequently utilized models in neurodegenerative research, and their use has advanced the understanding of the pathology of neurodegeneration over the past few decades. In Alzheimer's disease (AD), several pathogenic mutations have been described, all of which cause elevated levels of pathological hallmarks such as amyloid-beta (Aβ). Although the genetics of Alzheimer's disease is well known, familial AD only accounts for a small number of cases in the population, with the rest being sporadic AD, which contains no known mutations. Currently, most of the in vitro models used to study AD pathogenesis only examine the level of Aβ42 as a confirmation of successful model generation and only perform comparisons between wild-type APP and single mutants of the APP gene. Recent findings have shown that the Aβ42/40 ratio in cerebrospinal fluid (CSF) is a better diagnostic indicator for AD patients than is Aβ42 alone and that more extensive Aβ formation, such as accumulation of intraneuronal Aβ, Aβ plaques, soluble oligomeric Aβ (oAβ), and insoluble fibrillar Aβ (fAβ) occurs in TgCRND8 mice expressing a double-mutant form (Swedish and Indiana) of APP, later leading to greater progressive impairment of the brain. In this study, we generated SH-SY5Y cells stably transfected separately with wild-type APP, the Swedish mutation of APP, and the Swedish and Indiana mutations of APP and evaluated the APP expression as well as the Aβ42/40 ratio in those cells. The double-mutant form of APP (Swedish/Indiana) expressed markedly high levels of APP protein and showed a high Aβ2/40 ratio compared to wild-type and single-mutant cells.

  20. Enhancement of the point defect pinning effect in Mo-doped Bi2212 single crystals of reduced anisotropy

    CERN Document Server

    Han, S H; Dai, Y; Zhang, Y; Zhang, H; Zhao, Y

    2002-01-01

    High quality Bi sub 2 Sr sub 2 CaCu sub 2 sub - sub x Mo sub x O sub y (x = 0, 0.01 and 0.02) single crystals have been grown by a self-flux method in a horizontal temperature gradient and their flux pinning and irreversibility behaviour have been investigated. The irreversibility lines of the undoped and Mo-doped Bi2212 crystals have been greatly improved by reducing the anisotropy parameter gamma. However, this improvement is much more pronounced for Mo-doped crystals than for the undoped ones. The peak effect of magnetization loops also changes with both Mo-doping and gamma. The results provide strong evidence that the point defect pinning served by Mo is greatly enhanced when the anisotropy of the system is reduced.

  1. Revisiting single-point incremental forming and formability/failure diagrams by means of finite elements and experimentation

    DEFF Research Database (Denmark)

    Silva, M. B.; Skjødt, Martin; Bay, Niels

    2009-01-01

    the forming limits determined by the analytical framework with experimental values. It is shown that agreement between analytical, finite element, and experimental results is good, implying that the previously proposed analytical framework can be utilized to explain the mechanics of deformation......In a previously published work, the current authors presented an analytical framework, built upon the combined utilization of membrane analysis and ductile damage mechanics, that is capable of modelling the fundamentals of single-point incremental forming (SPIF) of metallic sheets. The analytical...... mode of deformation. The study of the morphology of the cracks combined with the experimentally observed suppression of neck formation enabled the authors to conclude that traditional forming limit curves are inapplicable for describing failure. Instead, fracture forming limit curves should be employed...

  2. High performance control strategy for single-phase three-level neutral-point-clamped traction four-quadrant converters

    DEFF Research Database (Denmark)

    Kejian, Song; Konstantinou, Georgios; Jing, Li

    2017-01-01

    multiple unit traction system is proposed in this study. The strategy is based on a multiple frequency tuned quasi-proportional resonant controller in the ac-side current loop and a multiple frequency tuned notch filter in the dc-link voltage loop. Under the typical supply voltage distortion present......Operational data from Chinese railways indicate a number of challenges for traction four-quadrant converter (4QC) control including low-order voltage and current harmonics and reference tracking. A control strategy for a single-phase three-level neutral-point-clamped 4QC employed in the electric...... in the network, the proposed control strategy improves the current tracking performance and reduces the line current harmonics, when compared with conventional control strategies. Experimental results, both under purely sinusoidal and distorted supply voltages, validate the effectiveness of the proposed control...

  3. Experimental and numerical study on optimization of the single point incremental forming of AINSI 304L stainless steel sheet

    Science.gov (United States)

    Saidi, B.; Giraud-Moreau, L.; Cherouat, A.; Nasri, R.

    2017-09-01

    AINSI 304L stainless steel sheets are commonly formed into a variety of shapes for applications in the industrial, architectural, transportation and automobile fields, it’s also used for manufacturing of denture base. In the field of dentistry, there is a need for personalized devises that are custom made for the patient. The single point incremental forming process is highly promising in this area for manufacturing of denture base. The single point incremental forming process (ISF) is an emerging process based on the use of a spherical tool, which is moved along CNC controlled tool path. One of the major advantages of this process is the ability to program several punch trajectories on the same machine in order to obtain different shapes. Several applications of this process exist in the medical field for the manufacturing of personalized titanium prosthesis (cranial plate, knee prosthesis...) due to the need of product customization to each patient. The objective of this paper is to study the incremental forming of AISI 304L stainless steel sheets for future applications in the dentistry field. During the incremental forming process, considerable forces can occur. The control of the forming force is particularly important to ensure the safe use of the CNC milling machine and preserve the tooling and machinery. In this paper, the effect of four different process parameters on the maximum force is studied. The proposed approach consists in using an experimental design based on experimental results. An analysis of variance was conducted with ANOVA to find the input parameters allowing to minimize the maximum forming force. A numerical simulation of the incremental forming process is performed with the optimal input process parameters. Numerical results are compared with the experimental ones.

  4. Investigating the use of multi-point coupling for single-sensor bearing estimation in one direction

    Science.gov (United States)

    Woolard, Americo G.; Phoenix, Austin A.; Tarazaga, Pablo A.

    2018-04-01

    Bearing estimation of radially propagating symmetric waves in solid structures typically requires a minimum of two sensors. As a test specimen, this research investigates the use of multi-point coupling to provide directional inference using a single-sensor. By this provision, the number of sensors required for localization can be reduced. A finite-element model of a beam is constructed with a symmetrically placed bipod that has asymmetric joint-stiffness properties. Impulse loading is applied at different points along the beam, and measurements are taken from the apex of the bipod. A technique is developed to determine the direction-of-arrival of the propagating wave. The accuracy when using the bipod with the developed technique is compared against results gathered without the bipod and measuring from an asymmetric location along the beam. The results show 92% accuracy when the bipod is used, compared to 75% when measuring without the bipod from an asymmetric location. A geometry investigation finds the best accuracy results when one leg of the bipod has a low stiffness and a large diameter relative to the other leg.

  5. Single-Point Incremental Forming of Two Biocompatible Polymers: An Insight into Their Thermal and Structural Properties

    Directory of Open Access Journals (Sweden)

    Luis Marcelo Lozano-Sánchez

    2018-04-01

    Full Text Available Sheets of polycaprolactone (PCL and ultra-high molecular weight polyethylene (UHMWPE were fabricated and shaped by the Single-Point Incremental Forming process (SPIF. The performance of these biocompatible polymers in SPIF was assessed through the variation of four main parameters: the diameter of the forming tool, the spindle speed, the feed rate, and the step size based on a Box–Behnken design of experiments of four variables and three levels. The design of experiments allowed us to identify the parameters that most affect the forming of PCL and UHMWPE. The study was completed by means of a deep characterization of the thermal and structural properties of both polymers. These properties were correlated to the performance of the polymers observed in SPIF, and it was found that the polymer chains are oriented as a consequence of the SPIF processing. Moreover, by X-ray diffraction it was proved that polymer chains behave differently on each surface of the fabricated parts, since the chains on the surface in contact with the forming tool are oriented horizontally, while on the opposite surface they are oriented in the vertical direction. The unit cell of UHMWPE is distorted, passing from an orthorhombic cell to a monoclinic due to the slippage between crystallites. This slippage between crystallites was observed in both PCL and UHMWPE, and was identified as an alpha star thermal transition located in the rubbery region between the glass transition and the melting point of each polymer.

  6. A Single Point Mutation within the Coding Sequence of Cholera Toxin B Subunit Will Increase Its Expression Yield

    OpenAIRE

    Bakhshi, Bita; Boustanshenas, Mina; Ghorbani, Masoud

    2014-01-01

    Background: Cholera toxin B subunit (CTB) has been extensively considered as an immunogenic and adjuvant protein, but its yield of expression is not satisfactory in many studies. The aim of this study was to compare the expression of native and mutant recombinant CTB (rCTB) in pQE vector. Methods: ctxB fragment from Vibrio cholerae O1 ATCC14035 containing the substitution of mutant ctxB for amino acid S128T was amplified by PCR and cloned in pGETM-T easy vector. It was then transformed to E. ...

  7. Mitigation of Critical Single Point Failure (SPF) Material - Laminac 4116 Binder Replacement Program for Parachute and Cluster Stars Illuminant Compositions for Hand Held Signals

    National Research Council Canada - National Science Library

    Lakshminarayanan, G. R; Chen, Gary; Ames, Richard; Lee, Wai T; Wejsa, James L

    2006-01-01

    Laminac 4116 binder has been identified as a single point failure (SPF) material since it is being produced by only one company and there is a possibility that the company may discontinue production due to low product demand...

  8. Modeling and simulation of the main metabolism in Escherichia coli and its several single-gene knockout mutants with experimental verification

    Directory of Open Access Journals (Sweden)

    McFadden Johnjoe

    2010-11-01

    Full Text Available Abstract Background It is quite important to simulate the metabolic changes of a cell in response to the change in culture environment and/or specific gene knockouts particularly for the purpose of application in industry. If this could be done, the cell design can be made without conducting exhaustive experiments, and one can screen out the promising candidates, proceeded by experimental verification of a select few of particular interest. Although several models have so far been proposed, most of them focus on the specific metabolic pathways. It is preferred to model the whole of the main metabolic pathways in Escherichia coli, allowing for the estimation of energy generation and cell synthesis, based on intracellular fluxes and that may be used to characterize phenotypic growth. Results In the present study, we considered the simulation of the main metabolic pathways such as glycolysis, TCA cycle, pentose phosphate (PP pathway, and the anapleorotic pathways using enzymatic reaction models of E. coli. Once intracellular fluxes were computed by this model, the specific ATP production rate, the specific CO2 production rate, and the specific NADPH production rate could be estimated. The specific ATP production rate thus computed was used for the estimation of the specific growth rate. The CO2 production rate could be used to estimate cell yield, and the specific NADPH production rate could be used to determine the flux of the oxidative PP pathway. The batch and continuous cultivations were simulated where the changing patterns of extracellular and intra-cellular metabolite concentrations were compared with experimental data. Moreover, the effects of the knockout of such pathways as Ppc, Pck and Pyk on the metabolism were simulated. It was shown to be difficult for the cell to grow in Ppc mutant due to low concentration of OAA, while Pck mutant does not necessarily show this phenomenon. The slower growth rate of the Ppc mutant was properly

  9. Dynamics of Pseudomonas aeruginosa azurin and its Cys3Ser mutant at single crystal gold surfaces investigated by cyclic voltammetry and atomic force microscopy

    DEFF Research Database (Denmark)

    Friis, Esben P.; Andersen, Jens Enevold Thaulov; Madsen, Lars Lithen

    1997-01-01

    Cyclic voltammetry of Pseudomonas aeruginosa azurin on polycrystalline gold is reversible (E0=360mV vs she;50mM ammonium acetate) but the voltammetric signals decay with time constants of about 3x10-3 s-1. No signal is observed for monocrystalline Au(111). Cys3Ser azurin is electrochemically...... into the solution, recovering the free Au(111) surface. The cyclic voltammetry and AFM data are consistent with time dependent adsorption of the azurins on gold via the disulphide bridge (wild-type) or free thiol group (Cys3Ser mutant)....

  10. Mutants of alfalfa mosaic virus

    International Nuclear Information System (INIS)

    Roosien, J.

    1983-01-01

    In this thesis the isolation and characterization of a number of mutants of alfalfa mosaic virus, a plant virus with a coat protein dependent genome, is described. Thermo-sensitive (ts) mutants were selected since, at least theoretically, ts mutations can be present in all virus coded functions. It was found that a high percentage of spontaneous mutants, isolated because of their aberrant symptoms, were ts. The majority of these isolates could grow at the non-permissive temperature in the presence of a single wild type (wt) component. To increase the mutation rate virus preparations were treated with several mutagens. After nitrous acid treatment or irradiation with ultraviolet light, an increase in the level of mutations was observed. UV irradiation was preferred since it did not require large amounts of purified viral components. During the preliminary characterization of potential ts mutants the author also obtained one structural and several symptom mutants which were analysed further (chapter 7, 8 and 9). The properties of the ts mutants are described in chapter 3-7. (Auth.)

  11. Evaluation of mixing downstream of tees in duct systems with respect to single point representative air sampling.

    Science.gov (United States)

    Kim, Taehong; O'Neal, Dennis L; Ortiz, Carlos

    2006-09-01

    Air duct systems in nuclear facilities must be monitored with continuous sampling in case of an accidental release of airborne radionuclides. The purpose of this work is to identify the air sampling locations where the velocity and contaminant concentrations fall below the 20% coefficient of variation required by the American National Standards Institute/Health Physics Society N13.1-1999. Experiments of velocity and tracer gas concentration were conducted on a generic "T" mixing system which included combinations of three sub ducts, one main duct, and air velocities from 0.5 to 2 m s (100 to 400 fpm). The experimental results suggest that turbulent mixing provides the accepted velocity coefficients of variation after 6 hydraulic diameters downstream of the T-junction. About 95% of the cases achieved coefficients of variation below 10% by 6 hydraulic diameters. However, above a velocity ratio (velocity in the sub duct/velocity in the main duct) of 2, velocity profiles were uniform in a shorter distance downstream of the T-junction as the velocity ratio went up. For the tracer gas concentration, the distance needed for the coefficients of variation to drop 20% decreased with increasing velocity ratio due to the sub duct airflow momentum. The results may apply to other duct systems with similar geometries and, ultimately, be a basis for selecting a proper sampling location under the requirements of single point representative sampling.

  12. Role of single-point mutations and deletions on transition temperatures in ideal proteinogenic heteropolymer chains in the gas phase.

    Science.gov (United States)

    Olivares-Quiroz, L

    2016-07-01

    A coarse-grained statistical mechanics-based model for ideal heteropolymer proteinogenic chains of non-interacting residues is presented in terms of the size K of the chain and the set of helical propensities [Formula: see text] associated with each residue j along the chain. For this model, we provide an algorithm to compute the degeneracy tensor [Formula: see text] associated with energy level [Formula: see text] where [Formula: see text] is the number of residues with a native contact in a given conformation. From these results, we calculate the equilibrium partition function [Formula: see text] and characteristic temperature [Formula: see text] at which a transition from a low to a high entropy states is observed. The formalism is applied to analyze the effect on characteristic temperatures [Formula: see text] of single-point mutations and deletions of specific amino acids [Formula: see text] along the chain. Two probe systems are considered. First, we address the case of a random heteropolymer of size K and given helical propensities [Formula: see text] on a conformational phase space. Second, we focus our attention to a particular set of neuropentapeptides, [Met-5] and [Leu-5] enkephalins whose thermodynamic stability is a key feature on their coupling to [Formula: see text] and [Formula: see text] receptors and the triggering of biochemical responses.

  13. Influence of Fiber Orientation on Single-Point Cutting Fracture Behavior of Carbon-Fiber/Epoxy Prepreg Sheets

    Directory of Open Access Journals (Sweden)

    Yingying Wei

    2015-10-01

    Full Text Available The purpose of this article is to investigate the influences of carbon fibers on the fracture mechanism of carbon fibers both in macroscopic view and microscopic view by using single-point flying cutting method. Cutting tools with three different materials were used in this research, namely, PCD (polycrystalline diamond tool, CVD (chemical vapor deposition diamond thin film coated carbide tool and uncoated carbide tool. The influence of fiber orientation on the cutting force and fracture topography were analyzed and conclusions were drawn that cutting forces are not affected by cutting speeds but significantly influenced by the fiber orientation. Cutting forces presented smaller values in the fiber orientation of 0/180° and 15/165° but the highest one in 30/150°. The fracture mechanism of carbon fibers was studied in different cutting conditions such as 0° orientation angle, 90° orientation angle, orientation angles along fiber direction, and orientation angles inverse to the fiber direction. In addition, a prediction model on the cutting defects of carbon fiber reinforced plastic was established based on acoustic emission (AE signals.

  14. Accurate mitochondrial DNA sequencing using off-target reads provides a single test to identify pathogenic point mutations.

    Science.gov (United States)

    Griffin, Helen R; Pyle, Angela; Blakely, Emma L; Alston, Charlotte L; Duff, Jennifer; Hudson, Gavin; Horvath, Rita; Wilson, Ian J; Santibanez-Koref, Mauro; Taylor, Robert W; Chinnery, Patrick F

    2014-12-01

    Mitochondrial disorders are a common cause of inherited metabolic disease and can be due to mutations affecting mitochondrial DNA or nuclear DNA. The current diagnostic approach involves the targeted resequencing of mitochondrial DNA and candidate nuclear genes, usually proceeds step by step, and is time consuming and costly. Recent evidence suggests that variations in mitochondrial DNA sequence can be obtained from whole-exome sequence data, raising the possibility of a comprehensive single diagnostic test to detect pathogenic point mutations. We compared the mitochondrial DNA sequence derived from off-target exome reads with conventional mitochondrial DNA Sanger sequencing in 46 subjects. Mitochondrial DNA sequences can be reliably obtained using three different whole-exome sequence capture kits. Coverage correlates with the relative amount of mitochondrial DNA in the original genomic DNA sample, heteroplasmy levels can be determined using variant and total read depths, and-providing there is a minimum read depth of 20-fold-rare sequencing errors occur at a rate similar to that observed with conventional Sanger sequencing. This offers the prospect of using whole-exome sequence in a diagnostic setting to screen not only all protein coding nuclear genes but also all mitochondrial DNA genes for pathogenic mutations. Off-target mitochondrial DNA reads can also be used to assess quality control and maternal ancestry, inform on ethnic origin, and allow genetic disease association studies not previously anticipated with existing whole-exome data sets.

  15. Comparison of plastic strains on AA5052 by single point incremental forming process using digital image processing

    Energy Technology Data Exchange (ETDEWEB)

    Mugendiran, V.; Gnanavelbabu, A. [Anna University, Chennai, Tamilnadu (India)

    2017-06-15

    In this study, a surface based strain measurement was used to determine the formability of the sheet metal. A strain measurement may employ manual calculation of plastic strains based on the reference circle and the deformed circle. The manual calculation method has a greater margin of error in the practical applications. In this paper, an attempt has been made to compare the formability by implementing three different theoretical approaches: Namely conventional method, least square method and digital based strain measurements. As the sheet metal was formed by a single point incremental process the etched circles get deformed into elliptical shapes approximately, image acquisition has been done before and after forming. The plastic strains of the deformed circle grids are calculated based on the non- deformed reference. The coordinates of the deformed circles are measured by various image processing steps. Finally the strains obtained from the deformed circle are used to plot the forming limit diagram. To evaluate the accuracy of the system, the conventional, least square and digital based method of prediction of the forming limit diagram was compared. Conventional method and least square method have marginal error when compared with digital based processing method. Measurement of strain based on image processing agrees well and can be used to improve the accuracy and to reduce the measurement error in prediction of forming limit diagram.

  16. PIV study of non-Marangoni surface flows in thin liquid films induced by single- and multi-point thermodes

    Science.gov (United States)

    Cui, Nai-Yi; Wang, Song-Po

    2018-03-01

    The non-Marangoni directional flows, which can occur in only very thin liquid films, have been studied using particle image velocimetry techniques. Single- and multi-point thermodes have been used in this study for generating the flows. The results show that the direction of these flows is governed by the variation trend of the thickness of the film and the shape of the temperature profile. A hot thermode always drives a thick-to-thin flow, whereas a cold thermode always drives a flow in the opposite direction. Increasing the temperature difference between the thermode and the ambience, or decreasing the thickness of the liquid film, can accelerate the flow speed. However, the flow speed cannot exceed an upper limit. When more than one thermode was used, different flow patterns, including thick-to-thin streams driven by hot thermodes and thin-to-thick streams driven by cold thermodes, could be formed. The experimental results strongly suggest that these flows were not driven by thermo-capillary forces but by a newly proposed thermo-dynamic mechanism.

  17. Profiling of the toxicity mechanisms of coated and uncoated silver nanoparticles to yeast Saccharomyces cerevisiae BY4741 using a set of its 9 single-gene deletion mutants defective in oxidative stress response, cell wall or membrane integrity and endocytosis.

    Science.gov (United States)

    Käosaar, Sandra; Kahru, Anne; Mantecca, Paride; Kasemets, Kaja

    2016-09-01

    The widespread use of nanosilver in various antibacterial, antifungal, and antiviral products warrants the studies of the toxicity pathways of nanosilver-enabled materials toward microbes and viruses. We profiled the toxicity mechanisms of uncoated, casein-coated, and polyvinylpyrrolidone-coated silver nanoparticles (AgNPs) using Saccharomyces cerevisiae wild-type (wt) and its 9 single-gene deletion mutants defective in oxidative stress (OS) defense, cell wall/membrane integrity, and endocytosis. The 48-h growth inhibition assay in organic-rich growth medium and 24-h cell viability assay in deionized (DI) water were applied whereas AgNO3, H2O2, and SDS served as positive controls. Both coated AgNPs (primary size 8-12nm) were significantly more toxic than the uncoated (~85nm) AgNPs. All studied AgNPs were ~30 times more toxic if exposed to yeast cells in DI water than in the rich growth medium: the IC50 based on nominal concentration of AgNPs in the growth inhibition test ranged from 77 to 576mg Ag/L and in the cell viability test from 2.7 to 18.7mg Ag/L, respectively. Confocal microscopy showed that wt but not endocytosis mutant (end3Δ) internalized AgNPs. Comparison of toxicity patterns of wt and mutant strains defective in OS defense and membrane integrity revealed that the toxicity of the studied AgNPs to S. cerevisiae was not caused by the OS or cell wall/membrane permeabilization. Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. Low-pressure, single-point grout injection for tank heel sludge mixing and in-situ immobilization

    International Nuclear Information System (INIS)

    Whyatt, G.A.; Hymas, C.R.

    1998-09-01

    This report describes tests conducted in an approximately 9-ft diameter test tank situated outside the 336 building in Hanford's 300 area. The tests were performed to measure the ability of jets of grout slurry to mobilize and mix simulated tank sludge. The technique is intended for in situ immobilization of tank waste heels. The current approach uses a single, rotated, larger-diameter nozzle driven at lower pressure. Due to the larger diameter, the potential for plugging is reduced and the effective radius around an injection point over which the jet is effective in mobilizing sludge from the tank bottom can be made larger. A total of three grout injection tests were conducted in a 9-ft diameter tank. In each case, a 2-in. layer of kaolin clay paste was placed on a dry tank floor to simulate a sludge heel. The clay was covered with 4 inches of water. The grout slurry, consisting of Portland cement, class F fly ash, and eater, was prepared and delivered by an offsite vendor. In the third test, the sludge in half of the tank was replaced by a layer of 20x50 mesh zeolite, and bentonite clay was added to the grout formulation. After injection, the grout was allowed to set and then the entire grout monolith was manually broken up and excavated using a jack hammer. Intact pieces of clay were visually apparent due to a sharp color contrast between the grout and clay. Remaining clay deposits were collected and weighed and suspended clay pieces within the monolith were photographed. The mobilization performance of the grout jets exceeded expectations

  19. Van Allen Probes Science Gateway: Single-Point Access to Long-Term Radiation Belt Measurements and Space Weather Nowcasting

    Science.gov (United States)

    Romeo, G.; Barnes, R. J.; Ukhorskiy, A. Y.; Sotirelis, T.; Stephens, G.

    2017-12-01

    The Science Gateway gives single-point access to over 4.5 years of comprehensive wave and particle measurements from the Van Allen Probes NASA twin-spacecraft mission. The Gateway provides a set of visualization and data analysis tools including: HTML5-based interactive visualization of high-level data products from all instrument teams in the form of: line plots, orbital content plots, dynamical energy spectra, L-shell context plots (including two-spacecraft plotting), FFT spectra of wave data, solar wind and geomagnetic indices data, etc.; download custom multi-instrument CDF data files of selected data products; publication quality plots of digital data; combined orbit predicts for mission planning and coordination including: Van Allen Probes, MMS, THEMIS, Arase (ERG), Cluster, GOES, Geotail, FIREBIRD; magnetic footpoint calculator for coordination with LEO and ground-based assets; real-time computation and processing of empirical magnetic field models - computation of magnetic ephemeris, computation of adiabatic invariants. Van Allen Probes is the first spacecraft mission to provide a nowcast of the radiation environment in the heart of the radiation belts, where the radiation levels are the highest and most dangerous for spacecraft operations. For this purpose, all instruments continuously broadcast a subset of their science data in real time. Van Allen Probes partners with four foreign institutions who operate ground stations that receive the broadcast: Korea (KASI), the Czech republic (CAS), Argentina (CONAE), and Brazil (INPE). The SpWx broadcast is then collected at APL and delivered to the community via the Science Gateway.

  20. Assessment of left ventricular function with single breath-hold highly accelerated cine MRI combined with guide-point modeling

    International Nuclear Information System (INIS)

    Heilmaier, Christina; Nassenstein, Kai; Nielles-Vallespin, Sonia; Zuehlsdorff, Sven; Hunold, Peter; Barkhausen, Joerg

    2010-01-01

    Purpose: To prospectively assess the performance of highly accelerated cine MRI in multi-orientations combined with a new guide-point modeling post-processing technique (GPM approach) for assessment of left ventricular (LV) function compared to the standard summation of slices method based on a stack of short axis views (SoS approach). Materials and methods: 33 consecutive patients were examined on a 1.5 T scanner with a standard steady state free precession (SSFP) sequence (TR, 3.0 ms; TE, 1.5 m; flip angle (FA), 60 o ; acceleration factor (AF), 2) analyzed with the SoS method and a highly accelerated, single breath-hold temporal parallel acquisition SSFP sequence (TR, 4.6 ms; TE, 1.1 ms; AF, 3) post-processed with the GPM method. LV function values were measured by two independent readers with different experience in cardiac MRI and compared by using the paired t-test and F-test. Inter- and intraobserver agreements were calculated using Bland-Altman-Plots. Results: Mean acquisition and post-processing time was significantly shorter with the GPM approach (15 s/3 min versus 360 s/6 min). For all LV function parameters interobserver agreement between the experienced and non-experienced reader was significantly improved when the GPM approach was used. However, end-diastolic and end-systolic volumes were larger for the GPM technique when compared to the SoS method (P 0.121). In both readers and for all parameters variances did not differ significantly (P ≥ 0.409) and the two approaches showed an excellent linear correlation (r > 0.951). Conclusion: Due to its accurate, fast and reproducible assessment of LV function parameters highly accelerated MRI combined with the GPM technique may become the technique of first choice for assessment of LV function in clinical routine.

  1. An elevated neutrophil-lymphocyte ratio is associated with adverse outcomes following single time-point paracetamol (acetaminophen) overdose: a time-course analysis.

    Science.gov (United States)

    Craig, Darren G; Kitto, Laura; Zafar, Sara; Reid, Thomas W D J; Martin, Kirsty G; Davidson, Janice S; Hayes, Peter C; Simpson, Kenneth J

    2014-09-01

    The innate immune system is profoundly dysregulated in paracetamol (acetaminophen)-induced liver injury. The neutrophil-lymphocyte ratio (NLR) is a simple bedside index with prognostic value in a number of inflammatory conditions. To evaluate the prognostic accuracy of the NLR in patients with significant liver injury following single time-point and staggered paracetamol overdoses. Time-course analysis of 100 single time-point and 50 staggered paracetamol overdoses admitted to a tertiary liver centre. Timed laboratory samples were correlated with time elapsed after overdose or admission, respectively, and the NLR was calculated. A total of 49/100 single time-point patients developed hepatic encephalopathy (HE). Median NLRs were higher at both 72 (P=0.0047) and 96 h after overdose (P=0.0041) in single time-point patients who died or were transplanted. Maximum NLR values by 96 h were associated with increasing HE grade (P=0.0005). An NLR of more than 16.7 during the first 96 h following overdose was independently associated with the development of HE [odds ratio 5.65 (95% confidence interval 1.67-19.13), P=0.005]. Maximum NLR values by 96 h were strongly associated with the requirement for intracranial pressure monitoring (Poverdose cohort, the NLR was not associated with adverse outcomes or death/transplantation either at admission or subsequently. The NLR is a simple test which is strongly associated with adverse outcomes following single time-point, but not staggered, paracetamol overdoses. Future studies should assess the value of incorporating the NLR into existing prognostic and triage indices of single time-point paracetamol overdose.

  2. Centrifugal multiplexing fixed-volume dispenser on a plastic lab-on-a-disk for parallel biochemical single-end-point assays.

    Science.gov (United States)

    La, Moonwoo; Park, Sang Min; Kim, Dong Sung

    2015-01-01

    In this study, a multiple sample dispenser for precisely metered fixed volumes was successfully designed, fabricated, and fully characterized on a plastic centrifugal lab-on-a-disk (LOD) for parallel biochemical single-end-point assays. The dispenser, namely, a centrifugal multiplexing fixed-volume dispenser (C-MUFID) was designed with microfluidic structures based on the theoretical modeling about a centrifugal circumferential filling flow. The designed LODs were fabricated with a polystyrene substrate through micromachining and they were thermally bonded with a flat substrate. Furthermore, six parallel metering and dispensing assays were conducted at the same fixed-volume (1.27 μl) with a relative variation of ±0.02 μl. Moreover, the samples were metered and dispensed at different sub-volumes. To visualize the metering and dispensing performances, the C-MUFID was integrated with a serpentine micromixer during parallel centrifugal mixing tests. Parallel biochemical single-end-point assays were successfully conducted on the developed LOD using a standard serum with albumin, glucose, and total protein reagents. The developed LOD could be widely applied to various biochemical single-end-point assays which require different volume ratios of the sample and reagent by controlling the design of the C-MUFID. The proposed LOD is feasible for point-of-care diagnostics because of its mass-producible structures, reliable metering/dispensing performance, and parallel biochemical single-end-point assays, which can identify numerous biochemical.

  3. Fixed Points of Single- and Set-Valued Mappings in Uniformly Convex Metric Spaces with No Metric Convexity

    Directory of Open Access Journals (Sweden)

    Rafa Espínola

    2010-01-01

    Full Text Available We study the existence of fixed points and convergence of iterates for asymptotic pointwise contractions in uniformly convex metric spaces. We also study the existence of fixed points for set-valued nonexpansive mappings in the same class of spaces. Our results do not assume convexity of the metric which makes a big difference when studying the existence of fixed points for set-valued mappings.

  4. A generic-tee-plenum mixing system for application to single point aerosol sampling in stacks and ducts.

    Science.gov (United States)

    Han, Taewon; O'Neal, Dennis L; Ortiz, Carlos A

    2007-01-01

    The ANSI/HPS-N13.1-1999 standard is based on the concept of obtaining a single point representative sample from a location where the velocity and contaminant profiles are relatively uniform. It is difficult to predict the level of mixing in an arbitrary stack or duct without experimental data to meet the ANSI/HPS N13.1-1999 requirements. The goal of this study was to develop experimental data for a range of conditions in "S" (S-shaped configuration) duct systems with different mixing elements and "S" systems having one or two mixing elements. Results were presented in terms of the coefficients of variation (COVs) for velocity, tracer gas, and 10-mum aerodynamic diameter (AD) aerosol particle profiles at different downstream locations for each mixing element. Five mixing elements were tested, including a 90 degrees elbow, a commercial static mixer, a Small-Horizontal Generic-Tee-Plenum (SH-GTP), a Small-Vertical Generic-Tee-Plenum (SV-GTP), and a Large-Horizontal Generic-Tee-Plenum (LH-GTP) system. The COVs for velocity, gas concentration, and aerosol particles for the three GTP systems were all determined to be less than 8%. Tests with two different sizes of GTPs were conducted, and the results showed the performance of the GTPs was relatively unaffected by either size or velocity as reflected by the Reynolds number. The pressure coefficients were 0.59, 0.57, and 0.65, respectively, for the SH-GTP, SV-GTP, and LH-GTP. The pressure drop for the GTPs was approximately twice that of the round elbow, but a factor of 5 less than a Type IV Air Blender. The GTP was developed to provide a sampling location less than 4-duct diameters downstream of a mixing element with low pressure drop condition. The object of the developmental effort was to provide a system that could be employed in new stack; however, the concept of GTPs could also be retrofitted onto existing system applications as well. Results from these tests show that the system performance is well within the ANSI

  5. X Marks the Spot: Creating and Managing a Single Service Point to Improve Customer Service and Maximize Resources

    Science.gov (United States)

    Venner, Mary Ann; Keshmiripour, Seti

    2016-01-01

    This article will describe how merging service points in an academic library is an opportunity to improve customer service and utilize staffing resources more efficiently. Combining service points provides libraries with the ability to create a more positive library experience for patrons by minimizing the ping-pong effect for assistance. The…

  6. Conversion of Human Steroid 5[beta]-Reductase (AKR1D1) into 3[beta]-Hydroxysteroid Dehydrogenase by Single Point Mutation E120H: Example of Perfect Enzyme Engineering

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Mo; Drury, Jason E.; Christianson, David W.; Penning, Trevor M. (UPENN)

    2012-10-10

    Human aldo-keto reductase 1D1 (AKR1D1) and AKR1C enzymes are essential for bile acid biosynthesis and steroid hormone metabolism. AKR1D1 catalyzes the 5{beta}-reduction of {Delta}{sup 4}-3-ketosteroids, whereas AKR1C enzymes are hydroxysteroid dehydrogenases (HSDs). These enzymes share high sequence identity and catalyze 4-pro-(R)-hydride transfer from NADPH to an electrophilic carbon but differ in that one residue in the conserved AKR catalytic tetrad, His120 (AKR1D1 numbering), is substituted by a glutamate in AKR1D1. We find that the AKR1D1 E120H mutant abolishes 5{beta}-reductase activity and introduces HSD activity. However, the E120H mutant unexpectedly favors dihydrosteroids with the 5{alpha}-configuration and, unlike most of the AKR1C enzymes, shows a dominant stereochemical preference to act as a 3{beta}-HSD as opposed to a 3{alpha}-HSD. The catalytic efficiency achieved for 3{beta}-HSD activity is higher than that observed for any AKR to date. High resolution crystal structures of the E120H mutant in complex with epiandrosterone, 5{beta}-dihydrotestosterone, and {Delta}{sup 4}-androstene-3,17-dione elucidated the structural basis for this functional change. The glutamate-histidine substitution prevents a 3-ketosteroid from penetrating the active site so that hydride transfer is directed toward the C3 carbonyl group rather than the {Delta}{sup 4}-double bond and confers 3{beta}-HSD activity on the 5{beta}-reductase. Structures indicate that stereospecificity of HSD activity is achieved because the steroid flips over to present its {alpha}-face to the A-face of NADPH. This is in contrast to the AKR1C enzymes, which can invert stereochemistry when the steroid swings across the binding pocket. These studies show how a single point mutation in AKR1D1 can introduce HSD activity with unexpected configurational and stereochemical preference.

  7. Dynamics of single photon transport in a one-dimensional waveguide two-point coupled with a Jaynes-Cummings system

    KAUST Repository

    Wang, Yuwen

    2016-09-22

    We study the dynamics of an ultrafast single photon pulse in a one-dimensional waveguide two-point coupled with a Jaynes-Cummings system. We find that for any single photon input the transmissivity depends periodically on the separation between the two coupling points. For a pulse containing many plane wave components it is almost impossible to suppress transmission, especially when the width of the pulse is less than 20 times the period. In contrast to plane wave input, the waveform of the pulse can be modified by controlling the coupling between the waveguide and Jaynes-Cummings system. Tailoring of the waveform is important for single photon manipulation in quantum informatics. © The Author(s) 2016.

  8. Allele-specific Gene Silencing of Mutant mRNA Restores Cellular Function in Ullrich Congenital Muscular Dystrophy Fibroblasts

    Directory of Open Access Journals (Sweden)

    Satoru Noguchi

    2014-01-01

    Full Text Available Ullrich congenital muscular dystrophy (UCMD is an inherited muscle disorder characterized clinically by muscle weakness, distal joint hyperlaxity, and proximal joint contractures. Sporadic and recessive mutations in the three collagen VI genes, COL6A1, COL6A2, and COL6A3, are reported to be causative. In the sporadic forms, a heterozygous point mutation causing glycine substitution in the triple helical domain has been identified in higher rate. In this study, we examined the efficacy of siRNAs, which target point mutation site, on specific knockdown toward transcripts from mutant allele and evaluated consequent cellular phenotype of UCMD fibroblasts. We evaluated the effect of siRNAs targeted to silence-specific COL6A1 alleles in UCMD fibroblasts, where simultaneous expression of both wild-type and mutant collagen VI resulted in defective collagen localization. Addition of mutant-specific siRNAs allowed normal extracellular localization of collagen VI surrounding fibroblasts, suggesting selective inhibition of mutant collagen VI. Targeting the single-nucleotide COL6A1 c.850G>A (p.G284R mutation responsible a sporadic autosomal dominant form of UCMD can potently and selectively block expression of mutant collagen VI. These results suggest that allele-specific knockdown of the mutant mRNA can potentially be considered as a therapeutic procedure in UCMD due to COL6A1 point mutations.

  9. FoCuS-point: software for STED fluorescence correlation and time-gated single photon counting

    DEFF Research Database (Denmark)

    Waithe, Dominic; Clausen, Mathias P; Sezgin, Erdinc

    2016-01-01

    microscopy (STED-FCS). Specifically, the use of gated detection has shown great potential for enhancing STED-FCS, but has also created a demand for software which is efficient and also implements the latest algorithms. Prior to this study, no open software has been available which would allow practical time...... to be established quickly and efficiently. AVAILABILITY AND IMPLEMENTATION: FoCuS-point is written in python and is available through the github repository: https://github.com/dwaithe/FCS_point_correlator. Furthermore, compiled versions of the code are available as executables which can be run directly in Linux...

  10. Phase transitions and multicritical points in the mixed spin-32 and spin-2 Ising system with a single-ion anisotropy

    International Nuclear Information System (INIS)

    Bobak, A.; Dely, J.

    2007-01-01

    The effect of a single-ion anisotropy on the phase diagram of the mixed spin-32 and spin-2 Ising system is investigated by the use of a mean-field theory based on the Bogoliubov inequality for the free energy. Topologically different kinds of phase diagrams are achieved by changing values of the parameter in the model Hamiltonian. Besides second-order transitions, lines of first-order transitions terminating either at a tricritical point or an isolated critical point, are found

  11. Promising rice mutants

    International Nuclear Information System (INIS)

    Hakim, L.; Azam, M.A.; Miah, A.J.; Mansur, M.A.; Akanda, H.R.

    1988-01-01

    Two induced mutants namely, Mut NS 1 (tall) and Mut NS 5 (semi-dwarf) derived from rice variety Nizersail were evaluated for various agronomic characters at four locations in Bangladesh. Both the mutants matured about three weeks earlier and yielded significantly higher than the parent variety Nizersail. (author). 3 tabs., 9 refs

  12. Mutant heterosis in rice

    International Nuclear Information System (INIS)

    1987-01-01

    In the variety TKM6 a high yielding semidwarf mutant has been induced. This TKM6 mutant was used in test crosses with a number of other varieties and mutants to examine the extent of heterosis of dwarfs in rice and to select superior crosses. An excerpt of the published data is given. It appears from the backcross of the mutant with its original variety, that an increase in number of productive tillers occurs in the hybrid, leading to a striking grain yield increase, while the semi-dwarf culm length (the main mutant character) reverts to the normal phenotype. In the cross with IR8 on the other hand, there is only a minimal increase in tiller number but a substantial increase in TGW leading to more than 30% yield increase over the better parent

  13. SU-E-T-263: Point Dose Variation Using a Single Ir-192 HDR Brachytherapy Plan for Two Treatments with a Single Tandem-Ovoid Insertion for Cervical Cancer

    Energy Technology Data Exchange (ETDEWEB)

    Liang, X; Morrill, S; Hardee, M; Han, E; Penagaricano, J; Zhang, X; Vaneerat, R [University of Arkansas Medical Science, Little Rock, AR (United States)

    2014-06-01

    Purpose: To evaluate the point dose variations between Ir-192 HDR treatments on two consecutive days using a single tandem-ovoid insertion without replanning in cervical cancer patients. Methods: This study includes eleven cervical cancer patients undergoing HDR brachytherapy with a prescribed dose of 28 Gy in 4 fractions. Each patient had two tandemovoid insertions one week apart. Each insertion was treated on consecutive days with rescanning and replanning prior to each treatment. To study the effect of no replanning for day 2 treatments, the day 1 plan dwell position and dwell time with decay were applied to the day 2 CT dataset. The point dose variations on the prescription point H (defined according to American Brachytherapy Society), and normal tissue doses at point B, bladder, rectum and vaginal mucosa (based on ICRU Report 38) were obtained. Results: Without replanning, the mean point H dose variation was 4.6 ± 10.7% on the left; 2.3 ± 2.9% on the right. The mean B point variation was 3.8 ± 4.9% on the left; 3.6 ± 4.7% on the right. The variation in the left vaginal mucosal point was 12.2 ± 10.7%; 9.5 ± 12.5% on the right; the bladder point 5.5 ± 7.4%; and the rectal point 7.9 ± 9.1%. Conclusion: Without replanning, there are variations both in the prescription point and the normal tissue point doses. The latter can vary as much as 10% or more. This is likely due to the steep dose gradient from brachytherapy compounded by shifts in the positions of the applicator in relationship to the patients anatomy. Imaging prior to each treatment and replanning ensure effective and safe brachytherapy are recommended.

  14. Comparison of single point normalized and modified Gate's method for measurement of glomerular filtration rate by nuclear medicine

    International Nuclear Information System (INIS)

    Fonseca, L.M.B. da; Fonseca, N.M.; Martins, E.; Pereira, E.

    1992-01-01

    Glomerular filtration rate was measured in 22 patients (mean age 30 years) using a noninvasive modified Gates method and the results were compared to the one point normalized technique. The values obtained were 74-11.7 and 82-11.09 ml/min., respectively. The advantages of the method are its reliability, easy adaptation to small laboratories and the avoidance of blood and urine sampling. (author)

  15. Bee venom acupuncture point injection for central post stroke pain: a preliminary single-blind randomized controlled trial.

    Science.gov (United States)

    Cho, Seung-Yeon; Park, Joo-Young; Jung, Woo-Sang; Moon, Sang-Kwan; Park, Jung-Mi; Ko, Chang-Nam; Park, Seong-Uk

    2013-06-01

    We investigated apipuncture, or acupuncture point injection with diluted bee venom, as a promising new treatment for central post stroke pain (CPSP). Bee venom, diluted to 0.005% in normal saline, was administered to the treatment group, and normal saline given to control group as twice-weekly injections for three weeks. The points were LI15, GB21, LI11, GB31, ST36 and GB39 of the affected side and the amount of injection was 0.05 ml at each point. Eight patients in each group were included in the analysis. After three weeks there were significant decreases in visual analogue pain scores compared with baseline in both groups and the treatment group improved more significantly than the control group (p = 0.009). Apipuncture significantly improved CPSP in this pilot trial. Further studies of its mechanisms and a larger and long-term follow-up trial will be needed to determine more definitely the efficacy of apipuncture and to elucidate duration of improvement. Copyright © 2013 Elsevier Ltd. All rights reserved.

  16. Enhancing the accuracy of GPS point positioning by converting the single frequency data to dual frequency data

    Directory of Open Access Journals (Sweden)

    Aly M. El-naggar

    2011-09-01

    Full Text Available The global positioning system (GPS has been used to support a wide variety of applications, such as high-accuracy positioning and navigation. Differential GPS techniques can largely eliminate common-mode errors between the reference and the rover GPS stations resulting from ionospheric and tropospheric refraction and delays, satellite and receiver clock biases, and orbital errors [1]. The ionospheric delay in the propagation of global positioning system (GPS signals is one of the main sources of error in GPS precise positioning and navigation. A dual-frequency GPS receiver can eliminate (to the first order the ionospheric delay through a linear combination of the L1 and L2 observations [2]. The most significant effect of ionospheric delay appear in case of using single frequency data. In this paper the single frequency data of concerned station are converted to dual frequency data by employing dual frequency data from 11 regional GPS stations distributed around it. Total electron content (TEC was calculated at every GPS station to produce the mathematical model of TEC which is a function of latitude (Φ and longitude (λ. By using this mathematical model the values of TEC and L2 can be predicted at the single frequency GPS station for each satellite, after that the comparison between predicted and observation values of TEC and L2 was performed. The estimation method and test results of the proposed method indicates that the difference between predicted and observation values is very small.

  17. First year's operation experience of the deepest SPM (Single Point Mooring) in the world

    Energy Technology Data Exchange (ETDEWEB)

    Millar, J.L.; Hughes, H.; Dyer, R.C.

    1979-01-01

    The British National Oil Corp. had a Single Anchor Leg Mooring (SALM) installed at the Thistle Field in Apr. 1977 for export loading of tankers pending completion of the Brent System 36 in. pipeline to Sullom-Voe, which became operational in Dec. 1978; the SALM will continue to be maintained in good condition as a standby alternative export outlet. Modification of the original three 80,000 dwt tankers during 1977 provided the original design capacity of an 80,000 dwt tanker, but with added clean ballast capacity to eliminate tank cleaning and treatment of dirty ballast water. A fully integrated tanker/SALM/platform instrumentation system monitors mooring loads, buoy and riser angles, and tanker status; the latter being radio telemetered to the nearby block platform. Operation of the system and necessary modifications are described.

  18. Extremely high genetic diversity in a single tumor points to prevalence of non-Darwinian cell evolution.

    Science.gov (United States)

    Ling, Shaoping; Hu, Zheng; Yang, Zuyu; Yang, Fang; Li, Yawei; Lin, Pei; Chen, Ke; Dong, Lili; Cao, Lihua; Tao, Yong; Hao, Lingtong; Chen, Qingjian; Gong, Qiang; Wu, Dafei; Li, Wenjie; Zhao, Wenming; Tian, Xiuyun; Hao, Chunyi; Hungate, Eric A; Catenacci, Daniel V T; Hudson, Richard R; Li, Wen-Hsiung; Lu, Xuemei; Wu, Chung-I

    2015-11-24

    The prevailing view that the evolution of cells in a tumor is driven by Darwinian selection has never been rigorously tested. Because selection greatly affects the level of intratumor genetic diversity, it is important to assess whether intratumor evolution follows the Darwinian or the non-Darwinian mode of evolution. To provide the statistical power, many regions in a single tumor need to be sampled and analyzed much more extensively than has been attempted in previous intratumor studies. Here, from a hepatocellular carcinoma (HCC) tumor, we evaluated multiregional samples from the tumor, using either whole-exome sequencing (WES) (n = 23 samples) or genotyping (n = 286) under both the infinite-site and infinite-allele models of population genetics. In addition to the many single-nucleotide variations (SNVs) present in all samples, there were 35 "polymorphic" SNVs among samples. High genetic diversity was evident as the 23 WES samples defined 20 unique cell clones. With all 286 samples genotyped, clonal diversity agreed well with the non-Darwinian model with no evidence of positive Darwinian selection. Under the non-Darwinian model, MALL (the number of coding region mutations in the entire tumor) was estimated to be greater than 100 million in this tumor. DNA sequences reveal local diversities in small patches of cells and validate the estimation. In contrast, the genetic diversity under a Darwinian model would generally be orders of magnitude smaller. Because the level of genetic diversity will have implications on therapeutic resistance, non-Darwinian evolution should be heeded in cancer treatments even for microscopic tumors.

  19. Papilla height in relation to the distance between bone crest and interproximal contact point at single-tooth implants: A systematic review.

    Science.gov (United States)

    Roccuzzo, Mario; Roccuzzo, Andrea; Ramanuskaite, Ausra

    2018-03-01

    The aim of this systematic review was to investigate the tooth-implant papilla formation in correlation with the distance between the interproximal bone level and the prosthetic contact point. A comprehensive search of the current literature (01/01/2000-01/01/2017) was performed to identify human trials that included 10 patients or more, with at least 12 months follow-up, in need of the replacement of one single tooth in the anterior maxillary region with an implant-supported single crown. To meet the inclusion criteria, studies had to provide both radiographic and clinical data regarding the distance between the interproximal bone level and the prosthetic contact point. The search yielded 136 records. After evaluation of abstracts and full texts, 12 papers were included in the final review, even though various reference points, for the comparison between the vertical distance and the papilla height, were used. The vertical distance between the interproximal bone level and prosthetic contact point ranged between 2 and 11 mm, and the partial or complete papilla fill (Jemt's score 2-3) ranged between 56.5% and 100% of cases. There is limited evidence that the vertical distance from the base of the interproximal contact point to the crestal bone level seems to affect the interproximal papilla height; that is, the lower is the distance the higher is the percentage of papilla fill. Complete embrasure fill between an implant restoration and the adjacent tooth seems to be correlated with the integrity of the periodontal ligament of the tooth. To reduce the risk of aesthetic failures, interproximal probing on the adjacent teeth should be encouraged before implant placement. © 2018 The Authors. Clinical Oral Implants Research Published by John Wiley & Sons Ltd.

  20. Performance evaluation of thermally treated graphite felt electrodes for vanadium redox flow battery and their four-point single cell characterization

    Science.gov (United States)

    Mazúr, P.; Mrlík, J.; Beneš, J.; Pocedič, J.; Vrána, J.; Dundálek, J.; Kosek, J.

    2018-03-01

    In our contribution we study the electrocatalytic effect of oxygen functionalization of thermally treated graphite felt on kinetics of electrode reactions of vanadium redox flow battery. Chemical and morphological changes of the felts are analysed by standard physico-chemical characterization techniques. A complex method four-point method is developed and employed for characterization of the felts in a laboratory single-cell. The method is based on electrochemical impedance spectroscopy and load curves measurements of positive and negative half-cells using platinum wire pseudo-reference electrodes. The distribution of ohmic and faradaic losses within a single-cell is evaluated for both symmetric and asymmetric electrode set-up with respect to the treatment conditions. Positive effect of oxygen functionalization is observed only for negative electrode, whereas kinetics of positive electrode reaction is almost unaffected by the treatment. This is in a contradiction to the results of typically employed cyclovoltammetric characterization which indicate that both electrodes are enhanced by the treatment to a similar extent. The developed four-point characterization method can be further used e.g., for the component screening and in-situ durability studies on single-cell scale redox flow batteries of various chemistries.

  1. A DAB Converter with Common-Point-Connected Winding Transformers Suitable for a Single-Phase 5-Level SST System

    Directory of Open Access Journals (Sweden)

    Hyeok-Jin Yun

    2018-04-01

    Full Text Available One of the main disadvantages of the multi-level solid-state transformer (SST system is the voltage imbalance on the output of the rectifier modules. This voltage imbalance can be caused by parameter mismatch of the active and passive components, different loads, and the floating structure of the high voltage DC-links. Some studies have been done to solve this voltage imbalance problem. A common way to avoid this imbalance is to balance the voltage of DC-links at the AC/DC conversion stage and balance the power between the modules at the DC/DC conversion stage. Most of these methods require a complex balancing controller or additional circuits. This paper proposes a novel dual active bridge (DAB converter specialized in power balancing in a single-phase 5-level SST system. The proposed DAB converter does not require any additional balancing controllers or techniques for power balancing. The performance of the proposed DAB converter was verified by simulation and experiments using a 3 kW 5-level SST prototype system.

  2. Nutritional analysis of gastric contents and body condition score at a single time point in feral horses in Australia.

    Science.gov (United States)

    Hampson, Brian A; Owens, Elizabeth; Watts, Kathryn A; Mills, Paul C; Pollitt, Christopher C; de Laat, Melody A

    2011-09-01

    To determine the impact of a free-choice diet on nutritional intake and body condition of feral horses. Cadavers of 41 feral horses from 5 Australian locations. Body condition score (BCS) was determined (scale of 1 to 9), and the stomach was removed from horses during postmortem examination. Stomach contents were analyzed for nutritional variables and macroelement and microelement concentrations. Data were compared among the locations and also compared with recommended daily intakes for horses. Mean BCS varied by location; all horses were judged to be moderately thin. The BCS for males was 1 to 3 points higher than that of females. Amount of protein in the stomach contents varied from 4.3% to 14.9% and was significantly associated with BCS. Amounts of water-soluble carbohydrate and ethanol-soluble carbohydrate in stomach contents of feral horses from all 5 locations were higher than those expected for horses eating high-quality forage. Some macroelement and microelement concentrations were grossly excessive, whereas others were grossly deficient. There was no evidence of ill health among the horses. Results suggested that the diet for several populations of feral horses in Australia appeared less than optimal. However, neither low BCS nor trace mineral deficiency appeared to affect survival of the horses. Additional studies on food sources in these regions, including analysis of water-soluble carbohydrate, ethanol-soluble carbohydrate, and mineral concentrations, are warranted to determine the provenance of such rich sources of nutrients. Determination of the optimal diet for horses may need revision.

  3. A γA-Crystallin Mouse Mutant Secc with Small Eye, Cataract and Closed Eyelid.

    Directory of Open Access Journals (Sweden)

    Man Hei Cheng

    Full Text Available Cataract is the most common cause of visual loss in humans. A spontaneously occurred, autosomal dominant mouse mutant Secc, which displayed combined features of small eye, cataract and closed eyelid was discovered in our laboratory. In this study, we identified the mutation and characterized the cataract phenotype of this novel Secc mutant. The Secc mutant mice have eyelids that remain half-closed throughout their life. The mutant lens has a significant reduction in size and with opaque spots clustered in the centre. Histological analysis showed that in the core region of the mutant lens, the fiber cells were disorganized and clefts and vacuoles were observed. The cataract phenotype was evident from new born stage. We identified the Secc mutation by linkage analysis using whole genome microsatellite markers and SNP markers. The Secc locus was mapped at chromosome 1 flanked by SNPs rs3158129 and rs13475900. Based on the chromosomal position, the candidate cataract locus γ-crystallin gene cluster (Cryg was investigated by sequencing. A single base deletion (299delG in exon 3 of Cryga which led to a frame-shift of amino acid sequence from position 91 was identified. As a result of this mutation, the sequences of the 3rd and 4th Greek-key motifs of the γA-crystallin are replaced with an unrelated C-terminal peptide of 75 residues long. Coincidentally, the point mutation generated a HindIII restriction site, allowing the identification of the CrygaSecc mutant allele by RFLP. Western blot analysis of 3-week old lenses showed that the expression of γ-crystallins was reduced in the CrygaSecc mutant. Furthermore, in cell transfection assays using CrygaSecc mutant cDNA expression constructs in 293T, COS-7 and human lens epithelial B3 cell lines, the mutant γA-crystallins were enriched in the insoluble fractions and appeared as insoluble aggregates in the transfected cells. In conclusion, we have demonstrated that the Secc mutation leads to the

  4. Single scan parameterization of space-variant point spread functions in image space via a printed array: the impact for two PET/CT scanners

    Energy Technology Data Exchange (ETDEWEB)

    Kotasidis, F A; Matthews, J C; Angelis, G I; Noonan, P J; Jackson, A [Imaging, Genomics and Proteomics, Wolfson Molecular Imaging Centre, MAHSC, University of Manchester, Manchester (United Kingdom); Price, P [Academic Department of Radiation Oncology, University of Manchester, Manchester (United Kingdom); Lionheart, W R [School of Mathematics, Alan Turing Building, University of Manchester, Manchester (United Kingdom); Reader, A J, E-mail: fotis.kotasidis@mmic.man.ac.uk [Brain Imaging Centre, Montreal Neurological Institute, McGill University, Montreal, QC (Canada)

    2011-05-21

    Incorporation of a resolution model during statistical image reconstruction often produces images of improved resolution and signal-to-noise ratio. A novel and practical methodology to rapidly and accurately determine the overall emission and detection blurring component of the system matrix using a printed point source array within a custom-made Perspex phantom is presented. The array was scanned at different positions and orientations within the field of view (FOV) to examine the feasibility of extrapolating the measured point source blurring to other locations in the FOV and the robustness of measurements from a single point source array scan. We measured the spatially-variant image-based blurring on two PET/CT scanners, the B-Hi-Rez and the TruePoint TrueV. These measured spatially-variant kernels and the spatially-invariant kernel at the FOV centre were then incorporated within an ordinary Poisson ordered subset expectation maximization (OP-OSEM) algorithm and compared to the manufacturer's implementation using projection space resolution modelling (RM). Comparisons were based on a point source array, the NEMA IEC image quality phantom, the Cologne resolution phantom and two clinical studies (carbon-11 labelled anti-sense oligonucleotide [{sup 11}C]-ASO and fluorine-18 labelled fluoro-l-thymidine [{sup 18}F]-FLT). Robust and accurate measurements of spatially-variant image blurring were successfully obtained from a single scan. Spatially-variant resolution modelling resulted in notable resolution improvements away from the centre of the FOV. Comparison between spatially-variant image-space methods and the projection-space approach (the first such report, using a range of studies) demonstrated very similar performance with our image-based implementation producing slightly better contrast recovery (CR) for the same level of image roughness (IR). These results demonstrate that image-based resolution modelling within reconstruction is a valid alternative to

  5. Mutant ribosomes can generate dominant kirromycin resistance.

    Science.gov (United States)

    Tubulekas, I; Buckingham, R H; Hughes, D

    1991-01-01

    Mutations in the two genes for EF-Tu in Salmonella typhimurium and Escherichia coli, tufA and tufB, can confer resistance to the antibiotic kirromycin. Kirromycin resistance is a recessive phenotype expressed when both tuf genes are mutant. We describe a new kirromycin-resistant phenotype dominant to the effect of wild-type EF-Tu. Strains carrying a single kirromycin-resistant tuf mutation and an error-restrictive, streptomycin-resistant rpsL mutation are resistant to high levels of kirromycin, even when the other tuf gene is wild type. This phenotype is dependent on error-restrictive mutations and is not expressed with nonrestrictive streptomycin-resistant mutations. Kirromycin resistance is also expressed at a low level in the absence of any mutant EF-Tu. These novel phenotypes exist as a result of differences in the interactions of mutant and wild-type EF-Tu with the mutant ribosomes. The restrictive ribosomes have a relatively poor interaction with wild-type EF-Tu and are thus more easily saturated with mutant kirromycin-resistant EF-Tu. In addition, the mutant ribosomes are inherently kirromycin resistant and support a significantly faster EF-Tu cycle time in the presence of the antibiotic than do wild-type ribosomes. A second phenotype associated with combinations of rpsL and error-prone tuf mutations is a reduction in the level of resistance to streptomycin. PMID:2050625

  6. In-vivo visualisation of the anatomical structures related to the acupuncture points Dai mai and Shen mai by MRI: A single-case pilot study

    International Nuclear Information System (INIS)

    Moncayo, Roy; Rudisch, Ansgar; Diemling, Markus; Kremser, Christian

    2007-01-01

    The concept of acupuncture point localisation in Traditional Chinese Medicine (TCM) is based on millenary practical experience. Modern imaging methods such as PET, MRI and SPECT have been used primary for the investigation of the mechanisms of action of acupuncture. In this pilot single-case study we have evaluated the technical possibilities for in-vivo imaging of the anatomical relations of acupuncture points using state of the art MRI. Preliminary experiments relating to the quality of acupuncture needles under the setting of MRI were done both with stainless steel and gold needles. In a second step, in-vivo imaging was carried out. A licensed acupuncture practitioner (RM) chose two points belonging to the so-called extraordinary vessels. In 2 sequential, separate procedures, he inserted himself gold acupuncture needles using a neutral technique (known as Ping Bu Ping Xie) into the Dai mai and Shen mai points, i.e. gall bladder 26 and bladder 62. Imaging was done on a Siemens Magnetom Avanto MR scanner using a head array and body coil. Mainly T1-weighted imaging sequences, as routinely used for patient exams, were used to obtain multi-slice images. In the preliminary experiments only acupuncture needles made of gold showed enough stability in order to be used for further imaging procedures. Using an onion and a banana as an object, further studies showed that the gold needles produced a void defect that corresponds to the tip of the inserted needle, while at the same time an artefactually increased diameter was observed. The in-vivo experiments showed that the Dai mai point was in relation to the abdominal internal oblique muscle. The Shen mai point artefact showed up close to the longus and brevis peroneal tendons at the fibular malleolus. Side effects related to heating or burning were not observed. Improved anatomical recognition was obtained using 3D-volume rendering techniques. Through an adequate choice of acupuncture material (gold needles) as well as of

  7. A sorghum (Sorghum bicolor mutant with altered carbon isotope ratio.

    Directory of Open Access Journals (Sweden)

    Govinda Rizal

    Full Text Available Recent efforts to engineer C4 photosynthetic traits into C3 plants such as rice demand an understanding of the genetic elements that enable C4 plants to outperform C3 plants. As a part of the C4 Rice Consortium's efforts to identify genes needed to support C4 photosynthesis, EMS mutagenized sorghum populations were generated and screened to identify genes that cause a loss of C4 function. Stable carbon isotope ratio (δ13C of leaf dry matter has been used to distinguishspecies with C3 and C4 photosynthetic pathways. Here, we report the identification of a sorghum (Sorghum bicolor mutant with a low δ13C characteristic. A mutant (named Mut33 with a pale phenotype and stunted growth was identified from an EMS treated sorghum M2 population. The stable carbon isotope analysis of the mutants showed a decrease of 13C uptake capacity. The noise of random mutation was reduced by crossing the mutant and its wildtype (WT. The back-cross (BC1F1 progenies were like the WT parent in terms of 13C values and plant phenotypes. All the BC1F2 plants with low δ13C died before they produced their 6th leaf. Gas exchange measurements of the low δ13C sorghum mutants showed a higher CO2 compensation point (25.24 μmol CO2.mol-1air and the maximum rate of photosynthesis was less than 5μmol.m-2.s-1. To identify the genetic determinant of this trait, four DNA pools were isolated; two each from normal and low δ13C BC1F2 mutant plants. These were sequenced using an Illumina platform. Comparison of allele frequency of the single nucleotide polymorphisms (SNPs between the pools with contrasting phenotype showed that a locus in Chromosome 10 between 57,941,104 and 59,985,708 bps had an allele frequency of 1. There were 211 mutations and 37 genes in the locus, out of which mutations in 9 genes showed non-synonymous changes. This finding is expected to contribute to future research on the identification of the causal factor differentiating C4 from C3 species that can be used

  8. Productive mutants of niger

    International Nuclear Information System (INIS)

    Misra, R.C.

    2001-01-01

    Seeds of six niger (Guizotia abyssinica Cass.) varieties ('GA-10', 'ONS-8', 'IGP-72', 'N-71', 'NB-9' and 'UN-4') were treated with 0.5, 0.75 and 1% ethyl methanesulphonate. After four generations of selection, 29 mutant lines were developed and those were evaluated from 1990-92 during Kharif (July to October) and Rabi (December to March) seasons. Average plant characteristics and yield data of four high yielding mutants along with 'IGP-76' (National Check), GA-10 (Zonal Check) and 'Semiliguda Local' (Local Check) are presented

  9. Isolation and characterisation of a dwarf rice mutant exhibiting defective gibberellins biosynthesis.

    Science.gov (United States)

    Ji, S H; Gururani, M A; Lee, J W; Ahn, B-O; Chun, S-C

    2014-03-01

    We have isolated a severe dwarf mutant derived from a Ds (Dissociation) insertion mutant rice (Oryza sativa var. japonica c.v. Dongjin). This severe dwarf phenotype, has short and dark green leaves, reduced shoot growth early in the seedling stage, and later severe dwarfism with failure to initiate flowering. When treated with bioactive GA3 , mutants are restored to the normal wild-type phenotype. Reverse transcription PCR analyses of 22 candidate genes related to the gibberellin (GA) biosynthesis pathway revealed that among 22 candidate genes tested, a dwarf mutant transcript was not expressed only in one OsKS2 gene. Genetic analysis revealed that the severe dwarf phenotype was controlled by recessive mutation of a single nuclear gene. The putative OsKS2 gene was a chromosome 4-located ent-kaurene synthase (KS), encoding the enzyme that catalyses an early step of the GA biosynthesis pathway. Sequence analysis revealed that osks2 carried a 1-bp deletion in the ORF region of OsKS2, which led to a loss-of-function mutation. The expression pattern of OsKS2 in wild-type cv Dongjin, showed that it is expressed in all organs, most prominently in the stem and floral organs. Morphological characteristics of the dwarf mutant showed dramatic modifications in internal structure and external morphology. We propose that dwarfism in this mutant is caused by a point mutation in OsKS2, which plays a significant role in growth and development of higher plants. Further investigation on OsKS2 and other OsKS-like proteins is underway and may yield better understanding of the putative role of OsKS in severe dwarf mutants. © 2013 German Botanical Society and The Royal Botanical Society of the Netherlands.

  10. A national assessment of underground natural gas storage: identifying wells with designs likely vulnerable to a single-point-of-failure

    Science.gov (United States)

    Michanowicz, Drew R.; Buonocore, Jonathan J.; Rowland, Sebastian T.; Konschnik, Katherine E.; Goho, Shaun A.; Bernstein, Aaron S.

    2017-05-01

    The leak of processed natural gas (PNG) from October 2015 to February 2016 from the Aliso Canyon storage facility, near Los Angeles, California, was the largest single accidental release of greenhouse gases in US history. The Interagency Task Force on Natural Gas Storage Safety and California regulators recently recommended operators phase out single-point-of-failure (SPF) well designs. Here, we develop a national dataset of UGS well activity in the continental US to assess regulatory data availability and uncertainty, and to assess the prevalence of certain well design deficiencies including single-point-of-failure designs. We identified 14 138 active UGS wells associated with 317 active UGS facilities in 29 states using regulatory and company data. State-level wellbore datasets contained numerous reporting inconsistencies that limited data concatenation. We identified 2715 active UGS wells across 160 facilities that, like the failed well at Aliso Canyon, predated the storage facility, and therefore were not originally designed for gas storage. The majority (88%) of these repurposed wells are located in OH, MI, PA, NY, and WV. Repurposed wells have a median age of 74 years, and the 2694 repurposed wells constructed prior to 1979 are particularly likely to exhibit design-related deficiencies. An estimated 210 active repurposed wells were constructed before 1917—before cement zonal isolation methods were utilized. These wells are located in OH, PA, NY, and WV and represent the highest priority related to potential design deficiencies that could lead to containment loss. This national baseline assessment identifies regulatory data uncertainties, highlights a potentially widespread vulnerability of the natural gas supply chain, and can aid in prioritization and oversight for high-risk wells and facilities.

  11. Robust mutant strain design by pessimistic optimization.

    Science.gov (United States)

    Apaydin, Meltem; Xu, Liang; Zeng, Bo; Qian, Xiaoning

    2017-10-03

    Flux Balance Analysis (FBA) based mathematical modeling enables in silico prediction of systems behavior for genome-scale metabolic networks. Computational methods have been derived in the FBA framework to solve bi-level optimization for deriving "optimal" mutant microbial strains with targeted biochemical overproduction. The common inherent assumption of these methods is that the surviving mutants will always cooperate with the engineering objective by overproducing the maximum desired biochemicals. However, it has been shown that this optimistic assumption may not be valid in practice. We study the validity and robustness of existing bi-level methods for strain optimization under uncertainty and non-cooperative environment. More importantly, we propose new pessimistic optimization formulations: P-ROOM and P-OptKnock, aiming to derive robust mutants with the desired overproduction under two different mutant cell survival models: (1) ROOM assuming mutants have the minimum changes in reaction fluxes from wild-type flux values, and (2) the one considered by OptKnock maximizing the biomass production yield. When optimizing for desired overproduction, our pessimistic formulations derive more robust mutant strains by considering the uncertainty of the cell survival models at the inner level and the cooperation between the outer- and inner-level decision makers. For both P-ROOM and P-OptKnock, by converting multi-level formulations into single-level Mixed Integer Programming (MIP) problems based on the strong duality theorem, we can derive exact optimal solutions that are highly scalable with large networks. Our robust formulations P-ROOM and P-OptKnock are tested with a small E. coli core metabolic network and a large-scale E. coli iAF1260 network. We demonstrate that the original bi-level formulations (ROOM and OptKnock) derive mutants that may not achieve the predicted overproduction under uncertainty and non-cooperative environment. The knockouts obtained by the

  12. On the structural affinity of macromolecules with different biological properties: Molecular dynamics simulations of a series of TEM-1 mutants

    Energy Technology Data Exchange (ETDEWEB)

    Giampaolo, Alessia Di [Dipartimento di Scienze Fisiche e Chimiche, Universita’ degli Studi di l’Aquila, Via Vetoio snc, 67100 Coppito (AQ) (Italy); Mazza, Fernando [Department of Health Sciences, Univ. of L’Aquila, 67010 L’Aquila (Italy); Daidone, Isabella [Dipartimento di Scienze Fisiche e Chimiche, Universita’ degli Studi di l’Aquila, Via Vetoio snc, 67100 Coppito (AQ) (Italy); Amicosante, Gianfranco; Perilli, Mariagrazia [Dipartimento di Scienze Cliniche Applicate e Biotecnologiche, Università degli Studi di l’Aquila, Via Vetoio snc, 67100 Coppito (AQ) (Italy); Aschi, Massimiliano, E-mail: massimiliano.aschi@univaq.it [Dipartimento di Scienze Fisiche e Chimiche, Universita’ degli Studi di l’Aquila, Via Vetoio snc, 67100 Coppito (AQ) (Italy)

    2013-07-12

    Highlights: •We have performed molecular dynamics simulations of TEM-1 mutants. •Mutations effects on the mechanical properties are considered. •Mutants do not significantly alter the average enzymes structure. •Mutants produce sharp alterations in enzyme conformational repertoire. •Mutants also produce changes in the active site volume. -- Abstract: Molecular Dynamics simulations have been carried out in order to provide a molecular rationalization of the biological and thermodynamic differences observed for a class of TEM β-lactamases. In particular we have considered the TEM-1(wt), the single point mutants TEM-40 and TEM-19 representative of IRT and ESBL classes respectively, and TEM-1 mutant M182T, TEM-32 and TEM-20 which differ from the first three for the additional of M182T mutation. Results indicate that most of the thermodynamic, and probably biological behaviour of these systems arise from subtle effects which, starting from the alterations of the local interactions, produce drastic modifications of the conformational space spanned by the enzymes. The present study suggests that systems showing essentially the same secondary and tertiary structure may differentiate their chemical–biological activity essentially (and probably exclusively) on the basis of the thermal fluctuations occurring in their physiological environment.

  13. A study on density functional theory of the effect of pressure on the formation and migration enthalpies of intrinsic point defects in growing single crystal Si

    Science.gov (United States)

    Sueoka, Koji; Kamiyama, Eiji; Kariyazaki, Hiroaki

    2012-05-01

    In 1982, Voronkov presented a model describing point defect behavior during the growth of single crystal Si from a melt and derived an expression to predict if the crystal was vacancy- or self-interstitial-rich. Recently, Vanhellemont claimed that one should take into account the impact of compressive stress introduced by the thermal gradient at the melt/solid interface by considering the hydrostatic pressure dependence of the formation enthalpy of the intrinsic point defects. To evaluate the impact of thermal stress more correctly, the pressure dependence of both the formation enthalpy (Hf) and the migration enthalpy (Hm) of the intrinsic point defects should be taken into account. Furthermore, growing single crystal Si is not under hydrostatic pressure but almost free of external pressure (generally in Ar gas under reduced pressure). In the present paper, the dependence of Hf and Hm on the pressure P, or in other words, the pressure dependence of the formation energy (Ef) and the relaxation volume (vf), is quantified by density functional theory calculations. Although a large number of ab initio calculations of the properties of intrinsic point defects have been published during the last years, calculations for Si crystals under pressure are rather scarce. For vacancies V, the reported pressure dependences of HfV are inconsistent. In the present study, by using 216-atom supercells with a sufficient cut-off energy and mesh of k-points, the neutral I and V are found to have nearly constant formation energies EfI and EfV for pressures up to 1 GPa. For the relaxation volume, vfI is almost constant while vfV decreases linearly with increasing pressure P. In case of the hydrostatic pressure Ph, the calculated formation enthalpy HfI and migration enthalpy HmI at the [110] dumbbell site are given by HfI = 3.425 - 0.057 × Ph (eV) and HmI = 0.981 - 0.039 × Ph (eV), respectively, with Ph given in GPa. The calculated HfV and HmV dependencies on Ph given by HfV = 3.543 - 0

  14. Single molecule TPM analysis of the catalytic pentad mutants of Cre and Flp site-specific recombinases: contributions of the pentad residues to the pre-chemical steps of recombination.

    Science.gov (United States)

    Fan, Hsiu-Fang; Cheng, Yong-Song; Ma, Chien-Hui; Jayaram, Makkuni

    2015-03-31

    Cre and Flp site-specific recombinase variants harboring point mutations at their conserved catalytic pentad positions were characterized using single molecule tethered particle motion (TPM) analysis. The findings reveal contributions of these amino acids to the pre-chemical steps of recombination. They suggest functional differences between positionally conserved residues in how they influence recombinase-target site association and formation of 'non-productive', 'pre-synaptic' and 'synaptic' complexes. The most striking difference between the two systems is noted for the single conserved lysine. The pentad residues in Cre enhance commitment to recombination by kinetically favoring the formation of pre-synaptic complexes. These residues in Flp serve a similar function by promoting Flp binding to target sites, reducing non-productive binding and/or enhancing the rate of assembly of synaptic complexes. Kinetic comparisons between Cre and Flp, and between their derivatives lacking the tyrosine nucleophile, are consistent with a stronger commitment to recombination in the Flp system. The effect of target site orientation (head-to-head or head-to-tail) on the TPM behavior of synapsed DNA molecules supports the selection of anti-parallel target site alignment prior to the chemical steps. The integrity of the synapse, whose establishment/stability is fostered by strand cleavage in the case of Flp but not Cre, appears to be compromised by the pentad mutations. © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

  15. Single molecule TPM analysis of the catalytic pentad mutants of Cre and Flp site-specific recombinases: contributions of the pentad residues to the pre-chemical steps of recombination

    Science.gov (United States)

    Fan, Hsiu-Fang; Cheng, Yong-Song; Ma, Chien-Hui; Jayaram, Makkuni

    2015-01-01

    Cre and Flp site-specific recombinase variants harboring point mutations at their conserved catalytic pentad positions were characterized using single molecule tethered particle motion (TPM) analysis. The findings reveal contributions of these amino acids to the pre-chemical steps of recombination. They suggest functional differences between positionally conserved residues in how they influence recombinase-target site association and formation of ‘non-productive’, ‘pre-synaptic’ and ‘synaptic’ complexes. The most striking difference between the two systems is noted for the single conserved lysine. The pentad residues in Cre enhance commitment to recombination by kinetically favoring the formation of pre-synaptic complexes. These residues in Flp serve a similar function by promoting Flp binding to target sites, reducing non-productive binding and/or enhancing the rate of assembly of synaptic complexes. Kinetic comparisons between Cre and Flp, and between their derivatives lacking the tyrosine nucleophile, are consistent with a stronger commitment to recombination in the Flp system. The effect of target site orientation (head-to-head or head-to-tail) on the TPM behavior of synapsed DNA molecules supports the selection of anti-parallel target site alignment prior to the chemical steps. The integrity of the synapse, whose establishment/stability is fostered by strand cleavage in the case of Flp but not Cre, appears to be compromised by the pentad mutations. PMID:25765648

  16. An experimental investigation on dynamics and heat transfer associated with a single droplet impacting on a hot surface above the Leidenfrost point temperature

    Energy Technology Data Exchange (ETDEWEB)

    Park, J.; Kim, H. [Kyung Hee Univ., Gyeonggi-do (Korea, Republic of). Dept. of Nuclear Engineering

    2016-07-15

    During large loss-of-coolant accidents in nuclear reactors, water splatters as the quench front propagates at the quenching surface, and many droplets of different sizes and velocities are generated and carried with the steam in the cooling channel. Heat transfer due to droplets striking an overheated fuel rod above the Leidenfrost point temperature is important for predicting the peak cladding temperature. This study investigated the dynamics and heat transfer characteristics when a single droplet at room temperature collided with a surface at 425 C experimentally, using synchronized high-speed video and infrared cameras. Various physical parameters related to heat transfer model development were measured, including the residence time, spreading diameter, local heat flux distribution, effective heat transfer area, average vapor film thickness, and total heat transfer per collision. The measured data were compared with the values of the physical parameters predicted by existing mechanistic models.

  17. A Generic Simulation Approach for the Fast and Accurate Estimation of the Outage Probability of Single Hop and Multihop FSO Links Subject to Generalized Pointing Errors

    KAUST Repository

    Ben Issaid, Chaouki

    2017-07-28

    When assessing the performance of the free space optical (FSO) communication systems, the outage probability encountered is generally very small, and thereby the use of nave Monte Carlo simulations becomes prohibitively expensive. To estimate these rare event probabilities, we propose in this work an importance sampling approach which is based on the exponential twisting technique to offer fast and accurate results. In fact, we consider a variety of turbulence regimes, and we investigate the outage probability of FSO communication systems, under a generalized pointing error model based on the Beckmann distribution, for both single and multihop scenarios. Selected numerical simulations are presented to show the accuracy and the efficiency of our approach compared to naive Monte Carlo.

  18. Comparison of a multi-breath-hold and a single breath-hold cine imaging approach for 4D guide-point modeling of the left ventricle

    International Nuclear Information System (INIS)

    Heilmaier, C.; Schlosser, T.; Nassenstein, K.; Bruder, O.

    2010-01-01

    Purpose: Guide-point modeling (GPM) enables reliable and time-efficient assessment of left ventricular (LV) volumes when using sequences that allow acquisition of short- and long-axis scans within a single breath-hold. Slice misalignment may influence GPM analysis of standard multi-breath-hold images due to image acquisition in different breath-holds. Thus, our study aimed to assess if such an approach allows for reliable volumetric calculations in the clinical routine. Materials and Methods: 52 patients were examined on a 1.5 T scanner with multi-breath-hold acquisitions on the standard short- and long-axis using an SSFP (TR 3 ms, TE 1.5 ms, FA 60 ) sequence and a TPAT accelerated SSFP (TR 4.6 msec, TE 1.1msec, FA 60 , acceleration factor 3) sequence that covered the LV in 3 short- and 2 long-axis slices within a single breath-hold. For both datasets GPM was used to assess LV volumes. In addition, LV parameters were calculated by applying the summation of slices (SoS) approach (standard of reference) with the short-axis views of the multi-breath-hold dataset. Results: The post-processing times were shorter with both GPM approaches (both, p 0.97). Conclusion: Cine short- and long-axis images that had been acquired in different breath-holds can be reliably evaluated by the GPM approach. (orig.)

  19. Visit of Peters Higgs at Point 2 ALICE Experiment - British theoretical physicist, He worked on proposals to unify the weak and the electromagnetic forces into a single electroweak theory, The Boson of Higgs.

    CERN Multimedia

    Mona Schweizer

    2008-01-01

    Visit of Peters Higgs at Point 2 ALICE Experiment - British theoretical physicist, He worked on proposals to unify the weak and the electromagnetic forces into a single electroweak theory, The Boson of Higgs.

  20. A forward genetic screen with a thalamocortical axon reporter mouse yields novel neurodevelopment mutants and a distinct emx2 mutant phenotype

    Directory of Open Access Journals (Sweden)

    Vock Vita M

    2011-01-01

    Full Text Available Abstract Background The dorsal thalamus acts as a gateway and modulator for information going to and from the cerebral cortex. This activity requires the formation of reciprocal topographic axon connections between thalamus and cortex. The axons grow along a complex multistep pathway, making sharp turns, crossing expression boundaries, and encountering intermediate targets. However, the cellular and molecular components mediating these steps remain poorly understood. Results To further elucidate the development of the thalamocortical system, we first created a thalamocortical axon reporter line to use as a genetic tool for sensitive analysis of mutant mouse phenotypes. The TCA-tau-lacZ reporter mouse shows specific, robust, and reproducible labeling of thalamocortical axons (TCAs, but not the overlapping corticothalamic axons, during development. Moreover, it readily reveals TCA pathfinding abnormalities in known cortical mutants such as reeler. Next, we performed an unbiased screen for genes involved in thalamocortical development using random mutagenesis with the TCA reporter. Six independent mutant lines show aberrant TCA phenotypes at different steps of the pathway. These include ventral misrouting, overfasciculation, stalling at the corticostriatal boundary, and invasion of ectopic cortical cell clusters. An outcross breeding strategy coupled with a genomic panel of single nucleotide polymorphisms facilitated genetic mapping with small numbers of mutant mice. We mapped a ventral misrouting mutant to the Emx2 gene, and discovered that some TCAs extend to the olfactory bulbs in this mutant. Mapping data suggest that other lines carry mutations in genes not previously known for roles in thalamocortical development. Conclusions These data demonstrate the feasibility of a forward genetic approach to understanding mammalian brain morphogenesis and wiring. A robust axonal reporter enabled sensitive analysis of a specific axon tract inside the

  1. Improved solubility of replication factor C (RFC) Walker A mutants.

    Science.gov (United States)

    Marzahn, Melissa R; Bloom, Linda B

    2012-06-01

    Protein insolubility often poses a significant problem during purification protocols and in enzyme assays, especially for eukaryotic proteins expressed in a recombinant bacterial system. The limited solubility of replication factor C (RFC), the clamp loader complex from Saccharomyces cerevisiae, has been previously documented. We found that mutant forms of RFC harboring a single point mutation in the Walker A motif were even less soluble than the wild-type complex. The addition of maltose at 0.75 M to the storage and assay buffers greatly increases protein solubility and prevents the complex from falling apart. Our analysis of the clamp loading reaction is dependent on fluorescence-based assays, which are environmentally sensitive. Using wt RFC as a control, we show that the addition of maltose to the reaction buffers does not affect fluorophore responses in the assays or the enzyme activity, indicating that maltose can be used as a buffer additive for further downstream analysis of these mutants. Copyright © 2012 Elsevier Inc. All rights reserved.

  2. Molecular dynamic simulation of wild type and mutants of the polymorphic amyloid NNQNTF segments of elk prion: structural stability and thermodynamic of association.

    Science.gov (United States)

    Berhanu, Workalemahu M; Masunov, Artëm E

    2011-09-01

    A hexapeptide with amino acid sequence NNQNTF from the elk prion protein forms amyloid fibrils. Here we use molecular dynamic simulations of the oligomers and their single point glycine mutants to study their stability. In an effort to probe the structural stability and association thermodynamic in a realistic environment, all wildtype of NNQNTF polymorphic forms with different size and their corresponding double layer 5 strands single point glycine mutants were subjected to a total of 500 ns of explicit-solvent molecular dynamics (MD) simulation. Our results show that the structural stability of the NNQNTF oligomers increases with increasing the number of β-strands for double layers. Our results also demonstrated that hydrophobic interaction is the principle driving force to stabilize the adjacent β-strands while the steric zipper is responsible for holding the neighboring β-sheet layers together. We used MM-PBSA approach free energy calculations to determine the role of nonpolar effects, electrostatics and entropy in binding. Nonpolar effects remained consistently more favorable in wild type and mutants reinforcing the importance of hydrophobic effects in protein-protein binding. While entropy systematically opposed binding in all cases, there was no observed trend in the entropy difference between wildtype and glycine mutant. Free energy decomposition shows residues situated at the interface were found to make favorable contributions to the peptide-peptide association. The study of the wild type and mutants in an explicit solvent may provide valuable insight for amyloid aggregation inhibitor design efforts. Copyright © 2011 Wiley Periodicals, Inc.

  3. Native Mutant Huntingtin in Human Brain

    Science.gov (United States)

    Sapp, Ellen; Valencia, Antonio; Li, Xueyi; Aronin, Neil; Kegel, Kimberly B.; Vonsattel, Jean-Paul; Young, Anne B.; Wexler, Nancy; DiFiglia, Marian

    2012-01-01

    Huntington disease (HD) is caused by polyglutamine expansion in the N terminus of huntingtin (htt). Analysis of human postmortem brain lysates by SDS-PAGE and Western blot reveals htt as full-length and fragmented. Here we used Blue Native PAGE (BNP) and Western blots to study native htt in human postmortem brain. Antisera against htt detected a single band broadly migrating at 575–850 kDa in control brain and at 650–885 kDa in heterozygous and Venezuelan homozygous HD brains. Anti-polyglutamine antisera detected full-length mutant htt in HD brain. There was little htt cleavage even if lysates were pretreated with trypsin, indicating a property of native htt to resist protease cleavage. A soluble mutant htt fragment of about 180 kDa was detected with anti-htt antibody Ab1 (htt-(1–17)) and increased when lysates were treated with denaturants (SDS, 8 m urea, DTT, or trypsin) before BNP. Wild-type htt was more resistant to denaturants. Based on migration of in vitro translated htt fragments, the 180-kDa segment terminated ≈htt 670–880 amino acids. If second dimension SDS-PAGE followed BNP, the 180-kDa mutant htt was absent, and 43–50 kDa htt fragments appeared. Brain lysates from two HD mouse models expressed native full-length htt; a mutant fragment formed if lysates were pretreated with 8 m urea + DTT. Native full-length mutant htt in embryonic HD140Q/140Q mouse primary neurons was intact during cell death and when cell lysates were exposed to denaturants before BNP. Thus, native mutant htt occurs in brain and primary neurons as a soluble full-length monomer. PMID:22375012

  4. Radiation induced early maturing mutants in barley

    International Nuclear Information System (INIS)

    Kumar, R.; Chauhan, S.V.S.; Sharma, R.P.

    1978-01-01

    In M 2 generation, two early maturing plants were screened from a single spike progeny of a plant obtained from 20 kR of gamma-ray irradiation of a six-rowed barley (Hordeum vulgare L. var. Jyoti). Their true breeding nature was confirmed in M 3 generation. These mutants flower and mature 38 and 22 days earlier than those of control. (auth.)

  5. Photorepair mutants of Arabidopsis

    International Nuclear Information System (INIS)

    Jiang, C.Z.; Yee, J.; Mitchell, D.L.; Britt, A.B.

    1997-01-01

    UV radiation induces two major DNA damage products, the cyclobutane pyrimidine dimer (CPD) and, at a lower frequency, the pyrimidine (6-4) pyrimidinone dimer (6-4 product). Although Escherichia coli and Saccharomyces cerevisiae produce a CPD-specific photolyase that eliminates only this class of dimer, Arabidopsis thaliana, Drosophila melanogaster, Crotalus atrox, and Xenopus laevis have recently been shown to photoreactivate both CPDs and 6-4 products. We describe the isolation and characterization of two new classes of mutants of Arabidopsis, termed uvr2 and uvr3, that are defective in the photoreactivation of CPDs and 6-4 products, respectively. We demonstrate that the CPD photolyase mutation is genetically linked to a DNA sequence encoding a type II (metazoan) CPD photolyase. In addition, we are able to generate plants in which only CPDs or 6-4 products are photoreactivated in the nuclear genome by exposing these mutants to UV light and then allowing them to repair one or the other class of dimers. This provides us with a unique opportunity to study the biological consequences of each of these two major UV-induced photoproducts in an intact living system

  6. Induction and characterization of Arabidopsis mutants by Ion beam

    International Nuclear Information System (INIS)

    Yoon, Y. H.; Choi, J. D.; Park, J. Y.; Lee, J. R.; Sohn, H. S.

    2008-03-01

    This study was conducted to search the proper conditions and times for irradiating proton beam to seeds generally used for induction of mutant. Arabidopsis as model plants has good characters that is a short generation time, producing a lot of seeds, sequenced genome, developed maker. This points were the best materials for plant breeding for this study. The data of inducing mutants of Arabidopsis is used to be applicate to crops have more longer generation that is the final goals of this study. The goals of this project were to inducing and characterizing arabidopsis mutants by the proton ion beam and γ-ray. As well as, the purpose of this study was securing more than 10 lines of arabidopsis mutants in this project and also to know the changed DNA structure of the mutants using the basic data for applying to the more study

  7. Isozyme differences in barley mutants

    International Nuclear Information System (INIS)

    AI-Jibouri, A.A.M.; Dham, K.M.

    1990-01-01

    Full text: Thirty mutants (M 11 ) of barley (Hordeum vulgare L.) induced by physical and chemical mutagens were analysed for isozyme composition using polyacrylamide gel electrophoresis. Results show that these mutants were different in the isozymes leucine aminopeptidase, esterase and peroxidase. The differences included the number of forms of each enzyme, relative mobility value and their intensity on the gel. Glutamate oxaloacetate transaminase isozyme was found in six molecular forms and these forms were similar in all mutants. (author)

  8. Relative effectiveness of kinetic analysis vs single point readings for classifying environmental samples based on community-level physiological profiles (CLPP)

    Science.gov (United States)

    Garland, J. L.; Mills, A. L.; Young, J. S.

    2001-01-01

    The relative effectiveness of average-well-color-development-normalized single-point absorbance readings (AWCD) vs the kinetic parameters mu(m), lambda, A, and integral (AREA) of the modified Gompertz equation fit to the color development curve resulting from reduction of a redox sensitive dye from microbial respiration of 95 separate sole carbon sources in microplate wells was compared for a dilution series of rhizosphere samples from hydroponically grown wheat and potato ranging in inoculum densities of 1 x 10(4)-4 x 10(6) cells ml-1. Patterns generated with each parameter were analyzed using principal component analysis (PCA) and discriminant function analysis (DFA) to test relative resolving power. Samples of equivalent cell density (undiluted samples) were correctly classified by rhizosphere type for all parameters based on DFA analysis of the first five PC scores. Analysis of undiluted and 1:4 diluted samples resulted in misclassification of at least two of the wheat samples for all parameters except the AWCD normalized (0.50 abs. units) data, and analysis of undiluted, 1:4, and 1:16 diluted samples resulted in misclassification for all parameter types. Ordination of samples along the first principal component (PC) was correlated to inoculum density in analyses performed on all of the kinetic parameters, but no such influence was seen for AWCD-derived results. The carbon sources responsible for classification differed among the variable types with the exception of AREA and A, which were strongly correlated. These results indicate that the use of kinetic parameters for pattern analysis in CLPP may provide some additional information, but only if the influence of inoculum density is carefully considered. c2001 Elsevier Science Ltd. All rights reserved.

  9. Point-of-Care Determination of Acetaminophen Levels with Multi-Hydrogen Bond Manipulated Single-Molecule Recognition (eMuHSiR).

    Science.gov (United States)

    Zhang, Yan; Huang, Zhongyuan; Wang, Letao; Wang, Chunming; Zhang, Changde; Wiese, Tomas; Wang, Guangdi; Riley, Kevin; Wang, Zhe

    2018-04-03

    This work aims to face the challenge of monitoring small molecule drugs accurately and rapidly for point-of-care (POC) diagnosis in current clinical settings. Overdose of acetaminophen (AP), a commonly used over the counter (OTC) analgesic drug, has been determined to be a major cause of acute liver failure in the US and the UK. However, there is no rapid and accurate detection method available for this drug in the emergency room. The present study examined an AP sensing strategy that relies on a previously unexplored strong interaction between AP and the arginine (Arg) molecule. It was found that as many as 4 hydrogen bonds can be formed between one Arg molecule and one AP molecule. By taking advantages of this structural selectivity and high tenability of hydrogen bonds, Arg, immobilized on a graphene surface via electrostatic interactions, was utilized to structurally capture AP. Interestingly, bonded AP still remained the perfect electrochemical activities. The extent of Arg-AP bonds was quantified using a newly designed electrochemical (EC) sensor. To verify the feasibility of this novel assay, based on multihydrogen bond manipulated single-molecule recognition (eMuHSiR), both pharmaceutical and serum sample were examined. In commercial tablet measurement, no significant difference was seen between the results of eMuHSiR and other standard methods. For measuring AP concentration in the mice blood, the substances in serum, such as sugars and fats, would not bring any interference to the eMuHSiR in a wide concentration range. This eMuHSiR method opens the way for future development of small molecule detection for the POC testing.

  10. Recommendations for collaborative paediatric research including biobanking in Europe: a Single Hub and Access point for paediatric Rheumatology in Europe (SHARE) initiative.

    Science.gov (United States)

    Kuemmerle-Deschner, Jasmin B; Hansmann, Sandra; Wulffraat, Nico M; Vastert, Sebastiaan J; Hens, Kristien; Anton, Jordi; Avcin, Tadej; Martini, Alberto; Koné-Paut, Isabelle; Uziel, Yosef; Ravelli, Angelo; Wouters, Carine; Shaw, David; Özen, Seza; Eikelberg, Andreas; Prakken, Berent J; Ruperto, Nicolino; Horneff, Gerd; Constantin, Tamas; Beresford, Michael W; Sikken, Marijn; Foster, Helen E; Haug, Iris; Schuller, Sabrina; Jägle, Christine; Benseler, Susanne M

    2018-03-01

    Innovative research in childhood rheumatic diseases mandates international collaborations. However, researchers struggle with significant regulatory heterogeneity; an enabling European Union (EU)-wide framework is missing. The aims of the study were to systematically review the evidence for best practice and to establish recommendations for collaborative research. The Paediatric Rheumatology European Single Hub and Access point for paediatric Rheumatology in Europe (SHARE) project enabled a scoping review and expert discussion, which then informed the systematic literature review. Published evidence was synthesised; recommendations were drafted. An iterative review process and consultations with Ethics Committees and European experts for ethical and legal aspects of paediatric research refined the recommendations. SHARE experts and patient representatives vetted the proposed recommendations at a consensus meeting using Nominal Group Technique. Agreement of 80% was mandatory for inclusion. The systematic literature review returned 1319 records. A total of 223 full-text publications plus 22 international normative documents were reviewed; 85 publications and 16 normative documents were included. A total of 21 recommendations were established including general principles (1-3), ethics (4-7), paediatric principles (8 and 9), consent to paediatric research (10-14), paediatric databank and biobank (15 and 16), sharing of data and samples (17-19), and commercialisation and third parties (20 and 21). The refined recommendations resulted in an agreement of >80% for all recommendations. The SHARE initiative established the first recommendations for Paediatric Rheumatology collaborative research across borders in Europe. These provide strong support for an urgently needed European framework and evidence-based guidance for its implementation. Such changes will promote research in children with rheumatic diseases. © Article author(s) (or their employer(s) unless otherwise

  11. Evaluation of tall rice mutant

    International Nuclear Information System (INIS)

    Hakim, L.; Azam, M.A.; Miah, A.J.; Mansur, M.A.; Akanda, H.R.

    1989-01-01

    One tall mutant (Mut NS1) of rice variety Nizersail was put to multilocation on-farm trial. It showed improvement over the parent in respect of by earlier maturity and higher grain yield at all locations and thus it appears as an improved mutant of Nizersail. (author). 6 refs

  12. Mitochondrial mutant cells are hypersensitive to ionizing radiation, phleomycin and mitomycin C

    Energy Technology Data Exchange (ETDEWEB)

    Kulkarni, Rohan; Reither, Adrian; Thomas, Robert A. [Department of Biological Sciences, Wayne State University, 5047 Gullen Mall, Suite 1370, Detroit, MI 48202 (United States); Tucker, James D., E-mail: jtucker@biology.biosci.wayne.edu [Department of Biological Sciences, Wayne State University, 5047 Gullen Mall, Suite 1370, Detroit, MI 48202 (United States)

    2009-04-26

    Mitochondrial DNA (mtDNA) is an important contributor to the ATP-generating oxidative phosphorylation complex. Single nucleotide mutations in mitochondrial genes involved in ATP synthesis result in a broad range of diseases. Leber optic atrophy and Leigh's syndrome are two such diseases arising from point mutations in the mitochondrial genome. Here, ionizing radiation, phleomycin and mitomycin C (MMC) were used to induce structural chromosomal aberrations in Leber's and Leigh's cells to investigate how these mitochondrial mutations affect the cell's DNA repair processes. Because of the energy deprivation that results from mitochondrial mutations, we hypothesized that these mutant cells would demonstrate hypersensitivity when exposed to oxidative and genotoxic stress and we also expected that these cells would not be able to repair nuclear DNA damage as efficiently as normal cells. As a consequence, these mutant cells are expected to show increased levels of DNA damage, longer cell cycle delays and increased levels of cell death. Following acute radiation exposure these mutant cells showed an increase in the number of chromosomal aberrations and decreased mitotic indices when compared with normal human lymphoblastoid cells with wild-type mtDNA. When exposed to phleomycin or MMC, the mitochondrial mutant cells again showed hypersensitivity and decreased mitotic indices compared to normal cells. These results suggest that Leber's and Leigh's cells have an impaired ability to cope with oxidative and genotoxic stress. These observations may help explain the role of ATP generation in understanding the enhanced sensitivity of mitochondrial mutant cells to cancer therapeutic agents and to adverse environmental exposure, suggesting that individuals with mtDNA mutations may be at a greater risk for cancer and other diseases that result from an accumulation of nuclear DNA damage.

  13. Feasibility of in vivo three-dimensional T2*mapping using dicarboxy-PROXYL and CW-EPR-based single-point imaging.

    Science.gov (United States)

    Kubota, Harue; Komarov, Denis A; Yasui, Hironobu; Matsumoto, Shingo; Inanami, Osamu; Kirilyuk, Igor A; Khramtsov, Valery V; Hirata, Hiroshi

    2017-06-01

    The aim of this study was to demonstrate the feasibility of in vivo three-dimensional (3D) relaxation time T 2 * mapping of a dicarboxy-PROXYL radical using continuous-wave electron paramagnetic resonance (CW-EPR) imaging. Isotopically substituted dicarboxy-PROXYL radicals, 3,4-dicarboxy-2,2,5,5-tetra( 2 H 3 )methylpyrrolidin-(3,4- 2 H 2 )-(1- 15 N)-1-oxyl ( 2 H, 15 N-DCP) and 3,4-dicarboxy-2,2,5,5-tetra( 2 H 3 )methylpyrrolidin-(3,4- 2 H 2 )-1-oxyl ( 2 H-DCP), were used in the study. A clonogenic cell survival assay was performed with the 2 H-DCP radical using squamous cell carcinoma (SCC VII) cells. The time course of EPR signal intensities of intravenously injected 2 H, 15 N-DCP and 2 H-DCP radicals were determined in tumor-bearing hind legs of mice (C3H/HeJ, male, n = 5). CW-EPR-based single-point imaging (SPI) was performed for 3D T 2 * mapping. 2 H-DCP radical did not exhibit cytotoxicity at concentrations below 10 mM. The in vivo half-life of 2 H, 15 N-DCP in tumor tissues was 24.7 ± 2.9 min (mean ± standard deviation [SD], n = 5). The in vivo time course of the EPR signal intensity of the 2 H, 15 N-DCP radical showed a plateau of 10.2 ± 1.2 min (mean ± SD) where the EPR signal intensity remained at more than 90% of the maximum intensity. During the plateau, in vivo 3D T 2 * maps with 2 H, 15 N-DCP were obtained from tumor-bearing hind legs, with a total acquisition time of 7.5 min. EPR signals of 2 H, 15 N-DCP persisted long enough after bolus intravenous injection to conduct in vivo 3D T 2 * mapping with CW-EPR-based SPI.

  14. Cardiac EASE (Ensuring Access and Speedy Evaluation) – the impact of a single-point-of-entry multidisciplinary outpatient cardiology consultation program on wait times in Canada

    Science.gov (United States)

    Bungard, Tammy J; Smigorowsky, Marcie J; Lalonde, Lucille D; Hogan, Terry; Doliszny, Katharine M; Gebreyesus, Ghirmay; Garg, Sipi; Archer, Stephen L

    2009-01-01

    BACKGROUND: Universal access to health care is valued in Canada but increasing wait times for services (eg, cardiology consultation) raise safety questions. Observations suggest that deficiencies in the process of care contribute to wait times. Consequently, an outpatient clinic was designed for Ensuring Access and Speedy Evaluation (Cardiac EASE) in a university group practice, providing cardiac consultative services for northern Alberta. Cardiac EASE has two components: a single-point-of-entry intake service (prospective testing using physician-approved algorithms and previsit triage) and a multidisciplinary clinic (staffed by cardiologists, nurse practitioners and doctoral-trained pharmacists). OBJECTIVES: It was hypothesized that Cardiac EASE would reduce the time to initial consultation and a definitive diagnosis, and also increase the referral capacity. METHODS: The primary and secondary outcomes were time from referral to initial consultation, and time to achieve a definitive diagnosis and management plan, respectively. A conventionally managed historical control group (three-month pre-EASE period in 2003) was compared with the EASE group (2004 to 2006). The conventional referral mechanism continued concurrently with EASE. RESULTS: A comparison between pre-EASE (n=311) and EASE (n=3096) revealed no difference in the mean (± SD) age (60±16 years), sex (55% and 52% men, respectively) or reason for referral, including chest pain (31% and 40%, respectively) and arrhythmia (27% and 29%, respectively). Cardiac EASE reduced the time to initial cardiac consultation (from 71±45 days to 33±19 days) and time to a definitive diagnosis (from 120±86 days to 51±58 days) (P<0.0001). The annual number of new referrals increased from 1512 in 2002 to 2574 in 2006 due to growth in the Cardiac EASE clinic. The number of patients seen through the conventional referral mechanism and their wait times remained constant during the study period. CONCLUSIONS: Cardiac EASE reduced

  15. The Swedish mutant barley collection

    International Nuclear Information System (INIS)

    1989-01-01

    Full text: The Swedish mutation research programme in barley began about 50 years ago and has mainly been carried out at Svaloev in co-operation with the institute of Genetics at the University of Lund. The collection has been produced from different Swedish high-yielding spring barley varieties, using the following mutagens: X-rays, neutrons, several organic chemical compounds such as ethyleneimine, several sulfonate derivatives and the inorganic chemical mutagen sodium azide. Nearly 10,000 barley mutants are stored in the Nordic Gene Bank and documented in databases developed by Udda Lundquist, Svaloev AB. The collection consists of the following nine categories with 94 different types of mutants: 1. Mutants with changes in the spike and spikelets; 2. Changes in culm length and culm composition; 3. Changes in growth types; 4. Physiological mutants; 5. Changes in awns; 6. Changes in seed size and shape; 7. Changes in leaf blades; 8. Changes in anthocyanin and colour; 9. Resistance to barley powdery mildew. Barley is one of the most thoroughly investigated crops in terms of induction of mutations and mutation genetics. So far, about half of the mutants stored at the Nordic Gene Bank, have been analysed genetically; They constitute, however, only a minority of the 94 different mutant types. The genetic analyses have given valuable insights into the mutation process but also into the genetic architecture of various characters. A number of mutants of two-row barley have been registered and commercially released. One of the earliest released, Mari, an early maturing, daylength neutral, straw stiff mutant, is still grown in Iceland. The Swedish mutation material has been used in Sweden, but also in other countries, such as Denmark, Germany, and USA, for various studies providing a better understanding of the barley genome. The collection will be immensely valuable for future molecular genetical analyses of clone mutant genes. (author)

  16. Mutant ribosomes can generate dominant kirromycin resistance.

    OpenAIRE

    Tubulekas, I; Buckingham, R H; Hughes, D

    1991-01-01

    Mutations in the two genes for EF-Tu in Salmonella typhimurium and Escherichia coli, tufA and tufB, can confer resistance to the antibiotic kirromycin. Kirromycin resistance is a recessive phenotype expressed when both tuf genes are mutant. We describe a new kirromycin-resistant phenotype dominant to the effect of wild-type EF-Tu. Strains carrying a single kirromycin-resistant tuf mutation and an error-restrictive, streptomycin-resistant rpsL mutation are resistant to high levels of kirromyci...

  17. Eradication of large solid tumors by gene therapy with a T cell receptor targeting a single cancer-specific point mutation

    Science.gov (United States)

    Leisegang, Matthias; Engels, Boris; Schreiber, Karin; Yew, Poh Yin; Kiyotani, Kazuma; Idel, Christian; Arina, Ainhoa; Duraiswamy, Jaikumar; Weichselbaum, Ralph R.; Uckert, Wolfgang; Nakamura, Yusuke; Schreiber, Hans

    2015-01-01

    Purpose Cancers usually contain multiple unique tumor-specific antigens produced by single amino acid substitutions (AAS) and encoded by somatic non-synonymous single nucleotide substitutions. We determined whether adoptively transferred T cells can reject large, well-established solid tumors when engineered to express a single type of T cell receptor (TCR) that is specific for a single AAS. Experimental Design By exome and RNA sequencing of an UV-induced tumor, we identified an AAS in p68 (mp68), a co-activator of p53. This AAS seemed to be an ideal tumor-specific neoepitope because it is encoded by a trunk mutation in the primary autochthonous cancer and binds with highest affinity to the MHC. A high-avidity mp68-specific TCR was used to genetically engineer T cells as well as to generate TCR-transgenic mice for adoptive therapy. Results When the neoepitope was expressed at high levels and by all cancer cells, their direct recognition sufficed to destroy intra-tumor vessels and eradicate large, long-established solid tumors. When the neoepitope was targeted as autochthonous antigen, T cells caused cancer regression followed by escape of antigen-negative variants. Escape could be thwarted by expressing the antigen at increased levels in all cancer cells or by combining T cell therapy with local irradiation. Therapeutic efficacies of TCR-transduced and TCR-transgenic T cells were similar. Conclusions Gene therapy with a single TCR targeting a single AAS can eradicate large established cancer but a uniform expression and/or sufficient levels of the targeted neoepitope or additional therapy are required to overcome tumor escape. PMID:26667491

  18. On the structural affinity of macromolecules with different biological properties: molecular dynamics simulations of a series of TEM-1 mutants.

    Science.gov (United States)

    Giampaolo, Alessia Di; Mazza, Fernando; Daidone, Isabella; Amicosante, Gianfranco; Perilli, Mariagrazia; Aschi, Massimiliano

    2013-07-12

    Molecular Dynamics simulations have been carried out in order to provide a molecular rationalization of the biological and thermodynamic differences observed for a class of TEM β-lactamases. In particular we have considered the TEM-1(wt), the single point mutants TEM-40 and TEM-19 representative of IRT and ESBL classes respectively, and TEM-1 mutant M182T, TEM-32 and TEM-20 which differ from the first three for the additional of M182T mutation. Results indicate that most of the thermodynamic, and probably biological behaviour of these systems arise from subtle effects which, starting from the alterations of the local interactions, produce drastic modifications of the conformational space spanned by the enzymes. The present study suggests that systems showing essentially the same secondary and tertiary structure may differentiate their chemical-biological activity essentially (and probably exclusively) on the basis of the thermal fluctuations occurring in their physiological environment. Copyright © 2013 Elsevier Inc. All rights reserved.

  19. Coproporphyrinogenase in a respiration-deficient mutant of yeast lacking all cytochromes and accumulating coproporphyrin.

    Science.gov (United States)

    Miyake, S; Sugimura, T

    1968-12-01

    In an earlier report, a respiration-deficient mutant of yeast which lacks all cytochromes and hemoproteins and accumulates coproporphyrin was described. This respiration-deficient mutant was temperature-sensitive and resulted from the single chromosomal gene(cyt). In this study, the activity of coproporphyrinogenase, catalyzing the conversion of coproporphyrinogen to protoporphyrinogen, was assayed in the cyt mutant and wild strains. Coproporphyrinogenase activity was 10 times higher in the cyt mutant than in the wild strains. Cells of the cyt mutant grown at 20 C had less activity than those grown at 35 C. The Michaelis constants, pH optima, and temperature activations of the enzymes of the cyt mutant and the wild strains were similar. The significance of the higher activity of this enzyme in the cyt mutant, in which this enzymatic step is apparently blocked in vivo, is discussed.

  20. Adjacent, distal, or combination of point-selective effects of acupuncture on temporomandibular joint disorders: A randomized, single-blind, assessor-blind controlled trial

    Directory of Open Access Journals (Sweden)

    Kyung-Won Kang

    2012-12-01

    Conclusion: Our results suggest that point-selective effects among adjacent, distal, or a combination of acupoints are hardly associated with pain intensity or palpation index in participants with TMDs. Larger sample size trials are required to overcome the shortcomings of the study.

  1. Genetic analysis of DNA repair in Aspergillus: evidence for different types of MMS-sensitive hyperrec mutants

    International Nuclear Information System (INIS)

    Kaefer, E.; Mayor, O.

    1986-01-01

    To identify genes which affect DNA repair and possibly recombination in Aspergillus nidulans, mutants hypersensitive to methyl methanesulphonate (MMS) were induced with ultraviolet light (UV) or γ-rays. To identify functional and epistatic groups, mutants from each uvs gene were tested for effects on recombination and mutation, and double mutant uvs strains were compared for UV survival to their component single mutant strains. (Auth.)

  2. Phenotypic and Genetic Analysis of ``chameleon,'' a Paramecium Mutant with an Enhanced Sensitivity to Magnesium

    OpenAIRE

    Preston, R. R.; Hammond, J. A.

    1997-01-01

    Three mutant strains of Paramecium tetraurelia with an enhanced sensitivity to magnesium have been isolated. These new ``Chameleon'' mutants result from partial- or codominant mutations at a single locus, Cha. Whereas the wild type responded to 5 mM Mg(2+) by swimming backward for 10-15 sec, Cha mutants responded with ~30 sec backward swimming. Electrophysiological analysis suggested that this behavior may be caused by slowing in the rate at which a Mg(2+)-specific ion conductance deactivates...

  3. Development of compact mutants in apple and sour cherry

    International Nuclear Information System (INIS)

    Zagaja, S.W.; Przybyla, A.; Machnik, B.

    1982-01-01

    During the period 1973 - 79 studies were conducted with the aim of developing compact mutants in apple and cherry cultivars and in apple vegetative rootstocks. During the investigations the effect of the dose of gamma rays on frequency of the mutants was studied. Attempts were also made to evolve a micropropagation technique adapted to propagate P 2 and P 22 apple rootstocks, as an aid in mutation breeding. Several mutants were produced in all the material studied, but none of them have yet reached a sufficient developmental stage to enable their complete assessment. On the basis of the results obtained so far the following conclusions can be drawn: higher doses of irradiation resulted in higher frequency of mutants in most apple cultivars and apple rootstocks; in sour cherries the effect of dose depended on the cultivars. Among V 1 shoots developed from sleeping buds on irradiated scion wood, compact mutants were found; their frequency, however, was about 60% lower than among V 1 shoots developed directly from irradiated dormant buds. In apple rootstocks A 2 and M 26 several dwarfed mutants were found; some of these produced thorny plants and some had lower rooting ability; both these characteristics are inferior from the practical point of view. Multiplication and rooting media for in vitro propagation of apple rootstocks, worked out for M 26, were found unsuitable for the rootstocks P 2 and P 22; modifications made in the growth substance composition of the above media enabled satisfactory propagation to be obtained. (author)

  4. A NOVEL SINGLE IMPEDANCE NETWORK BASED NEUTRAL POINT CLAMPED SEVEN LEVEL THREE PHASE INVETER WITH REDUCED CLAMPING DIODES FOR REGENERATIVE APPLICATIONS

    OpenAIRE

    T. A. Raghavendiran; C.L.Kuppuswamy

    2012-01-01

    A Multilevel Inverter (MLI) can eliminate the need for the step-up transformer and reduce the harmonics produced by the inverter. This paper presents Neutral Point Clamped (NPC) seven level inverter with less number of clamping diodes which is suitable for regenerative loads such as three phase induction motor with regenerative braking. To reduce the stress across the main switches, diodes are clamped anti-parallel to the main switches. In Earlier configurations, the numbers of clamping diode...

  5. Ab initio analysis of a vacancy and a self-interstitial near single crystal silicon surfaces: Implications for intrinsic point defect incorporation during crystal growth from a melt

    Energy Technology Data Exchange (ETDEWEB)

    Kamiyama, Eiji; Sueoka, Koji [Department of Communication Engineering, Okayama Prefectural University, 111 Kuboki, Soja, Okayama 719-1197 (Japan); Vanhellemont, Jan [Department of Solid State Sciences, Ghent University, Krijgslaan 281-S1, Gent 9000 (Belgium)

    2012-10-15

    The microscopic model of the Si (001) crystal surface was investigated by first principles calculations to clarify the behavior of intrinsic point defects near crystal surfaces. A c(4 x 2) structure model was used to describe the crystal surface in contact with vacuum. The calculations show lower formation energy near the surface and the existence of formation energy differences between the surface and the bulk for both types of intrinsic point defects. The tetrahedral (T)-site and the dumbbell (DB)-site, in which a Si atom is captured from the surface and forms a self-interstitial, are found as stable sites near the third atomic layer. The T-site has a barrier of 0.48 eV, whereas the DB-site has no barrier for the interstitial to penetrate into the crystal from the vacuum. Si atoms in a melt can migrate and reach at the third layer during crystal growth when bulk diffusion coefficient is used. Therefore, the melt/solid interface is always a source of intrinsic point defects. (Copyright copyright 2012 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

  6. The analysis of dyes in ball point pen inks on single paper fibres using laser desorption ionisation time of flight mass spectrometry (LDI-TOFMS).

    Science.gov (United States)

    Matthews, Broderick; Walker, G Stewart; Kobus, Hilton; Pigou, Paul; Bird, Carolyne; Smith, Glyn

    2011-06-15

    An important requisite for the forensic analysis of inks on documents is that damage to the document is avoided or minimised. This paper describes a technique for dye identification in ballpoint pen inks using LDI-TOFMS on single ink bearing paper fibres and its application to a case. A single ink bearing paper fibre can be prised from the surface of the document under a stereo microscope and presented to the instrument for analysis without further treatment. This sampling process causes imperceptible damage to the surface of the document. Clear mass spectrometric identification of the ink dyes is obtained. A case example is provided to illustrate the practical application of the technique. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

  7. Enhanced longevity in tau mutant Syrian hamsters, Mesocricetus auratus

    NARCIS (Netherlands)

    Oklejewicz, Malgorzata; Daan, Serge

    The single-gene mutation tau in the Syrian hamster shortens the circadian period by about 20% in the homozygous mutant and simultaneously increases the mass-specific metabolic rate by about 20%. Both effects might be expected to lead to a change in longevity. To test such expectations, the life span

  8. Is auxin involved in the induction of genetic instability in barley homeotic double mutants?

    Science.gov (United States)

    Šiukšta, Raimondas; Vaitkūnienė, Virginija; Rančelis, Vytautas

    2018-02-01

    The triggers of genetic instability in barley homeotic double mutants are tweaky spike -type mutations associated with an auxin imbalance in separate spike phytomeres. Barley homeotic tweaky spike;Hooded (tw;Hd) double mutants are characterized by an inherited instability of spike and flower development, which is absent in the single parental constituents. The aim of the present study was to show that the trigger of genetic instability in the double mutants is the tw mutations, which are associated with an auxin imbalance in the developing spikes. Their pleiotropic effects on genes related to spike/flower development may cause the genetic instability of double mutants. The study of four double-mutant groups composed of different mutant alleles showed that the instability arose only if the mutant allele tw was a constituent of the double mutants. Application of auxin inhibitors and 2,4-dichlorophenoxyacetic acid (2,4-D) demonstrated the relationship of the instability of the double mutants and the phenotype of the tw mutants to auxin imbalance. 2,4-D induced phenocopies of the tw mutation in wild-type plants and rescued the phenotypes of three allelic tw mutants. The differential display (dd-PCR) method allowed the identification of several putative candidate genes in tw that may be responsible for the initiation of instability in the double mutants by pleiotropic variations of their expression in the tw mutant associated with auxin imbalance in the developing spikes. The results of the present study linked the genetic instability of homeotic double mutants with an auxin imbalance caused by one of the constituents (tw). The genetic instability of the double mutants in relation to auxin imbalance was studied for the first time. A matrocliny on instability expression was also observed.

  9. An SVM-Based Classifier for Estimating the State of Various Rotating Components in Agro-Industrial Machinery with a Vibration Signal Acquired from a Single Point on the Machine Chassis

    Directory of Open Access Journals (Sweden)

    Ruben Ruiz-Gonzalez

    2014-11-01

    Full Text Available The goal of this article is to assess the feasibility of estimating the state of various rotating components in agro-industrial machinery by employing just one vibration signal acquired from a single point on the machine chassis. To do so, a Support Vector Machine (SVM-based system is employed. Experimental tests evaluated this system by acquiring vibration data from a single point of an agricultural harvester, while varying several of its working conditions. The whole process included two major steps. Initially, the vibration data were preprocessed through twelve feature extraction algorithms, after which the Exhaustive Search method selected the most suitable features. Secondly, the SVM-based system accuracy was evaluated by using Leave-One-Out cross-validation, with the selected features as the input data. The results of this study provide evidence that (i accurate estimation of the status of various rotating components in agro-industrial machinery is possible by processing the vibration signal acquired from a single point on the machine structure; (ii the vibration signal can be acquired with a uniaxial accelerometer, the orientation of which does not significantly affect the classification accuracy; and, (iii when using an SVM classifier, an 85% mean cross-validation accuracy can be reached, which only requires a maximum of seven features as its input, and no significant improvements are noted between the use of either nonlinear or linear kernels.

  10. Effect of ribosome-targeting antibiotics on streptomycin-resistant Mycobacterium mutants in the rpsL gene.

    Science.gov (United States)

    Pelchovich, Gidi; Zhuravlev, Alina; Gophna, Uri

    2013-08-01

    Streptomycin (Sm) was the first antibiotic used against Mycobacterium tuberculosis, the aetiological agent of tuberculosis (TB). However, point mutations in the rpsL gene can generate resistance to Sm, which is why spontaneous resistance to this antibiotic emerges so rapidly during treatment. Here we examine the interaction between Sm resistance and sensitivity to other ribosome-targeting antibiotics. Levels of resistance of rpsL mutants to the ribosome-affecting antibiotics chloramphenicol, tetracycline, gentamicin and erythromycin were tested, both singly and in combination. For this purpose, Mycobacterium smegmatis was used, which is commonly used in laboratory experiments as a model for TB. Generally, Sm-resistant mutants were as sensitive to the ribosome-affecting antibiotics as the wild-type strain. Combinations of different ribosome-affecting antibiotics were occasionally more potent than either of the single drugs, with better inhibition of both wild-type and mutant strains. Combining different ribosome-affecting drugs could represent an additional strategy in treating mycobacterial infections, including those resistant to newer drugs such as isoniazid or ethambutol. Copyright © 2013 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.

  11. Targeting ESR1-Mutant Breast Cancer

    Science.gov (United States)

    2015-09-01

    we have developed models of mutant ER driven cancer in which to characterize gene expression. Specifically, tetracycline inducible MCF7 cells that...expression profiling. Figure 1: Degradation of WT and mutant ER with ARN810. MCF7 cells stably transfected with tet-inducible WT and mutant...mutant ER. MCF7 cells stably transfected with tet- inducible WT and mutant ER are treated for 24 hours with estradiol and gene expression profiling

  12. Tipping Point

    Medline Plus

    Full Text Available ... en español Blog About OnSafety CPSC Stands for Safety The Tipping Point Home > 60 Seconds of Safety (Videos) > The Tipping Point The Tipping Point by ... danger death electrical fall furniture head injury product safety television tipover tv Watch the video in Adobe ...

  13. The effect of correlated and point defects on the vortex lattice melting transition in single crystal YBa2Cu3O7-δ

    International Nuclear Information System (INIS)

    Kwok, W.K.; Fleshler, S.; Welp, U.; Downey, J.; Crabtree, G.W.; Fendrich, J. Giapintzakis, J.

    1993-08-01

    The vortex melting transition T m in several untwinned and twinned crystals measured resistively in fields up to 8 Tesla. A Lindemann criterion for vortex lattice melting is obtained in addition to a sharp hysteresis in the magnetoresistance at B m supporting a first order phase transition. The anisotropy of twin boundary pinning and its reduction of the ''kink'' in ρ(T) associated with the first order melting transition is discussed in samples with very dilute twin boundaries. We also report on direct suppression of melting transition by intrinsic pinning for H parallel ab and by electron-irradiation-induced point defects

  14. Cost-effectiveness of lumbar discectomy and single-level fusion for spondylolisthesis: experience with the NeuroPoint-SD registry.

    Science.gov (United States)

    Mummaneni, Praveen V; Whitmore, Robert G; Curran, Jill N; Ziewacz, John E; Wadhwa, Rishi; Shaffrey, Christopher I; Asher, Anthony L; Heary, Robert F; Cheng, Joseph S; Hurlbert, R John; Douglas, Andrea F; Smith, Justin S; Malhotra, Neil R; Dante, Stephen J; Magge, Subu N; Kaiser, Michael G; Abbed, Khalid M; Resnick, Daniel K; Ghogawala, Zoher

    2014-06-01

    There is significant practice variation and uncertainty as to the value of surgical treatments for lumbar spine disorders. The authors' aim was to establish a multicenter registry to assess the efficacy and costs of common lumbar spinal procedures by using prospectively collected outcomes. An observational prospective cohort study was completed at 13 academic and community sites. Patients undergoing single-level fusion for spondylolisthesis or single-level lumbar discectomy were included. The 36-Item Short Form Health Survey (SF-36) and Oswestry Disability Index (ODI) data were obtained preoperatively and at 1, 3, 6, and 12 months postoperatively. Power analysis estimated a sample size of 160 patients: lumbar disc (125 patients) and lumbar listhesis (35 patients). The quality-adjusted life year (QALY) data were calculated using 6-dimension utility index scores. Direct costs and complication costs were estimated using Medicare reimbursement values from 2011, and indirect costs were estimated using the human capital approach with the 2011 US national wage index. Total costs equaled $14,980 for lumbar discectomy and $43,852 for surgery for lumbar spondylolisthesis. There were 198 patients enrolled over 1 year. The mean age was 46 years (49% female) for lumbar discectomy (n = 148) and 58.1 years (60% female) for lumbar spondylolisthesis (n = 50). Ten patients with disc herniation (6.8%) and 1 with listhesis (2%) required repeat operation at 1 year. The overall 1-year follow-up rate was 88%. At 30 days, both lumbar discectomy and single-level fusion procedures were associated with significant improvements in ODI, visual analog scale, and SF-36 scores (p = 0.0002), which persisted at the 1-year evaluation (p spondylolisthesis. One-year cost per QALY was obtained, and this cost per QALY is expected to improve further by 2 years. This work sets the stage for real-world analysis of the value of health interventions.

  15. PedonnanceofE3rly MatUring MutantS Derived from ''SuPa'~ Rice ...

    African Journals Online (AJOL)

    Vienna, Austria in 1994. The dry seeds were in-adiated with gamma rays using three doses (170, 210. --iifid 24OC;Y).frOm C.obalt 60 (lCO) in order shorten the plant height and maturity period. From the resulting mutant. PoPulations ortgindtiriifroni modified single seed descent method, five Jery early maturing lines plus the ...

  16. Wild Accessions and Mutant Resources

    DEFF Research Database (Denmark)

    Kawaguchi, Masayoshi; Sandal, Niels Nørgaard

    2014-01-01

    Lotus japonicus, Lotus burttii, and Lotus filicaulis are species of Lotus genus that are utilized for molecular genetic analysis such as the construction of a linkage map and QTL analysis. Among them, a number of mutants have been isolated from two wild accessions: L. japonicus Gifu B-129...

  17. components in induced sorghum mutants

    African Journals Online (AJOL)

    (1984) evaluated induced mutation and hybridisation methods for producing genetic variability in 15 quantitative characters of sorghum. Their results showed large variability in grain yield, plant maturity, plant height and panicles length. Selected mutants with favorable properties can be directly combined in varietal hybrids.

  18. Development and Positioning Accuracy Assessment of Single-Frequency Precise Point Positioning Algorithms by Combining GPS Code-Pseudorange Measurements with Real-Time SSR Corrections

    Directory of Open Access Journals (Sweden)

    Miso Kim

    2017-06-01

    Full Text Available We have developed a suite of real-time precise point positioning programs to process GPS pseudorange observables, and validated their performance through static and kinematic positioning tests. To correct inaccurate broadcast orbits and clocks, and account for signal delays occurring from the ionosphere and troposphere, we applied State Space Representation (SSR error corrections provided by the Seoul Broadcasting System (SBS in South Korea. Site displacements due to solid earth tide loading are also considered for the purpose of improving the positioning accuracy, particularly in the height direction. When the developed algorithm was tested under static positioning, Kalman-filtered solutions produced a root-mean-square error (RMSE of 0.32 and 0.40 m in the horizontal and vertical directions, respectively. For the moving platform, the RMSE was found to be 0.53 and 0.69 m in the horizontal and vertical directions.

  19. Indy mutants: live long and prosper

    Directory of Open Access Journals (Sweden)

    Stewart eFrankel

    2012-02-01

    Full Text Available Indy encodes the fly homologue of a mammalian transporter of di and tricarboxylatecomponents of the Krebs cycle. Reduced expression of fly Indy or two of the C. elegansIndy homologs leads to an increase in life span. Fly and worm tissues that play key roles inintermediary metabolism are also the places where Indy genes are expressed. One of themouse homologs of Indy (mIndy is mainly expressed in the liver. It has been hypothesizedthat decreased INDY activity creates a state similar to caloric restriction (CR. Thishypothesis is supported by the physiological similarities between Indy mutant flies on highcalorie food and control flies on CR, such as increased physical activity and decreases inweight, egg production, triglyceride levels, starvation resistance, and insulin signaling. Inaddition, Indy mutant flies undergo changes in mitochondrial biogenesis also observed inCR animals. Recent findings with mIndy knockout mice support and extend the findingsfrom flies. mIndy-/- mice display an increase in hepatic mitochondrial biogenesis, lipidoxidation and decreased hepatic lipogenesis. When mIndy-/- mice are fed high calorie foodthey are protected from adiposity and insulin resistance. These findings point to INDY as apotential drug target for the treatment of metabolic syndrome, type 2 diabetes and obesity.

  20. UV and gamma-ray sensitivity of meiosis-deficient mutants in Podospora anserina

    International Nuclear Information System (INIS)

    Simonet, J.M.

    1976-01-01

    Two mutants, mei1 and mei2, were isolated by screening for deficiencies occurring in the meiotic process. The sensitivity of mei1 and mei2 mutant strains to UV irradiation showed a significant increase as compared with that of the wild-type stock, hwhereas the sensitivity to γ-rays remained unchanged. The double-mutant strains were no more sensitive than each single mutant. The data indicate that both mei1 and mei2 loci are probably involved in the same pathway of excision-repair of UV-induced lesions

  1. Diverse point mutations in the human glucose-6-phosphate dehydrogenase gene cause enzyme deficiency and mild or severe hemolytic anemia

    Energy Technology Data Exchange (ETDEWEB)

    Vulliamy, T.J.; D' Urso, M.; Battistuzzi, G.; Estrada, M.; Foulkes, N.S.; Martini, G.; Calabro, V.; Poggi, V.; Giordano, R.; Town, M.; Luzzatto, L.; Persico, M.G. (Royal Postgraduate Medical School, London (England))

    1988-07-01

    Glucose-6-phosphate dehydrogenase deficiency is a common genetic abnormality affecting an estimated 400 million people worldwide. Clinical and biochemical analyses have identified many variants exhibiting a range of phenotypes, which have been well characterized from the hematological point of view. However, until now, their precise molecular basis has remained unknown. The authors have cloned and sequenced seven mutant G6PD alleles. In the nondeficient polymorphic African variant G6PD A they have found a single point mutation. The other six mutants investigated were all associated with enzyme deficiency. The mutations observed show a striking predominance of C {yields} T transitions, with CG doublets involved in four of seven cases. Thus, diverse point mutations may account largely for the phenotypic heterogeneity of G6PD deficiency.

  2. Induced High Lysine Mutants in Barley

    DEFF Research Database (Denmark)

    Doll, Hans; Køie, B.; Eggum, B. O.

    1974-01-01

    variety. Comparisons of six high lysine mutants with the parent variety showed that grain yield and seed size of the mutants are reduced between 10 and 30 per cent. However, the most promising mutant had the lowest reduction in grain yield, and the absolute lysine yield of this mutant was some 30 per cent...... above that of the parent variety. Feeding tests with rats revealed substantial increases in the biological value of the high lysine mutant protein. Also the net protein utilization was improved but less so because of a somewhat reduced digestibility of the mutant protein....

  3. Structure-Guided, Single-Point Modifications in the Phosphinic Dipeptide Structure Yield Highly Potent and Selective Inhibitors of Neutral Aminopeptidases

    Energy Technology Data Exchange (ETDEWEB)

    Vassiliou, Stamatia; Węglarz-Tomczak, Ewelina; Berlicki, Łukasz; Pawełczak, Małgorzata; Nocek, Bogusław; Mulligan, Rory; Joachimiak, Andrzej; Mucha, Artur

    2014-10-09

    Seven crystal structures of alanyl aminopeptidase from Neisseria meningitides (the etiological agent of meningitis, NmAPN) complexed with organophosphorus compounds were resolved to determine the optimal inhibitor-enzyme interactions. The enantiomeric phosphonic acid analogs of Leu and hPhe, which correspond to the P1 amino acid residues of well-processed substrates, were used to assess the impact of the absolute configuration and the stereospecific hydrogen bond network formed between the aminophosphonate polar head and the active site residues on the binding affinity. For the hPhe analog, an imperfect stereochemical complementarity could be overcome by incorporating an appropriate P1 side chain. The constitution of P1'-extended structures was rationally designed and the lead, phosphinic dipeptide hPhePψ[CH2]Phe, was modified in a single position. Introducing a heteroatom/heteroatom-based fragment to either the P1 or P1' residue required new synthetic pathways. The compounds in the refined structure were low nanomolar and subnanomolar inhibitors of N. meningitides, porcine and human APNs, and the reference leucine aminopeptidase (LAP). The unnatural phosphinic dipeptide analogs exhibited a high affinity for monozinc APNs associated with a reasonable selectivity versus dizinc LAP. Another set of crystal structures containing the NmAPN dipeptide ligand were used to verify and to confirm the predicted binding modes; furthermore, novel contacts, which were promising for inhibitor development, were identified, including a π-π stacking interaction between a pyridine ring and Tyr372.

  4. Accurate approach for determining fresh-water carbonate (H2CO3(*)) alkalinity, using a single H3PO4 titration point.

    Science.gov (United States)

    Birnhack, Liat; Sabach, Sara; Lahav, Ori

    2012-10-15

    A new, simple and accurate method is introduced for determining H(2)CO(3)(*) alkalinity in fresh waters dominated by the carbonate weak-acid system. The method relies on a single H(3)PO(4) dosage and two pH readings (acidic pH value target: pH~4.0). The computation algorithm is based on the concept that the overall alkalinity mass of a solution does not change upon the addition of a non-proton-accepting species. The accuracy of the new method was assessed batch-wise with both synthetic and actual tap waters and the results were compared to those obtained from two widely used alkalinity analysis methods (titration to pH~4.5 and the Gran titration method). The experimental results, which were deliberately obtained with simple laboratory equipment (glass buret, general-purpose pH electrode, magnetic stirrer) proved the method to be as accurate as the conventional methods at a wide range of alkalinity values (20-400 mg L(-1) as CaCO(3)). Analysis of the relative error attained in the proposed method as a function of the target (acidic) pH showed that at the range 4.0pHalkalinity measurement is also described. Copyright © 2012 Elsevier B.V. All rights reserved.

  5. SAAFEC: Predicting the Effect of Single Point Mutations on Protein Folding Free Energy Using a Knowledge-Modified MM/PBSA Approach.

    Science.gov (United States)

    Getov, Ivan; Petukh, Marharyta; Alexov, Emil

    2016-04-07

    Folding free energy is an important biophysical characteristic of proteins that reflects the overall stability of the 3D structure of macromolecules. Changes in the amino acid sequence, naturally occurring or made in vitro, may affect the stability of the corresponding protein and thus could be associated with disease. Several approaches that predict the changes of the folding free energy caused by mutations have been proposed, but there is no method that is clearly superior to the others. The optimal goal is not only to accurately predict the folding free energy changes, but also to characterize the structural changes induced by mutations and the physical nature of the predicted folding free energy changes. Here we report a new method to predict the Single Amino Acid Folding free Energy Changes (SAAFEC) based on a knowledge-modified Molecular Mechanics Poisson-Boltzmann (MM/PBSA) approach. The method is comprised of two main components: a MM/PBSA component and a set of knowledge based terms delivered from a statistical study of the biophysical characteristics of proteins. The predictor utilizes a multiple linear regression model with weighted coefficients of various terms optimized against a set of experimental data. The aforementioned approach yields a correlation coefficient of 0.65 when benchmarked against 983 cases from 42 proteins in the ProTherm database. the webserver can be accessed via http://compbio.clemson.edu/SAAFEC/.

  6. High genetic homogeneity points to a single introduction event responsible for invasion of Cotton leaf curl Multan virus and its associated betasatellite into China.

    Science.gov (United States)

    Du, Zhenguo; Tang, Yafei; He, Zifu; She, Xiaoman

    2015-10-07

    Cotton leaf curl Multan virus (CLCuMuV) is a Whitefly Transmitted Geminivirus (WTG) endemic to the India subcontinent and is notorious as a causal agent of cotton leaf curl disease (CLCuD), a major constraint to cotton production in south Asia. We found CLCuMuV infecting Hibiscus rosa-sinensis in Guangzhou, China in 2006. The spread and evolution of the invading CLCuMuV were monitored in the following nine years. CLCuMuV spread rapidly in the last nine years and became established in Southern China. It infects at least five malvaceous plant species, H. rosa-sinensis, H. esculentus, Malvaiscus arboreus, Gossypium hirsutum and H. cannabinus. Complete nucleotide sequences of 34 geographically and/or temporally distinct CLCuMuV isolates were determined and analyzed together with six other publicly available genomes of CLCuMuV occurring in China. The 40 CLCuMuV isolates were found to share > 99 % nucleotide sequence identity with each other. In all cases tested, the CLCuMuVs were associated with a CLCuMuB. The 36 CLCuMuBs (30 sequenced by us) shared > 98 % nucleotide sequence identity. The high genetic homogeneity of CLCuMuV and CLCuMuB in China suggests the establishment of them from a single founder event.

  7. Tipping Point

    Medline Plus

    Full Text Available ... OnSafety CPSC Stands for Safety The Tipping Point Home > 60 Seconds of Safety (Videos) > The Tipping Point ... 24 hours a day. For young children whose home is a playground, it’s the best way to ...

  8. Fixed Points

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education; Volume 5; Issue 5. Fixed Points - From Russia with Love - A Primer of Fixed Point Theory. A K Vijaykumar. Book Review Volume 5 Issue 5 May 2000 pp 101-102. Fulltext. Click here to view fulltext PDF. Permanent link:

  9. Normal aging modulates the neurotoxicity of mutant huntingtin.

    Directory of Open Access Journals (Sweden)

    Elsa Diguet

    Full Text Available Aging likely plays a role in neurodegenerative disorders. In Huntington's disease (HD, a disorder caused by an abnormal expansion of a polyglutamine tract in the protein huntingtin (Htt, the role of aging is unclear. For a given tract length, the probability of disease onset increases with age. There are mainly two hypotheses that could explain adult onset in HD: Either mutant Htt progressively produces cumulative defects over time or "normal" aging renders neurons more vulnerable to mutant Htt toxicity. In the present study, we directly explored whether aging affected the toxicity of mutant Htt in vivo. We studied the impact of aging on the effects produced by overexpression of an N-terminal fragment of mutant Htt, of wild-type Htt or of a beta-Galactosidase (beta-Gal reporter gene in the rat striatum. Stereotaxic injections of lentiviral vectors were performed simultaneously in young (3 week and old (15 month rats. Histological evaluation at different time points after infection demonstrated that the expression of mutant Htt led to pathological changes that were more severe in old rats, including an increase in the number of small Htt-containing aggregates in the neuropil, a greater loss of DARPP-32 immunoreactivity and striatal neurons as assessed by unbiased stereological counts.The present results support the hypothesis that "normal" aging is involved in HD pathogenesis, and suggest that age-related cellular defects might constitute potential therapeutic targets for HD.

  10. Development of Bacillus subtilis mutants to produce tryptophan in pigs.

    Science.gov (United States)

    Bjerre, Karin; Cantor, Mette D; Nørgaard, Jan V; Poulsen, Hanne D; Blaabjerg, Karoline; Canibe, Nuria; Jensen, Bent B; Stuer-Lauridsen, Birgitte; Nielsen, Bea; Derkx, Patrick M F

    2017-02-01

    To generate tryptophan-overproducing Bacillus subtilis strains for in situ use in pigs, to reduce the feed cost for farmers and nitrogen pollution. A novel concept has been investigated-to generate B. subtilis strains able to produce tryptophan (Trp) in situ in pigs. Mutagenesis by UV was combined with selection on Trp and purine analogues in an iterative process. Two mutants from different wild types were obtained, mutant 1 (M1) produced 1 mg Trp/l and mutant 2 (M2) 14 mg Trp/l. Genome sequence analysis revealed that M1 had three single nuclear polymorphisms (SNPs) and M2 had two SNPs compared to the wild type strains. In both mutants SNPs were found in genes regulating tryptophan synthesis. Reverse transcription PCR confirmed up-regulation of the tryptophan synthesis genes in both mutants, the expression was up to 3 times higher in M2 than in M1. Tryptophan-excreting B. subtilis strains were obtained with UV-mutagenesis and analogue selection and can be used in animal feed applications.

  11. Rapid MR assessment of left ventricular systolic function after acute myocardial infarction using single breath-hold cine imaging with the temporal parallel acquisition technique (TPAT) and 4D guide-point modelling analysis of left ventricular function

    International Nuclear Information System (INIS)

    Eberle, Holger C.; Jensen, Christoph J.; Sabin, Georg V.; Naber, Christoph K.; Bruder, Oliver; Nassenstein, Kai; Schlosser, Thomas

    2010-01-01

    We compared four-dimensional guide-point modelling left ventricular function analysis (4DVF) results of cine images in four short-axis and two long-axis slices acquired in a single breath-hold, obtained with the temporal parallel acquisition technique (TPAT), with standard left ventricular function (LVF) analysis results determined by the summation of discs method, in patients who had recently suffered myocardial infarction. Despite wall motion abnormalities, 4DVF yields results for left ventricular ejection fractions and end-diastolic and end-systolic volumes that are in excellent agreement with standard LVF analysis results in these patients. A shortened cardiac magnetic resonance (CMR) protocol using single breath-hold cine image acquisition could facilitate the assessment of left ventricular function soon after myocardial infarction in critically ill patients who are unable to comply with the multiple breath-holds required for standard LVF analysis. (orig.)

  12. Mutant genes in pea breeding

    International Nuclear Information System (INIS)

    Swiecicki, W.K.

    1990-01-01

    Full text: Mutations of genes Dpo (dehiscing pods) and A (anthocyanin synthesis) played a role in pea domestication. A number of other genes were important in cultivar development for 3 types of usage (dry seeds, green vegetable types, fodder), e.g. fn, fna, le, p, v, fas and af. New genes (induced and spontaneous), are important for present ideotypes and are registered by the Pisum Genetics Association (PGA). Comparison of a pea variety ideotype with the variation available in gene banks shows that breeders need 'new' features. In mutation induction experiments, genotype, mutagen and method of treatment (e.g. combined or fractionated doses) are varied for broadening the mutation spectrum and selecting more genes of agronomic value. New genes are genetically analysed. In Poland, some mutant varieties with the gene afila were registered, controlling lodging by a shorter stem and a higher number of internodes. Really non-lodging pea varieties could strongly increase seed yield. But the probability of detecting a major gene for lodging resistance is low. Therefore, mutant genes with smaller influence on plant architecture are sought, to combine their effect by crossing. Promising seem to be the genes rogue, reductus and arthritic as well as a number of mutant genes not yet genetically identified. The gene det for terminal inflorescence - similarly to Vicia faba - changes plant development. Utilisation of assimilates and ripening should be better. Improvement of harvest index should give higher seed yield. A number of genes controlling disease resistance are well known (eg. Fw, Fnw, En, mo and sbm). Important in mass screening of resistance are closely linked gene markers. Pea gene banks collect respective lines, but mutants induced in highly productive cultivars would be better. Inducing gene markers sometimes seems to be easier than transfer by crossing. Mutation induction in pea breeding is probably more important because a high number of monogenic features are

  13. Saint Louis Encephalitis Temperature-Sensitive Mutants.

    Science.gov (United States)

    1981-01-01

    group II contains two mutants and group III contains three mutants. Examination of the ability of mutants to grow at 300C, 40 C or 37 C indicates that...importance. Information from studies with Poliovirus has shown that the use of live attenuated virus vaccines result in longer lasting, more effec- tive...sensitive mutant to grow at internal body temperature may have a significant effect on the ability of the virus to induce a protective immune response or

  14. Advantages of conducting in-situ U-Pb age dating of multiple U-bearing minerals from a single complex: Case in point - the Oka Carbonatite Complex

    Science.gov (United States)

    Chen, W.; Simonetti, A.

    2012-12-01

    sample. The niocalite for a carbonatite sample Oka153 defines a bimodal age distribution, with weighted average 206Pb/238U ages of 110.1 ±5.0 and 133.2 ±6.1 Ma. Apatite from the same sample also records a similar bimodal age distribution of 111.4 ±2.8 and 126.9 ±1.8 Ma. The combined in situ U-Pb dating results for apatite, pervoskite, niocalite from Oka clearly support a protracted history of magmatic activity (~30 Myr) for this carbonatite complex. Of importance, the U-Pb results from this study clearly indicate the significance of conducting a thorough geochronological investigation rather than defining the age of any one alkaline complex solely on the basis of a single radiometric age determination.

  15. A double-mutant collection targeting MAP kinase related genes in Arabidopsis for studying genetic interactions.

    Science.gov (United States)

    Su, Shih-Heng; Krysan, Patrick J

    2016-12-01

    Mitogen-activated protein kinase cascades are conserved in all eukaryotes. In Arabidopsis thaliana there are approximately 80 genes encoding MAP kinase kinase kinases (MAP3K), 10 genes encoding MAP kinase kinases (MAP2K), and 20 genes encoding MAP kinases (MAPK). Reverse genetic analysis has failed to reveal abnormal phenotypes for a majority of these genes. One strategy for uncovering gene function when single-mutant lines do not produce an informative phenotype is to perform a systematic genetic interaction screen whereby double-mutants are created from a large library of single-mutant lines. Here we describe a new collection of 275 double-mutant lines derived from a library of single-mutants targeting genes related to MAP kinase signaling. To facilitate this study, we developed a high-throughput double-mutant generating pipeline using a system for growing Arabidopsis seedlings in 96-well plates. A quantitative root growth assay was used to screen for evidence of genetic interactions in this double-mutant collection. Our screen revealed four genetic interactions, all of which caused synthetic enhancement of the root growth defects observed in a MAP kinase 4 (MPK4) single-mutant line. Seeds for this double-mutant collection are publicly available through the Arabidopsis Biological Resource Center. Scientists interested in diverse biological processes can now screen this double-mutant collection under a wide range of growth conditions in order to search for additional genetic interactions that may provide new insights into MAP kinase signaling. © 2016 The Authors The Plant Journal © 2016 John Wiley & Sons Ltd.

  16. Tipping Point

    Medline Plus

    Full Text Available ... Point by CPSC Blogger September 22, 2009 appliance child Childproofing CPSC danger death electrical fall furniture head ... see news reports about horrible accidents involving young children and furniture, appliance and tv tip-overs. The ...

  17. Molecular alterations underlying the spontaneous and γ-ray-induced point mutations at the white locus of Drosophila Melanogaster

    International Nuclear Information System (INIS)

    Aleksandrova, M.V.; Lapidus, I.L.; Aleksandrov, I.D.; Karpovskij, A.L.

    1996-01-01

    The white locus in D.Melanogaster was selected as a target gene for the study of the mutational spectra of spontaneously arising and radiation-induced gene mutations in a whole organism. Analysis of 6 spontaneous and 73 γ-ray-induced white mutations by a combination of cytological, genetic and molecular techniques revealed that on the chromosomal and genetic levels all spontaneous mutations showed themselves to be point mutants. The share of such mutants among all heritable radiation-induced gene mutations is about 40%, whereas the rest ones are due to exchange breaks (8%) as well as multilocus, single-locus or partial-locus (intragenic) deletions (52%). The DNAs from 4 spontaneous and 17 γ-ray-induced point mutants were analysed by Southern blot-hybridization. The three spontaneous and 7 radiation mutants showed an altered DNA sequence at the left (distal) half of the white gene due to insertion or DNA rearrangement. The rest (58%) of the radiation-induced point mutations did not indicate any alternations in this part of the gene as detected by this technique and probes employed. 15 refs., 3 figs., 1 tab

  18. An extra early mutant of pigeonpea

    International Nuclear Information System (INIS)

    Ravikesavan, R.; Kalaimagal, T.; Rathnaswamy, R.

    2001-01-01

    The redgram (Cajanus cajan (L.) Huth) variety 'Prabhat DT' was gamma irradiated with 100, 200, 300 and 400 Gy doses. Several mutants have been identified viz., extra early mutants, monostem mutants, obcordifoliate mutants and bi-stigmatic mutants. The extra early mutant was obtained when treated with 100 Gy dose. The mutant was selfed and forwarded from M 2 to M 4 generation. In the M 4 generation the mutant line was raised along with the parental variety. Normal cultural practices were followed and the biometrical observations were recorded. It was observed that for the characters viz., total number of branches per plant, number of pods per plants, seeds per pod, 100 seed weight and seed yield per plant there was no difference between the mutant and parent variety. Whereas, regarding the days to flowering and maturity the mutants were earlier than the parents. The observation was recorded from two hundred plants each. The mutant gives the same yield in 90 days as that of the parent variety in 107 days, which make it an economic mutant

  19. Investigación de la unión soldada entre el vástago y las placas de las cuchillas calzadas // Investigation of the welded joint between plates and tipped single-point lathe tools

    Directory of Open Access Journals (Sweden)

    M. Jacas Cabrera

    2000-01-01

    Full Text Available El presente trabajo está dirigido al incremento de los niveles de producción y calidad, específicamente en la línea defabricación de cuchillas calzadas para torno en el centro fabril “Miguel Saavedra” HERRAMIX.En este caso se realizó un análisis para la sustitución de las pastillas de soldar, fabricadas por CIME por nuevastrimetálicas.En el mismo se determinaron los tiempos de calentamiento necesarios para realizar la soldadura, en las máquinas deinducción (TBCHE, así como los valores de resistencia al cizallamiento de los calzos una vez soldados.Palabras claves: Cuchillas de punta, calzos metalo-ceramicos, placas trimetálicas_____________________________________________________________________Abstract:The present work is directed to increase the production and quality levels of tipped single-point lathe tools at “MiguelSaavedra” plant HERRAMIX.This work deals with a study about the substitution of brazing-pads made by CIME for tri-metallic new ones.The induction brazing heating time’s necessaries at induction machines (TBCHE as well as shear-stress values at the tips afterwelding are determined.Key words: tip single point lathe tool, metal ceramic plate, trimetallic pads.

  20. IDH-mutant glioma specific association of rs55705857 located at 8q24.21 involves MYC deregulation.

    Science.gov (United States)

    Oktay, Yavuz; Ülgen, Ege; Can, Özge; Akyerli, Cemaliye B; Yüksel, Şirin; Erdemgil, Yiğit; Durası, I Melis; Henegariu, Octavian Ioan; Nanni, E Paolo; Selevsek, Nathalie; Grossmann, Jonas; Erson-Omay, E Zeynep; Bai, Hanwen; Gupta, Manu; Lee, William; Turcan, Şevin; Özpınar, Aysel; Huse, Jason T; Sav, M Aydın; Flanagan, Adrienne; Günel, Murat; Sezerman, O Uğur; Yakıcıer, M Cengiz; Pamir, M Necmettin; Özduman, Koray

    2016-06-10

    The single nucleotide polymorphism rs55705857, located in a non-coding but evolutionarily conserved region at 8q24.21, is strongly associated with IDH-mutant glioma development and was suggested to be a causal variant. However, the molecular mechanism underlying this association has remained unknown. With a case control study in 285 gliomas, 316 healthy controls, 380 systemic cancers, 31 other CNS-tumors, and 120 IDH-mutant cartilaginous tumors, we identified that the association was specific to IDH-mutant gliomas. Odds-ratios were 9.25 (5.17-16.52; 95% CI) for IDH-mutated gliomas and 12.85 (5.94-27.83; 95% CI) for IDH-mutated, 1p/19q co-deleted gliomas. Decreasing strength with increasing anaplasia implied a modulatory effect. No somatic mutations were noted at this locus in 114 blood-tumor pairs, nor was there a copy number difference between risk-allele and only-ancestral allele carriers. CCDC26 RNA-expression was rare and not different between the two groups. There were only minor subtype-specific differences in common glioma driver genes. RNA sequencing and LC-MS/MS comparisons pointed to significantly altered MYC-signaling. Baseline enhancer activity of the conserved region specifically on the MYC promoter and its further positive modulation by the SNP risk-allele was shown in vitro. Our findings implicate MYC deregulation as the underlying cause of the observed association.

  1. Expression of an amino-terminal BRCA1 deletion mutant causes a dominant growth inhibition in MCF10A cells.

    Science.gov (United States)

    You, Fanglei; Chiba, Natsuko; Ishioka, Chikashi; Parvin, Jeffrey D

    2004-07-29

    Expression of deletion mutants of the breast and ovarian cancer-specific tumor suppressor protein, BRCA1, in the mammary epithelial cell line MCF10A revealed a powerful growth suppressive effect by a mutant that has the amino-terminal 302 amino acids deleted (DeltaN-BRCA1). The growth suppression is associated with an increase in apoptosis and amplification in centrosome number. The growth inhibitory effect of DeltaN-BRCA1 was not observed in cervical epithelial HeLa cells, suggesting that the phenotypes of BRCA1 mutant proteins differ depending on the cell line being tested. An internal domain, including BRCA1 residues 303-1292, caused the suppression of MCF10A cell growth, and the amino terminus of BRCA1 autoinhibited the growth suppression. Single point mutations that disrupted the amino-terminal RING domain of BRCA1 caused significant suppression of growth in MCF10A cells. These results suggest that the proper function of the RING domain, likely to be ubiquitin ligase function, is important in regulating the growth of the mammary epithelial cell line and in autoregulating the powerful internal growth-inhibiting domain of the BRCA1 tumor suppressor.

  2. uvsI mutants defective in UV mutagenesis define a fourth epistatic group of uvs genes in Aspergillus.

    Science.gov (United States)

    Chae, S K; Kafer, E

    1993-01-01

    Three UV-sensitive mutations of A. nidulans, uvsI, uvsJ and uvsA, were tested for epistatic relationships with members of the previously established groups, here called the "UvsF", "UvsC", and "UvsB" groups. uvsI mutants are defective for spontaneous and induced reversion of certain point mutations and differ also for other properties from previously analyzed uvs types. They are very sensitive to the killing effects of UV-light and 4-NQO (4-nitro-quinoline-N-oxide) but not to MMS (methylmethane sulfonate). When double- and single-mutant uvs strains were compared for sensitivity to these three agents, synergistic or additive effects were found for uvsI with all members of the three groups. The uvsI gene may therefore represent a fourth epistatic group, possibly involved in mutagenic repair. On the other hand, uvsJ was clearly epistatic with members of the UvsF group and fitted well into this group also by phenotype. The uvsA gene was tentatively assigned to the UvsC group. uvsA showed epistatic interactions with uvsC in all tests, and like UvsC-group mutants is UV-sensitive mainly in dividing cells. However, the uvsA mutation does not cause the defects in recombination and UV mutagenesis typical for this group.

  3. Toll-like receptor 4 mutant and null mice retain morphine-induced tolerance, hyperalgesia, and physical dependence.

    Directory of Open Access Journals (Sweden)

    Theresa Alexandra Mattioli

    Full Text Available The innate immune system modulates opioid-induced effects within the central nervous system and one target that has received considerable attention is the toll-like receptor 4 (TLR4. Here, we examined the contribution of TLR4 in the development of morphine tolerance, hyperalgesia, and physical dependence in two inbred mouse strains: C3H/HeJ mice which have a dominant negative point mutation in the Tlr4 gene rendering the receptor non-functional, and B10ScNJ mice which are TLR4 null mutants. We found that neither acute antinociceptive response to a single dose of morphine, nor the development of analgesic tolerance to repeated morphine treatment, was affected by TLR4 genotype. Likewise, opioid induced hyperalgesia and opioid physical dependence (assessed by naloxone precipitated withdrawal were not altered in TLR4 mutant or null mice. We also examined the behavioural consequence of two stereoisomers of naloxone: (- naloxone, an opioid receptor antagonist, and (+ naloxone, a purported antagonist of TLR4. Both stereoisomers of naloxone suppressed opioid induced hyperalgesia in wild-type control, TLR4 mutant, and TLR4 null mice. Collectively, our data suggest that TLR4 is not required for opioid-induced analgesic tolerance, hyperalgesia, or physical dependence.

  4. Enhancers of Conidiation Mutants in Aspergillus Nidulans

    OpenAIRE

    Gems, D. H.; Clutterbuck, A. J.

    1994-01-01

    Mutants at a number of loci, designated sthenyo, have been isolated as enhancers of the oligoconidial mutations at the medA locus. Two loci have been mapped: sthA on linkage group I, and sthB on linkage group V. Two probable alleles have been identified at each locus but two further mutants were unlinked to either sthA or sthB. Neither sthA nor sthB mutants have conspicuous effects on morphology on their own, nor could the sthA1 sthB2 double mutant be distinguished from wild type. Mutants at ...

  5. Dwarf mutant of rice variety Seratus Malam

    International Nuclear Information System (INIS)

    Mugiono, P. S.; Soemanggono, A.M.R.

    1989-01-01

    Full text: Seeds of 'Seratus Malam', a local tall upland variety with long panicles and high yield potential were irradiated with 10-50 krad gamma rays in 1983. From 50,000 M 2 plants, 130 semidwarf mutants and 1 dwarf mutant were selected. The dwarf mutant M-362 was obtained from the 10 krad treatment. The mutant shows about 50% reduction in plant height, but also in number of productive tillers. Thus the yield per plant is also significantly less. However, the mutant gene is not allelic to DGWG and therefore may be useful in cross breeding. (author)

  6. X-ray-sensitive mutants of Chinese hamster ovary cell line

    International Nuclear Information System (INIS)

    Jeggo, P.A.; Kemp, L.M.

    1983-01-01

    A standard technique of microbial genetics, which involves the transfer of cells from single colonies by means of sterile toothpicks, has been adapted to somatic cell genetics. Its use has been demonstrated in the isolation of X-ray-sensitive mutants of CHO cells. 9000 colonies have been tested and 6 appreciably X-ray-sensitive mutants were isolated. (D 10 values 5-10-fold of wild-type D 10 value.) A further 6 mutants were obtained which showed a slight level of sensitivity (D 10 values less than 2-fold of wild-type D 10 value). The 6 more sensitive mutants were also sensitive to bleomycin, a chemotherapeutic agent inducing X-ray-like damage. Cross-sensitivity to UV-irradiation and treatment with the alkylating agents, MMS, EMS and MNNG, was investigated for these mutants. Some sensitivity to these other agents was observed, but in all cases it was less severe than the level of sensitivity to X-irradiation. Each mutant showed a different overall response to the spectrum of agents examined and these appear to represent new mutant phenotypes derived from cultured mammalian cell lines. One mutant strain, xrs-7, was cross-sensitive to all the DNA-damaging agents, but was proficient in the repair of single-strand breaks. (Auth.)

  7. Induction of single flower mutant in Hibiscus cv. 'Alipur Beauty'

    International Nuclear Information System (INIS)

    Banerji, B.K.; Datta, S.K.

    1986-01-01

    Stem cuttings of Hibicus cv. 'Alipur Beauty' were irradiated with 0, 1.0, 2.0 Krad of gamma rays in 60 Co source and planted just after irradiation. Data were recorded on days to sprout, number of sprouts, sprouting percentage, survival, plant height, leaf number and size, abnormal leaves and plants, floral abnormalities and induction of somatic mutations in flower colour/form. Some of the characters like plant height, leaf number, abnormal leaves and plants and leaf size were recorded after 60 and 120 days. 7 refs. (author)

  8. Isolation of a Defective Prion Mutant from Natural Scrapie.

    Directory of Open Access Journals (Sweden)

    Ilaria Vanni

    2016-11-01

    Full Text Available It is widely known that prion strains can mutate in response to modification of the replication environment and we have recently reported that prion mutations can occur in vitro during amplification of vole-adapted prions by Protein Misfolding Cyclic Amplification on bank vole substrate (bvPMCA. Here we exploited the high efficiency of prion replication by bvPMCA to study the in vitro propagation of natural scrapie isolates. Although in vitro vole-adapted PrPSc conformers were usually similar to the sheep counterpart, we repeatedly isolated a PrPSc mutant exclusively when starting from extremely diluted seeds of a single sheep isolate. The mutant and faithful PrPSc conformers showed to be efficiently autocatalytic in vitro and were characterized by different PrP protease resistant cores, spanning aa ∼155-231 and ∼80-231 respectively, and by different conformational stabilities. The two conformers could thus be seen as different bona fide PrPSc types, putatively accounting for prion populations with different biological properties. Indeed, once inoculated in bank vole the faithful conformer was competent for in vivo replication while the mutant was unable to infect voles, de facto behaving like a defective prion mutant. Overall, our findings confirm that prions can adapt and evolve in the new replication environments and that the starting population size can affect their evolutionary landscape, at least in vitro. Furthermore, we report the first example of "authentic" defective prion mutant, composed of brain-derived PrPC and originating from a natural scrapie isolate. Our results clearly indicate that the defective mutant lacks of some structural characteristics, that presumably involve the central region ∼90-155, critical for infectivity but not for in vitro replication. Finally, we propose a molecular mechanism able to account for the discordant in vitro and in vivo behavior, suggesting possible new paths for investigating the molecular

  9. PNRI mutant variety: Cordyline 'Afable'

    International Nuclear Information System (INIS)

    Aurigue, Fernando B.

    2012-01-01

    Cordyline 'Afable', registered by the Philippine Nuclear Research Institute as NSIC 2009 Or-83, is an induced mutant developed from Cordyline 'Kiwi' by treating stem cuttings with acute gamma radiation from a Cobalt-60 source. The new mutant is identical to Cordyline 'Kiwi' in growth habit but differs in foliage color, and exhibits field resistance to Phytophthora sp., a fungus that causes leaf blight and rot in Ti plants. Results of this mutation breeding experiment showed that leaf color was altered by gamma irradiation and resistance to fungal diseases was improved. It also demonstrated how mutations that occur in nature may be generated artificially. Propagation of cordyline 'Afable' is true-to-type by vegetative propagation methods, such as separation of suckers and offshoots, shoot tip cutting, and top cutting. Aside from landscaping material, terrarium or dish-garden plant, it is ideal as containerized plant for indoor and outdoor use. The leaves or shoots may be harvested as cut foliage for flower arrangements. (author)

  10. Binding affinity and adhesion force of organophosphate hydrolase enzyme with soil particles related to the isoelectric point of the enzyme.

    Science.gov (United States)

    Islam, Shah Md Asraful; Yeasmin, Shabina; Islam, Md Saiful; Islam, Md Shariful

    2017-07-01

    The binding affinity of organophosphate hydrolase enzyme (OphB) with soil particles in relation to the isoelectric point (pI) was studied. Immobilization of OphB with soil particles was observed by confocal microscopy, Fourier transform infrared spectroscopy (FT-IR), and Atomic force microscopy (AFM). The calculated pI of OphB enzyme was increased from 8.69 to 8.89, 9.04 and 9.16 by the single, double and triple mutant of OphB enzyme, respectively through the replacement of negatively charged aspartate with positively charged histidine. Practically, the binding affinity was increased to 5.30%, 11.50%, and 16.80% for single, double and triple mutants, respectively. In contrast, enzyme activity of OphB did not change by the mutation of the enzyme. On the other hand, adhesion forces were gradually increased for wild type OphB enzyme (90 pN) to 96, 100 and 104 pN for single, double and triple mutants of OphB enzyme, respectively. There was an increasing trend of binding affinity and adhesion force by the increase of isoelectric point (pI) of OphB enzyme. Copyright © 2017 Elsevier Inc. All rights reserved.

  11. Gamma ray induced mutants in Coleus

    International Nuclear Information System (INIS)

    Vasudevan, K.; Jos, J.S.

    1988-01-01

    The germplasm collection of Chinese potato (Coleus parviflorus Benth) contains almost no variation for yield contributing traits. The crop does not produce seeds. Treatment of underground tubers with 1 kR, 2 kR, 3 kR and 4 kR gamma rays resulted in 50 morphologically different mutants which are maintained as mutant clones. In the M 1 V 1 generation, suspected mutant sprouts, were carefully removed and grown separately. The most interesting mutant types are the following: (i) erect mutant with spoon shaped light green leaves, 30 cm long inflorescences against 20 cm in the control, cylindrical tubers measuring ca. 7.0 cm long and 3 cm girth against 4 cm and 2.5 cm in the control (ii) early mutants 1 and 2, one having less leaf serration, the other having light green small leaves and dwarf type (iii) fleshy leaf mutant, dark green, thick and smooth leaves. Control plants spread almost in 1 m 2 area and bear tubers from the nodes of branches. In the early mutants tuber formation is mainly restricted to the base of the plant, which makes harvest easier. The crop usually matures within 150 - 160 days, the early mutants are ready for harvest 100 days after planting. As the mutants are less spreading, the yield could be increased by closer spacing

  12. Coxsackievirus mutants that can bypass host factor PI4KIIIβ and the need for high levels of PI4P lipids for replication.

    Science.gov (United States)

    van der Schaar, Hilde M; van der Linden, Lonneke; Lanke, Kjerstin H W; Strating, Jeroen R P M; Pürstinger, Gerhard; de Vries, Erik; de Haan, Cornelis A M; Neyts, Johan; van Kuppeveld, Frank J M

    2012-11-01

    RNA viruses can rapidly mutate and acquire resistance to drugs that directly target viral enzymes, which poses serious problems in a clinical context. Therefore, there is a growing interest in the development of antiviral drugs that target host factors critical for viral replication, since they are unlikely to mutate in response to therapy. We recently demonstrated that phosphatidylinositol-4-kinase IIIβ (PI4KIIIβ) and its product phosphatidylinositol-4-phosphate (PI4P) are essential for replication of enteroviruses, a group of medically important RNA viruses including poliovirus (PV), coxsackievirus, rhinovirus, and enterovirus 71. Here, we show that enviroxime and GW5074 decreased PI4P levels at the Golgi complex by directly inhibiting PI4KIIIβ. Coxsackievirus mutants resistant to these inhibitors harbor single point mutations in the non-structural protein 3A. These 3A mutations did not confer compound-resistance by restoring the activity of PI4KIIIβ in the presence of the compounds. Instead, replication of the mutant viruses no longer depended on PI4KIIIβ, since their replication was insensitive to siRNA-mediated depletion of PI4KIIIβ. The mutant viruses also did not rely on other isoforms of PI4K. Consistently, no high level of PI4P could be detected at the replication sites induced by the mutant viruses in the presence of the compounds. Collectively, these findings indicate that through specific single point mutations in 3A, CVB3 can bypass an essential host factor and lipid for its propagation, which is a new example of RNA viruses acquiring resistance against antiviral compounds, even when they directly target host factors.

  13. Sharing mutants and experimental information prepublication using FgMutantDb (https://scabusa.org/FgMutantDb).

    Science.gov (United States)

    Baldwin, Thomas T; Basenko, Evelina; Harb, Omar; Brown, Neil A; Urban, Martin; Hammond-Kosack, Kim E; Bregitzer, Phil P

    2018-02-02

    There is no comprehensive storage for generated mutants of Fusarium graminearum or data associated with these mutants. Instead, researchers relied on several independent and non-integrated databases. FgMutantDb was designed as a simple spreadsheet that is accessible globally on the web that will function as a centralized source of information on F. graminearum mutants. FgMutantDb aids in the maintenance and sharing of mutants within a research community. It will serve also as a platform for disseminating prepublication results as well as negative results that often go unreported. Additionally, the highly curated information on mutants in FgMutantDb will be shared with other databases (FungiDB, Ensembl, PhytoPath, and PHI-base) through updating reports. Here we describe the creation and potential usefulness of FgMutantDb to the F. graminearum research community, and provide a tutorial on its use. This type of database could be easily emulated for other fungal species. Published by Elsevier Inc.

  14. Gamma ray induced high yielding dwarf mutant in Vigna radiata L. Wilczek

    International Nuclear Information System (INIS)

    Pande, Kalpana; Raghuvanshi, S.S.

    1988-01-01

    Single and combined treatments with gamma rays and EMS were tried on V. radiata (L. Wilczek) variety K 851. Seeds were exposed to 10, 20 and 30 krad gamma rays. One set of each dose was treated with 0.25% EMS solution (pH 7 at 30 deg. C) for 6 hours. The M 2 generation was screened for mutants. A dwarf mutant with signs of higher yield was observed in the 20 krad plot. The mutant exceeded the parent variety in several agronomic traits and was true breeding in M 3 . Crossing with the control confirmed its monogenic and recessive character. Significant increase was found in number of pods/plant and number of seeds/pod leading to higher seed yield. The mutant took only 54 days to mature. This early mutant is very interesting for double/triple cropping and may help to bridge the widening gap between pulse production and consumption in India

  15. Scalable single point power extraction for compact mobile and stand-alone solar harvesting power sources based on fully printed organic photovoltaic modules and efficient high voltage DC/DC conversion

    DEFF Research Database (Denmark)

    Garcia Valverde, Rafael; Villarejo, José A.; Hösel, Markus

    2015-01-01

    Patterns for fully printed polymer solar cells are presented that inherently enable scaling of the power output with single point electrical energy connection is presented. Connection is made to only one end of the printed foil that can be rolled out for light energy harvesting. The power level...... (AM1.5G, 1000 W m−2). As a demonstration we present a scalable fully integrated and compact power unit for mobile applications comprising solar energy harvesting OPV modules, power conversion and storage. Applications possible include electrical charging of mobile devices, illumination using LED lamps...... takes place in a HVDC–DC converter that is tailored specifically for operation with polymer solar cells by regulation on the input side. The system charges a lithium-polymer battery thus enabling storage of 82 Wh for a printed OPV foil measuring 0.305 m×9 m having a nominal power output of at least 15 W...

  16. Abnormal grooming activity in Dab1(scm) (scrambler) mutant mice.

    Science.gov (United States)

    Strazielle, C; Lefevre, A; Jacquelin, C; Lalonde, R

    2012-07-15

    Dab1(scm) mutant mice, characterized by cell ectopias and degeneration in cerebellum, hippocampus, and neocortex, were compared to non-ataxic controls for different facets of grooming caused by brief water immersions, as well as some non-grooming behaviors. Dab1(scm) mutants were strongly affected in their quantitative functional parameters, exhibiting higher starting latencies before grooming relative to non-ataxic littermates of the A/A strain, fewer grooming bouts, and grooming components of shorter duration, with an unequal regional distribution targeting almost totally the rostral part (head washing and forelimb licking) of the animal. Only bouts of a single grooming element were preserved. The cephalocaudal order of grooming elements appeared less disorganized, mutant and control mice initiating the grooming with head washing and forelimb licking prior to licking posterior parts. However, mutants differed from controls in that all their bouts were incomplete but uninterrupted, although intergroup difference for percentage of the incorrect transitions was not significant. In contrast to grooming, Dab1(scm) mice ambulated for a longer time. During walking episodes, they exhibited more body scratching than controls, possibly to compensate for the lack of licking different body parts. In conjunction with studies with other ataxic mice, these results indicate that the cerebellar cortex affects grooming activity and is consequently involved in executing various components, but not in its sequential organization, which requires other brain regions such as cerebral cortices or basal ganglia. Copyright © 2012 Elsevier B.V. All rights reserved.

  17. Is It Possible to Detect Activated Brown Adipose Tissue in Humans Using Single-Time-Point Infrared Thermography under Thermoneutral Conditions? Impact of BMI and Subcutaneous Adipose Tissue Thickness.

    Science.gov (United States)

    Gatidis, Sergios; Schmidt, Holger; Pfannenberg, Christina A; Nikolaou, Konstantin; Schick, Fritz; Schwenzer, Nina F

    2016-01-01

    To evaluate the feasibility to detect activated brown adipose tissue (BAT) using single-time-point infrared thermography of the supraclavicular skin region under thermoneutral conditions. To this end, infrared thermography was compared with 18-F-FDG PET, the current reference standard for the detection of activated BAT. 120 patients were enrolled in this study. After exclusion of 18 patients, 102 patients (44 female, 58 male, mean age 58±17 years) were included for final analysis. All patients underwent a clinically indicated 18F-FDG-PET/CT examination. Immediately prior to tracer injection skin temperatures of the supraclavicular, presternal and jugular regions were measured using spatially resolved infrared thermography at room temperature. The presence of activated BAT was determined in PET by typical FDG uptake within the supraclavicular adipose tissue compartments. Local thickness of supraclavicular subcutaneous adipose tissue (SCAT) was measured on CT. Measured skin temperatures were statistically correlated with the presence of activated BAT and anthropometric data. Activated BAT was detected in 9 of 102 patients (8.8%). Local skin temperature of the supraclavicular region was significantly higher in individuals with active BAT compared to individuals without active BAT. However, after statistical correction for the influence of BMI, no predictive value of activated BAT on skin temperature of the supraclavicular region could be observed. Supraclavicular skin temperature was significantly negatively correlated with supraclavicular SCAT thickness. We conclude that supraclavicular SCAT thickness influences supraclavicular skin temperature and thus makes a specific detection of activated BAT using single-time-point thermography difficult. Further studies are necessary to evaluate the possibility of BAT detection using alternative thermographic methods, e.g. dynamic thermography or MR-based thermometry taking into account BMI as a confounding factor.

  18. Is It Possible to Detect Activated Brown Adipose Tissue in Humans Using Single-Time-Point Infrared Thermography under Thermoneutral Conditions? Impact of BMI and Subcutaneous Adipose Tissue Thickness.

    Directory of Open Access Journals (Sweden)

    Sergios Gatidis

    Full Text Available To evaluate the feasibility to detect activated brown adipose tissue (BAT using single-time-point infrared thermography of the supraclavicular skin region under thermoneutral conditions. To this end, infrared thermography was compared with 18-F-FDG PET, the current reference standard for the detection of activated BAT.120 patients were enrolled in this study. After exclusion of 18 patients, 102 patients (44 female, 58 male, mean age 58±17 years were included for final analysis. All patients underwent a clinically indicated 18F-FDG-PET/CT examination. Immediately prior to tracer injection skin temperatures of the supraclavicular, presternal and jugular regions were measured using spatially resolved infrared thermography at room temperature. The presence of activated BAT was determined in PET by typical FDG uptake within the supraclavicular adipose tissue compartments. Local thickness of supraclavicular subcutaneous adipose tissue (SCAT was measured on CT. Measured skin temperatures were statistically correlated with the presence of activated BAT and anthropometric data.Activated BAT was detected in 9 of 102 patients (8.8%. Local skin temperature of the supraclavicular region was significantly higher in individuals with active BAT compared to individuals without active BAT. However, after statistical correction for the influence of BMI, no predictive value of activated BAT on skin temperature of the supraclavicular region could be observed. Supraclavicular skin temperature was significantly negatively correlated with supraclavicular SCAT thickness.We conclude that supraclavicular SCAT thickness influences supraclavicular skin temperature and thus makes a specific detection of activated BAT using single-time-point thermography difficult. Further studies are necessary to evaluate the possibility of BAT detection using alternative thermographic methods, e.g. dynamic thermography or MR-based thermometry taking into account BMI as a confounding factor.

  19. Single point mutations in the helicase domain of the NS3 protein enhance dengue virus replicative capacity in human monocyte-derived dendritic cells and circumvent the type I interferon response.

    Science.gov (United States)

    Silveira, G F; Strottmann, D M; de Borba, L; Mansur, D S; Zanchin, N I T; Bordignon, J; dos Santos, C N Duarte

    2016-01-01

    Dengue is the most prevalent arboviral disease worldwide. The outcome of the infection is determined by the interplay of viral and host factors. In the present study, we evaluated the cellular response of human monocyte-derived DCs (mdDCs) infected with recombinant dengue virus type 1 (DV1) strains carrying a single point mutation in the NS3hel protein (L435S or L480S). Both mutated viruses infect and replicate more efficiently and produce more viral progeny in infected mdDCs compared with the parental, non-mutated virus (vBACDV1). Additionally, global gene expression analysis using cDNA microarrays revealed that the mutated DVs induce the up-regulation of the interferon (IFN) signalling and pattern recognition receptor (PRR) canonical pathways in mdDCs. Pronounced production of type I IFN were detected specifically in mdDCs infected with DV1-NS3hel-mutated virus compared with mdDCs infected with the parental virus. In addition, we showed that the type I IFN produced by mdDCs is able to reduce DV1 infection rates, suggesting that cytokine function is effective but not sufficient to mediate viral clearance of DV1-NS3hel-mutated strains. Our results demonstrate that single point mutations in subdomain 2 have important implications for adenosine triphosphatase (ATPase) activity of DV1-NS3hel. Although a direct functional connection between the increased ATPase activity and viral replication still requires further studies, these mutations speed up viral RNA replication and are sufficient to enhance viral replicative capacity in human primary cell infection and circumvent type I IFN activity. This information may have particular relevance for attenuated vaccine protocols designed for DV. © 2015 British Society for Immunology.

  20. Mutants for plant height in hexaploid triticale

    International Nuclear Information System (INIS)

    Reddy, V.R.K.; Gupta, P.K.

    1988-01-01

    Full text: Four hexaploid triticale varieties namely Beagle, Coorong, TL 419 and Welsh were subjected to gamma rays (100 Gy, 200 Gy, 300 Gy) and to aqueous solution of EMS (0.5%, (8h, 12h, 16h). In all four varieties, three types of mutants for plant height were observed: Semidwarf - the mutant plants are 20-25 cm shorter than the shortest plant in the control. Dwarf - mutant plants grow up to 40-60 cm. Stunted - mutant plants grow up to 10-20 cm. The segregation pattern suggests that semidwarf mutants are quantitatively inherited, showing continuous segregation in M 3 , M 4 and M 5 , whereas dwarf and stunted are monogenic recessive. They showed true breeding in M 3 and later generations. The semi-dwarf, dwarf and stunted mutants can be used as initial material for development of new varieties with short straw and resistance to lodging. (author)

  1. Studies on cytological, physiological and genetic characteristics in somatic mutant strains of Sugi (Cryptomeria japonica D. Don)

    International Nuclear Information System (INIS)

    Maeta, T.; Somegou, M.; Nakahira, K.; Miyazaki, Y.; Kondo, T.

    1982-01-01

    From microscopic observation of the pollen of induced mutant strains in Sugi (Cryptomeria japonica D. Don), it was found that there were large differences in pollen fertility among the mutant strains, and that it deviated year to year from the mother plants. The large differences in frequency of sterile pollen among mutant strains depended on the genetic characteristics of each mutant strain. Higher frequencies of sterile pollen were observed at the terminal part of branchlets in some mutant strains, and this was considered to be induced by the lateness of flower-bud formation at low temperature conditions in late summer. Delayed formation and gibberellic acid treatment applied for flower induction resulted in low fertility and abnormality of pollen in mutant strains. Chromosome aberration in mutant strains was caused either by gamma irradiation or by some mutational events that responded to environmental conditions. In the former case, aberration might have been maintained for a long period through vegetative propagation. Some of the irregularities were due to mitotic cell division, because cells with micronuclei at the pacytene stage in pollen mother cells and with fragments at MI were observed. Somatic mutability of Kuma-sugi mutants after re-irradiation was investigated. From waxless mutants morphological somatic mutations, which have fat or stout stems and thick and short needles, were frequently produced, whereas from morphological mutants the lowest somatic mutation frequency was induced. In some mutant strains higher rooting ability than the mother plants was found, and the possibility of character improvement was pointed out. (author)

  2. Gamma ray induced mutants in Colocasia

    International Nuclear Information System (INIS)

    Vasudevan, K.; Jos, J.S.

    1988-01-01

    Presented are selected treatments with 250 r, 500 r and 1000 r gamma rays Colocasia mutants with changes in morphological and yield characters. Results from a preliminary yield trial of four mutants with its control variety C 9 are presented. The mutant's characteristics are (i) erect and narrow leaf (ii) cup shaped leaf, dwarf, matures within 120 days against 180 days in control (iii) narrow and thicker leaves, colour of lamina chalky and pale green (iv) vigorous

  3. GALT protein database, a bioinformatics resource for the management and analysis of structural features of a galactosemia-related protein and its mutants.

    Science.gov (United States)

    d'Acierno, Antonio; Facchiano, Angelo; Marabotti, Anna

    2009-06-01

    We describe the GALT-Prot database and its related web-based application that have been developed to collect information about the structural and functional effects of mutations on the human enzyme galactose-1-phosphate uridyltransferase (GALT) involved in the genetic disease named galactosemia type I. Besides a list of missense mutations at gene and protein sequence levels, GALT-Prot reports the analysis results of mutant GALT structures. In addition to the structural information about the wild-type enzyme, the database also includes structures of over 100 single point mutants simulated by means of a computational procedure, and the analysis to each mutant was made with several bioinformatics programs in order to investigate the effect of the mutations. The web-based interface allows querying of the database, and several links are also provided in order to guarantee a high integration with other resources already present on the web. Moreover, the architecture of the database and the web application is flexible and can be easily adapted to store data related to other proteins with point mutations. GALT-Prot is freely available at http://bioinformatica.isa.cnr.it/GALT/.

  4. Procalcitonin for selecting the antibiotic regimen in outpatients with low-risk community-acquired pneumonia using a rapid point-of-care testing: A single-arm clinical trial

    Science.gov (United States)

    Masiá, Mar; Padilla, Sergio; Ortiz de la Tabla, Victoria; González, Matilde; Bas, Cristina; Gutiérrez, Félix

    2017-01-01

    Objective We aimed to assess the role of procalcitonin (PCT) to guide the initial selection of the antibiotic regimen for low-risk community-acquired pneumonia (CAP). Methods A single-arm clinical trial was conducted including outpatients with CAP and Pneumonia Severity Index risk classes I-II. Antimicrobial selection was based on the results of PCT measured with a rapid point-of-care testing. According to serum PCT levels, patients were assigned to two treatment strategies: oral azithromycin if PCT was Atypical organisms were more common in patients given azithromycin (18.5% vs 8.1%, respectively). The majority (93%) of patients with atypical pneumonia had low PCT levels. Clinical cure rates were 95.8% in the azithromycin group, 94.6% in the levofloxacin group, and 94.4% in the historical control group. No 30-day mortality or recurrences were observed, and the 3-year rates of recurrence and mortality were very low in both groups. Adverse events occurrence was also infrequent. Conclusion A PCT-guided strategy with a rapid point-of-care testing safely allowed selecting empirical narrow-spectrum antibiotics in outpatients with CAP. Trial registration The study is registered with ClinicalTrials.gov, number NCT02600806 PMID:28426811

  5. Key Components of Different Plant Defense Pathways Are Dispensable for Powdery Mildew Resistance of the Arabidopsis mlo2 mlo6 mlo12 Triple Mutant

    Directory of Open Access Journals (Sweden)

    Hannah Kuhn

    2017-06-01

    Full Text Available Loss of function mutations of particular plant MILDEW RESISTANCE LOCUS O (MLO genes confer durable and broad-spectrum penetration resistance against powdery mildew fungi. Here, we combined genetic, transcriptomic and metabolomic analyses to explore the defense mechanisms in the fully resistant Arabidopsis thaliana mlo2 mlo6 mlo12 triple mutant. We found that this genotype unexpectedly overcomes the requirement for indolic antimicrobials and defense-related secretion, which are critical for incomplete resistance of mlo2 single mutants. Comparative microarray-based transcriptome analysis of mlo2 mlo6 mlo12 mutants and wild type plants upon Golovinomyces orontii inoculation revealed an increased and accelerated accumulation of many defense-related transcripts. Despite the biotrophic nature of the interaction, this included the non-canonical activation of a jasmonic acid/ethylene-dependent transcriptional program. In contrast to a non-adapted powdery mildew pathogen, the adapted powdery mildew fungus is able to defeat the accumulation of defense-relevant indolic metabolites in a MLO protein-dependent manner. We suggest that a broad and fast activation of immune responses in mlo2 mlo6 mlo12 plants can compensate for the lack of single or few defense pathways. In addition, our results point to a role of Arabidopsis MLO2, MLO6, and MLO12 in enabling defense suppression during invasion by adapted powdery mildew fungi.

  6. Key Components of Different Plant Defense Pathways Are Dispensable for Powdery Mildew Resistance of the Arabidopsismlo2 mlo6 mlo12Triple Mutant.

    Science.gov (United States)

    Kuhn, Hannah; Lorek, Justine; Kwaaitaal, Mark; Consonni, Chiara; Becker, Katia; Micali, Cristina; Ver Loren van Themaat, Emiel; Bednarek, Paweł; Raaymakers, Tom M; Appiano, Michela; Bai, Yuling; Meldau, Dorothea; Baum, Stephani; Conrath, Uwe; Feussner, Ivo; Panstruga, Ralph

    2017-01-01

    Loss of function mutations of particular plant MILDEW RESISTANCE LOCUS O ( MLO ) genes confer durable and broad-spectrum penetration resistance against powdery mildew fungi. Here, we combined genetic, transcriptomic and metabolomic analyses to explore the defense mechanisms in the fully resistant Arabidopsis thaliana mlo2 mlo6 mlo12 triple mutant. We found that this genotype unexpectedly overcomes the requirement for indolic antimicrobials and defense-related secretion, which are critical for incomplete resistance of mlo2 single mutants. Comparative microarray-based transcriptome analysis of mlo2 mlo6 mlo12 mutants and wild type plants upon Golovinomyces orontii inoculation revealed an increased and accelerated accumulation of many defense-related transcripts. Despite the biotrophic nature of the interaction, this included the non-canonical activation of a jasmonic acid/ethylene-dependent transcriptional program. In contrast to a non-adapted powdery mildew pathogen, the adapted powdery mildew fungus is able to defeat the accumulation of defense-relevant indolic metabolites in a MLO protein-dependent manner. We suggest that a broad and fast activation of immune responses in mlo2 mlo6 mlo12 plants can compensate for the lack of single or few defense pathways. In addition, our results point to a role of Arabidopsis MLO2, MLO6, and MLO12 in enabling defense suppression during invasion by adapted powdery mildew fungi.

  7. Efficacy and Safety of Single Botulinum Toxin Type A (Botox®) Injection for Relief of Upper Trapezius Myofascial Trigger Point: A Randomized, Double-Blind, Placebo-Controlled Study.

    Science.gov (United States)

    Kwanchuay, Photsawee; Petchnumsin, Thavatchai; Yiemsiri, Pichet; Pasuk, Nakkamol; Srikanok, Wannarat; Hathaiareerug, Chanasak

    2015-12-01

    Botulinum toxin injection has been applied for pain relief in various chronic pain syndromes. Recently, systematic review studies reported inconclusive effects of Botulinum toxin in myofascial pain management. The present study aimed to demonstrate the efficacy and safety of Botulinum toxin type A (BTxA) (Botox®) injection for pain reduction in myofascial trigger point (MTrP) of the upper trapezius muscle. Thirty-three patients with 48 MTrP on the upper trapezius muscles over three months with moderate to severe pain intensity diagnosed at physical medicine and rehabilitation outpatient department were recruited between December 2011 and March 2012. Eligible patients were blinded and randomly injected with single 0.2 ml (20 IU) of BTxA for 24 MTrP and 0.2 ml of 0.9% NaCl solution for 24 MTrP at the most tender trigger point on the upper trapezius muscle. All patients were advised for stretching exercise and ergonomic adaptation throughout the study. At 3- and 6-week after injections, visual analogue scale (VAS), the pressure pain threshold (PPT), and reported adverse effects were measured. Both BTxA and control groups demonstrated statistically significant differences in VAS reduction and increased PPT after 3 weeks and 6 weeks compared with before treatment. There were no statistically significant differences in VAS reduction from baseline between the two groups at 3- and 6-week after treatment. A statistically significant difference in improvement of PPT from baseline and 6-week after BTxA injection compared with 0.9% NaCl group was shown (1.0 ± 0.9 and 0.5 ± 0.7, p = 0.036). There was mild degree side-effects that spontaneous resolved within one week in both groups without significant difference in percentage. No severe adverse effects were reported during the study. The efficacy in VAS reduction of a single 20 IU of Botulinum toxin type A (Botox®) injection was not different from 0.9% NaCl for myofascial trigger point at the upper trapezius muscle. However

  8. Determining mutant spectra of three RNA viral samples using ultra-deep sequencing

    Energy Technology Data Exchange (ETDEWEB)

    Chen, H

    2012-06-06

    RNA viruses have extremely high mutation rates that enable the virus to adapt to new host environments and even jump from one species to another. As part of a viral transmission study, three viral samples collected from naturally infected animals were sequenced using Illumina paired-end technology at ultra-deep coverage. In order to determine the mutant spectra within the viral quasispecies, it is critical to understand the sequencing error rates and control for false positive calls of viral variants (point mutantations). I will estimate the sequencing error rate from two control sequences and characterize the mutant spectra in the natural samples with this error rate.

  9. A chilling sensitive mutant of Arabidopsis with altered steryl-ester metabolism

    International Nuclear Information System (INIS)

    Hugly, S.; McCourt, P.; Somerville, C.; Browse, J.; Patterson, G.W.

    1990-01-01

    A chilling-sensitive mutant of Arabidopsis thaliana was isolated and subjected to genetic, physiological, and biochemical analysis. The chilling-sensitive nature of the mutant line is due to a single recessive nuclear mutation at a locus designated chs1. In contrast to wild-type plants, which are not adversely affected by low temperatures, the chs1 mutant is killed by several days of exposure to temperatures below 18 degree C. Following exposure to chilling temperatures, the mutant displays two common symptoms of chilling injury - leaf chlorosis and electrolyte leakage. In these respects, the physiological response of the mutant to low temperatures mimics the response observed in some naturally occurring chilling sensitive species. The biochemical basis of chilling sensitivity was explored by examining the pattern of incorporation of 14 CO 2 into soluble metabolites and lipids in wild-type and mutant plants. The only difference observed between the mutant and wild type was that following low temperature treatment, the mutant accumulated 10-fold more radioactivity in a specific class of neutral lipids which were identified by a variety of criteria to be steryl-esters. The accumulation of radioactivity in the steryl-ester fraction occurs 24 hours before there is any visible evidence of chilling injury

  10. Use of virion DNA as a cloning vector for the construction of mutant and recombinant herpesviruses.

    OpenAIRE

    Duboise, S M; Guo, J; Desrosiers, R C; Jung, J U

    1996-01-01

    We have developed improved procedures for the isolation of deletion mutant, point mutant, and recombinant herpesvirus saimiri. These procedures take advantage of the absence of NotI and AscI restriction enzyme sites within the viral genome and use reporter genes for the identification of recombinant viruses. Genes for secreted engineered alkaline phosphatase and green fluorescent protein were placed under simian virus 40 early promoter control and flanked by NotI and AscI restriction sites. W...

  11. Response of the pearly eye melon fly Bactrocera cucurbitae (Coquillett)(Diptera:Tephritidae) mutant to host-associated visual cues

    Science.gov (United States)

    We report on a pearly eye mutant (PEM) line generated from a single male Bactrocera cucurbitae collected in Kapoho, Hawaii. Crossing experiments with colony wild-type flies indicate that the locus controlling this trait is autosomal and the mutant allele is recessive. Experiments with females to ass...

  12. A new mutant, White larva, of the mosquito Toxorhynchites splendens: genetics and cannibalism.

    Science.gov (United States)

    Horio, M; Tsukamoto, M; Miyagi, I

    1990-09-01

    A strain of a new body-color mutant, white larva (wl), was established from a field-collected wild-type strain of Toxorhynchites splendens. The mutant can be distinguished from the wild type in both the larval and pupal stages, but not in the adult. Crossing experiments confirmed its mode of inheritance to be a single recessive system. This is the first visible mutant found in Tx. splendens. Larvae of the wl phenotype seem to be recognized as prey by other individuals in mass larvae rearing.

  13. Electric signals of light excited bacteriorhodopsin mutant D96N.

    Science.gov (United States)

    Tóth-Boconádi, R; Taneva, S G; Keszthelyi, L

    2001-12-31

    The study of mutant D96N played an important role in understanding proton translocation by light driven bacteriorhodopsin. Our measurement of photoelectric current for single and double flash illumination revealed new details of the photocycle of this mutant. With double flash excitation we found an intermediate absorbing near the wavelength of the ground state of bacteriorhodopsin (bR) but pumping in the opposite direction. This intermediate has the same lifetime as the species described by Zimányi et al. [Proc. Natl. Acad. Sci. USA 96 (1999) 4414-4419] and was assigned to early recovery of a fraction of the ground state after excitation. Because the electric response does not reconcile with that of the ground state, we tentatively assign it to the L intermediate or to an intermediate similar in absorption to bR (bR').

  14. Mutant of Japanese pear resistant to Black Spot Disease

    International Nuclear Information System (INIS)

    Sanada, T.; Nishida, T.; Ikeda, F.

    1987-01-01

    Full text: Nijisseike is one of the leading cultivars of Japanese pear (Pyrus serotinea Rehd.), but susceptible to black spot disease. Farmers try to prevent this disease by wrapping the fruit with a paper bag and by repeated spraying of fungicides. The disease is caused by a Japanese pear pathotype of Alternaria alternata (Fr.) Keissler. Susceptibility is controlled by a single dominant gene. In 1962, grafted trees of this cultivar were planted at a distance between 53 and 93 m from the 60 Co source in the gamma-field (daily dose 15-4 rad). One branch on a tree planted at 53 m was detected as resistant in 1981. Under field conditions, black spots were observed on many fruits and leaves of the original trees by natural infection in early July, however, they were not observed on the mutant. To examine the resistance of the mutant, artificial inoculations were made using spores of the pathogen and the host specific toxin produced by germinating spores. When some drops of the spore suspension are placed on leaves, the formation of black spots depends upon the leaf age. In a resistant cv. as Chojuro, black spot symptoms are formed only when inoculated on young leaves. An intermediate reaction was observed in the mutant, whereas the original Nijisseiki showed severe symptoms. When inoculation was made on matured fruit skins, no black spot was formed on the mutant just like on the resistant cv. Chojuro, while many small black spots were formed and grew into large spots overlapping each other on the susceptible cv. Nijisseiki. In case of the crude toxin inoculation (4-0.04 ppm) of cv. Nijisseiki black spots were formed on the surface of the susceptible fruit skin, and necrotic lesions at the cut end of detached small pieces of leaves, although reaction on fruit skins was weaker compared with inoculation by spores. However, no symptoms were observed from the toxin application on the mutant and the resistant cv. Chojuro. That the resistance of the mutant is classified as

  15. Flocculation phenomenon of a mutant flocculent Saccharomyces ...

    African Journals Online (AJOL)

    The flocculation mechanism of a stable mutant flocculent yeast strain Saccharomyces cerevisiae KRM-1 was quantitatively investigated for potential industrial interest. It was found that the mutant flocculent strain was NewFlo phenotype by means of sugar inhibition test. The flocculation was completely inhibited by treatment ...

  16. Generation and characterization of pigment mutants of ...

    African Journals Online (AJOL)

    Compared to the wild CC-124, these mutants are characterized by a decrease in chlorophyll a & b content and an increase in carotenoids. The lowest decrease in chlorophyll a was 3 to 4 folds, while the highest increase in carotenoids was 2 to 4 folds. The result of bio-test, using the resulting pigment mutant of C. reinhardtii ...

  17. Los mutantes de la escuela

    Directory of Open Access Journals (Sweden)

    Diego Armando Jaramillo-Ocampo

    2013-01-01

    Full Text Available El presente artículo muestra los resultados parciales del estudio “Juegos en el recreo escolar: un escenario para la formación ciudadana”, cuya pretensión fue comprender los imaginarios sociales de juego en el recreo escolar y su relación con la convivencia social desde la proximidad del enfoque de complementariedad y el diseño de investigación emergente, planteado por Murcia y Jaramillo (2008. Se presentan los desarrollos logrados en dos categorías centrales del estudio: el patio y el cuerpo; dos categorías que mutan constantemente como entidades vivas en la escuela, hacia la configuración de sujetos que reconocen en el otro y lo otro su posibilidad. La escuela viva, donde es posible “ser en relación con”… se reduce a un espacio temporal y físico, limitado por la campana, “el recreo”. El texto muestra, desde la voz de los actores, esa vida que se da y se quita en la escuela y que se posiciona como una más de las imposiciones normalizadas para controlar. Reconoce, finalmente, una propuesta desde la posibilidad que estos dos mutantes propician para una escuela libre y dinámica.

  18. Mutants dissecting development and behaviour in drosophila

    International Nuclear Information System (INIS)

    Joshi, Adita; Chandrashekaran, Shanti; Sharma, R.P.

    2005-01-01

    We have traced in this paper the progress in Drosophila genetics research from the 1960s, at the IARI, spearheaded by the visionary insight of M. S. Swaminathan. The work started with the study of indirect effect of radiation and the synergistic interaction of physical and chemical mutagens on chromosomal and genetic changes. This paved the way for the study of single gene mutants in dissecting developmental and behavioural processes. New genes discovered by us have been shown to encode conserved cell signalling molecules controlling developmental and behavioural pathways. With the complete sequencing of the Drosophila genome, in the year 2000, mounting evidence for the homology between Drosophila and human genes controlling genetic disorders became available. This has led to the fly becoming an indispensable tool for studying human diseases as well as a model to test for drugs and pharmaceuticals against human diseases and complex behavioural processes. For example wingless in Drosophila belongs to the conserved Wnt gene family and aberrant WNT signalling is linked to a range of human diseases, most notably cancer. Inhibition as well as activation of WNT signalling form the basis of an effective therapy for some cancers as well as several other clinical conditions. Recent experiments have shown that WNTs might also normally participate in self-renewal, proliferation or differentiation of stem cells and altering WNT signalling might be beneficial to the use of stem cells for therapeutic means. Likewise, the stambhA mutant of Drosophila which was discovered for its temperature-dependent paralytic behaviour is the fly homologue of Phospholipase Cβ. Phospholipase C mediated G protein signalling plays a central role in vital processes controlling epilepsy, vision, taste, and olfaction in animals. Proteins of the G-signalling pathway are of intense research interest since many human diseases involve defects in G-protein signalling pathways. In fact, approximately 50

  19. Modification and Validation of the Triglyceride-to-HDL Cholesterol Ratio as a Surrogate of Insulin Sensitivity in White Juveniles and Adults without Diabetes Mellitus: The Single Point Insulin Sensitivity Estimator (SPISE).

    Science.gov (United States)

    Paulmichl, Katharina; Hatunic, Mensud; Højlund, Kurt; Jotic, Aleksandra; Krebs, Michael; Mitrakou, Asimina; Porcellati, Francesca; Tura, Andrea; Bergsten, Peter; Forslund, Anders; Manell, Hannes; Widhalm, Kurt; Weghuber, Daniel; Anderwald, Christian-Heinz

    2016-09-01

    The triglyceride-to-HDL cholesterol (TG/HDL-C) ratio was introduced as a tool to estimate insulin resistance, because circulating lipid measurements are available in routine settings. Insulin, C-peptide, and free fatty acids are components of other insulin-sensitivity indices but their measurement is expensive. Easier and more affordable tools are of interest for both pediatric and adult patients. Study participants from the Relationship Between Insulin Sensitivity and Cardiovascular Disease [43.9 (8.3) years, n = 1260] as well as the Beta-Cell Function in Juvenile Diabetes and Obesity study cohorts [15 (1.9) years, n = 29] underwent oral-glucose-tolerance tests and euglycemic clamp tests for estimation of whole-body insulin sensitivity and calculation of insulin sensitivity indices. To refine the TG/HDL ratio, mathematical modeling was applied including body mass index (BMI), fasting TG, and HDL cholesterol and compared to the clamp-derived M-value as an estimate of insulin sensitivity. Each modeling result was scored by identifying insulin resistance and correlation coefficient. The Single Point Insulin Sensitivity Estimator (SPISE) was compared to traditional insulin sensitivity indices using area under the ROC curve (aROC) analysis and χ(2) test. The novel formula for SPISE was computed as follows: SPISE = 600 × HDL-C(0.185)/(TG(0.2) × BMI(1.338)), with fasting HDL-C (mg/dL), fasting TG concentrations (mg/dL), and BMI (kg/m(2)). A cutoff value of 6.61 corresponds to an M-value smaller than 4.7 mg · kg(-1) · min(-1) (aROC, M:0.797). SPISE showed a significantly better aROC than the TG/HDL-C ratio. SPISE aROC was comparable to the Matsuda ISI (insulin sensitivity index) and equal to the QUICKI (quantitative insulin sensitivity check index) and HOMA-IR (homeostasis model assessment-insulin resistance) when calculated with M-values. The SPISE seems well suited to surrogate whole-body insulin sensitivity from inexpensive fasting single-point blood draw and BMI

  20. Procalcitonin for selecting the antibiotic regimen in outpatients with low-risk community-acquired pneumonia using a rapid point-of-care testing: A single-arm clinical trial.

    Directory of Open Access Journals (Sweden)

    Mar Masiá

    Full Text Available We aimed to assess the role of procalcitonin (PCT to guide the initial selection of the antibiotic regimen for low-risk community-acquired pneumonia (CAP.A single-arm clinical trial was conducted including outpatients with CAP and Pneumonia Severity Index risk classes I-II. Antimicrobial selection was based on the results of PCT measured with a rapid point-of-care testing. According to serum PCT levels, patients were assigned to two treatment strategies: oral azithromycin if PCT was <0.5 ng/ml, or levofloxacin if levels were ≥0.5 ng/ml. Primary outcome was clinical cure rate. Short-term and long-term outcomes were assessed. Results were compared with those of a historical standard-of-care control-group treated in our centre.Of 253 subjects included, 216 (85.4% were assigned to azithromycin. Pneumococcal infection was diagnosed in 26 (12% and 21 (56.8% patients allocated to azithromycin and levofloxacin groups, respectively. No patients in the azithromycin group developed bacteraemia. Atypical organisms were more common in patients given azithromycin (18.5% vs 8.1%, respectively. The majority (93% of patients with atypical pneumonia had low PCT levels. Clinical cure rates were 95.8% in the azithromycin group, 94.6% in the levofloxacin group, and 94.4% in the historical control group. No 30-day mortality or recurrences were observed, and the 3-year rates of recurrence and mortality were very low in both groups. Adverse events occurrence was also infrequent.A PCT-guided strategy with a rapid point-of-care testing safely allowed selecting empirical narrow-spectrum antibiotics in outpatients with CAP.The study is registered with ClinicalTrials.gov, number NCT02600806.

  1. Examining the virulence of Candida albicans transcription factor mutants using Galleria mellonella and mouse infection models

    Directory of Open Access Journals (Sweden)

    Sara eAmorim-Vaz

    2015-05-01

    Full Text Available The aim of the present study was to identify C. albicans transcription factors (TF involved in virulence. Although mice are considered the gold-standard model to study fungal virulence, mini-host infection models have been increasingly used. Here, barcoded TF mutants were first screened in mice by pools of strains and fungal burdens quantified in kidneys. Mutants of unannotated genes which generated a kidney fungal burden significantly different from that of wild-type were selected and individually examined in G. mellonella. In addition, mutants that could not be detected in mice were also tested in G. mellonella. Only 25 % of these mutants displayed matching phenotypes in both hosts, highlighting a significant discrepancy between the two models. To address the basis of this difference (pool or host effects, a set of 19 mutants tested in G. mellonella were also injected individually into mice. Matching fungal burden phenotypes were observed in 50 % of the cases, highlighting the bias due to host effects. In contrast, 33.4 % concordance was observed between pool and single strain infections in mice, thereby highlighting the bias introduced by the pool effect. After filtering the results obtained from the two infection models, mutants for MBF1 and ZCF6 were selected. Independent marker-free mutants were subsequently tested in both hosts to validate previous results. The MBF1 mutant showed impaired infection in both models, while the ZCF6 mutant was only significant in mice infections. The two mutants showed no obvious in vitro phenotypes compared with the wild-type, indicating that these genes might be specifically involved in in vivo adaptation.

  2. Evolution and clinical impact of co-occurring genetic alterations in advanced-stage EGFR-mutant lung cancers. | Office of Cancer Genomics

    Science.gov (United States)

    A widespread approach to modern cancer therapy is to identify a single oncogenic driver gene and target its mutant-protein product (for example, EGFR-inhibitor treatment in EGFR-mutant lung cancers). However, genetically driven resistance to targeted therapy limits patient survival. Through genomic analysis of 1,122 EGFR-mutant lung cancer cell-free DNA samples and whole-exome analysis of seven longitudinally collected tumor samples from a patient with EGFR-mutant lung cancer, we identified critical co-occurring oncogenic events present in most advanced-stage EGFR-mutant lung cancers.

  3. Studies on induced partially resistant mutants of barley against powdery mildew

    International Nuclear Information System (INIS)

    Roebbelen, G.; Abdel-Hafez, A.G.; Reinhold, M.; Kwon, H.J.; Neuhaus-Steinmetz, J.P.; Heun, M.

    1983-01-01

    After mutagenic seed treatment of three partially resistant cultivars of spring barley with EMS and NaN 3 , 45 mutants in a first and 16 in a second experiment were selected in the M 2 -M 4 generations. The screening was done alternatively under natural infection in the field or controlled infection with a single pathotype in the greenhouse. These mutants exhibited a higher resistance and a higher susceptibility, respectively, than the initial cultivars Asse, Bomi and Vada. Some mutants expressed their altered resistance behaviour particularly during certain stages of development. High-level resistance was conditioned by mutation in the ml-o locus in three cases. For several Bomi mutants pathotype specificity with and without reversed ranking was proven as well as pathotype non-specificity in comparison with the reaction of the original cultivar. In 14 cases studied the inheritance of the involved mutants was monogenic recessive. The laevigatum locus responsible for the intermediate mildew resistance of Bomi was not affected by the mutations. Detection of groups of allelic mutants showed that there are at least two regions in the barley genome in which mutations for mildew resistance can occur rather frequently. In total, the past ten years of this mutation research have given convincing evidence that the strategies of mutant screening applied have yielded promising new material both for breeding and for progress in basic understanding of host-pathogen interactions. (author)

  4. A new Arabidopsis mutant induced by ion beams affects flavonoid synthesis with spotted pigmentation in testa

    International Nuclear Information System (INIS)

    Tanaka, A.; Tano, S.; Chantes, T.; Yokota, Y.; Shikazono, N.; Watanabe, H.

    1997-01-01

    A new stable mutant of Arabidopsis thaliana with a spotted pigment in the seed coat, named anthocyanin spotted testa (ast), was induced by carbon ion irradiation. The spotted pigmentation of ast mutant was observed in immature seeds from 1-2 days after flowering (DAF), at the integument of the ovule, and spread as the seed coat formed. Anthocyanin accumulation was about 6 times higher in ast mutant than in the wild-type at 6 DAF of the immature seeds, but was almost the same in mature dry seeds. A higher anthocyanin accumulation was not observed in the seedlings, leaves or floral buds of ast mutant compared with the wild-type, which suggests that a high accumulation of anthocyanins is specific to the seed coat of the immature ast seeds. Reciprocal crosses between ast mutant and the wild-type indicated that ast is a single recessive gene mutation and segregates as a delayed inheritance. The results of crossing with tt7 and ttg mutants also confirmed that the AST gene is probably a regulatory locus that controls flavonoid biosynthesis. A mapping analysis revealed that the gene is located on chromosome I and is closely linked to the SSLP DNA marker nga280 with a distance of 3.2 cM. AST has been registered as a new mutant of Arabidopsis

  5. Decreased uv mutagenesis in cdc8, a DNA replication mutant of Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    Prakash, L.; Hinkle, D.; Prakash, S.

    1978-01-01

    A DNA replication mutant of yeast, cdc8, was found to decrease uv-induced reversion of lys2-1, arg4-17, tryl and ural. This effect was observed with all three alleles of cdc8 tested. Survival curves obtained following uv irradiation in cdc8 rad double mutants show that cdc8 is epistatic to rad6, as well as to rad1; cdc8 rad51 double mutants seem to be more sensitive than the single mutants. Since uv-induced reversion in cdc8 rad1 and cdc8 rad51 double mutants is like that of the cdc8 single mutants, we conclude that CDC8 plays a direct role in error-prone repair. To test whether CDC8 codes for a DNA polymerase, we have purified both DNA polymerase I and DNA polymerase II from cdc8 and CDC+ cells. The purified DNA polymerases from cdc8 were no more heat labile than those from CDC+, suggesting that CDC8 is not a structural gene for either enzyme

  6. Decreased uv mutagenesis in cdc8, a DNA replication mutant of Saccharomyces cerevisiae

    Energy Technology Data Exchange (ETDEWEB)

    Prakash, L.; Hinkle, D.; Prakash, S.

    1978-01-01

    A DNA replication mutant of yeast, cdc8, was found to decrease uv-induced reversion of lys2-1, arg4-17, tryl and ural. This effect was observed with all three alleles of cdc8 tested. Survival curves obtained following uv irradiation in cdc8 rad double mutants show that cdc8 is epistatic to rad6, as well as to rad1; cdc8 rad51 double mutants seem to be more sensitive than the single mutants. Since uv-induced reversion in cdc8 rad1 and cdc8 rad51 double mutants is like that of the cdc8 single mutants, we conclude that CDC8 plays a direct role in error-prone repair. To test whether CDC8 codes for a DNA polymerase, we have purified both DNA polymerase I and DNA polymerase II from cdc8 and CDC+ cells. The purified DNA polymerases from cdc8 were no more heat labile than those from CDC+, suggesting that CDC8 is not a structural gene for either enzyme.

  7. Purification of Escherichia coli L-asparaginase mutants by a native polyacrylamide gel electrophoresis.

    Science.gov (United States)

    Wei, Yujun; Chen, Jianhua; Jia, Ruibo; Wang, Min; Wu, Wutong

    2008-07-01

    The antigenicity of L-asaparaginase (L-ASP) has been problematic for the treatment of leukemia for many years. In order to establish a relationship between the antigenic epitope of L-asparaginase and its antigenicity, several L-asparaginase mutants (mL-ASPs) are constructed and expressed. To effectively purify these enzyme mutants for further investigation, a native preparative polyacrylamide gel electrophoresis is developed. The simplicity and reproducibility of this approach permits the purification of different mutants from the crude enzyme extracts, with a sufficient activity to perform immunological and biological studies. Furthermore, the newly developed method is efficient and cost-effective compared with other methods, such as column chromatography and affinity chromatography. As a result, the enzyme mutants with specific activity of 300 approximately 400 U/mg are obtained by the single-step purification with a high degree of purity.

  8. X-rays sensitive mammalian cell mutant

    International Nuclear Information System (INIS)

    Utsumi, Hiroshi

    1982-01-01

    A phenomenon that in x-ray-sensitive mammalian-cell mutants, cellular death due to x-ray radiation was not increased by caffeine, but on the contrary, the dead cells were resuscitated by it was discussed. The survival rate of mutant cells increased by caffein in a low concentration. This suggested that caffeine may have induced some mechanism to produce x-ray resistant mutant cells. Postirradiation treatment with caffeine increased considerably the survival rate of the mutant cells, and this suggested the existence of latent caffeine-sensitive potentially lethal damage repair system. This system, after a few hours, is thought to be substituted by caffeine-resistant repair system which is induced by caffeine, and this may be further substituted by x-ray-resistant repair system. The repair system was also induced by adenine. (Ueda, J.)

  9. Semi-dwarf mutants for rice improvement

    International Nuclear Information System (INIS)

    Othman, Ramli; Osman, Mohammad; Ibrahim, Rusli

    1990-01-01

    Full text: MARDI and the National University of Malaysia embarked on a programme to induce resistance against blast in rice in 1978. MARDI also obtained semi dwarf mutants of cvs 'Mahsuri', 'Muda', 'Pongsu seribu' and 'Jarum Mas', which are under evaluation. The popular local rice variety 'Manik' was subjected to gamma irradiation (15-40 krad) and 101 promising semidwarf mutants have been obtained following selection in M 2 -M 6 . 29 of them show grain yields of 6.0-7.3 t/ha, compared with 5.7t for 'Manik'. Other valuable mutants were found showing long grain, less shattering, earlier maturity, and glutinous endosperm. One mutant, resistant to brown plant hopper yields 6.3t/ha. (author)

  10. Commercialization Of Orchid Mutants For Floriculture Industry

    International Nuclear Information System (INIS)

    Sakinah Ariffin; Zaiton Ahmad

    2014-01-01

    Orchids are the main contributors to cut flower industry in Malaysia with an existing good market and a huge business potential. Orchid industry has been established in Malaysia since 1960s but only started to develop and expand since 1980s. Continuous development of new orchid varieties is essential to meet customers' demands. Orchid mutagenesis research using gamma irradiation at Malaysian Nuclear Agency has successfully generated a number of new orchid varieties with commercial potentials. Therefore, Nuclear Malaysia has collaborated with an industrial partner, Hexagon Green Sdn Bhd (HGSB), to carry out commercialization research on these mutants under a Technofund project entitled 'Pre-Commercialization of Mutant Orchids for Cut Flowers Industry' from July 2011 to July 2014. Through this collaboration, Dendrobium orchid mutant plants developed by Nuclear Malaysia were transferred to HGSB's commercial orchid nursery at Bukit Changgang Agrotechnology Park, Banting, Selangor, for mass-propagation. The activities include evaluations on plant growth performance, flower quality, post harvest and market potential of these mutants. Mutants with good field performance have been identified and filed for Plant Variety Protection (PVP) with Department of Agriculture Malaysia. This paper describes outputs from this collaboration and activities undertaken in commercializing these mutants. (author)

  11. Comparison of a coq7 deletion mutant with other respiration-defective mutants in fission yeast.

    Science.gov (United States)

    Miki, Risa; Saiki, Ryoichi; Ozoe, Yoshihisa; Kawamukai, Makoto

    2008-11-01

    Among the steps in ubiquinone biosynthesis, that catalyzed by the product of the clk-1/coq7 gene has received considerable attention because of its relevance to life span in Caenorhabditis elegans. We analyzed the coq7 ortholog (denoted coq7) in Schizosaccharomyces pombe, to determine whether coq7 has specific roles that differ from those of other coq genes. We first confirmed that coq7 is necessary for the penultimate step in ubiquinone biosynthesis, from the observation that the deletion mutant accumulated the ubiquinone precursor demethoxyubiquinone-10 instead of ubiquinone-10. The coq7 mutant displayed phenotypes characteristic of other ubiquinone-deficient Sc. pombe mutants, namely, hypersensitivity to hydrogen peroxide, a requirement for antioxidants for growth on minimal medium, and an elevated production of sulfide. To compare these phenotypes with those of other respiration-deficient mutants, we constructed cytochrome c (cyc1) and coq3 deletion mutants. We also assessed accumulation of oxidative stress in various ubiquinone-deficient strains and in the cyc1 mutant by measuring mRNA levels of stress-inducible genes and the phosphorylation level of the Spc1 MAP kinase. Induction of ctt1, encoding catalase, and apt1, encoding a 25 kDa protein, but not that of gpx1, encoding glutathione peroxidase, was indistinguishable in four ubiquinone-deficient mutants, indicating that the oxidative stress response operates at similar levels in the tested strains. One new phenotype was observed, namely, loss of viability in stationary phase (chronological life span) in both the ubiquinone-deficient mutant and in the cyc1 mutant. Finally, Coq7 was found to localize in mitochondria, consistent with the possibility that ubiquinone biosynthesis occurs in mitochondria in yeasts. In summary, our results indicate that coq7 is required for ubiquinone biosynthesis and the coq7 mutant is not distinguishable from other ubiquinone-deficient mutants, except that its phenotypes are more

  12. Construction of Comprehensive Dosage-Matching Core Histone Mutant Libraries forSaccharomyces cerevisiae.

    Science.gov (United States)

    Jiang, Shuangying; Liu, Yan; Wang, Ann; Qin, Yiran; Luo, Maoguo; Wu, Qingyu; Boeke, Jef D; Dai, Junbiao

    2017-12-01

    Saccharomyces cerevisiae contains two genes for each core histone, which are presented as pairs under the control of a divergent promoter, i.e. , HHT1-HHF1 , HHT2-HHF2 , HTA1-HTB1 and HTA2-HTB2 HHT1-HHF1 , and HHT2-HHF2 encode histone H3 and H4 with identical amino acid sequences but under the control of differently regulated promoters. Previous mutagenesis studies were carried out by deleting one pair and mutating the other one. Here, we present the design and construction of three additional libraries covering HTA1-HTB1 , HTA2-HTB2 , and HHT1-HHF1 respectively. Together with the previously described library of HHT2-HHF2 mutants, a systematic and complete collection of mutants for each of the eight core S. cerevisiae histone genes becomes available. Each designed mutant was incorporated into the genome, generating three more corresponding libraries of yeast strains. We demonstrated that, although, under normal growth conditions, strains with single-copy integrated histone genes lacked phenotypes, in some growth conditions, growth deficiencies were observed. Specifically, we showed that addition of a second copy of the mutant histone gene could rescue the lethality in some previously known mutants that cannot survive with a single copy. This resource enables systematic studies of function of each nucleosome residue in plasmid, single-copy, and double-copy integrated formats. Copyright © 2017 by the Genetics Society of America.

  13. Single gaze gestures

    DEFF Research Database (Denmark)

    Møllenbach, Emilie; Lilholm, Martin; Gail, Alastair

    2010-01-01

    This paper examines gaze gestures and their applicability as a generic selection method for gaze-only controlled interfaces. The method explored here is the Single Gaze Gesture (SGG), i.e. gestures consisting of a single point-to-point eye movement. Horizontal and vertical, long and short SGGs were...

  14. Development of high yielding mutants in lentil

    International Nuclear Information System (INIS)

    Rajput, M.A.; Sarwar, G.; Siddiqui, K.A.

    2001-01-01

    Full text: Lentil (Lens culinaris Medik.) locally known as Masoor, is the second most important rabi pulse crop, after chickpea, in Pakistan. It is cultivated on an area of over 63,400 ha, which constitutes about 4.83% of the total area under pulses. The annual production of the crop is 28,200 tones with an average yield of 445 kg/ha. Yield at the national level is very low, about one-half of the world's yield, which is mainly due to non-availability of high yield potential genotypes. Keeping in view the importance of mutants in developing a large number of new varieties, an induced mutations programme was initiated at AEARC, Tandojam during 1987-88, to develop high yielding varieties in lentil. For this, seeds of two lentil varieties, 'Masoor-85' and 'ICARDA-8' had been irradiated with gamma-rays ranging from 100-600 Gy in NIAB, Faisalabad during 1990. Selections were made in M2 on the basis of earliness, plant height, branches/plant and 100 grain weight. After confirming these mutants in M3 they were promoted in station yield trials and studied continuously for three consecutive years (1993- 1995). Overall results revealed that these mutants have consistent improvement of earliness in flowering and maturity. Plant height also increased in all mutant lines except AEL 23/40/91 where reduction in this attribute was observed as compared to parent variety. Mutant lines AEL 49/20/91 and AEL 13/30/91 showed improvement in 100 grain weight. The improvement of some agronomic characters enhanced the yield of mutant lines in comparison to parent varieties (Masoor-85 and ICARDA-8). The diversity in yield over the respective parents was computed from 6.94 to 60.12%. From these encouraging results it is hoped that mutant lines like AEL 12/30/91 and AEL 49/20/91 may serve as potential lentil genotypes in future. (author)

  15. Officially released mutant varieties in China

    International Nuclear Information System (INIS)

    Liu, L.; Van Zanten, L.; Shu, Q.Y.; Maluszynski, M.

    2004-01-01

    The use of mutation techniques for crop improvement in China has a long and well-established tradition of more than 50 years. As the result of intensive research in many institutes dealing with application of nuclear technologies more than 620 cultivars of 44 crop species have been released. Numerous mutant varieties have been grown on a large scale bringing significant economic impact, sustaining crop production and greatly contributing to increase of food production also in stress prone areas of the country. However, there is still missing information not only on the number of mutant varieties released in particular crop species but also on mutagens applied, selection approaches and on the use of mutants in cross breeding. Numerous Chinese scientists collected and systematized this information. Results of their work were often published in local scientific journals in the Chinese language and as such were unavailable to breeders from other countries. Having this in mind, we requested Dr. Liu Luxiang, the Director of the Department of Plant Mutation Breeding and Genetics, Institute for Application of Atomic Energy, Chinese Academy of Agricultural Sciences in Beijing to help us in finding as much information as possible on mutant varieties officially released in China. The data has been collected in close collaboration with his colleagues from various institutions all over the country and then evaluated, edited and prepared for publication by our team responsible for the FAO/IAEA Database of Officially Released Mutant Varieties. We would like to thank all Chinese colleagues who contributed to this list of Chinese mutant varieties. We hope that this publication will stimulate plant breeders in China to collect more information on released mutant varieties and especially on the use of mutated genes in cross breeding. (author)

  16. Computational identification of adaptive mutants using the VERT system

    Directory of Open Access Journals (Sweden)

    Winkler James

    2012-04-01

    Full Text Available Background Evolutionary dynamics of microbial organisms can now be visualized using the Visualizing Evolution in Real Time (VERT system, in which several isogenic strains expressing different fluorescent proteins compete during adaptive evolution and are tracked using fluorescent cell sorting to construct a population history over time. Mutations conferring enhanced growth rates can be detected by observing changes in the fluorescent population proportions. Results Using data obtained from several VERT experiments, we construct a hidden Markov-derived model to detect these adaptive events in VERT experiments without external intervention beyond initial training. Analysis of annotated data revealed that the model achieves consensus with human annotation for 85-93% of the data points when detecting adaptive events. A method to determine the optimal time point to isolate adaptive mutants is also introduced. Conclusions The developed model offers a new way to monitor adaptive evolution experiments without the need for external intervention, thereby simplifying adaptive evolution efforts relying on population tracking. Future efforts to construct a fully automated system to isolate adaptive mutants may find the algorithm a useful tool.

  17. Hepatitis B surface gene 145 mutant as a minor population in hepatitis B virus carriers

    Directory of Open Access Journals (Sweden)

    Komatsu Haruki

    2012-01-01

    Full Text Available Abstract Background Hepatitis B virus (HBV can have mutations that include the a determinant, which causes breakthrough infection. In particular, a single mutation at amino acid 145 of the surface protein (G145 is frequently reported in the failure of prophylactic treatment. The aim of this study was to evaluate the frequency of the a determinant mutants, especially the G145 variant, in Japan, where universal vaccination has not been adopted. Methods The present study was a retrospective study. The study cohorts were defined as follows: group 1, children with failure to prevent mother-to-child transmission despite immunoprophylaxis (n = 18, male/female = 8/10, age 1-14 years; median 6 years; group 2, HBV carriers who had not received vaccination or hepatitis B immunoglobulin (n = 107, male/female = 107, age 1-52 years; median 16 years. To detect the G145R and G145A mutants in patients, we designed 3 probes for real-time PCR. We also performed direct sequencing and cloning of PCR products. Results By mutant-specific real-time PCR, one subject (5.6% was positive for the G145R mutant in group 1, while the G145 mutant was undetectable in group 2. The a determinant mutants were detected in one (5.6% of the group 1 subjects and 10 (9.3% of the group 2 subjects using direct sequencing, but direct sequencing did not reveal the G145 mutant as a predominant strain in the two groups. However, the subject who was positive according to the mutant-specific real-time PCR in group 1 had overlapped peaks at nt 587 in the electropherogram. In group 2, 11 patients had overlapped peaks at nt 587 in the electropherogram. Cloning of PCR products allowed detection of the G145R mutant as a minor strain in 7 (group 1: 1 subject, group 2: 6 subjects of 12 subjects who had overlapped peaks at nt 587 in the electropherogram. Conclusions The frequency of the a determinant mutants was not high in Japan. However, the G145R mutant was often present as a minor population in

  18. Agronomically valuable mutant lines of castor

    International Nuclear Information System (INIS)

    Bokhan, I.K.

    1990-01-01

    Dry seeds of four castor varieties (VNIIMK 165-improved, VNIIMK 18, Chervonnaya and Antika) were treated with six chemical mutagens, N-nitroso-N-methyl urea (NMU), N-nitroso-N-ethyl urea (NEU), dimethyl sulphate (DMS), diethyl sulphate (DES), ethylenimine (EI) and 1,4-bis-diazoacetyl-butane (DAB) in various doses during 18 hours. About 40,000 plants were studied in M 2 and 80 types of mutations were found, including a number of valuable mutants: short-stemmed, semi-dwarf, dwarf, early maturing, with female and interspersed types of racemes, highly productive etc. Based on trials in M 3 -M 4 , on small plots with two or three replications, the superior mutant lines were identified. The best mutants are presented in the table. Early maturation is very important for growing castor in the USSR, as it is the predecessor of winter wheat in crop rotation. The mutants M2-323 and Ml-83 are of great value as they show early maturation and high yield. Their productivity is mainly conditioned by a high percentage of interspersed plants. The reduction of plant height is of great importance for the successful combine harvesting of castor. Mutant lines M2-119 and Ml-284 characterised by low plant height and high yield are very interesting in this respect. The obtained initial material will be used in further breeding work

  19. Plasmodium berghei: in vivo generation and selection of karyotype mutants and non-gametocyte producer mutants

    NARCIS (Netherlands)

    Janse, C. J.; Ramesar, J.; van den Berg, F. M.; Mons, B.

    1992-01-01

    We previously reported that karyotype and gametocyte-producer mutants spontaneously arose during in vivo asexual multiplication of Plasmodium berghei. Here we studied the rate of selection of these mutants in vivo. Gametocyte production and karyotype pattern were established at regular intervals

  20. Trehalose, glycogen and ethanol metabolism in the gcr1 mutant of Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Seker, Tamay; Hamamci, H.

    2003-01-01

    Since Gcr1p is pivotal in controlling the transcription of glycolytic enzymes and trehalose metabolism seems to be one of the control points of glycolysis, we examined trehalose and glycogen synthesis in response to 2 % glucose pulse during batch growth in gcr1 (glucose regulation-1) mutant lacking...

  1. The research progress on plant mutant germplasm resources in China

    International Nuclear Information System (INIS)

    He Cexi; Ji Linzhen; Zhao Shirong

    1991-07-01

    Mutants induced by nuclear radiation or other mutagens are new artificial germplasm resources. Some mutants have been applied in plant breeding and great achievements have been reached. The status and progress on the collection, identification and utilization of mutants in China are introduced. A proposal for developing mutant germplasm resources with good agronomic characters is suggested

  2. High Persister Mutants in Mycobacterium tuberculosis.

    Directory of Open Access Journals (Sweden)

    Heather L Torrey

    Full Text Available Mycobacterium tuberculosis forms drug-tolerant persister cells that are the probable cause of its recalcitrance to antibiotic therapy. While genetically identical to the rest of the population, persisters are dormant, which protects them from killing by bactericidal antibiotics. The mechanism of persister formation in M. tuberculosis is not well understood. In this study, we selected for high persister (hip mutants and characterized them by whole genome sequencing and transcriptome analysis. In parallel, we identified and characterized clinical isolates that naturally produce high levels of persisters. We compared the hip mutants obtained in vitro with clinical isolates to identify candidate persister genes. Genes involved in lipid biosynthesis, carbon metabolism, toxin-antitoxin systems, and transcriptional regulators were among those identified. We also found that clinical hip isolates exhibited greater ex vivo survival than the low persister isolates. Our data suggest that M. tuberculosis persister formation involves multiple pathways, and hip mutants may contribute to the recalcitrance of the infection.

  3. Cooperative induction of apoptosis in NRAS mutant melanoma by inhibition of MEK and ROCK

    NARCIS (Netherlands)

    Vogel, Celia J.; Smit, Marjon A.; Maddalo, Gianluca; Possik, Patricia A.; Sparidans, Rolf W.; van der Burg, Sjoerd H.; Verdegaal, Els M.; Heck, Albert J R; Samatar, Ahmed A.; Beijnen, Jos H.; Altelaar, A. F Maarten; Peeper, Daniel S.

    2015-01-01

    Summary: No effective targeted therapy is currently available for NRAS mutant melanoma. Experimental MEK inhibition is rather toxic and has only limited efficacy in clinical trials. At least in part, this is caused by the emergence of drug resistance, which is commonly seen for single agent

  4. Cytosine hypomethylation at CHG and CHH sites in the pleiotropic mutants of Mendelian inheritance in Catharanthus roseus.

    Science.gov (United States)

    Kumari, Renu; Yadav, Gitanjali; Sharma, Vishakha; Sharma, Vinay; Kumar, Sushil

    2013-12-01

    The 5S and 18S rDNA sequences of Catharanthus roseus cv 'Nirmal' (wild type) and its leafless inflorescence (lli), evergreen dwarf (egd) and irregular leaf lamina (ill) single mutants and lli egd, lli ill and egd ill double mutants were characterized. The lli, egd and ill mutants of Mendelian inheritance bore the names after their most conspicuous morphological feature(s). They had been chemically induced and isolated for their salt tolerance. The double mutants were isolated as morphological segregants from crosses between single mutants. The morphological features of the two parents accompanied salt tolerance in the double mutants. All the six mutants were hypomethylated at repeat sequences, upregulated and downregulated for many genes and carried pleiotropic alterations for several traits. Here the 5S and 18S rDNAs of C. roseus were found to be relatively low in cytosine content. Cytosines were preponderantly in CG context (53%) and almost all of them were methylated (97%). The cytosines in CHH and CHG (where H = A, T or C) contexts were largely demethylated (92%) in mutants. The demethylation was attributable to reduced expression of RDR2 and DRM2 led RNA dependant DNA methylation and CMT3 led maintenance methylation pathways. Mutants had gained some cytosines by substitution of C at T sites. These perhaps arose on account of errors in DNA replication, mediated by widespread cytosine demethylation at CHG and CHH sites. It was concluded that the regulation of cytosine ethylation mechanisms was disturbed in the mutants. ILL, EGD and LLI genes were identified as the positive regulators of other genes mediating the RdDM and CMT3 pathways, for establishment and maintenance of cytosine methylation in C. roseus.

  5. An Ethylmethane Sulfonate Mutant Resource in Pre-Green Revolution Hexaploid Wheat

    Science.gov (United States)

    Dhaliwal, Amandeep K.; Mohan, Amita; Sidhu, Gaganjot; Maqbool, Rizwana; Gill, Kulvinder S.

    2015-01-01

    Mutagenesis is a powerful tool used for studying gene function as well as for crop improvement. It is regaining popularity because of the development of effective and cost efficient methods for high-throughput mutation detection. Selection for semi-dwarf phenotype during green revolution has reduced genetic diversity including that for agronomically desirable traits. Most of the available mutant populations in wheat (Triticum aestivum L.) were developed in post-green revolution cultivars. Besides the identification and isolation of agronomically important alleles in the mutant population of pre-green revolution cultivar, this population can be a vital resource for expanding the genetic diversity for wheat breeding. Here we report an Ethylmethane Sulfonate (EMS) generated mutant population consisting of 4,180 unique mutant plants in a pre-green revolution spring wheat cultivar ‘Indian’. Released in early 1900s, ‘Indian’ is devoid of any known height-reducing mutations. Unique mutations were captured by proceeding with single M2 seed from each of the 4,180 M1 plants. Mutants for various phenotypic traits were identified by detailed phenotyping for altered morphological and agronomic traits on M2 plants in the greenhouse and M3 plants in the field. Of the 86 identified mutants, 75 (87%) were phenotypically stable at the M4 generation. Among the observed phenotypes, variation in plant height was the most frequent followed by the leaf morphology. Several mutant phenotypes including looped peduncle, crooked plant morphology, ‘gritty’ coleoptiles, looped lower internodes, and burnt leaf tips are not reported in other plant species. Considering the extent and diversity of the observed mutant phenotypes, this population appears to be a useful resource for the forward and reverse genetic studies. This resource is available to the scientific community. PMID:26678261

  6. An Ethylmethane Sulfonate Mutant Resource in Pre-Green Revolution Hexaploid Wheat.

    Science.gov (United States)

    Dhaliwal, Amandeep K; Mohan, Amita; Sidhu, Gaganjot; Maqbool, Rizwana; Gill, Kulvinder S

    2015-01-01

    Mutagenesis is a powerful tool used for studying gene function as well as for crop improvement. It is regaining popularity because of the development of effective and cost efficient methods for high-throughput mutation detection. Selection for semi-dwarf phenotype during green revolution has reduced genetic diversity including that for agronomically desirable traits. Most of the available mutant populations in wheat (Triticum aestivum L.) were developed in post-green revolution cultivars. Besides the identification and isolation of agronomically important alleles in the mutant population of pre-green revolution cultivar, this population can be a vital resource for expanding the genetic diversity for wheat breeding. Here we report an Ethylmethane Sulfonate (EMS) generated mutant population consisting of 4,180 unique mutant plants in a pre-green revolution spring wheat cultivar 'Indian'. Released in early 1900s, 'Indian' is devoid of any known height-reducing mutations. Unique mutations were captured by proceeding with single M2 seed from each of the 4,180 M1 plants. Mutants for various phenotypic traits were identified by detailed phenotyping for altered morphological and agronomic traits on M2 plants in the greenhouse and M3 plants in the field. Of the 86 identified mutants, 75 (87%) were phenotypically stable at the M4 generation. Among the observed phenotypes, variation in plant height was the most frequent followed by the leaf morphology. Several mutant phenotypes including looped peduncle, crooked plant morphology, 'gritty' coleoptiles, looped lower internodes, and burnt leaf tips are not reported in other plant species. Considering the extent and diversity of the observed mutant phenotypes, this population appears to be a useful resource for the forward and reverse genetic studies. This resource is available to the scientific community.

  7. An Ethylmethane Sulfonate Mutant Resource in Pre-Green Revolution Hexaploid Wheat.

    Directory of Open Access Journals (Sweden)

    Amandeep K Dhaliwal

    Full Text Available Mutagenesis is a powerful tool used for studying gene function as well as for crop improvement. It is regaining popularity because of the development of effective and cost efficient methods for high-throughput mutation detection. Selection for semi-dwarf phenotype during green revolution has reduced genetic diversity including that for agronomically desirable traits. Most of the available mutant populations in wheat (Triticum aestivum L. were developed in post-green revolution cultivars. Besides the identification and isolation of agronomically important alleles in the mutant population of pre-green revolution cultivar, this population can be a vital resource for expanding the genetic diversity for wheat breeding. Here we report an Ethylmethane Sulfonate (EMS generated mutant population consisting of 4,180 unique mutant plants in a pre-green revolution spring wheat cultivar 'Indian'. Released in early 1900s, 'Indian' is devoid of any known height-reducing mutations. Unique mutations were captured by proceeding with single M2 seed from each of the 4,180 M1 plants. Mutants for various phenotypic traits were identified by detailed phenotyping for altered morphological and agronomic traits on M2 plants in the greenhouse and M3 plants in the field. Of the 86 identified mutants, 75 (87% were phenotypically stable at the M4 generation. Among the observed phenotypes, variation in plant height was the most frequent followed by the leaf morphology. Several mutant phenotypes including looped peduncle, crooked plant morphology, 'gritty' coleoptiles, looped lower internodes, and burnt leaf tips are not reported in other plant species. Considering the extent and diversity of the observed mutant phenotypes, this population appears to be a useful resource for the forward and reverse genetic studies. This resource is available to the scientific community.

  8. Out-of-plane integration of a multimode optical fiber for single particle/cell detection at multiple points on a microfluidic device with applications to particle/cell counting, velocimetry, size discrimination and the analysis of single cell lysate injections.

    Science.gov (United States)

    Sadeghi, Jalal; Patabadige, Damith E W; Culbertson, Anne H; Latifi, Hamid; Culbertson, Christopher T

    2016-12-20

    In this paper a single particle/cell-tracking microfluidic device that integrates an out-of-plane multimode optical fiber (OP-MMF) is reported. This OP-MMF is used to generate three excitation light-lines and three detection spots using only one excitation source and one detector. It takes advantage of an optical tunneling mode to create two excitation lines in a microfluidic channel emanating from a single fiber end. This method was used to accurately count particles/cells and perform velocity measurements and size discrimination. The velocity and size distributions of 5, 7, and 10 μm fluorescently labeled polystyrene beads were determined using the OP-MMF. Additionally, this method was used to analyze cell lysates with the third excitation line in the separation channel. The OP-MMF setup accurately detected an intact cell twice ∼2 mm prior to lysis, determined its velocity, and detected the injected cell lysate 3 mm downstream of the injection point in the separation channel. Using this setup, the velocity of cells entering the lysis intersection and the absolute migration times of fluorescently labeled analytes injected into the separation channel were determined in an automated fashion. This method enabled us to determine a lysing/injection efficiency coefficient (K) using signals from the injected lysate signal and from the intact cell before lysing. K provided a reliable measurement of the amount of cell lysate that was injected into the separation channel. The approach reported here could be used in the future to track particles, cells or droplets in a variety of existing microfluidic devices without the need for multiplexed masks, layers, bulky optical elements or complex optical designs.

  9. Efficacy of hepatitis B vaccine against antiviral drug-resistant hepatitis B virus mutants in the chimpanzee model.

    Science.gov (United States)

    Kamili, Saleem; Sozzi, Vitini; Thompson, Geoff; Campbell, Katie; Walker, Christopher M; Locarnini, Stephen; Krawczynski, Krzysztof

    2009-05-01

    Hepatitis B virus (HBV) mutants resistant to treatment with nucleoside or nucleotide analogs and those with the ability to escape from HBV-neutralizing antibody have the potential to infect HBV-vaccinated individuals. To address this potential serious public health challenge, we tested the efficacy of immunity induced by a commercial hepatitis B vaccine against a tissue culture-derived, clonal HBV polymerase mutant in HBV seronegative chimpanzees. The polymerase gene mutant contained a combination of three mutations (rtV173L, rtL180M, rtM204V), two of which resulted in changes to the overlapping viral envelope of the hepatitis B surface antigen (sE164D, sI195M). Prior to the HBV mutant challenge of vaccinated chimpanzees, we established virologic, serologic, and pathologic characteristics of infections resulting from intravenous inoculation of the HBV polymerase gene mutant and the sG145R vaccine-escape surface gene mutant. Cloning and sequencing experiments determined that the three mutations in the polymerase gene mutant remained stable and that the single mutation in the surface gene mutant reverted to the wild-type sequence. Immunological evidence of HBV replication was observed in the vaccinated chimpanzees after challenge with the polymerase gene mutant as well as after rechallenge with serum-derived wild-type HBV (5,000 chimpanzee infectious doses administered intravenously), despite robust humoral and cellular anti-HBV immune responses after hepatitis B vaccination. Our data showing successful experimental infection by HBV mutants despite the presence of high anti-HBs levels considered protective in the vaccinated host are consistent with clinical reports on breakthrough infection in anti-HBs-positive patients infected with HBV mutants. In the absence of a protective humoral immunity, adaptive cellular immune responses elicited by infection may limit HBV replication and persistence.

  10. Mutant DNA quantification by digital PCR can be confounded by heating during DNA fragmentation.

    Science.gov (United States)

    Kang, Qing; Parkin, Brian; Giraldez, Maria D; Tewari, Muneesh

    2016-04-01

    Digital PCR (dPCR) is gaining popularity as a DNA mutation quantification method for clinical specimens. Fragmentation prior to dPCR is required for non-fragmented genomic DNA samples; however, the effect of fragmentation on DNA analysis has not been well-studied. Here we evaluated three fragmentation methods for their effects on dPCR point mutation assay performance. Wild-type (WT) human genomic DNA was fragmented by heating, restriction digestion, or acoustic shearing using a Covaris focused-ultrasonicator. dPCR was then used to determine the limit of blank (LoB) by quantifying observed WT and mutant allele counts of the proto-oncogenes KRAS and BRAF in the WT DNA sample. DNA fragmentation by heating to 95°C, while the simplest and least expensive method, produced a high background mutation frequency for certain KRAS mutations relative to the other methods. This was due to heat-induced mutations, specifically affecting dPCR assays designed to interrogate guanine to adenine (G>A) mutations. Moreover, heat-induced fragmentation overestimated gene copy number, potentially due to denaturation and partition of single-stranded DNA into different droplets. Covaris acoustic shearing and restriction enzyme digestion showed similar LoBs and gene copy number estimates to one another. It should be noted that moderate heating, commonly used in genomic DNA extraction protocols, did not significantly increase observed KRAS mutation counts.

  11. Whole-genome sequencing of mutants with increased resistance against the two-peptide bacteriocin plantaricin JK reveals a putative receptor and potential docking site.

    Science.gov (United States)

    Ekblad, Bie; Nissen-Meyer, Jon; Kristensen, Tom

    2017-01-01

    By whole-genome sequencing of resistant mutants, a putative receptor for plantaricin JK, a two-peptide bacteriocin produced by some Lactobacillus plantarum strains, was identified in Lactobacillus plantarum NCFB 965 and Weissella viridescens NCFB 1655. The receptors of the two species had 66% identical amino acid sequences and belong to the amino acid-polyamine-organocation (APC) transporter protein family. The resistant mutants contained point mutations in the protein-encoding gene resulting in either premature stop codons, leading to truncated versions of the protein, or single amino acid substitutions. The secondary structure of the W. viridescens protein was predicted to contain 12 transmembrane (TM) helices, a core structure shared by most members of the APC protein family. The single amino acid substitutions that resulted in resistant strains were located in a confined region of the protein that consists of TM helix 10, which is predicted to be part of an inner membrane pore, and an extracellular loop between TM helix 11 and 12. By use of template-based modeling a 3D structure model of the protein was obtained, which visualizes this mutational hotspot region and further strengthen the hypothesis that it represents a docking site for plantaricin JK.

  12. Combined MEK and ERK inhibition overcomes therapy-mediated pathway reactivation in RAS mutant tumors.

    Science.gov (United States)

    Merchant, Mark; Moffat, John; Schaefer, Gabriele; Chan, Jocelyn; Wang, Xi; Orr, Christine; Cheng, Jason; Hunsaker, Thomas; Shao, Lily; Wang, Stephanie J; Wagle, Marie-Claire; Lin, Eva; Haverty, Peter M; Shahidi-Latham, Sheerin; Ngu, Hai; Solon, Margaret; Eastham-Anderson, Jeffrey; Koeppen, Hartmut; Huang, Shih-Min A; Schwarz, Jacob; Belvin, Marcia; Kirouac, Daniel; Junttila, Melissa R

    2017-01-01

    Mitogen-activated protein kinase (MAPK) pathway dysregulation is implicated in >30% of all cancers, rationalizing the development of RAF, MEK and ERK inhibitors. While BRAF and MEK inhibitors improve BRAF mutant melanoma patient outcomes, these inhibitors had limited success in other MAPK dysregulated tumors, with insufficient pathway suppression and likely pathway reactivation. In this study we show that inhibition of either MEK or ERK alone only transiently inhibits the MAPK pathway due to feedback reactivation. Simultaneous targeting of both MEK and ERK nodes results in deeper and more durable suppression of MAPK signaling that is not achievable with any dose of single agent, in tumors where feedback reactivation occurs. Strikingly, combined MEK and ERK inhibition is synergistic in RAS mutant models but only additive in BRAF mutant models where the RAF complex is dissociated from RAS and thus feedback productivity is disabled. We discovered that pathway reactivation in RAS mutant models occurs at the level of CRAF with combination treatment resulting in a markedly more active pool of CRAF. However, distinct from single node targeting, combining MEK and ERK inhibitor treatment effectively blocks the downstream signaling as assessed by transcriptional signatures and phospho-p90RSK. Importantly, these data reveal that MAPK pathway inhibitors whose activity is attenuated due to feedback reactivation can be rescued with sufficient inhibition by using a combination of MEK and ERK inhibitors. The MEK and ERK combination significantly suppresses MAPK pathway output and tumor growth in vivo to a greater extent than the maximum tolerated doses of single agents, and results in improved anti-tumor activity in multiple xenografts as well as in two Kras mutant genetically engineered mouse (GEM) models. Collectively, these data demonstrate that combined MEK and ERK inhibition is functionally unique, yielding greater than additive anti-tumor effects and elucidates a highly

  13. Nicotinamide ribosyl uptake mutants in Haemophilus influenzae.

    Science.gov (United States)

    Herbert, Mark; Sauer, Elizabeta; Smethurst, Graeme; Kraiss, Anita; Hilpert, Anna-Karina; Reidl, Joachim

    2003-09-01

    The gene for the nicotinamide riboside (NR) transporter (pnuC) was identified in Haemophilus influenzae. A pnuC mutant had only residual NR uptake and could survive in vitro with high concentrations of NR, but could not survive in vivo. PnuC may represent a target for the development of inhibitors for preventing H. influenzae disease.

  14. Ethanol production using engineered mutant E. coli

    Science.gov (United States)

    Ingram, Lonnie O.; Clark, David P.

    1991-01-01

    The subject invention concerns novel means and materials for producing ethanol as a fermentation product. Mutant E. coli are transformed with a gene coding for pyruvate decarboxylase activity. The resulting system is capable of producing relatively large amounts of ethanol from a variety of biomass sources.

  15. Avirulent mutants of Macrophomina phaseolina and Aspergillus ...

    Indian Academy of Sciences (India)

    albino colonies out of the above ten were moderately viru- lent compared to wild type strain. They were sclerotia forming and produced phaseolinone in culture. A brown variant, designated Muv 21, and two other albino mutants, designated Muv 19 and Muv 20, were avirulent and pro- duced no phaseolinone in culture.

  16. Induced mutants for cereal grain protein improvement

    International Nuclear Information System (INIS)

    1982-01-01

    Out of 17 papers and one summary presented, six dealing with the genetic improvement of seed protein using ionizing radiations fall within the INIS subject scope. Other topics discussed were non-radiation induced mutants used for cereal grain protein improvement

  17. Mutant PTEN in Cancer : Worse Than Nothing

    NARCIS (Netherlands)

    Leslie, Nick R; den Hertog, Jeroen

    2014-01-01

    Tumor suppressors block the development of cancer and are often lost during tumor development. Papa et al. show that partial loss of normal PTEN tumor suppressor function can be compounded by additional disruption caused by the expression of inactive mutant PTEN protein. This has significant

  18. Generation and characterization of pigment mutants of ...

    African Journals Online (AJOL)

    The result of bio-test, using the resulting pigment mutant of C. reinhardtii 124y-1 showed that mutagenic activity was observed significantly in both Tekeli River and Pavlodar Oil Refinery in Kazakhstan; the waste water of the Pavlodar Oil Refinery had high-toxicity while the water of the Tekeli River had medium-toxicity.

  19. Crystal Structures of Wild-type and Mutant Methicillin-resistant Staphylococcus aureus Dihydrofolate Reductase Reveal an Alternative Conformation of NADPH that may be Linked to Trimethoprim Resistance

    Energy Technology Data Exchange (ETDEWEB)

    Frey, K.; Liu, J; Lombardo, M; Bolstad, D; Wright, D; Anderson, A

    2009-01-01

    Both hospital- and community-acquired Staphylococcus aureus infections have become major health concerns in terms of morbidity, suffering and cost. Trimethoprim-sulfamethoxazole (TMP-SMZ) is an alternative treatment for methicillin-resistant S. aureus (MRSA) infections. However, TMP-resistant strains have arisen with point mutations in dihydrofolate reductase (DHFR), the target for TMP. A single point mutation, F98Y, has been shown biochemically to confer the majority of this resistance to TMP. Using a structure-based approach, we have designed a series of novel propargyl-linked DHFR inhibitors that are active against several trimethoprim-resistant enzymes. We screened this series against wild-type and mutant (F98Y) S. aureus DHFR and found that several are active against both enzymes and specifically that the meta-biphenyl class of these inhibitors is the most potent. In order to understand the structural basis of this potency, we determined eight high-resolution crystal structures: four each of the wild-type and mutant DHFR enzymes bound to various propargyl-linked DHFR inhibitors. In addition to explaining the structure-activity relationships, several of the structures reveal a novel conformation for the cofactor, NADPH. In this new conformation that is predominantly associated with the mutant enzyme, the nicotinamide ring is displaced from its conserved location and three water molecules complete a network of hydrogen bonds between the nicotinamide ring and the protein. In this new position, NADPH has reduced interactions with the inhibitor. An equilibrium between the two conformations of NADPH, implied by their occupancies in the eight crystal structures, is influenced both by the ligand and the F98Y mutation. The mutation induced equilibrium between two NADPH-binding conformations may contribute to decrease TMP binding and thus may be responsible for TMP resistance.

  20. Equations for arithmetic pointed tori

    NARCIS (Netherlands)

    Sijsling, J.R.

    2010-01-01

    In 1983, Kisao Takeuchi enumerated all 71 arithmetic (1;e)-groups. This is a special set of discrete subgroups of SL(2,R) of finite covolume and signature (1;e). The corresponding quotients of the upper half plane (called (1;e)-curves) have genus equal to 1 and a single elliptic point of order e.

  1. Phanerochaete mutants with enhanced ligninolytic activity

    International Nuclear Information System (INIS)

    Kakar, S.N.; Perez, A.; Gonzales, J.

    1994-01-01

    In addition to lignin, the white rot fungus Phanerochaete chrysosporium has the ability to degrade a wide spectrum of recalcitrant organo pollutants in soils and aqueous media. Most of the organic compounds are degraded under ligninolytic conditions with the involvement of the extracellular enzymes, lignin peroxidases, and manganese-dependent peroxidases, which are produced as secondary metabolites triggered by conditions of nutrient starvation (e.g., nitrogen limitation). The fungus and its enzymes can thus provide alternative technologies for bioremediation, bio pulping, bio bleaching, and other industrial applications. The efficiency and effectiveness of the fungus can be enhanced by increasing production and secretion of the important enzymes in large quantities and as primary metabolites under enriched conditions. One way this can be achieved is through isolation of mutants that are deregulated, or are hyper producers or super secretors of key enzymes under enriched conditions. Through UV-light and γ-ray mutagenesis, we have isolated a variety of mutants, some of which produce key enzymes of the ligninolytic system under high-nitrogen growth conditions. One of the mutants, 76UV, produced 272 U of lignin peroxidases enzyme activity/L after 9 d under high nitrogen (although the parent strain does not produce this enzyme under these conditions). The mutant and the parent strains produced up to 54 and 62 U/L, respectively, of the enzyme activity under low nitrogen growth conditions during this period. In some experiments, the mutant showed 281 U/L of enzyme activity under high nitrogen after 17 d

  2. Address Points - Allegheny County Address Points 201601

    Data.gov (United States)

    NSGIC Education | GIS Inventory — This dataset contains Address Points in Allegheny County. The Address Points were created by GDR for the Allegheny County CAD project, October 2008. Data is updated...

  3. Structural dataset for the PPARγ V290M mutant

    Directory of Open Access Journals (Sweden)

    Ana C. Puhl

    2016-06-01

    Full Text Available Loss-of-function mutation V290M in the ligand-binding domain of the peroxisome proliferator activated receptor γ (PPARγ is associated with a ligand resistance syndrome (PLRS, characterized by partial lipodystrophy and severe insulin resistance. In this data article we discuss an X-ray diffraction dataset that yielded the structure of PPARγ LBD V290M mutant refined at 2.3 Å resolution, that allowed building of 3D model of the receptor mutant with high confidence and revealed continuous well-defined electron density for the partial agonist diclofenac bound to hydrophobic pocket of the PPARγ. These structural data provide significant insights into molecular basis of PLRS caused by V290M mutation and are correlated with the receptor disability of rosiglitazone binding and increased affinity for corepressors. Furthermore, our structural evidence helps to explain clinical observations which point out to a failure to restore receptor function by the treatment with a full agonist of PPARγ, rosiglitazone.

  4. Bacillus Pumilus Cyanide Dihydratase Mutants with Higher Catalytic Activity

    Directory of Open Access Journals (Sweden)

    Mary A Crum

    2016-08-01

    Full Text Available Cyanide degrading nitrilases are noted for their potential to detoxify industrial wastewater contaminated with cyanide. However, such application would benefit from an improvement to characteristics such as their catalytic activity and stability. Following error-prone PCR for random mutagenesis, several cyanide dihydratases mutants from Bacillus pumilus were isolated based on improved catalysis. Four point mutations, K93R, D172N, A202T and E327K were characterized and their effects on kinetics, thermostability and pH tolerance were studied. K93R and D172N increased the enzyme’s thermostability whereas E327K mutation had a less pronounced effect on stability. The D172N mutation also increased the affinity of the enzyme for its substrate at pH 7.7 but lowered its kcat. However, the A202T mutation, located in the dimerization or the A surface, destabilized the protein and abolished its activity. No significant effect on activity at alkaline pH was observed for any of the purified mutants. These mutations help confirm the model of CynD and are discussed in the context of the protein-protein interfaces leading to the protein quaternary structure.

  5. Normal DNA ligase activity in a γ-ray-sensitive Chinese hamster mutant

    International Nuclear Information System (INIS)

    Stamato, T.D.; Hu, J.

    1987-01-01

    A Chinese hamster cell mutant (XR-1) was previously described that is extremely deficient in the repair of double-strand DNA breaks produced by γ-irradiation during the sensitive G 1 -early-S period and somewhat deficient in repair of γ-ray-induced single-strand DNA breaks. To determine whether a deficiency in DNA ligase activity might underlie the biochemical defect, protein extracts from mutant and parental cells were examined for their ability to ligate single- and double-strand breaks in DNA. The kinetics of ligation of single 5'-phosphate-3'-hydroxyl breaks in double-stranded DNA were the same in protein extracts from both cells. After separation of protein extracts by gel-filtration chromatography, the percentage of activity in the large and small molecular forms of DNA ligase was also similar in the two cells. Finally, protein extracts prepared from exponentially growing or G 1 -synchronized mutant and parental cells were equal in their ability to ligate blunt-end DNA substrates. These data suggest that a deficiency in DNA ligase is not the cause of the repair defect in the XR-1 mutant cell. (Auth.)

  6. Access to Liver Transplantation in Different ABO-Blood Groups and "Exceptions Points" in a Model for End-Stage Liver Disease Allocation System: A Brazilian Single-Center Study.

    Science.gov (United States)

    Martino, R B; Waisberg, D R; Dias, A P M; Inoue, V B S; Arantes, R M; Haddad, L B P; Rocha-Santos, V; Pinheiro, R S N; Nacif, L S; D'Albuquerque, L A C

    2018-04-01

    In the Model for End-Stage Liver Disease (MELD) system, patients with "MELD exceptions" points may have unfair privilege in the competition for liver grafts. Furthermore, organ distribution following identical ABO blood types may also result in unjust organ allocation. The aim of this study was to investigate access to liver transplantation in a tertiary Brazilian center, regarding "MELD exceptions" situations and among ABO-blood groups. A total of 465 adult patients on the liver waitlist from August 2015 to August 2016 were followed up until August 2017. Patients were divided into groups according to ABO-blood type and presence of "exceptions points." No differences in outcomes were observed among ABO-blood groups. However, patients from B and AB blood types spent less time on the list than patients from A and O groups (median, 46, 176, 415, and 401 days, respectively; P = .03). "Exceptions points" were granted for 141 patients (30.1%), hepatocellular carcinoma being the most common reason (52.4%). Patients with "exceptions points" showed higher transplantation rate, lower mortality on the list, and lower delta-MELD than non-exceptions patients (56.7% vs 19.1% [P blood types, despite shorter time on the waitlist for AB and B groups. The current MELD exception system provides advantages for candidates with "exception points," resulting in superior outcomes compared with those without exceptions. Copyright © 2018 Elsevier Inc. All rights reserved.

  7. Relative Critical Points

    Directory of Open Access Journals (Sweden)

    Debra Lewis

    2013-05-01

    Full Text Available Relative equilibria of Lagrangian and Hamiltonian systems with symmetry are critical points of appropriate scalar functions parametrized by the Lie algebra (or its dual of the symmetry group. Setting aside the structures – symplectic, Poisson, or variational – generating dynamical systems from such functions highlights the common features of their construction and analysis, and supports the construction of analogous functions in non-Hamiltonian settings. If the symmetry group is nonabelian, the functions are invariant only with respect to the isotropy subgroup of the given parameter value. Replacing the parametrized family of functions with a single function on the product manifold and extending the action using the (coadjoint action on the algebra or its dual yields a fully invariant function. An invariant map can be used to reverse the usual perspective: rather than selecting a parametrized family of functions and finding their critical points, conditions under which functions will be critical on specific orbits, typically distinguished by isotropy class, can be derived. This strategy is illustrated using several well-known mechanical systems – the Lagrange top, the double spherical pendulum, the free rigid body, and the Riemann ellipsoids – and generalizations of these systems.

  8. Tipping point leadership.

    Science.gov (United States)

    Kim, W Chan; Mauborgne, Renée

    2003-04-01

    When William Bratton was appointed police commissioner of New York City in 1994, turf wars over jurisdiction and funding were rife and crime was out of control. Yet in less than two years, and without an increase in his budget, Bratton turned New York into the safest large city in the nation. And the NYPD was only the latest of five law-enforcement agencies Bratton had turned around. In each case, he succeeded in record time despite limited resources, a demotivated staff, opposition from powerful vested interests, and an organization wedded to the status quo. Bratton's turnarounds demonstrate what the authors call tipping point leadership. The theory of tipping points hinges on the insight that in any organization, fundamental changes can occur quickly when the beliefs and energies of a critical mass of people create an epidemic movement toward an idea. Bratton begins by overcoming the cognitive hurdles that block organizations from recognizing the need for change. He does this by putting managers face-to-face with operational problems. Next, he manages around limitations on funds, staff, or equipment by concentrating resources on the areas that are most in need of change and that have the biggest payoffs. He meanwhile solves the motivation problem by singling out key influencers--people with disproportionate power due to their connections or persuasive abilities. Finally, he closes off resistance from powerful opponents. Not every CEO has the personality to be a Bill Bratton, but his successes are due to much more than his personality. He relies on a remarkably consistent method that any manager looking to turn around an organization can use to overcome the forces of inertia and reach the tipping point.

  9. Expression profiling of S. pombe acetyltransferase mutants identifies redundant pathways of gene regulation

    Directory of Open Access Journals (Sweden)

    Wright Anthony PH

    2010-01-01

    Full Text Available Abstract Background Histone acetyltransferase enzymes (HATs are implicated in regulation of transcription. HATs from different families may overlap in target and substrate specificity. Results We isolated the elp3+ gene encoding the histone acetyltransferase subunit of the Elongator complex in fission yeast and characterized the phenotype of an Δelp3 mutant. We examined genetic interactions between Δelp3 and two other HAT mutants, Δmst2 and Δgcn5 and used whole genome microarray analysis to analyze their effects on gene expression. Conclusions Comparison of phenotypes and expression profiles in single, double and triple mutants indicate that these HAT enzymes have overlapping functions. Consistent with this, overlapping specificity in histone H3 acetylation is observed. However, there is no evidence for overlap with another HAT enzyme, encoded by the essential mst1+ gene.

  10. A thermo-sensitive purple leaf rice mutant--PLM12 and its genetical study

    International Nuclear Information System (INIS)

    Wu Guanting; Wang Xianyu; Jin Wei

    2001-01-01

    PLM12 was a thermo-sensitive purple leaf mutant selected from Indica rice variety Luqingzao 1 treated with pingyangmycin in combination with γ-rays, and for display of its mutant character, a relatively high temperature was required. Compared with its original parent, many major agronomic traits of PLM12 changed to varied extents. Based on spikelet cutting experiment, it was believed that significant decreases in number of filled grains per panicle, fertility, and 1000-grain weight in PLM12 resulted mainly from a great decline in photosynthetic capacity and serious lack of photosynthate in purple leaves. It was indicated by genetic analysis that expression of the mutant phenotype in PLM12 was conditioned by a single recessive major nuclear gene and modified by several minor genes

  11. Origins and spread of pfdhfr mutant alleles in Plasmodium falciparum.

    Science.gov (United States)

    Mita, Toshihiro

    2010-06-01

    The emergence and spread of Plasmodium falciparum parasite resistant to sulfadoxine and pyrimethamine (SP) poses a serious public health problem. Resistance is caused by point mutations in dihydrofolate reductase (pfdhfr) and dihydropteroate synthase (pfdhps), the two key enzymes in the folate biosynthetic pathway. The use of microsatellite markers flanking pfdhfr has recently shown that the invasion of limited resistant lineages may explain the widespread SP resistance in many endemic regions. In Africa, however, multiple indigenous origins of pfdhfr triple mutants have been demonstrated. More new independent lineages and routes of geographical spread of resistance may be found by further molecular evolutionary analyses using samples from various endemic regions. Here, I review recent studies about the history of SP usage and the evolution and spread of resistant lineages while addressing the technical issue of microsatellite analysis. Copyright (c) 2009 Elsevier B.V. All rights reserved.

  12. Photosynthesis, Light Use Efficiency, and Yield of Reduced-Chlorophyll Soybean Mutants in Field Conditions

    Science.gov (United States)

    Slattery, Rebecca A.; VanLoocke, Andy; Bernacchi, Carl J.; Zhu, Xin-Guang; Ort, Donald R.

    2017-01-01

    Reducing chlorophyll (chl) content may improve the conversion efficiency of absorbed photosynthetically active radiation into biomass and therefore yield in dense monoculture crops by improving light penetration and distribution within the canopy. The effects of reduced chl on leaf and canopy photosynthesis and photosynthetic efficiency were studied in two reportedly robust reduced-chl soybean mutants, Y11y11 and y9y9, in comparison to the wild-type (WT) “Clark” cultivar. Both mutants were characterized during the 2012 growing season whereas only the Y11y11 mutant was characterized during the 2013 growing season. Chl deficiency led to greater rates of leaf-level photosynthesis per absorbed photon early in the growing season when mutant chl content was ∼35% of the WT, but there was no effect on photosynthesis later in the season when mutant leaf chl approached 50% of the WT. Transient benefits of reduced chl at the leaf level did not translate to improvements in canopy-level processes. Reduced pigmentation in these mutants was linked to lower water use efficiency, which may have dampened any photosynthetic benefits of reduced chl, especially since both growing seasons experienced significant drought conditions. These results, while not confirming our hypothesis or an earlier published study in which the Y11y11 mutant significantly outyielded the WT, do demonstrate that soybean significantly overinvests in chl. Despite a >50% chl reduction, there was little negative impact on biomass accumulation or yield, and the small negative effects present were likely due to pleiotropic effects of the mutation. This outcome points to an opportunity to reinvest nitrogen and energy resources that would otherwise be used in pigment-proteins into increasing biochemical photosynthetic capacity, thereby improving canopy photosynthesis and biomass production. PMID:28458677

  13. Design and characterization of genetically engineered zebrafish aquaporin-3 mutants highly permeable to the cryoprotectant ethylene glycol

    Directory of Open Access Journals (Sweden)

    Lubzens Esther

    2011-04-01

    Full Text Available Abstract Background Increasing cell membrane permeability to water and cryoprotectants is critical for the successful cryopreservation of cells with large volumes. Artificial expression of water-selective aquaporins or aquaglyceroporins (GLPs, such as mammalian aquaporin-3 (AQP3, enhances cell permeability to water and cryoprotectants, but it is known that AQP3-mediated water and solute permeation is limited and pH dependent. To exploit further the possibilities of using aquaporins in cryobiology, we investigated the functional properties of zebrafish (Danio rerio GLPs. Results Water, glycerol, propylene glycol and ethylene glycol permeability of zebrafish Aqp3a, -3b, -7, -9a, -9b, -10a and -10b, and human AQP3, was examined. Expression in Xenopus laevis oocytes indicated that the permeability of DrAqp3a and -3b to ethylene glycol was higher than for glycerol or propylene glycol under isotonic conditions, unlike other zebrafish GLPs and human AQP3, which were more permeable to glycerol. In addition, dose-response experiments and radiolabeled ethylene glycol uptake assays suggested that oocytes expressing DrAqp3b were permeated by this cryoprotectant more efficiently than those expressing AQP3. Water and ethylene glycol transport through DrAqp3a and -3b were, however, highest at pH 8.5 and completely abolished at pH 6.0. Point mutations in the DrAqp3b amino acid sequence rendered two constructs, DrAqp3b-T85A showing higher water and ethylene glycol permeability at neutral and alkaline pH, and DrAqp3b-H53A/G54H/T85A, no longer inhibited at acidic pH but less permeable than the wild type. Finally, calculation of permeability coefficients for ethylene glycol under concentration gradients confirmed that the two DrAqp3b mutants were more permeable than wild-type DrAqp3b and/or AQP3 at neutral pH, resulting in a 2.6- to 4-fold increase in the oocyte intracellular concentration of ethylene glycol. Conclusion By single or triple point mutations in the Dr

  14. Mutants induction in sweet potato by irradiation of Co60 and selection, propagation, evaluation of desirable mutants

    International Nuclear Information System (INIS)

    Lu Shuyun

    1987-11-01

    Cuttings of the sweet potato cultivar Xu-18 were exposed to gamma rays in 1983. This cultivar is susceptible to ''sweet potato black rot'' caused by Ceratocystis fimbriata E11. et Halst. and leading to serious post-harvest losses of tubers. The irradiated material was vegetatively propagated to M 1 V 3 . Mutant selection was undertaken on single plants in M 1 V 2 and on clonal progenies in M 1 V 3 . Two promising clonal lines namely ''12-11-8'' and ''09-56'', possessing much better resistance to black rot than the original cultivar ''Xu-18'' were identified in 1985 and 1986. Their yield is comparable to the original cultivar. The mutant clones are now to be tested in different locations, where the disease is endemic. If the results of these tests are positive, the clones will be released as improved cultivars to the farmers. The experiment has been very successful and results of practical significance were obtained within the short period of only 4 years. 4 refs, 1 fig, 3 tabs

  15. A large-scale genetic screen for mutants with altered salicylic acid accumulation in Arabidopsis

    Directory of Open Access Journals (Sweden)

    Yezhang eDing

    2015-01-01

    Full Text Available Salicylic acid (SA is a key defense signal molecule against biotrophic and hemibiotrophic pathogens in plants, but how SA is synthesized in plant cells still remains elusive. Identification of new components involved in pathogen-induced SA accumulation would help address this question. To this end, we performed a large-scale genetic screen for mutants with altered SA accumulation during pathogen infection in Arabidopsis using a bacterial biosensor Acinetobacter sp. ADPWH_lux-based SA quantification method. A total of 35,000 M2 plants in the npr1-3 mutant background have been individually analyzed for the bacterial pathogen Pseudomonas syringae pv. maculicola (Psm ES4326-induced SA accumulation. Among the mutants isolated, 19 had SA levels lower than npr1 (sln and two exhibited increased SA accumulation in npr1 (isn. Complementation tests revealed that seven of the sln mutants are new alleles of eds5/sid1, two are sid2/eds16 alleles, one is allelic to pad4, and the remaining seven sln and two isn mutants are new non-allelic SA accumulation mutants. Interestingly, a large group of mutants (in the npr1-3 background, in which Psm ES4326-induced SA levels were similar to those in the wild-type Columbia plants, were identified, suggesting that the signaling network fine-tuning pathogen-induced SA accumulation is complex. We further characterized the sln1 single mutant and found that Psm ES4326-induced defense responses were compromised in this mutant. These defense response defects could be rescued by exogenous SA, suggesting that SLN1 functions upstream of SA. The sln1 mutation was mapped to a region on the north arm of chromosome I, which contains no known genes regulating pathogen-induced SA accumulation, indicating that SLN1 likely encodes a new regulator of SA biosynthesis. Thus, the new sln and isn mutants identified in this genetic screen are valuable for dissecting the molecular mechanisms underlying pathogen-induced SA accumulation in plants.

  16. Molecular defect of isovaleryl-CoA dehydrogenase in the skunk mutant of silkworm, Bombyx mori.

    Science.gov (United States)

    Urano, Kei; Daimon, Takaaki; Banno, Yutaka; Mita, Kazuei; Terada, Tohru; Shimizu, Kentaro; Katsuma, Susumu; Shimada, Toru

    2010-11-01

    The isovaleric acid-emanating silkworm mutant skunk (sku) was first studied over 30 years ago because of its unusual odour and prepupal lethality. Here, we report the identification and characterization of the gene responsible for the sku mutant. Because of its specific features and symptoms similar to human isovaleryl-CoA dehydrogenase (IVD) deficiency, also known as isovaleric acidaemia, IVD dysfunction in silkworms was predicted to be responsible for the phenotype of the sku mutant. Linkage analysis revealed that the silkworm IVD gene (BmIVD) was closely linked to the odorous phenotype as expected, and a single amino acid substitution (G376V) was found in BmIVD of the sku mutant. To investigate the effect of the G376V substitution on BmIVD function, wild-type and sku-type recombinants were constructed with a baculovirus expression system and the subsequent enzyme activity of sku-type BmIVD was shown to be significantly reduced compared with that of wild-type BmIVD. Molecular modelling suggested that this reduction in the enzyme activity may be due to negative effects of G376V mutation on FAD-binding or on monomer-monomer interactions. These observations strongly suggest that BmIVD is responsible for the sku locus and that the molecular defect in BmIVD causes the characteristic smell and prepupal lethality of the sku mutant. To our knowledge, this is, aside from humans, the first characterization of IVD deficiency in metazoa. Considering that IVD acts in the third step of leucine degradation and the sku mutant accumulates branched-chain amino acids in haemolymph, this mutant may be useful in the investigation of unique branched-chain amino acid catabolism in insects. © 2010 The Authors Journal compilation © 2010 FEBS.

  17. Effects of Water Content Levels on Yield Components of Some Peanut Mutant Lines (Arachis Hypogaea L)

    International Nuclear Information System (INIS)

    Carkum; Kumala Dewi; Parno; Sobrizal

    2004-01-01

    Study about effects of water content levels on yield components of peanut mutant lines has been conducted. Mutant lines tested were A20/3/PsJ, B30/12/10 and D25/3/2. The original varieties/lines of the mutants were, AH1781Si, Kidang and Pelanduk was included as control. The treatments for levels of water application were 100%, 75%, 50% and 25%, where 100% water available means a difference between water content at field capacity and at content in permanent wilting point. Based on the data of parameters observed, to interaction between varieties/lines and water content levels was not found except for dry matter weight. For this parameter, the highest dry matter was observed when B30/12/10 mutant line was watered by 100% water available. Nevertheless, based on relative values of number of mature pod, dry pod weight and 50 nut weight the A20/3/PsJ mutant line seems tolerant to drought. (author)

  18. Targeting Gpr52 lowers mutant HTT levels and rescues Huntington's disease-associated phenotypes.

    Science.gov (United States)

    Song, Haikun; Li, Hexuan; Guo, Shimeng; Pan, Yuyin; Fu, Yuhua; Zhou, Zijian; Li, Zhaoyang; Wen, Xue; Sun, Xiaoli; He, Bingqing; Gu, Haifeng; Zhao, Quan; Wang, Cen; An, Ping; Luo, Shouqing; Hu, Youhong; Xie, Xin; Lu, Boxun

    2018-03-28

    Lowering the levels of disease-causing proteins is an attractive treatment strategy for neurodegenerative disorders, among which Huntington's disease is an appealing disease for testing this strategy because of its monogenetic nature. Huntington's disease is mainly caused by cytotoxicity of the mutant HTT protein with an expanded polyglutamine repeat tract. Lowering the soluble mutant HTT may reduce its downstream toxicity and provide potential treatment for Huntington's disease. This is hard to achieve by small-molecule compound drugs because of a lack of effective targets. Here we demonstrate Gpr52, an orphan G protein-coupled receptor, as a potential Huntington's disease drug target. Knocking-out Gpr52 significantly reduces mutant HTT levels in the striatum and rescues Huntington's disease-associated behavioural phenotypes in a knock-in Huntington's disease mouse model expressing endogenous mutant Htt. Importantly, a novel Gpr52 antagonist E7 reduces mutant HTT levels and rescues Huntington's disease-associated phenotypes in cellular and mouse models. Our study provides an entry point for Huntington's disease drug discovery by targeting Gpr52.

  19. Hair defects in Hoxc13 mutant mice.

    Science.gov (United States)

    Godwin, A R; Capecchi, M R

    1999-12-01

    Hox genes encode transcription factors that are important during normal embryonic development of diverse organisms including vertebrates. In mammals, Hox genes are responsible for conferring regional identity in embryonic tissues, including the limb bud, the neural tube, the presomitic mesoderm and the intestinal tract. Recent studies have demonstrated expression of Hox genes in skin and hair follicles, suggesting potential functions for these genes in epidermal appendages. These studies are reviewed here with emphasis on Hoxc13, as Hoxc13 mutants are the first Hox mutants to demonstrate overt hair defects. In addition, because Hoxc13 does not show regionally restricted expression in the skin, as demonstrated for other Hox genes, the potentially different roles of Hoxc13 versus other Hox genes in the skin are discussed.

  20. PNRI mutant variety: sansevieria 'Sword of Ibe'

    International Nuclear Information System (INIS)

    Aurigue, Fernando B.

    2011-01-01

    Sansevieria 'Sword of Ibe,' registered by the Philippine Nuclear Research Institute as NSIC 2008 Or-66, is a chlorophyll mutant of Sansevieria trifasciata 'Moonshine' developed by treating its suckers or shoots arising from a rhizome with acute gamma radiation from a Cobalt-60 source. The new mutant is identical in growth habit and vigor to Sansevieria 'Moonshine,' also known as Moonglow. Results of this mutation breeding experiment showed that leaf color and flowering were altered by gamma irradiation without changing the other characteristics of the plant. Propagation is true-to-type by separation of sucker and top cutting. The plant is recommended for use as landscaping material and as pot plant for indoor and outdoor use. The leaves may be harvested as cut foliage for Japanese flower arrangements. (author)

  1. A Meiotic Uv-Sensitive Mutant That Causes Deletion of Duplications in Neurospora

    Science.gov (United States)

    Newmeyer, Dorothy; Galeazzi, Donna R.

    1978-01-01

    The meiotic-3 (mei-3) mutant of Neurospora crassa has several effects: (1) When homozygous, it almost completely blocks meiosis and ascospore formation, (2) it is sensitive to UV, (3) its growth is inhibited by histidine and, (4) it increases the instability of nontandem duplications. This was shown for duplications produced by five different rearrangements and was demonstrated by two different criteria. The effects on meiosis and duplication instability are expressed strongly at 25°; the effects on sensitivity to UV and to histidine are expressed strongly at 38.5° but only slightly at 25°. Nevertheless, all four effects were shown to be due to a single gene. mei-3 is not allelic with previously reported UV-sensitive mutants.—Two other results were obtained that are not necessarily due to mei-3: (1) A cross involving mei-3 produced a new unlinked meiotic mutant, mei-4, which is not sensitive to UV or histidine, and (2) a burst of several new mutants occurred in a different mei-3 stock, including a partial revertant of mei-3.—mei-3 has previously been shown to cause frequent complete loss of a terminal duplicate segment, beginning exactly at the original rearrangement breakpoint. Possible mechanisms are discussed by which a UV-sensitive mutant could cause such precise deletions. PMID:17248837

  2. Genetic Analysis and Mapping of TWH Gene in Rice Twisted Hull Mutant

    Directory of Open Access Journals (Sweden)

    Jin-bo LI

    2009-03-01

    Full Text Available A mutant with twisted hulls was found in a breeding population of rice (Oryza sativa L.. The mutant shows less grain weight and inferior grain quality in addition to twisted hulls. Genetic analysis indicated that the phenotype of mutant was controlled by a single recessive gene (temporarily designated as TWH. To map the TWH gene, an F2 population was generated by crossing the twh mutant to R725, an indica rice variety with normal hulls. For bulked segregant analysis, the bulk of mutant plants was prepared by mixing equal amount of plant tissue from 10 twisted-hull plants and the bulk of normal plants was obtained by pooling equal amount tissue of 10 normal-hull plants. Two hundred and seven pairs of simple sequence repeat (SSR primers, which are distributed on 12 rice chromosomes, were used for polymorphism analysis of the parents and the two bulks. The TWH locus was initially mapped close to the SSR marker RM526 on chromosome 2. Therefore, further mapping was performed using 50 pairs of SSR primers around the marker RM526. The TWH was delimited between the SSR markers RM14128 and RM208 on the long arm of chromosome 2 at the genetic distances of 1.4 cM and 2.7 cM, respectively. These results provide the foundation for further fine mapping, cloning and functional analysis of the TWH gene.

  3. Detection of a true breeding homeotic gene mutant Pps-1 with partially petaloid sepals in opium poppy (Papaver somniferum L.) and its genetic behavior.

    Science.gov (United States)

    Dhawan, Om Prakash; Dubey, Mukesh Kumar; Khanuja, Suman Preet Singh

    2007-01-01

    A spontaneous true breeding homeotic gene mutant Pps-1 with distinct partial petaloid sepals was detected in the population of downy mildew (DM)-resistant elite accession I-14 during our studies for the identification of disease resistance sources in opium poppy. The trait was found to be stable and inherited truly in the subsequent generations. Genetic studies were carried out through systematic reciprocal crosses with the parental wild-type genotype I-14, and segregation pattern of phenotypic characteristics in F(1) and F(2) populations clearly indicated single recessive nuclear gene control of the mutant character. The studies have demonstrated that the mutant phenotype is due to mutations at the Pps-1 locus that possibly corresponds to B-class function (according to ABC model) with negative control function. The mutant Pps-1 being single-whorl homeotic mutant might greatly help in providing insight into mechanisms of flower development in opium poppy.

  4. Recombination-deficient mutant of Streptococcus faecalis

    International Nuclear Information System (INIS)

    Yagi, Y.; Clewell, D.B.

    1980-01-01

    An ultraviolet radiation-sensitive derivative of Streptococcus faecalis strain JH2-2 was isolated and found to be deficient in recombination, using a plasmid-plasmid recombination system. The strain was sensitive to chemical agents which interact with deoxyribonucleic acid and also underwent deoxyribonucleic acid degradation after ultraviolet irradiation. Thus, the mutant has properties similar to those of recA strains of Escherichia coli

  5. Characterization of a Legionella micdadei mip mutant

    DEFF Research Database (Denmark)

    O'Connell, W A; Bangsborg, Jette Marie; Cianciotto, N P

    1995-01-01

    The pathogenesis of Legionella micdadei is dependent upon its ability to infect alveolar phagocytes. To better understand the basis of intracellular infection by this organism, we examined the importance of its Mip surface protein. In Legionella pneumophila, Mip promotes infection of both human m...... into the phagocyte. Similarly, the mutant was less able to parasitize Hartmannella amoebae. Taken together, these data argue that Mip specifically potentiates intracellular growth by L. micdadei....

  6. Selection of hyperadherent mutants in Pseudomonas putida biofilms

    DEFF Research Database (Denmark)

    Yousef-Coronado, Fatima; Soriano, María Isabel; Yang, Liang

    2011-01-01

    transposon Pseudomonas putida KT2440 mutants showing increased biofilm formation, and the detailed characterization of one of them. This mutant exhibits a complex phenotype, including altered colony morphology, increased production of extracellular polymeric substances and enhanced swarming motility, along...

  7. Whole-genome analysis of herbicide-tolerant mutant rice generated by Agrobacterium-mediated gene targeting.

    Science.gov (United States)

    Endo, Masaki; Kumagai, Masahiko; Motoyama, Ritsuko; Sasaki-Yamagata, Harumi; Mori-Hosokawa, Satomi; Hamada, Masao; Kanamori, Hiroyuki; Nagamura, Yoshiaki; Katayose, Yuichi; Itoh, Takeshi; Toki, Seiichi

    2015-01-01

    Gene targeting (GT) is a technique used to modify endogenous genes in target genomes precisely via homologous recombination (HR). Although GT plants are produced using genetic transformation techniques, if the difference between the endogenous and the modified gene is limited to point mutations, GT crops can be considered equivalent to non-genetically modified mutant crops generated by conventional mutagenesis techniques. However, it is difficult to guarantee the non-incorporation of DNA fragments from Agrobacterium in GT plants created by Agrobacterium-mediated GT despite screening with conventional Southern blot and/or PCR techniques. Here, we report a comprehensive analysis of herbicide-tolerant rice plants generated by inducing point mutations in the rice ALS gene via Agrobacterium-mediated GT. We performed genome comparative genomic hybridization (CGH) array analysis and whole-genome sequencing to evaluate the molecular composition of GT rice plants. Thus far, no integration of Agrobacterium-derived DNA fragments has been detected in GT rice plants. However, >1,000 single nucleotide polymorphisms (SNPs) and insertion/deletion (InDels) were found in GT plants. Among these mutations, 20-100 variants might have some effect on expression levels and/or protein function. Information about additive mutations should be useful in clearing out unwanted mutations by backcrossing. © The Author 2014. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists.

  8. Multivariate analysis for selecting apple mutants

    International Nuclear Information System (INIS)

    Faedi, W.; Bagnara, G.L.; Rosati, P.; Cecchini, M.

    1992-01-01

    The mutlivariate analysis of four year records on several vegetative and productive traits of twenty-one apple mutants (3 of 'Jonathan', 3 of 'Ozark Gold', 14 of 'Mollie's Delicious', 1 of 'Neipling's Early Stayman)' induced by gamma radiations showed that observation of some traits of one-year-old shoots is the most efficient way to reveal compact growing apple mutants. In particular, basal cross-section area, total length and leaf area resulted the most appropriate parameters, while internode length together with conopy height and width are less appropriate. The most interesting mutants we found are: one of 'Mollie's Delicious for the best balance among tree and fruit traits and for high skin color; one of 'Neipling's Early Stayman' with an earlier and more extensively red colored apple than the original clone. (author)

  9. Probiotic features of Lactobacillus plantarum mutant strains.

    Science.gov (United States)

    Bove, Pasquale; Gallone, Anna; Russo, Pasquale; Capozzi, Vittorio; Albenzio, Marzia; Spano, Giuseppe; Fiocco, Daniela

    2012-10-01

    In this study, the probiotic potential of Lactobacillus plantarum wild-type and derivative mutant strains was investigated. Bacterial survival was evaluated in an in vitro system, simulating the transit along the human oro-gastro-intestinal tract. Interaction with human gut epithelial cells was studied by assessing bacterial adhesive ability to Caco-2 cells and induction of genes involved in innate immunity. L. plantarum strains were resistant to the combined stress at the various steps of the simulated gastrointestinal tract. Major decreases in the viability of L. plantarum cells were observed mainly under drastic acidic conditions (pH ≤ 2.0) of the gastric compartment. Abiotic stresses associated to small intestine poorly affected bacterial viability. All the bacterial strains significantly adhered to Caco-2 cells, with the ΔctsR mutant strain exhibiting the highest adhesion. Induction of immune-related genes resulted higher upon incubation with heat-inactivated bacteria rather than with live ones. For specific genes, a differential transcriptional pattern was observed upon stimulation with different L. plantarum strains, evidencing a possible role of the knocked out bacterial genes in the modulation of host cell response. In particular, cells from Δhsp18.55 and ΔftsH mutants strongly triggered immune defence genes. Our study highlights the relevance of microbial genetic background in host-probiotic interaction and might contribute to identify candidate bacterial genes and molecules involved in probiosis.

  10. Grain product of 34 soya mutant lines

    International Nuclear Information System (INIS)

    Salmeron E, J.; Mastache L, A. A.; Valencia E, F.; Diaz V, G. E.; Cervantes S, T.; De la Cruz T, E.; Garcia A, J. M.; Falcon B, T.; Gatica T, M. A.

    2009-01-01

    This work was development with the objective of obtaining information of the agronomic behavior of 34 soya mutant lines (R 4 M 18 ) for human consumption and this way to select the 2 better lines. The genetic materials were obtained starting from the variety ISAAEG-B M2 by means of the application of recurrent radiation with Co 60 gammas, to a dose of 350 Gray for the first two generations and both later to 200 Gray and selection during 17 cycles, being obtained the 34 better lines mutants with agronomic characteristic wanted and good flavor. The obtained results were that the mutant lines L 25 and L 32 produced the major quantity in branches/plant number with 7.5 and 7.25, pods/plant number with 171.25 and 167, grains/plant number with 350.89 and 333.07 and grain product (ton/ha) to 15% of humidity 5.15 and 4.68 ton/ha, respectively. (Author)

  11. Agrobacterium rhizogenes mutants that fail to bind to plant cells.

    OpenAIRE

    Crews, J L; Colby, S; Matthysse, A G

    1990-01-01

    Transposon insertion mutants of Agrobacterium rhizogenes were screened to obtain mutant bacteria that failed to bind to carrot suspension culture cells. A light microscope binding assay was used. The bacterial isolates that were reduced in binding to carrot cells were all avirulent on Bryophyllum diagremontiana leaves and on carrot root disks. The mutants did not appear to be altered in cellulose production. The composition of the medium affected the ability of the parent and mutant bacteria ...

  12. Point mutations in segment I-S6 render voltage-gated Na+ channels resistant to batrachotoxin.

    Science.gov (United States)

    Wang, S Y; Wang, G K

    1998-03-03

    Batrachotoxin (BTX) is a steroidal alkaloid that causes Na+ channels to open persistently. This toxin has been used widely as a tool for studying Na+ channel gating processes and for estimating Na+ channel density. In this report we used point mutations to identify critical residues involved in BTX binding and to examine if such mutations affect channel gating. We show that a single asparagine --> lysine substitution of the rat muscle Na+ channel alpha-subunit, mu1-N434K, renders the channel completely insensitive to 5 microM BTX when expressed in mammalian cells. This mutant channel nonetheless displays normal current kinetics with minimal changes in gating properties. Another substitution, mu1-N434A, yields a partial BTX-sensitive mutant. Unlike wild-type currents, the BTX-modified mu1-N434A currents continue to undergo fast and slow inactivation as if the inactivation processes remain functional. This finding implies that the mu1-N434 residue upon binding with BTX is critical for subsequent changes on gating; alanine at the mu1-434 position apparently diminishes the efficacy of BTX on eliminating Na+ channel inactivation. Mutants of two adjacent residues, mu1-I433K and mu1-L437K, also were found to exhibit the identical BTX-resistant phenotype. We propose that the mu1-I433, mu1-N434, and mu1-L437 residues in transmembrane segment I-S6 probably form a part of the BTX receptor.

  13. DNA sequence analysis of HPRT- mutants induced in human lymphoblastoid cells adapted to ionizing radiation

    International Nuclear Information System (INIS)

    Rigaud, O.; Laquerbe, A.; Moustacchi, E.

    1995-01-01

    Radioadaptation to the mutagenic effect of ionizing radiation by pre-exposure of human cells to a low dose has been shown to decrease the proportion of HPRT - mutants of the deletion type. To determine whether point mutations would be affected by the adaptive treatment, the molecular nature of mutations induced after exposure to low, high or low plus high doses was established. DNA sequencing of 38 point mutants which still expressed mRNA was performed using reverse transcription/polymerase chain reaction amplification. Under all conditions, base substitutions were the most common mutational event (range 72-80%), the remainder being frameshift and small deletions. The types and proportions of base changes did not appear to be differentially modified. A clustering of mutations was observed in exon 8, independently of the radiation protocol. About 40% of the mutants exhibited incorrect splicing of mRNA. The lack of striking modifications between the different molecular spectra of point mutations suggests that low-dose pre-exposure does not affect the production and/or the processing of lesions leading to point mutations. Thus the highly significant effect triggered by the low dose is the preferential reduction of deletion-type mutations. In view of the actual small data set, definitive conclusions will be drawn only when our observations are confirmed or can be generalized to human endogenous loci other than the HPRT locus, which is particularly prone to the recovery of deletion-type mutations. 37 refs., 1 fig., 5 tabs

  14. Evaluation of soybean mutants evolved from gamma irradiation

    International Nuclear Information System (INIS)

    Naseri Tafti, M.; Yousefi, F.; Rezazadeh, M.; Sabzi, H.; Ojani, R.

    2003-01-01

    Pure early soybean mutants evolved through mutagenesis (Co-60) from cultivar Clark irradiated with doses 100 Gy, 150 Gy and 250 Gy (absorbed dose) were evaluated for agronomic al traits and compared with two commercial cultivars; Clark and Williams in two regions, Karaj and Alishtar. Experimental design was conducted in a simple lattice (7 m x 7 m) with two replications. A significant statistical difference in yield existed at 1 and 5 percent level among mutants lines and between mutants - Williams and mutants - Clark, respectively in Karaj. The mutant line number 47 placed itself at the top of the list with yield of 4782 Kg/hect., followed by mutant line number 38 with 4722 Kg/hect. A number of mutant lines matured between 10 to 12 days earlier than the commercial soybean cultivars used as checks in the experiment. In Alishtar seed yield of mutant lines compared to the cultivar Williams showed a significant difference at 5% level. The highest seed yield of 3147 Kg/hect. belonged to the mutant line 47 which also matured two weeks earlier compared to the cultivar Clark. The compound analysis of seed yield in Karaj and Alishtar showed superiority of 15 mutant lines over the cultivar Clark and 36 mutant lines over the cultivar Williams. The mutant line number 18 producing seed yield of 3643 Kg/hect. ranks first in the list while, it matured earlier than both check cultivars, Clark and Williams

  15. Isolation and characterization of gallium resistant Pseudomonas aeruginosa mutants

    NARCIS (Netherlands)

    García-Contreras, R; Lira-Silva, E; Jasso-Chávez, R; Hernández-González, I.L.; Maeda, T.; Hashimoto, T.; Boogerd, F.C.; Sheng, L; Wood, TK; Moreno-Sánchez, R

    2013-01-01

    Pseudomonas aeruginosa PA14 cells resistant to the novel antimicrobial gallium nitrate (Ga) were developed using transposon mutagenesis and by selecting spontaneous mutants. The mutants showing the highest growth in the presence of Ga were selected for further characterization. These mutants showed

  16. Biological changes in Barley mutants resistant to powdery mildew disease

    International Nuclear Information System (INIS)

    Amer, I. M.; Fahim, M. M.; Moustafa, N. A.

    2012-12-01

    physiological studies showed that all kinds of chlorophyll (a), (b) and (a + b) content in infected plant were decreased while, the carotenes pigment were increased. Infection generally reduced total sugars content of all resistant mutants. Infected resistant mutant showed more phenols content and peroxidase, polyphenoloxidase activities than healthy ones of the mutants. (Author)

  17. Human GLTP and mutant forms of ACD11 suppress cell death in the Arabidopsis acd11 mutant

    DEFF Research Database (Denmark)

    Petersen, Nikolaj H T; McKinney, Lea V; Pike, Helen

    2008-01-01

    The Arabidopsis acd11 mutant exhibits runaway, programmed cell death due to the loss of a putative sphingosine transfer protein (ACD11) with homology to mammalian GLTP. We demonstrate that transgenic expression in Arabidopsis thaliana of human GLTP partially suppressed the phenotype of the acd11...... null mutant, resulting in delayed programmed cell death development and plant survival. Surprisingly, a GLTP mutant form impaired in glycolipid transfer activity also complemented the acd11 mutants. To understand the relationship between functional complementarity and transfer activity, we generated...... site-specific mutants in ACD11 based on homologous GLTP residues required for glycolipid transfer. We show that these ACD11 mutant forms are impaired in their in vitro transfer activity of sphingolipids. However, transgenic expression of these mutant forms fully complemented acd11 mutant cell death...

  18. New non detrimental DNA binding mutants of the Escherichia coli initiator protein DnaA

    DEFF Research Database (Denmark)

    Asklund, Marlene; Atlung, Tove

    2004-01-01

    The initiator protein DnaA has several unique DNA-binding features. It binds with high affinity as a monomer to the nonamer DnaA box. In the ATP form, DnaA binds cooperatively to the low-affinity ATP-DnaA boxes, and to single-stranded DNA in the 13mer region of the origin. We have carried out...... an extensive mutational analysis of the DNA-binding domain of the Escherichia coli DnaA protein using mutagenic PCR. We analyzed mutants exhibiting more or less partial activity by selecting for complementation of a dnaA(Ts) mutant strain at different expression levels of the new mutant proteins. The selection...... gave rise to 30 single amino acid substitutions and, including double substitutions, more than 100 mutants functional in initiation of chromosome replication were characterized. The analysis indicated that all regions of the DNA-binding domain are involved in DNA binding, but the most important amino...

  19. TagSmart: analysis and visualization for yeast mutant fitness data measured by tag microarrays

    Directory of Open Access Journals (Sweden)

    Xie Dan

    2007-04-01

    Full Text Available Abstract Background A nearly complete collection of gene-deletion mutants (96% of annotated open reading frames of the yeast Saccharomyces cerevisiae has been systematically constructed. Tag microarrays are widely used to measure the fitness of each mutant in a mutant mixture. The tag array experiments can have a complex experimental design, such as time course measurements and drug treatment with multiple dosages. Results TagSmart is a web application for analysis and visualization of Saccharomyces cerevisiae mutant fitness data measured by tag microarrays. It implements a robust statistical approach to assess the concentration differences among S. cerevisiae mutant strains. It also provides an interactive environment for data analysis and visualization. TagSmart has the following advantages over previously described analysis procedures: 1 it is user-friendly software rather than merely a description of analytical procedure; 2 It can handle complicated experimental designs, such as multiple time points and treatment with multiple dosages; 3 it has higher sensitivity and specificity; 4 It allows users to mask out "bad" tags in the analysis. Two biological tests were performed to illustrate the performance of TagSmart. First, we generated titration mixtures of mutant strains, in which the relative concentration of each strain was controlled. We used tag microarrays to measure the numbers of tag copies in each titration mixture. The data was analyzed with TagSmart and the result showed high precision and recall. Second, TagSmart was applied to a dataset in which heterozygous deletion strain mixture pools were treated with a new drug, Cincreasin. TagSmart identified 53 mutant strains as sensitive to Cincreasin treatment. We individually tested each identified mutant, and found 52 out of the 53 predicted mutants were indeed sensitive to Cincreasin. Conclusion TagSmart is provided "as is" to analyze tag array data produced by Affymetrix and Agilent

  20. [Ribosome engineering of streptomyces sp. FJ3 from Three Gorges reservoir area and metabolic product of the selected mutant strain].

    Science.gov (United States)

    Hai, Le; Huang, Yuqi; Liao, Guojian; Hu, Changhua

    2011-07-01

    To explore new resource from inactive actinomycete strains, we screened resistant mutant strains by ribosome engineering, and analyzed the products derived from the selected mutant strains. Three Gorges reservoir area-derived actinomycete strains including BD20, FJ3, WZ20 and FJ5 were used as initial strains, which showed no-antibacterial activities. The streptomycin-resistant (str(R)) mutants and rifampicin-resistant (rif(R)) mutants were screened by single colony isolation on streptomycin-containing plates and rifampicin-containing plates according to the method for obtaining drug-resistant mutants in ribosome engineering. The four initial strains and their str(R)-mutants and rif(R)-mutants were fermented in a liquid medium with the same composition. Mutants with anti-Staphylococcus aureus activity were obtained by paper chromatography. The components of fermentation broth were analyzed by high performance liquid chromatography (HPLC) and high performance liquid chromatography-mass spectrometry (LC-MS). Furthermore, FJ3 strain was identified by 16S rDNA and morphology. The minimal inhibitory concentration (MIC) of streptomycin and rifampicin for FJ3 was: 0.5 microg/mL and 110 microg/mL, respectively. Twenty-four strR-mutant strains and 20 rif(R)-mutant strains of FJ3 mutant strains were selected for bioassay. The result of the antibacterial activity screening demonstrated that six strains inhibited bacteria. Two strains (FJ3-2 and FJ3-6) were screened from the streptomycin-resistance mutants of inactive strain FJ3. The result of bioassay showed that the fermentation broth of FJ3-2 and FJ3-6 exhibited obvious anti-Staphylococcus aureus activity. The assay of paper chromatography showed that the active substance may be nucleic acid class antibiotic via using solvent system Doskochilova. Moreover, the results of HPLC and LC-MS exhibited that this substance may be thiolutin. Ribosome engineering for changing the secondary metabolic function of the inactive wild

  1. High yielding mutants of blackgram variety 'PH-25'

    International Nuclear Information System (INIS)

    Misra, R.C.; Mohapatra, B.D.; Panda, B.S.

    2001-01-01

    Seeds of blackgram (Vigna mungo L.) variety 'PH-5' were treated with chemical mutagens ethyl methanesulfonate (EMS), nitrosoguanidine (NG), maleic hydrazide (MH) and sodium azide (NaN 3 ), each at 3 different concentrations. Thirty six mutant lines developed from mutagenic treatments along with parent varieties were tested in M 4 generation. The mutants showed wide variation in most of the traits and multivariante D 2 analysis showed genetic divergence among themselves. Twenty of the thirty mutants showed genetic divergence from parent. Ten selected high yielding mutants were tested in M 5 . Yield and other productive traits of five high yielding mutants in M 4 and M 5 are presented

  2. Using of AFLP to evaluate gamma-irradiated amaranth mutants

    Directory of Open Access Journals (Sweden)

    Labajová Mária

    2013-01-01

    Full Text Available To determine which of several gamma-irradiated mutants of amaranth Ficha cultivar and K-433 hybrid are most genetically similar to their non-irradiated control genotypes, we performed amplified fragment length polymorphism (AFLP based analysis. A total of 40 selective primer combinations were used in reported analyses. First analyses of gamma-irradiated amaranth mutant lines were done used the AFLP. In the study, primers with the differentiation ability for all analysed mutant lines are reported. The very specific changes in the mutant lines´ non-coding regions based on AFLP length polymorphism were analysed. Mutant lines of the Ficha cultivar (C15, C26, C27, C82, C236 shared a genetic dissimilarity of 0,11 and their ISSR profiles are more similar to the Ficha than those of K-433 hybrid mutant lines. The K-433 mutant lines (D54, D279, D282 shared genetic dissimilarity of 0,534 but are more distinct to their control plant as a whole, as those of the Ficha mutant lines. Different AFLP fingerprints patters of the mutant lines when compared to the Ficha cultivar and K-433 hybrid AFLP profiles may be a consequence of the complex response of the intergenic space of mutant lines to the gamma-radiance. Although a genetic polymorphism was detected within accessions, the AFLP markers successfully identified all the accessions. The AFLP results are discussed by a combination of biochemical characteristics of mutant lines and their control genotypes.

  3. Distribution of soluble amino acids in maize endosperm mutants

    Directory of Open Access Journals (Sweden)

    Toro Alejandro Alberto

    2003-01-01

    Full Text Available For human nutrition the main source of vegetable proteins are cereal and legume seeds. The content of total soluble amino acids in mature endosperm of wild-type, opaque and floury maize (Zea mays L. mutants were determined by HPLC. The total absolute concentration of soluble amino acids among the mutants varied depending on the mutant. The o11 and o13 mutants exhibited the highest average content, whereas o10, fl3 and fl1 exhibited the lowest average content. In general, the mutants exhibited similar concentrations of total soluble amino acids when compared to the wild-type lines, with the clear exception of mutants o11 and fl1, with the o11 mutant exhibiting a higher concentration of total soluble amino acids when compared to its wild-type counterpart W22 and the fl1 mutant a lower concentration when compared to its wild-type counterpart Oh43. For methionine, the mutants o2 and o11 and wild-type Oh43 exhibited the highest concentrations of this amino acid. Significant differences were not observed between mutants for other amino acids such as lysine and threonine. The high lysine concentrations obtained originally for these mutants may be due to the amino acids incorporated into storage proteins, but not those present in the soluble form.

  4. Cell-cycle kinetics and ultraviolet light survival in UV-1, a Chinese hamster ovary cell mutant defective in post-replication recovery

    International Nuclear Information System (INIS)

    Collins, A.

    1982-01-01

    UV-I, an ultraviolet-sensitive mutant of CHO-KI, is abnormally slow to recover from the inhibition of DNA synthesis caused by u.v. irradiation. When synchronized UV-I cells are irradiated in G 1 , their movement into S phase is unaltered, but thymidine incorporation is depressed. When irradiated in S phase, again incorporation is more depressed, and S phase suffers a greater delay in UV-I than in the parent cell. UV-I and its parent have similar capacities for excision repair of u.v.-induced damage inflicted in G 1 , and so enter S phase with similar amounts of unrepaired damage. The single-cell survival was measured after irradiation at different points in the cell cycle. The mutant and parent cells have similar values of D 0 (mean lethal dose) except in mitosis, when the parent cell shows markedly greater resistance to u.v. irradiation. Dsub(q) (quasi-threshold dose) is fairly constant for the parent cell, but in UV-I it falls to a minimum in S phase. The responses of UV-I to u.v. irradiation are generally consistent with its known defect in post-replication recovery, i.e. the ability to join up the abnormally small DNA fragments synthesized on a u.v.-damaged template. (author)

  5. Combined Targeting of JAK2 and Bcl-2/Bcl-xL to Cure Mutant JAK2-Driven Malignancies and Overcome Acquired Resistance to JAK2 Inhibitors

    Directory of Open Access Journals (Sweden)

    Michaela Waibel

    2013-11-01

    Full Text Available To design rational therapies for JAK2-driven hematological malignancies, we functionally dissected the key survival pathways downstream of hyperactive JAK2. In tumors driven by mutant JAK2, Stat1, Stat3, Stat5, and the Pi3k and Mek/Erk pathways were constitutively active, and gene expression profiling of TEL-JAK2 T-ALL cells revealed the upregulation of prosurvival Bcl-2 family genes. Combining the Bcl-2/Bcl-xL inhibitor ABT-737 with JAK2 inhibitors mediated prolonged disease regressions and cures in mice bearing primary human and mouse JAK2 mutant tumors. Moreover, combined targeting of JAK2 and Bcl-2/Bcl-xL was able to circumvent and overcome acquired resistance to single-agent JAK2 inhibitor treatment. Thus, inhibiting the oncogenic JAK2 signaling network at two nodal points, at the initiating stage (JAK2 and the effector stage (Bcl-2/Bcl-xL, is highly effective and provides a clearly superior therapeutic benefit than targeting just one node. Therefore, we have defined a potentially curative treatment for hematological malignancies expressing constitutively active JAK2.

  6. Myofascial trigger point pain.

    Science.gov (United States)

    Jaeger, Bernadette

    2013-01-01

    Myofascial trigger point pain is an extremely prevalent cause of persistent pain disorders in all parts of the body, not just the head, neck, and face. Features include deep aching pain in any structure, referred from focally tender points in taut bands of skeletal muscle (the trigger points). Diagnosis depends on accurate palpation with 2-4 kg/cm2 of pressure for 10 to 20 seconds over the suspected trigger point to allow the referred pain pattern to develop. In the head and neck region, cervical muscle trigger points (key trigger points) often incite and perpetuate trigger points (satellite trigger points) and referred pain from masticatory muscles. Management requires identification and control of as many perpetuating factors as possible (posture, body mechanics, psychological stress or depression, poor sleep or nutrition). Trigger point therapies such as spray and stretch or trigger point injections are best used as adjunctive therapy.

  7. Rational points on varieties

    CERN Document Server

    Poonen, Bjorn

    2017-01-01

    This book is motivated by the problem of determining the set of rational points on a variety, but its true goal is to equip readers with a broad range of tools essential for current research in algebraic geometry and number theory. The book is unconventional in that it provides concise accounts of many topics instead of a comprehensive account of just one-this is intentionally designed to bring readers up to speed rapidly. Among the topics included are Brauer groups, faithfully flat descent, algebraic groups, torsors, étale and fppf cohomology, the Weil conjectures, and the Brauer-Manin and descent obstructions. A final chapter applies all these to study the arithmetic of surfaces. The down-to-earth explanations and the over 100 exercises make the book suitable for use as a graduate-level textbook, but even experts will appreciate having a single source covering many aspects of geometry over an unrestricted ground field and containing some material that cannot be found elsewhere. The origins of arithmetic (o...

  8. A mutant gene that increases gibberellin production in Brassica

    International Nuclear Information System (INIS)

    Rood, S.B.; Williams, P.H.; Pearce, D.; Pharis, R.P.; Murofushi, Noboru; Mander, L.N.

    1990-01-01

    A single gene mutant (elongated internode [ein/ein]) with accelerated shoot elongation was identified from a rapid cycling line of Brassica rapa. Relative to normal plants, mutant plants had slightly accelerated floral development, greater stem dry weights, and particularly, increased internode and inflorescence elongation. The application of the triazole plant growth retardant, paclobutrazol, inhibited shoot elongation, returning ein to a more normal phenotype. Conversely, exogenous gibberellin A 3 (GA 3 ) can convert normal genotypes to a phenotype resembling ein. The content of endogenous GA 1 and GA 3 were estimated by gas chromatography-selected ion monitoring using [ 2 H]GA 1 as a quantitative internal standard and at day 14 were 1.5- and 12.1-fold higher per stem, respectively, in ein than in normal plants, although GA concentrations were more similar. The endogenous levels of GA 20 and GA 1 , and the rate of GA 19 metabolism were simultaneously analyzed. Levels of GA 1 and GA 20 were 4.6- and 12.9-fold higher, respectively, and conversions to GA 20 and GA 1 were 8.3 and 1.3 times faster in ein than normal plants. Confirming the enhanced rate of GA 1 biosynthesis in ein, the conversion of [ 3 H]GA 20 to [ 3 H] GA 1 was also faster in ein than in the normal genotype. Thus, the ein allele results in accelerated GA 1 biosynthesis and an elevated content of endogenous GAs, including the dihydroxylated GAs A 1 and A 3

  9. An ordered, nonredundant library of Pseudomonas aeruginosa strain PA14 transposon insertion mutants

    OpenAIRE

    Liberati, Nicole T.; Urbach, Jonathan M.; Miyata, Sachiko; Lee, Daniel G.; Drenkard, Eliana; Wu, Gang; Villanueva, Jacinto; Wei, Tao; Ausubel, Frederick M.

    2006-01-01

    Random transposon insertion libraries have proven invaluable in studying bacterial genomes. Libraries that approach saturation must be large, with multiple insertions per gene, making comprehensive genome-wide scanning difficult. To facilitate genome-scale study of the opportunistic human pathogen Pseudomonas aeruginosa strain PA14, we constructed a nonredundant library of PA14 transposon mutants (the PA14NR Set) in which nonessential PA14 genes are represented by a single transposon insertio...

  10. Induction of drought tolerant mutants of rice

    International Nuclear Information System (INIS)

    El-Hissewy, A.A.; Abd Allah, A.

    2001-01-01

    The ultimate goal of crop breeding is to develop varieties with a high yield potential and desirable agronomic characteristics. In Egypt, the most important qualities sought by breeders have been high yield potential, resistance to major diseases and insects, and improved grain and eating quality. However, breeding efforts should concentrate on varieties with the potential to minimize yield losses under unfavorable conditions such as drought, and to maximize yields when conditions are favorable. Rice (Oryza sativa L.) in Egypt is completely irrigated and a significant portion of the rice cultivated area is subject to water deficit resulting from an inadequate or insufficient irrigation supply. Drought tolerance is a complex trait in that it results from the interaction of histological and physiological characters of plant with environmental factors, both above-ground and under-ground. Accordingly, root characters are closely related to drought tolerance. Little attention has been paid in Egyptian breeding programs to root characters and their relation to shoot characters. Furthermore, induced mutations are considered as one of the most important methods to induce useful mutants, especially with improved root characters, to overcome the drought problem. The present investigation aimed to study the effect of different doses of gamma rays on several characters of three Egyptian rice varieties, i.e. 'Giza 171', 'Giza 175' and 'Giza 176' and to induce one or more mutants possessing drought tolerance

  11. Flower morphology of Dendrobium Sonia mutants

    International Nuclear Information System (INIS)

    Sakinah Ariffin; Azhar Mohamad; Affrida Abu Hassan; Zaiton Ahmad; Mohd Nazir Basiran

    2010-01-01

    Dendrobium Sonia is a commercial hybrid which is popular as cut flower and potted plant in Malaysia. Variability in flower is important for new variety to generate more demands and choices in selection. Mutation induction is a tool in creating variability for new flower color and shape. In vitro cultures of protocorm-like bodies (PLBs) were exposed to gamma ray at dose 35 Gy. Phenotypic characteristics of the flower were observed at fully bloomed flower with emphasis on shape and color. Approximately 2000 regenerated irradiated plants were observed and after subsequent flowering, 100 plants were finally selected for further evaluation. Most of the color and shape changes are expressed in different combinations of petal, sepal and lip of the flower. In this work, 11 stable mutants were found different at flower phenotype as compared to control. Amongst these, four mutant varieties with commercial potential has been named as Dendrobium 'SoniaKeenaOval', Dendrobium 'SoniaKeenaRadiant', Dendrobium 'SoniaKeenaHiengDing' and Dendrobium 'Sonia KeenaAhmadSobri'. In this paper, variations in flower morphology and flower color were discussed, giving emphasis on variations in flower petal shape. (author)

  12. Pinched flow fractionation devices for detection of single nucleotide polymorphisms

    DEFF Research Database (Denmark)

    Larsen, A.V.; Poulsen, L.; Birgens, H.

    2008-01-01

    We demonstrate a new and flexible micro fluidic based method for genotyping single nucleotide polymorphisms ( SNPs). The method relies on size separation of selectively hybridized polystyrene microspheres in a micro fluidic pinched flow fractionation (PFF) device. The micro fluidic PFF devices...... and 5.6 mu m were functionalized with biotin-labeled oligonucleotides for the detection of a mutant (Mt) or wild-type (Wt) DNA sequence in the HBB gene, respectively. Hybridization to functionalized beads was performed with fluorescent targets comprising synthetic DNA oligonucleotides or amplified RNA......, synthesized using human DNA samples from individuals with point mutations in the HBB gene. Following a stringent wash, the beads were separated in a PFF device and the fluorescent signal from the beads was analyzed. Patients being wildtypes, heterozygotes or mutated respectively for the investigated mutation...

  13. Impedance analysis of acupuncture points and pathways

    International Nuclear Information System (INIS)

    Teplan, Michal; Kukucka, Marek; Ondrejkovicová, Alena

    2011-01-01

    Investigation of impedance characteristics of acupuncture points from acoustic to radio frequency range is addressed. Discernment and localization of acupuncture points in initial single subject study was unsuccessfully attempted by impedance map technique. Vector impedance analyses determined possible resonant zones in MHz region.

  14. Impedance analysis of acupuncture points and pathways

    Science.gov (United States)

    Teplan, Michal; Kukučka, Marek; Ondrejkovičová, Alena

    2011-12-01

    Investigation of impedance characteristics of acupuncture points from acoustic to radio frequency range is addressed. Discernment and localization of acupuncture points in initial single subject study was unsuccessfully attempted by impedance map technique. Vector impedance analyses determined possible resonant zones in MHz region.

  15. Allosteric Mutant IDH1 Inhibitors Reveal Mechanisms for IDH1 Mutant and Isoform Selectivity

    Energy Technology Data Exchange (ETDEWEB)

    Xie, Xiaoling; Baird, Daniel; Bowen, Kimberly; Capka, Vladimir; Chen, Jinyun; Chenail, Gregg; Cho, YoungShin; Dooley, Julia; Farsidjani, Ali; Fortin, Pascal; Kohls, Darcy; Kulathila, Raviraj; Lin, Fallon; McKay, Daniel; Rodrigues, Lindsey; Sage, David; Touré, B. Barry; van der Plas, Simon; Wright, Kirk; Xu, Ming; Yin, Hong; Levell, Julian; Pagliarini, Raymond A. (Novartis)

    2017-03-01

    Oncogenic IDH1 and IDH2 mutations contribute to cancer via production of R-2-hydroxyglutarate (2-HG). Here, we characterize two structurally distinct mutant- and isoform-selective IDH1 inhibitors that inhibit 2-HG production. Both bind to an allosteric pocket on IDH1, yet shape it differently, highlighting the plasticity of this site. Oncogenic IDH1R132H mutation destabilizes an IDH1 “regulatory segment,” which otherwise restricts compound access to the allosteric pocket. Regulatory segment destabilization in wild-type IDH1 promotes inhibitor binding, suggesting that destabilization is critical for mutant selectivity. We also report crystal structures of oncogenic IDH2 mutant isoforms, highlighting the fact that the analogous segment of IDH2 is not similarly destabilized. This intrinsic stability of IDH2 may contribute to observed inhibitor IDH1 isoform selectivity. Moreover, discrete residues in the IDH1 allosteric pocket that differ from IDH2 may also guide IDH1 isoform selectivity. These data provide a deeper understanding of how IDH1 inhibitors achieve mutant and isoform selectivity.

  16. Serrated leaf mutant in mungbean (Vigna radiata (L) Wilczek)

    International Nuclear Information System (INIS)

    Malik, I.A.; Ghulam, Sarwar; Yousaf, Ali; Saleem, M.

    1988-01-01

    Dry dormant seeds of mungbean (Vigna radiata (L) Wilczek) were treated with gamma rays (15, 30 and 60 kR). The serrated leaf mutation was noticed in M 2 of cultivar Pak 32 treated with 60 kR. Cf 14 plants, 3 showed the altered leaf structure and the others were normal. The feature of this mutant was the deep serration of leaflet margins. The mutant had large thick leaflets with prominent venation. The mutant bred true in the M 3 and successive generation. Details of the morphological characteristics of the mutant are presented. The mutant exhibited slower growth particularly during the early stages of development, flowered later and attained shorter height. There was an increase in the number of pods, in seed weight and in seed protein content, but number of seed per pod was considerably reduced. The seed coat colour showed a change from green to yellowish green. In the mutant's flowers the stamina were placed much below the stigma level and the stigma sometimes protruded the corolla. Outcrossing of 4% recorded in some of the mutant lines revealed a reduced cleistogamy. The low number of seeds per pod in the mutant could be due to reduced pollen fertility. The mutant behaved as monogenic recessive. The symbols SL/sl are proposed for this allelic pair. The mutant may have use as a green manure crop because of its large foliage and for the breeders as a genetic marker

  17. Forward genetic screen for auxin-deficient mutants by cytokinin.

    Science.gov (United States)

    Wu, Lei; Luo, Pan; Di, Dong-Wei; Wang, Li; Wang, Ming; Lu, Cheng-Kai; Wei, Shao-Dong; Zhang, Li; Zhang, Tian-Zi; Amakorová, Petra; Strnad, Miroslav; Novák, Ondřej; Guo, Guang-Qin

    2015-07-06

    Identification of mutants with impairments in auxin biosynthesis and dynamics by forward genetic screening is hindered by the complexity, redundancy and necessity of the pathways involved. Furthermore, although a few auxin-deficient mutants have been recently identified by screening for altered responses to shade, ethylene, N-1-naphthylphthalamic acid (NPA) or cytokinin (CK), there is still a lack of robust markers for systematically isolating such mutants. We hypothesized that a potentially suitable phenotypic marker is root curling induced by CK, as observed in the auxin biosynthesis mutant CK-induced root curling 1 / tryptophan aminotransferase of Arabidopsis 1 (ckrc1/taa1). Phenotypic observations, genetic analyses and biochemical complementation tests of Arabidopsis seedlings displaying the trait in large-scale genetic screens showed that it can facilitate isolation of mutants with perturbations in auxin biosynthesis, transport and signaling. However, unlike transport/signaling mutants, the curled (or wavy) root phenotypes of auxin-deficient mutants were significantly induced by CKs and could be rescued by exogenous auxins. Mutants allelic to several known auxin biosynthesis mutants were re-isolated, but several new classes of auxin-deficient mutants were also isolated. The findings show that CK-induced root curling provides an effective marker for discovering genes involved in auxin biosynthesis or homeostasis.

  18. Neurobehavioral Mutants Identified in an ENU Mutagenesis Project

    Energy Technology Data Exchange (ETDEWEB)

    Cook, Melloni N. [University of Memphis; Dunning, Jonathan P [University of Memphis; Wiley, Ronald G [Vanderbilt University and Veterans Administration, Nashville, TN; Chesler, Elissa J [ORNL; Johnson, Dabney K [ORNL; Goldowitz, Daniel [University of Tennessee Health Science Center, Memphis

    2007-01-01

    We report on a behavioral screening test battery that successfully identified several neurobehavioral mutants among a large-scale ENU-mutagenized mouse population. Large numbers of ENU mutagenized mice were screened for abnormalities in central nervous system function based on abnormal performance in a series of behavior tasks. We developed and employed a high-throughput screen of behavioral tasks to detect behavioral outliers. Twelve mutant pedigrees, representing a broad range of behavioral phenotypes, have been identified. Specifically, we have identified two open field mutants (one displaying hyper-locomotion, the other hypo-locomotion), four tail suspension mutants (all displaying increased immobility), one nociception mutant (displaying abnormal responsiveness to thermal pain), two prepulse inhibition mutants (displaying poor inhibition of the startle response), one anxiety-related mutant (displaying decreased anxiety in the light/dark test), and one learning and memory mutant (displaying reduced response to the conditioned stimulus) These findings highlight the utility of a set of behavioral tasks used in a high throughput screen to identify neurobehavioral mutants. Further analysis (i.e., behavioral and genetic mapping studies) of mutants is in progress with the ultimate goal of identification of novel genes and mouse models relevant to human disorders as well as the identification of novel therapeutic targets.

  19. A switch I mutant of Cdc42 exhibits decreased conformational freedom

    Science.gov (United States)

    Chandrashekar, Reena; Salem, Omar; Krizova, Hana; McFeeters, Robert; Adams, Paul D.

    2011-01-01

    Cdc42 is a Ras-related small G-protein, and functions as a molecular switch in signal transduction pathways linked with cell growth and differentiation. It is controlled by cycling between GTP-bound (active) and GDP-bound (inactive) forms. Nucleotide binding and hydrolysis are modulated by interactions with effectors and/or regulatory proteins. These interactions are centralized in two relatively flexible “Switch” regions as characterized by internal dynamics on multiple timescales (Loh et al., (2001) Biochemistry 40, 4590–4600), and this flexibility may be essential for protein interactions. In the Switch I region, Thr35 seems critical for function, as it is completely invariant in Ras-related proteins. To investigate the importance of conformational flexibility in Switch I of Cdc42, we mutated threonine to alanine, determined the solution structure and characterized the backbone dynamics of the single-point mutant protein, Cdc42(T35A). Backbone dynamics data suggests that the mutation changes the timescale of the internal motions of several residues, with several resonances appearing not discernable in Cdc42 wild type (Adams and Oswald (2007) Biomolecular NMR Assignments 1, 225–227). The mutation does not appear to affect the thermal stability of Cdc42, and chymotrypsin digestion data further suggests that changes in conformational flexibility in Switch I slow proteolytic cleavage relative to wild type. In-vitro binding assays show reduced binding of Cdc42(T35A), relative to wild type, to a GTPase binding protein that inhibits GTP hydrolysis in Cdc42. These results suggest that the mutation of T35 leads to the loss of conformational freedom in Switch I that could affect effector/regulatory protein interactions. PMID:21667996

  20. Single ventricle cardiac defect

    International Nuclear Information System (INIS)

    Eren, B.; Turkmen, N.; Fedakar, R.; Cetin, V.

    2010-01-01

    Single ventricle heart is defined as a rare cardiac abnormality with a single ventricle chamber involving diverse functional and physiological defects. Our case is of a ten month-old baby boy who died shortly after admission to the hospital due to vomiting and diarrhoea. Autopsy findings revealed cyanosis of finger nails and ears. Internal examination revealed; large heart, weighing 60 grams, single ventricle, without a septum and upper membranous part. Single ventricle is a rare pathology, hence, this paper aims to discuss this case from a medico-legal point of view. (author)

  1. Single gene deletions of mrpA to mrpG and mrpE point mutations affect activity of the Mrp Na+/H+ antiporter of alkaliphilic Bacillus and formation of hetero-oligomeric Mrp complexes.

    Science.gov (United States)

    Morino, Masato; Natsui, Shinsuke; Swartz, Talia H; Krulwich, Terry A; Ito, Masahiro

    2008-06-01

    Mrp antiporters catalyze secondary Na(+)(Li(+))/H(+) antiport and/or K(+)/H(+) antiport that is physiologically important in diverse bacteria. An additional capacity for anion flux has been observed for a few systems. Mrp is unique among antiporters in that it requires all six or seven hydrophobic gene products (MrpA to MrpG) of the mrp operon for full antiporter activity, but MrpE has been reported to be dispensable. Here, the membrane complexes formed by Mrp proteins were examined using a cloned mrp operon from alkaliphilic Bacillus pseudofirmus OF4. The operon was engineered so that the seven Mrp proteins could be detected in single samples. Membrane extracts of an antiporter-deficient Escherichia coli strain expressing this construct were analyzed by blue native-sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Mrp complexes of two sizes were identified containing all seven Mrp proteins. Studies of the single nonpolar mrp gene deletions in the construct showed that a subcomplex of MrpA, MrpB, MrpC, and MrpD was formed in the absence of MrpE, MrpF, or MrpG. By contrast, MrpE, MrpF, and MrpG were not observed in membranes lacking MrpA, MrpB, MrpC, or MrpD. Although MrpA and MrpD have been hypothesized to be the antiporter proteins, the MrpA-to-D complex was inactive. Every Mrp protein was required for an activity level near that of the wild-type Na(+)/H(+) antiporter, but a very low activity level was observed in the absence of MrpE. The introduction of an MrpE(P114G) mutation into the full Mrp complex led to antiport activity with a greatly increased apparent K(m) value for Na(+). The results suggested that interactions among the proteins of heterooligomeric Mrp complexes strongly impact antiporter properties.

  2. Short-Term Effects of Kinesio Taping and Cross Taping Application in the Treatment of Latent Upper Trapezius Trigger Points: A Prospective, Single-Blind, Randomized, Sham-Controlled Trial

    Directory of Open Access Journals (Sweden)

    Tomasz Halski

    2015-01-01

    Full Text Available Kinesio taping (KT may be a new treatment in patients with myofascial trigger points (MTrPs. A new method available for taping practitioners is cross taping (CT. The main objective was to determine how CT, KT, and medical adhesive tape (sham group affect the subjective assessment of resting bioelectrical activity and pain of the upper trapezius muscle (UT in patients with MTrPs. 105 volunteers were recruited to participate. The primary outcome was resting bioelectrical activity of UT muscle as assessed by surface electromyography (sEMG in each group and pain intensity on a visual analog scale (VAS. Assessments were collected before and after intervention and after the 24-hours follow-up. No significant differences were observed in bioelectrical activity of UT between pre-, post-, and follow-up results. In three groups patients had significantly lower pain VAS score after the intervention (CT—p<0.001, KT—p<0.001, and sham—p<0.01. The Kruskal-Wallis ANOVA showed no significant differences in almost all measurements between groups. The application of all three types of tapes does not influence the resting bioelectrical activity of UT muscle and may not lead to a reduction in muscle tone in the case of MTrPs.

  3. Novel Synthesis and Phenotypic Analysis of Mutant Clouds for Hepatitis E Virus Genotype 1.

    Science.gov (United States)

    Agarwal, Shubhra; Baccam, Prasith; Aggarwal, Rakesh; Veerapu, Naga Suresh

    2018-02-15

    Many RNA viruses exist as an ensemble of genetically diverse, replicating populations known as a mutant cloud. The genetic diversity (cloud size) and composition of this mutant cloud may influence several important phenotypic features of the virus, including its replication capacity. We applied a straightforward, bacterium-free approach using error-prone PCR coupled with reverse genetics to generate infectious mutant RNA clouds with various levels of genetic diversity from a genotype 1 strain of hepatitis E virus (HEV). Cloning and sequencing of a genomic fragment encompassing 70% of open reading frame 1 ( ORF1 ) or of the full genome from variants in the resultant clouds showed the occurrence of nucleotide mutations at a frequency on the order of 10 -3 per nucleotide copied and the existence of marked genetic diversity, with a high normalized Shannon entropy value. The mutant clouds showed transient replication in cell culture, while wild-type HEV did not. Cross-sectional data from these cell cultures supported the existence of differential effects of clouds of various sizes and compositions on phenotypic characteristics, such as the replication level of (+)-RNA progeny, the amounts of double-stranded RNA (a surrogate for the rate of viral replication) and ORF1 protein, and the expression of interferon-stimulated genes. Since mutant cloud size and composition influenced the viral phenotypic properties, a better understanding of this relationship may help to provide further insights into virus evolution and prediction of emerging viral diseases. IMPORTANCE Several biological or practical limitations currently prevent the study of phenotypic behavior of a mutant cloud in vitro We developed a simple and rapid method for synthesizing mutant clouds of hepatitis E virus (HEV), a single-stranded (+)-RNA [ss(+) RNA] virus, with various and controllable levels of genetic diversity, which could then be used in a cell culture system to study the effects of cloud size and

  4. Properties of adenyl cyclase and cyclic adenosine 3',5'-monophosphate receptor protein-deficient mutants of Escherichia coli

    International Nuclear Information System (INIS)

    Kumar, S.

    1976-01-01

    Several spontaneous cya and crp mutants of Escherichia coli have been selected as clones simultaneously resistant to phage lambda and nalidixic acid and characterized. Both cya and crp mutants have been found to grow as cocci with increased doubling times. They have increased resistance to some mutagens (methylmethanesulfonate, ultraviolet light, gamma rays), antibiotics (nalidixic acid, ampicillin), phages (lambda, T6), sublethal heat and hypotonic shock, and decreased resistance to neutral detergents (sodium dodecyl sulfate, sodium deoxycholate), a protein synthesis inhibitor (streptomycin), and a respiratory inhibitor (sodium azide). The nature of changes in cell parameters indicate fundamental alterations in the envelope structure of the cya and crp mutant cells. The new cya and crp mutants have been found to be multiply carbohydrate negative and nonmotile in conformity with similar previously isolated mutants. Studies of revertants and phi 80 cya + and phi 80 cya transductants indicated that the pleiotropic phenotype is related to a single mutational event at the cya or the crp locus in the mutants

  5. MASTR: A Technique for Mosaic Mutant Analysis with Spatial and Temporal Control of Recombination Using Conditional Floxed Alleles in Mice

    Directory of Open Access Journals (Sweden)

    Zhimin Lao

    2012-08-01

    Full Text Available Mosaic mutant analysis, the study of cellular defects in scattered mutant cells in a wild-type environment, is a powerful approach for identifying critical functions of genes and has been applied extensively to invertebrate model organisms. A highly versatile technique has been developed in mouse: MASTR (mosaic mutant analysis with spatial and temporal control of recombination, which utilizes the increasing number of floxed alleles and simultaneously combines conditional gene mutagenesis and cell marking for fate analysis. A targeted allele (R26MASTR was engineered; the allele expresses a GFPcre fusion protein following FLP-mediated recombination, which serves the dual function of deleting floxed alleles and marking mutant cells with GFP. Within 24 hr of tamoxifen administration to R26MASTR mice carrying an inducible FlpoER transgene and a floxed allele, nearly all GFP-expressing cells have a mutant allele. The fate of single cells lacking FGF8 or SHH signaling in the developing hindbrain was analyzed using MASTR, and it was revealed that there is only a short time window when neural progenitors require FGFR1 for viability and that granule cell precursors differentiate rapidly when SMO is lost. MASTR is a powerful tool that provides cell-type-specific (spatial and temporal marking of mosaic mutant cells and is broadly applicable to developmental, cancer, and adult stem cell studies.

  6. Genetic analysis of the induced mutants of rice resistant to bacterial leaf blight

    International Nuclear Information System (INIS)

    Nakai, H.

    1990-01-01

    Full text: Seeds of the rice cultivar 'Harebare', which is susceptible to bacterial leaf blight (BLB), were treated with thermal neutrons, gamma-rays, ethyleneimine and ethylmethane-sulfonate. In the M2, plants with better resistance to BLB were identified through inoculation at the seedling and the flag leaf stages with an isolate (T7174) of the Japanese differential race I. Several mutant lines resistant to BLB were selected through tests of the M 3 or M 4 lines derived from selected resistant M 2 plants. The frequency of resistant mutants was significantly higher after the thermal neutron treatment than after treatments with other mutagens. Two mutants, which originated from the neutron treatment, showing a highly quantitative resistance to multiple BLB races were analysed for gene(s) for resistance. The resistance of one of them (M41) to the Japanese races I, II, III, IV, and V was found to be conditioned by a single recessive gene. Three other recessive genes for resistance are known, but their reaction to differential races is different. Therefore, this gene was thought to be new and was tentatively designated as xa-nm(t). The resistance of another mutant (M57) was found to be polygenically inherited. (author)

  7. Genetic control of some morphological mutants in sunflower [Helianthus annuus L.

    International Nuclear Information System (INIS)

    Nabipour, A.; Sarrafi, A.; Yazdi-Samadi, B.

    2004-01-01

    Inheritance study of induced mutants is an important tool in genetic and breeding programs. Sunflower is one of the most important oil crops for which mutant collection is meager. Seeds of sunflower line AS-613 were irradiated with gamma rays and mutant phenotypes were traced until M4 generation. In M5 generation, the following traits were studied: dwarfing, branching, leaf shape, albinism, rosette, lack of apex and alternative leaves. In most cases, the mutated characters were controlled by a single recessive gene, while in two cases they were controlled by two recessive genes. In M5 progenies, segregation for two albino, one alternative leaves, one dwarfism, 5 branching, one rosette, 2 lacks of apex and 5 leaf shape mutants was recorded. Amongst five cases of branching, one was controlled by two recessive genes, where at least one homozygote recessive locus was necessary for branching. In one case, the lack of apex was controlled by two recessive genes and even only one dominant allele could provoke the normal plant [it

  8. Phenotypic and genetic analysis of "Chameleon," a paramecium mutant with an enhanced sensitivity to magnesium.

    Science.gov (United States)

    Preston, R R; Hammond, J A

    1997-07-01

    Three mutant strains of Paramecium tetraurelia with an enhanced sensitivity to magnesium have been isolated. These new "Chameleon" mutants result from partial- or codominant mutations at a single locus, Cha. Whereas the wild type responded to 5 mM Mg2+ by swimming backward for 10-15 sec, Cha mutants responded with approximately 30 sec backward swimming. Electrophysiological analysis suggested that this behavior may be caused by slowing in the rate at which a Mg(2+)-specific ion conductance deactivates following membrane excitation. This would be consistent with an observed increase in the sensitivity of Cha mutants to nickel poisoning, since Ni2+ is also able to enter the cell via this pathway. More extensive behavioral analysis showed that Cha cells also overresponded to Na+, but there was no evidence for a defect in intracellular Ca2+ homeostasis that might account for a simultaneous enhancement of both the Mg2+ and Na+ conductances. The possibility that the Cha locus may encode a specific regulator of the Mg(2+)- and Na(+)-permeabilities is considered.

  9. api, A novel Medicago truncatula symbiotic mutant impaired in nodule primordium invasion.

    Science.gov (United States)

    Teillet, Alice; Garcia, Joseph; de Billy, Françoise; Gherardi, Michèle; Huguet, Thierry; Barker, David G; de Carvalho-Niebel, Fernanda; Journet, Etienne-Pascal

    2008-05-01

    Genetic approaches have proved to be extremely useful in dissecting the complex nitrogen-fixing Rhizobium-legume endosymbiotic association. Here we describe a novel Medicago truncatula mutant called api, whose primary phenotype is the blockage of rhizobial infection just prior to nodule primordium invasion, leading to the formation of large infection pockets within the cortex of noninvaded root outgrowths. The mutant api originally was identified as a double symbiotic mutant associated with a new allele (nip-3) of the NIP/LATD gene, following the screening of an ethylmethane sulphonate-mutagenized population. Detailed characterization of the segregating single api mutant showed that rhizobial infection is also defective at the earlier stage of infection thread (IT) initiation in root hairs, as well as later during IT growth in the small percentage of nodules which overcome the primordium invasion block. Neither modulating ethylene biosynthesis (with L-alpha-(2-aminoethoxyvinylglycine or 1-aminocyclopropane-1-carboxylic acid) nor reducing ethylene sensitivity in a skl genetic background alters the basic api phenotype, suggesting that API function is not closely linked to ethylene metabolism or signaling. Genetic mapping places the API gene on the upper arm of the M. truncatula linkage group 4, and epistasis analyses show that API functions downstream of BIT1/ERN1 and LIN and upstream of NIP/LATD and the DNF genes.

  10. The End of Points

    Science.gov (United States)

    Feldman, Jo

    2018-01-01

    Have teachers become too dependent on points? This article explores educators' dependency on their points systems, and the ways that points can distract teachers from really analyzing students' capabilities and achievements. Feldman argues that using a more subjective grading system can help illuminate crucial information about students and what…

  11. Temperature sensitive riboflavin mutants of Penicillium vermiculatum Dangeard

    International Nuclear Information System (INIS)

    Mitra, J.; Chaudhari, K.L.

    1974-01-01

    Two temperature sensitive UV induced riboflavin mutants rib 1 and rib 6 have been physiologically and genetically characterized. The two mutants behave differently with regard to their temperature sensitivity. The rib 1 mutant exhibits a leaky growth in minimal medium between 15 0 C and 30 0 C but grows well when the medium is supplemented with riboflavin. At 35 0 C the growth response of the mutant is at its max. and at 40 0 C and below 15 0 C it ceases to grow. The rib 6 mutant which is red brown in colour shows wild type character at temp. below 25 0 C in minimal medium but requires riboflavin at 30 0 C and above. Heterokaryotic analysis revealed the nonallelic nature of the two temperature mutants. Genetic tests of allelic relationship between riboflavin markers by crossing were also done. (author)

  12. Mutants of Cercospora kikuchii altered in cercosporin synthesis and pathogenicity

    Energy Technology Data Exchange (ETDEWEB)

    Upchurch, R.G.; Walker, D.C.; Rollins, J.A.; Ehrenshaft, M.; Daub, M.E. (North Carolina State Univ., Raleigh (United States))

    1991-10-01

    The authors have obtained spontaneous and UV-induced stable mutants, altered in the synthesis of cercosporin, of the fungal soybean pathogen Cercospora kikuchii. The mutants were isolated on the basis of colony color on minimal medium. The UV-induced mutants accumulated, at most, 2% of wild-type cercosporin levels on all media tested. In contrast, cercosporin accumulation by the spontaneous mutants was strongly medium regulated, occurring only on potato dextrose medium but at concentrations comparable to those produced by the wild-type strain. UV-induced mutants unable to synthesize cercosporin on any medium were unable to incite lesions when inoculated onto the soybean host. Cercosporin was reproducibly isolated from all inoculated leaves showing lesions. Although cercosporin involvement in disease has been indirectly suggested by many previous studies, this is the first report in which mutants blocked in cercosporin synthesis have been used to demonstrate that cercosporin is a crucial pathogenicity factor for this fungal genus.

  13. Identification of the second mutation of BADH2 gene derived from rice mutant lines induced by gamma rays

    International Nuclear Information System (INIS)

    I Ishak

    2016-01-01

    The BADH2 gene acts as suppressor of 2-acetyl-1-pyrolline (2AP) biosynthesis in plants. 2AP is the volatile compound which provides fragrance in rice. Biosynthesis of 2AP occurs when BADH2 loses its function as suppressor gene. Aromatic rice cultivars naturally incur mutation of BADH2 gene at 8 bp. In this experiment, aromatic mutant rice lines derived from irradiation of Sintanur cultivar by gamma rays with dose of 100 Gy were studied in molecular level. These mutant lines were characterized at the M10 plantgeneration under the assumption that genetically these aromatic mutant rice lines were homozygotic. Several primers related to aroma in rice have been used for polymerase chain reaction (PCR) in a thermal cycler instrument. Gel electrophoreses were carried out using 1.5% agarose in TAE buffer. DNA fragments at 254 bp and 355 bp (base pair) were taken and amplified by primer for nucleotide sequencing of these fragments. Molecular identification and characterization after electrophoresis showed that the mutant line from AR1020 can be differentiated from AR.1080 at 254 bp. Nucleotide sequence data from of these DNA fragments showed that point mutations (deletions and substitutions) occurred at the BADH2 gene in exon 7; those are called second mutation and were caused by gamma rays effects. The Sintanur variety was used as check cultivar and its DNA sequence was compared to that of the AR.1020 mutant line. The results from both DNA sequences (from cv. Sintanur and AR.1020) derived from fragments at 254 bp show that point mutations occurred within exon 7 and earlier stop codon occurred in the AR.1020 mutant rice line. Further, the use of EA primer in PCR resulted in detection of deletion and substitution of nucleotides in the AR.1020 mutant line. (author)

  14. Hoxc13 mutant mice lack external hair.

    Science.gov (United States)

    Godwin, A R; Capecchi, M R

    1998-01-01

    Hox genes are usually expressed temporally and spatially in a colinear manner with respect to their positions in the Hox complex. Consistent with the expected pattern for a paralogous group 13 member, early embryonic Hoxc13 expression is found in the nails and tail. Hoxc13 is also expressed in vibrissae, in the filiform papillae of the tongue, and in hair follicles throughout the body; a pattern that apparently violates spatial colinearity. Mice carrying mutant alleles of Hoxc13 have been generated by gene targeting. Homozygotes have defects in every region in which gene expression is seen. The most striking defect is brittle hair resulting in alopecia (hairless mice). One explanation for this novel role is that Hoxc13 has been recruited for a function common to hair, nail, and filiform papilla development.

  15. Google: a narrativa de uma marca mutante

    Directory of Open Access Journals (Sweden)

    Elizete de Azevedo Kreutz

    2010-01-01

    Full Text Available As marcas mutantes já fazem parte de nossa realidade, embora ainda não totalmente percebidas e/ou aceitas como tal. O presente artigo busca refletir sobre a relevância dessas novas estratégias de comunicação e branding, identificando suas principais características. Para isso, utilizamos o método de estudo de caso, o Google, ancorado nos métodos de pesquisa bibliográfica e de internet. A escolha foi intencional, posto que a organização é referência em sua categoria, mecanismo de busca, e reflete essa estratégia comunicacional contemporânea. Como resultado, as informações obtidas nos possibilitam compreender essa tendência de comportamento de marca que busca a interação com seus públicos.

  16. Studies on mutant breeding of Hibiscus syriacus

    Energy Technology Data Exchange (ETDEWEB)

    Song, Hi Sup; Kim, Jin Kyu; Lee, Ki Un; Kim, Young Taik

    1997-01-01

    Hibiscus has been known as a national flower of Korea. Hibiscus has such a characteristic of self-incompatibility that all the plant exist as natural hybrids and have heterogeneous genes. Many domestic 91 varieties of Hibiscus syriacus were collected. Radiosensitivity of H. Syriacus irradiated with {gamma}-ray was investigated in plant cuttings. The plant height was reduced by 45% in 5KR irradiated group, compared to control group. The radiation dose of 5KR could be recommended for mutation breeding of Hibiscus cuttings. Radiosensitivity of {gamma}-ray irradiated Hibiscus seed were investigated. The germination rate, survival rate and plant height was better in the 4KR irradiation plot than control. The radiation dose of 10{approx}12KR are recommended for mutation breeding of Hibiscus. Promising mutant lines were selected form the varieties of Hwarang, Wolsan no. 176, Ilpyondansim, Emille, Hanol, Yongkwang, Saeyongkwang, Chungmu, Imjinhong, Arang, Hungdansim-1 and Hongdansim-2. (author). 66 refs., 16 tabs., 13 figs.

  17. Studies on mutant breeding of Hibiscus syriacus

    International Nuclear Information System (INIS)

    Song, Hi Sup; Kim, Jin Kyu; Lee, Ki Un; Kim, Young Taik.

    1997-01-01

    Hibiscus has been known as a national flower of Korea. Hibiscus has such a characteristic of self-incompatibility that all the plant exist as natural hybrids and have heterogeneous genes. Many domestic 91 varieties of Hibiscus syriacus were collected. Radiosensitivity of H. Syriacus irradiated with γ-ray was investigated in plant cuttings. The plant height was reduced by 45% in 5KR irradiated group, compared to control group. The radiation dose of 5KR could be recommended for mutation breeding of Hibiscus cuttings. Radiosensitivity of γ-ray irradiated Hibiscus seed were investigated. The germination rate, survival rate and plant height was better in the 4KR irradiation plot than control. The radiation dose of 10∼12KR are recommended for mutation breeding of Hibiscus. Promising mutant lines were selected form the varieties of Hwarang, Wolsan no. 176, Ilpyondansim, Emille, Hanol, Yongkwang, Saeyongkwang, Chungmu, Imjinhong, Arang, Hungdansim-1 and Hongdansim-2. (author). 66 refs., 16 tabs., 13 figs

  18. Winter barley mutants created in the Ukraine

    International Nuclear Information System (INIS)

    Zayats, O.M.

    2001-01-01

    Full text: Increasing fodder and protein production is one of the objectives of the development of agriculture in Ukraine. Higher productivity of fodder crops, due to new highly productive varieties, is the means to meet this aim. Winter barley is an important crop for fodder purposes. The climate of the Ukraine is favourable for growing this crop. The areas used for the growth of winter barley are however, small (500,000-550,000 ha) and there is a shortage of good quality varieties. The main aim of the work was therefore to create new varieties of highly productive winter barley, of good quality. The new varieties and mutation lines of winter barley were created under the influence of water solutions of N-nitroso-N-methylurea (NMH - 0,012, 0,005%), N-nitroso-N-ethylurea (NEH - 0,05; 0.025; 0,012%) ethyleneimine (EI - 0,02; 0,01; 0,005%) on winter barley seeds of the varieties of local and foreign selections. On the basis of many years of investigations (1984-94) the following mutations were described: hard-grained, winter-hardiness, earliness, middle-maturity, late-maturity, wide and large leaves, narrow leaves, multinodal, great number of leaves, great number of flowers, strong stem (lodging resistant), tallness, semi-dwarfness, dwarfness, and high productivity. Particularly valuable are mutants with high productivity of green bulk. Their potential yield is 70 t/ha. As a result of the work two varieties of winter barley 'Shyrokolysty' and 'Kormovy' were released into the State register of plant varieties of the Ukraine. The other valuable mutant genotypes are used in cross breeding programmes. (author)

  19. Temperature-sensitive glutamate dehydrogenase mutants of Salmonella typhimurium.

    OpenAIRE

    Dendinger, S M; Brenchley, J E

    1980-01-01

    Mutants of Salmonella typhimurium defective in glutamate dehydrogenase activity were isolated in parent strains lacking glutamate synthase activity by localizcd mutagenesis or by a general mutagenesis combined with a cycloserine enrichment for glutamate auxotrophs. Two mutants with temperature-sensitive phenotypes had glutamate dehydrogenase activities that were more thermolabile than that of an isogenic control strain. Eight other mutants had less than 10% of the wild-type glutamate dehydrog...

  20. Study on ionizing radiosensitivity of respiratory deficiency yeast mutants

    International Nuclear Information System (INIS)

    Mao Shuhong; Chinese Academy of Sciences, Beijing; Jin Genming; Wei Zengquan; Xie Hongmei

    2006-01-01

    The radiosensitivity of respiratory deficiency yeast mutants has been studied in this work. The mutants which were screened from the yeasts after ionizing irradiation were irradiated with 12 C 6+ at different doses. Because of the great change in its mitochondria and mitochondrial DNA, the respiratory deficiency yeast mutants show radio-sensitivity at dose less than 1 Gy and radioresistance at doses higher than 1 Gy. (authors)

  1. Defective Glycinergic Synaptic Transmission in Zebrafish Motility Mutants

    OpenAIRE

    Hirata, Hiromi; Carta, Eloisa; Yamanaka, Iori; Harvey, Robert J.; Kuwada, John Y.

    2010-01-01

    Glycine is a major inhibitory neurotransmitter in the spinal cord and brainstem. Recently, in vivo analysis of glycinergic synaptic transmission has been pursued in zebrafish using molecular genetics. An ENU mutagenesis screen identified two behavioral mutants that are defective in glycinergic synaptic transmission. Zebrafish bandoneon (beo) mutants have a defect in glrbb, one of the duplicated glycine receptor (GlyR) β subunit genes. These mutants exhibit a loss of glycinergic synaptic ...

  2. Chloroplast genes in Chlamydomonas affecting organelle ribosomes. Genetic and biochemical analysis of analysis of antibiotic-resistant mutants at several gene loci.

    Science.gov (United States)

    Conde, M F; Boynton, J E; Gillham, N W; Harris, E H; Tingle, C L; Wang, W L

    1975-10-03

    Six chloroplast gene mutants of Chlamydomonas reinhardtii resistant to spectinomycin, erythromycin, or streptomycin have been assessed for antibiotic resistance of their chloroplast ribosomes. Four of these mutations clearly confer high levels of antibiotic resistance on the chloroplast ribosomes both in vivo. Although one mutant resistant to streptomycin and one resistant to spectinomycin have chloroplast ribosomes as sensitive to antibiotics as those of wild type in vivo, these mutations can be shown to alter the wildtype sensitivity of chloroplast ribosomes in polynucleotide-directed amino acid incorporation in vitro. Genetic analysis of these six chloroplast mutants and three similar mutants (Sager, 1972), two of which have been shown to affect chloroplast ribosomes (Mets and Bogorad, 1972; Schlanger and Sager, 1974), indicates that in Chlamydomonas at least three chloroplast gene loci can affect streptomycin resistance of chloroplast ribosomes and that two can affect erythromycin resistance. The three spectinomycin-resistant mutants examined appear to be alleles at a single chloroplast gene locus, but may represent mutations at two different sites within the same gene. Unlike wild type, the streptomycin and spectinomycin resistant mutants which have chloroplast ribosomes sensitive to antibiotics in vivo, grow well in the presence of antibiotic by respiring exogenously supplied acetate as a carbon source, and have normal levels of cytochrome oxidase activity and cyanide-sensitive respiration. We conclude that mitochondrial protein synthesis in these mutants is resistant to these antibiotics, whereas in wild type it is sensitive. To explain the behavior of these two chloroplast gene mutants as well as other one-step mutants which are resistant both photosynthetically and when respiring acetate in the dark, we have postulated that a mutation in a single chloroplast gene may result in alteration of both chloroplast and mitochondrial ribosomes. Mitochondrial

  3. Mutation induction in γ-irradiated primary human bronchial epithelial cells and molecular analysis of the HPRT- mutants

    International Nuclear Information System (INIS)

    Suzuki, Keiji; Hei, Tom K.

    1996-01-01

    We have examined various radiobiological parameters using commercially-available primary normal human bronchial epithelial (NHBE) cells, which can be subcultured more than 20 population doublings, and have established the mutation system in order to characterize the molecular changes in γ-irradiated primary cells. The survival curve, obtained after irradiation of cells with 137 Cs γ-rays, indicates that the D 0 , D q , and n values are 1.34 Gy, 1.12 Gy, and 2.3, respectively. The induction of HPRT - mutation was dose-dependent and the mutant fraction increased in a non-linear fashion. Since the doubling number of NHBE cells is limited, DNA was extracted directly from the single mutant colonies and alteration in the HPRT gene locus was analyzed using multiplex PCR technique. Among spontaneous mutants, the proportion with total and partial deletions of the gene was 10.0% (2/20) and 60.0% (12/20), respectively, while 30.0% (6/20) did not have any detectable changes in the nine exons examined. On the other hand, the fraction of total deletion increased by more than 2-fold among mutants induced by γ-rays in that 26.3% (10/38) of them showed the total gene deletions. Twenty-five out of 38 γ-induced mutants (65.8%) had partial deletions and 3 mutants (7.9%) had no detectable alteration. The present results showed that γ-irradiation efficiently induced HPRT gene mutation in primary human epithelial cells and that most of the induced mutants suffered larger deletions compared to that observed in spontaneous mutants. This system provides a useful tool for determination of mutagenicity and understanding the molecular mechanisms of environmental carcinogens in primary human bronchial cells

  4. Assessment of the Toxicity of CuO Nanoparticles by Using Saccharomyces cerevisiae Mutants with Multiple Genes Deleted

    Science.gov (United States)

    Bao, Shaopan; Lu, Qicong; Dai, Heping; Zhang, Chao

    2015-01-01

    To develop applicable and susceptible models to evaluate the toxicity of nanoparticles, the antimicrobial effects of CuO nanoparticles (CuO-NPs) on various Saccharomyces cerevisiae (S. cerevisiae) strains (wild type, single-gene-deleted mutants, and multiple-gene-deleted mutants) were determined and compared. Further experiments were also conducted to analyze the mechanisms associated with toxicity using copper salt, bulk CuO (bCuO), carbon-shelled copper nanoparticles (C/Cu-NPs), and carbon nanoparticles (C-NPs) for comparisons. The results indicated that the growth inhibition rates of CuO-NPs for the wild-type and the single-gene-deleted strains were comparable, while for the multiple-gene deletion mutant, significantly higher toxicity was observed (P CuO-NPs to yeast cells was compared with the toxicities of copper salt and bCuO, we concluded that the toxicity of CuO-NPs should be attributed to soluble copper rather than to the nanoparticles. The striking difference in adverse effects of C-NPs and C/Cu-NPs with equivalent surface areas also proved this. A toxicity assay revealed that the multiple-gene-deleted mutant was significantly more sensitive to CuO-NPs than the wild type. Specifically, compared with the wild-type strain, copper was readily taken up by mutant strains when cell permeability genes were knocked out, and the mutants with deletions of genes regulated under oxidative stress (OS) were likely producing more reactive oxygen species (ROS). Hence, as mechanism-based gene inactivation could increase the susceptibility of yeast, the multiple-gene-deleted mutants should be improved model organisms to investigate the toxicity of nanoparticles. PMID:26386067

  5. Induction and selection of citrus mutant by gamma-irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Kim, In Jung; Oh, Seung Kyu; Lee, Hyo Yeon [Jeju National University, Jeju (Korea, Republic of)

    2010-09-15

    We have subjected to gamma-irradiation to citrus buds and then grafted onto mature citrus tree. Mutant citrus branch lines have been induced. As a result of first selection, we found the several mutant lines showing interesting phenotypes such as higher sugar content. We have selected several branches showing good qualities such as higher sweetness and/or lower acidity. Some branch lines showed over 13 .deg. Brix sugar content and below 0.9% acidity. Other mutant branch lines showed the changes of shape, size, peel thickness, and fiber contents or distribution of fruits. The results suggest that gamma-irradiation is an effective tool for induction of citrus mutant lines.

  6. Seed protein and nitrogen fixation in chickpea mutant variety Hyprosola

    International Nuclear Information System (INIS)

    Schroeder, H.E.; Gibson, A.H.; Oram, R.N.; Shaikh, M.A.Q.

    1989-01-01

    Full text: 'Hyprosola' is a high yielding, high protein mutant cultivar obtained after gamma irradiation from the variety 'Faridpur-1'. The mutant yields 45 % more protein per unit area. The essential amino acid index is unchanged. It is likely that the high nutritional value in 'Hyprosola' seed protein arises from an increase in the albumin:globulin ratio. Nitrogen fixation rates of the mutant during the first 7 weeks of growth were found to be similar to 'Faridpur-1'. Under field conditions, the mutant may be able to nodulate more rapidly and more extensively than the parent variety. (author)

  7. Sphingolipid synthesis deficiency in a mutant of Bacteroides levii

    Energy Technology Data Exchange (ETDEWEB)

    Brumleve, B.; Lev, M.

    1986-05-01

    Bacteroides levii, an anaerobic bacterium, synthesizes two sphingolipids; the sphingomyelin analogue, ceramide phosphorylethanolamine (CPE), and also ceramide phosphorylglycerol (CPG). The first enzyme in the sphingolipid pathway, 3-ketodihydro-sphingosine (3KDS) synthase, has been partially purified previously. To study subsequent steps in the pathways, mutants defective in sphingolipid synthesis were derived by ethyl methanesulfonate and nitrosoguanidine mutagenesis. Extracts of the mutant, 1075BB, show synthase activity although the cells do not synthesize CPE or CPG. The mutant differs from the wild type in that: (1) synthase activity was much diminished in the mutant, (2) sphingolipid synthesis does not occur in the mutant as evidenced by the absence of spots at sites where CPE and CPG migrate following two-dimensional thin layer chromatography, (3) incorporation of uniformly-labelled (/sup 14/C)serine carbon or (/sup 14/C)3KDS into sphingolipids was not observed in the mutant, (4) following incubation with (/sup 14/C)3KDS, radioactivity corresponding to dihydrosphingosine (DHS) and ceramide were observed in the mutant; no (/sup 14/C)DHS was detected in the wild type, and (5) enhanced incorporation of (/sup 14/C)serine carbon into two lipids not containing phosphorus was found in the mutant. The authors conclude, therefore, that this mutant, 1075BB, has a metabolic block at the terminal biosynthetic steps of sphingolipid synthesis.

  8. Isoenzymes performance of some rice varieties and their mutants

    International Nuclear Information System (INIS)

    Winarno, Ermin; Suliwarno, Ambyah; Ismachin, M.

    1992-01-01

    Isoenzymes performance of some rice varieties and their mutants. Genetics studies on alcohol dehydrogenase, malic enzyme, peroxidase, acid phosphase, and aminopeptidase isoenzymes were carried out on several groups of rice varieties and their mutant lines. The first groups consisted of Atomita I, Pelita I/1, A227/5, Mudgo, TN-1, and IR-26. The second group was Cisadane variety and its five mutants, namely OBS 18, OBS 208, OBS 297, OBS 306, and OBS 330. The third group was mutants line 627-10-3 and its mutants, namely 1063, 1066, 1067, 1076, and 1090. Isoenzymes extracts of the rice leaves were fractionated using polyacrylamide gel disc electrophoresis. The pattern of acid phosphate isoenzyme shows the specific character of rice mutants susceptible to brown plant hopper biotype 1. The gene(s) controlling malic enzyme in Cisadane's mutants is (are) estimated more resistant toward gamma irradiation than gene(s) responsible for controlling the other enzymes. Generally, the isoenzymes zymograms show that gene(s) controlling the mutants enzyme have undergone mutation. This case is shown by the changes of Rm value, as well as the amount and intensity of mutants bands. (authors). 7 refs., 7 figs

  9. Conditional lethal mutants of bacteriophage T4 unable to grow on a streptomycin resistant mutant of Escherichia coli

    Energy Technology Data Exchange (ETDEWEB)

    Childs, J.D.

    1977-01-01

    Sixteen conditional lethal mutants of bacteriophage T4D have been isolated which grow on Escherichia coli CR63 (a su/sup +/ streptomycin-sensitive K12 strain) but are restricted by CR/s (a streptomycin-resistant derivative of CR63). These mutants have been given the prefix str. Four of these mutants are amber and 12 appear to be missense. Eleven of the 12 missense mutants appear to be ''pseudo-amber'' (i.e., they are restricted by a su/sup -/ E. coli B strain but not by a su/sup -/ K12 strain); the other missense mutant was not restricted by either B or K12. The str mutations mapped in 12 different genes. Most were clustered in a region of early genes (gene 56 to gene 47). Fifty-eight amber and 10 ''pseudo-amber'' mutants isolated previously for their inability to grow on E. coli B were tested for restriction by CR/s. All the amber mutants grew normally on CR/s, whereas all 10 ''pseudo-amber'' mutants were restricted by CR/s. This implies that the phenotype of the ''pseudo-amber'' mutants is the result of a ribosomal difference between the permissive host CR63 and the restrictive hosts B and CR/s. These str mutants should prove to be useful alternatives to amber mutants for genetic and biochemical studies of bacteriophage T4 and for studies of the E. coli ribosome. It should be possible to isolate similar mutants in other bacteriophages provided that streptomycin resistant hosts are available.

  10. Thermal Isomerization Mechanism in Dronpa and Its Mutants.

    Science.gov (United States)

    Smyrnova, Daryna; Zinovjev, Kirill; Tuñón, Iñaki; Ceulemans, Arnout

    2016-12-22

    The photoswitching speed of the reversibly switchable fluorescent proteins (RSFPs) from the family of green fluorescent proteins (GFPs) changes upon mutation which is of direct importance for various high-resolution techniques. Dronpa is one of the most used RSFPs. Its point mutants rsFastLime (Dronpa V157G) and rsKame (Dronpa V157L) exhibit a striking difference in their photoswitching speed. Here the QM/MM on-the-fly string method is used in order to explore the details of the thermal isomerization mechanism. The four principal ways in which isomerization may occur have been scrutinized for each of the three proteins. It has been shown that thermal isomerization occurs via a one-bond-flip mechanism in all three proteins, although, in rsKame, where the chromophore is constrained more, the activation free energy difference between hula-twist and one-bond-flip is significantly smaller. Functional mode analysis has been applied to examine the motions of the amino acids during the isomerization. It clearly identifies the importance of Val/Leu 157 as well as the amino acids in the α-helix during the isomerization.

  11. Theoretical Analysis of Allosteric and Operator Binding for Cyclic-AMP Receptor Protein Mutants

    Science.gov (United States)

    Einav, Tal; Duque, Julia; Phillips, Rob

    2018-02-01

    Allosteric transcription factors undergo binding events both at their inducer binding sites as well as at distinct DNA binding domains, and it is often difficult to disentangle the structural and functional consequences of these two classes of interactions. In this work, we compare the ability of two statistical mechanical models - the Monod-Wyman-Changeux (MWC) and the Koshland-N\\'emethy-Filmer (KNF) models of protein conformational change - to characterize the multi-step activation mechanism of the broadly acting cyclic-AMP receptor protein (CRP). We first consider the allosteric transition resulting from cyclic-AMP binding to CRP, then analyze how CRP binds to its operator, and finally investigate the ability of CRP to activate gene expression. In light of these models, we examine data from a beautiful recent experiment that created a single-chain version of the CRP homodimer, thereby enabling each subunit to be mutated separately. Using this construct, six mutants were created using all possible combinations of the wild type subunit, a D53H mutant subunit, and an S62F mutant subunit. We demonstrate that both the MWC and KNF models can explain the behavior of all six mutants using a small, self-consistent set of parameters. In comparing the results, we find that the MWC model slightly outperforms the KNF model in the quality of its fits, but more importantly the parameters inferred by the MWC model are more in line with structural knowledge of CRP. In addition, we discuss how the conceptual framework developed here for CRP enables us to not merely analyze data retrospectively, but has the predictive power to determine how combinations of mutations will interact, how double mutants will behave, and how each construct would regulate gene expression.

  12. Characterization of a Thermo-Inducible Chlorophyll-Deficient Mutant in Barley

    Directory of Open Access Journals (Sweden)

    Rong Wang

    2017-11-01

    Full Text Available Leaf color is an important trait for not only controlling crop yield but also monitoring plant status under temperature stress. In this study, a thermo-inducible chlorophyll-deficient mutant, named V-V-Y, was identified from a gamma-radiated population of the barley variety Vlamingh. The leaves of the mutant were green under normal growing temperature but turned yellowish under high temperature in the glasshouse experiment. The ratio of chlorophyll a and chlorophyll b in the mutant declined much faster in the first 7–9 days under heat treatment. The leaves of V-V-Y turned yellowish but took longer to senesce under heat stress in the field experiment. Genetic analysis indicated that a single nuclear gene controlled the mutant trait. The mutant gene (vvy was mapped to the long arm of chromosome 4H between SNP markers 1_0269 and 1_1531 with a genetic distance of 2.2 cM and a physical interval of 9.85 Mb. A QTL for grain yield was mapped to the same interval and explained 10.4% of the yield variation with a LOD score of 4. This QTL is coincident with the vvy gene interval that is responsible for the thermo-inducible chlorophyll-deficient trait. Fine mapping, based on the barley reference genome sequence, further narrowed the vvy gene to a physical interval of 0.428 Mb with 11 annotated genes. This is the first report of fine mapping a thermo-inducible chlorophyll-deficient gene in barley.

  13. Optimized production of Serratia marcescens B742 mutants for preparing chitin from shrimp shells powders.

    Science.gov (United States)

    Zhang, Hongcai; Fang, Jiyang; Deng, Yun; Zhao, Yanyun

    2014-08-01

    To improve the deproteinization (DP) efficacy of shrimp shell powders (SSP) for preparing chitin, Serratia marcescens B742 mutants were prepared using 2% diethyl sulfate (DES), UV-irradiation, and/or microwave heating treatments. Both single-stage and multi-stage mutations were investigated for optimizing S. marcescens B742 mutation conditions. Under the optimized mutation conditions (2% DES treatment for 30min plus successive 20min UV-irradiation), the protease and chitosanase activity produced by mutant S. marcescens B742 was 240.15 and 170.6mU/mL, respectively, as compared with 212.58±1.51 and 83.75±6.51mU/mL, respectively, by wild S. marcescens B742. DP efficacy of SSP by mutant S. marcescens B742 reached 91.4±4.6% after 3d of submerged fermentation instead of 83.4±4.7% from the wild S. marcescens B742 after 4d of submerged fermentation. Molecular mass of chitosanase and protease was 41.20 and 47.10kDa, respectively, and both enzymes were verified by mass spectrometry analysis. The chitosanase from both wild and mutant S. marcescens B742 was activated by sodium dodecyl sulfate (SDS), Tween 20, Tween 40, and Triton-100, and the protease and chitosanase were strongly inhibited by ethylenediaminetetraacetic acid (EDTA). These results suggested that S. marcescens B742 mutants can be used in the biological production of chitin through deproteinization of SSP. Copyright © 2014 Elsevier B.V. All rights reserved.

  14. Flexibility in Anaerobic Metabolism as Revealed in a Mutant of Chlamydomonas reinhardtii Lacking Hydrogenase Activity

    Energy Technology Data Exchange (ETDEWEB)

    Dubini, A.; Mus, F.; Seibert, M.; Grossman, A. R.; Posewitz, M. C.

    2009-03-13

    The green alga Chlamydomonas reinhardtii has a network of fermentation pathways that become active when cells acclimate to anoxia. Hydrogenase activity is an important component of this metabolism, and we have compared metabolic and regulatory responses that accompany anaerobiosis in wild-type C. reinhardtii cells and a null mutant strain for the HYDEF gene (hydEF-1 mutant), which encodes an [FeFe] hydrogenase maturation protein. This mutant has no hydrogenase activity and exhibits elevated accumulation of succinate and diminished production of CO2 relative to the parental strain during dark, anaerobic metabolism. In the absence of hydrogenase activity, increased succinate accumulation suggests that the cells activate alternative pathways for pyruvate metabolism, which contribute to NAD(P)H reoxidation, and continued glycolysis and fermentation in the absence of O2. Fermentative succinate production potentially proceeds via the formation of malate, and increases in the abundance of mRNAs encoding two malateforming enzymes, pyruvate carboxylase and malic enzyme, are observed in the mutant relative to the parental strain following transfer of cells from oxic to anoxic conditions. Although C. reinhardtii has a single gene encoding pyruvate carboxylase, it has six genes encoding putative malic enzymes. Only one of the malic enzyme genes, MME4, shows a dramatic increase in expression (mRNA abundance) in the hydEF-1 mutant during anaerobiosis. Furthermore, there are marked increases in transcripts encoding fumarase and fumarate reductase, enzymes putatively required to convert malate to succinate. These results illustrate the marked metabolic flexibility of C. reinhardtii and contribute to the development of an informed model of anaerobic metabolism in this and potentially other algae.

  15. Metaharmonic Lattice Point Theory

    CERN Document Server

    Freeden, Willi

    2011-01-01

    Metaharmonic Lattice Point Theory covers interrelated methods and tools of spherically oriented geomathematics and periodically reflected analytic number theory. The book establishes multi-dimensional Euler and Poisson summation formulas corresponding to elliptic operators for the adaptive determination and calculation of formulas and identities of weighted lattice point numbers, in particular the non-uniform distribution of lattice points. The author explains how to obtain multi-dimensional generalizations of the Euler summation formula by interpreting classical Bernoulli polynomials as Green

  16. Flat Coalgebraic Fixed Point Logics

    Science.gov (United States)

    Schröder, Lutz; Venema, Yde

    Fixed point logics are widely used in computer science, in particular in artificial intelligence and concurrency. The most expressive logics of this type are the μ-calculus and its relatives. However, popular fixed point logics tend to trade expressivity for simplicity and readability, and in fact often live within the single variable fragment of the μ-calculus. The family of such flat fixed point logics includes, e.g., CTL, the *-nesting-free fragment of PDL, and the logic of common knowledge. Here, we extend this notion to the generic semantic framework of coalgebraic logic, thus covering a wide range of logics beyond the standard μ-calculus including, e.g., flat fragments of the graded μ-calculus and the alternating-time μ-calculus (such as ATL), as well as probabilistic and monotone fixed point logics. Our main results are completeness of the Kozen-Park axiomatization and a timed-out tableaux method that matches ExpTime upper bounds inherited from the coalgebraic μ-calculus but avoids using automata.

  17. Dwarf Rice Mutant Derived from 0.2 kGy Gamma Rays Irradiated Seeds of Atomita 4 Variety

    International Nuclear Information System (INIS)

    Sobrizal; Sutisna Sanjaya; Carkum; Mohamad Ismachin

    2004-01-01

    Dwarf rice mutant was obtained when Atomita 4 seeds were irradiated by 0.2 kGy gamma rays. The results of segregation analyses in F2 populations and F3 lines derived from reciprocal crosses of mutant and Atomita 4 suggested that the dwarf was controlled by a single recessive gene. This gene was not located on rice cytoplasmic genome but on nuclear genome. The gene for dwarf obtained in this study tentatively could be assumed as a new finding until the allelic relationships with other dwarf genes are verified. (author)

  18. AMPA receptor flip/flop mutants affecting deactivation, desensitization, and modulation by cyclothiazide, aniracetam, and thiocyanate.

    Science.gov (United States)

    Partin, K M; Fleck, M W; Mayer, M L

    1996-11-01

    AMPA receptor GluRA subunits with mutations at position 750, a residue shown previously to control allosteric regulation by cyclothiazide, were analyzed for modulation of deactivation and desensitization by cyclothiazide, aniracetam, and thiocyanate. Point mutations from Ser to Asn, Ala, Asp, Gly, Gln, Met, Cys, Thr, Leu, Val, and Tyr were constructed in GluRAflip. The last four of these mutants were not functional; S750D was active only in the presence of cyclothiazide, and the remaining mutants exhibited altered rates of deactivation and desensitization for control responses to glutamate, and showed differential modulation by cyclothiazide and aniracetam. Results from kinetic analysis are consistent with aniracetam and cyclothiazide acting via distinct mechanisms. Our experiments demonstrate for the first time the functional importance of residue 750 in regulating intrinsic channel-gating kinetics and emphasize the biological significance of alternative splicing in the M3-M4 extracellular loop.

  19. Analysis of a brittle-culm mutant of rice (Oryza sativa) induced bay gamma rays

    International Nuclear Information System (INIS)

    Doat, Jacqueline; Marie, R.

    1977-01-01

    An unexpected ''brittle-culm'' mutant has been screened in the progeny of the rice cultivar ''Balilla 28'' after a seed treatment by gamma rays from a Cobalt-60 source. This property proved hereditable and true-breeding. It does not affect the high resistance to lodging of rice plants. Important difference were pointed out between control and mutant lines in cellulose content and 1 p. cent NaOH extracts: ''brittle-culm'' straw contains less cellulose and shows a degradation of glucid coupounds. The brittleness of plant tissues appears to be correlated with a partial depolymerization of cellulose, associated with a possible transformation from alpha- to beta- or gamma-cellulose [fr

  20. Genetics of dwarfness in induced mutants of hexaploid triticale and its response to exogenous GA3

    International Nuclear Information System (INIS)

    Reddy, V.D.; Reddy, G.M.

    1991-01-01

    Genetics of dwarfism in two induced mutant (d 1 and d 2 ) of hexaploid triticale, DTS 330, revealed that this trait is governed by single recessive gene. Both d 1 and d 2 were allelic to each other and d 1 was dominant over d 2 . Both d 1 , d 2 and their F 1 showed no response to exogenous GA 3 , whereas, DTS 330, d 1 x DTS 330 and d 2 x DTS 330 were responsive. The endogenous levels of GA 3 were more in the dwarf mutants than control, suggesting that dwarfness in these may be due to a partial block in the GA utilizing mechanism, rather than a block in GA biosynthesis. (author). 5 refs., 2 tabs

  1. Cp/Heph mutant mice have iron-induced neurodegeneration diminished by deferiprone

    Science.gov (United States)

    Zhao, Liangliang; Hadziahmetovic, Majda; Wang, Chenguang; Xu, Xueying; Song, Ying; Jinnah, H.A.; Wodzinska, Jolanta; Iacovelli, Jared; Wolkow, Natalie; Krajacic, Predrag; Weissberger, Alyssa Cwanger; Connelly, John; Spino, Michael; Lee, Michael K.; Connor, James; Giasson, Benoit; Harris, Z. Leah; Dunaief, Joshua L.

    2016-01-01

    Brain iron accumulates in several neurodegenerative diseases and can cause oxidative damage, but mechanisms of brain iron homeostasis are incompletely understood. Patients with mutations in the cellular iron-exporting ferroxidase ceruloplasmin (Cp) have brain iron accumulation causing neurodegeneration. Here, we assessed the brains of mice with combined mutation of Cp and its homolog hephaestin. Compared to single mutants, brain iron accumulation was accelerated in double mutants in the cerebellum, substantia nigra, and hippocampus. Iron accumulated within glia, while neurons were iron deficient. There was loss of both neurons and glia. Mice developed ataxia and tremor, and most died by 9 months. Treatment with the oral iron chelator deferiprone diminished brain iron levels, protected against neuron loss, and extended lifespan. Ferroxidases play important, partially overlapping roles in brain iron homeostasis by facilitating iron export from glia, making iron available to neurons. PMID:26303407

  2. Isolation and genetic analysis of Aspergillus niger mutants with reduced extracellular glucoamylase

    International Nuclear Information System (INIS)

    Valent, G.U.; Calil, M.R.; Bonatelli Junior, R.

    1992-01-01

    Mutants with impaired production of extracellular glucoamylase were isolated at a high frequency (2% of survivors) from an Aspergillus niger strain treated with UV light. These were designated as low glucoamylase producers (lgp, up to 30% of the parental yield) and medium producers (mgp, a 35 to 50% decrease in enzyme level). All the mutants were shown to be recessive; one strain segregated two unlinked genes. Complementation tests, and segregation from heterozygous diploid, suggested at least three to four unlinked genes, each able to impair glucoamylase production. There is evidence of a single structural gene for glucoamylase in A. niger. Therefore, as production of extracellular enzymes is normally the final result of several steps at intracellular and membrane levels, including regulation of enzyme synthesis, we suggest intergenic interaction that controls extracellular enzyme accumulation and that mutation in any of these genes would result in impaired production. (author)

  3. Simultaneous analysis of multiple Mycobacterium tuberculosis knockdown mutants in vitro and in vivo.

    Directory of Open Access Journals (Sweden)

    Antje Blumenthal

    2010-12-01

    Full Text Available Mycobacterium tuberculosis (Mtb represents one of the most persistent bacterial threats to human health and new drugs are needed to limit its impact. Conditional knockdown mutants can help validate new drug targets, but the analysis of individual mutants is laborious and time consuming. Here, we describe quantitative DNA tags (qTags and their use to simultaneously analyze conditional Mtb knockdown mutants that allowed silencing the glyoxylate and methylcitrate cycles (via depletion of isocitrate lyase, ICL, the serine protease Rv3671c, and the core subunits of the mycobacterial proteasome, PrcB and PrcA. The impact of gene silencing in multi-strain cultures was determined by measuring the relative abundance of mutant-specific qTags with real-time PCR. This achieved accurate quantification over a broad range of qTag abundances and depletion of ICL, Rv3671c, or PrcBA resulted in the expected impairment of growth of Mtb with butyrate as the primary carbon source, survival during oxidative stress, acid stress and starvation. The impact of depleting ICL, Rv3671c, or PrcBA in multi-strain mouse infections was analyzed with two approaches. We first measured the relative abundance of mutant-specific qTags in total chromosomal DNA isolated from bacteria that were recovered from infected lungs on agar plates. We then developed a two-step amplification procedure, which allowed us to measure the abundances of individual mutants directly in infected lung tissue. Both strategies confirmed that inactivation of Rv3671c and PrcBA severely reduced persistence of Mtb in mice. The multi-strain infections furthermore suggested that silencing ICL not only prevented growth of Mtb during acute infections but also prevented survival of Mtb during chronic infections. Analyses of the ICL knockdown mutant in single-strain infections confirmed this and demonstrated that silencing of ICL during chronic infections impaired persistence of Mtb to the extent that the pathogen

  4. Photosynthetic characterization of a rolled leaf mutant of rice ( Oryza ...

    African Journals Online (AJOL)

    A new rolling leaf rice mutant was identified which showed an apparently straighter longitudinal shape normal transverse rolling characters at all developing stages. The chlorophyll contents per fresh weight of this mutant leaves were lower than those of wild-type. The electron transfer rate (ETR) and photochemical ...

  5. Complementation of sweet corn mutants: a method for grouping ...

    Indian Academy of Sciences (India)

    Maize endosperm mutant genes that affect quality of sweet corn can be grouped in two classes. One group of mutants namely brittle1 (bt1), brittle2 (bt2) and shrunken2 (sh2) .... significant influence on yield improvement, efficient test- ing of hybrids, and increasing the probability of identify- ing desirable hybrids (Tracy 1990).

  6. Screening of in vitro derived mutants of banana against nematodes ...

    African Journals Online (AJOL)

    The rest of the mutants namely Ro Im V4 6-1-2 and Si Im V4 6-2-5 were found to be susceptible to nematodes. The resistant and moderately resistant mutants of banana could be further used in breeding programmes as well as being recognized as potential cultivars of commerce. Key words: Banana, nematode, resistance, ...

  7. Isolation and characterization of stable mutants of Streptomyces ...

    Indian Academy of Sciences (India)

    2016-08-26

    Aug 26, 2016 ... Daunorubicin and its derivative doxorubicin are antitumour anthracycline antibiotics produced by Streptomyces peucetius. In this study we report isolation of stable mutants of S. peucetius blocked in different steps of the daunorubicin biosynthesis pathway. Mutants were screened on the basis of colony ...

  8. Development of Database Software with Plant Mutant Resources

    International Nuclear Information System (INIS)

    Namgoong, Won; Lee, M. J.; Kim, J. D.; Ma, N. K.

    2007-03-01

    In this research, mutants induced by nuclear radiation are developed information computerised system. The status and progress on the collection, identification and utilization of mutants in Korea are introduced. And it was produced home page, manual, test record, construction of system

  9. Lifespan and Glucose Metabolism in Insulin Receptor Mutant Mice

    Directory of Open Access Journals (Sweden)

    Takahiko Shimizu

    2011-01-01

    Full Text Available Insulin/insulin-like growth factor type 1 signaling regulates lifespan and resistance to oxidative stress in worms, flies, and mammals. In a previous study, we revealed that insulin receptor (IR mutant mice, which carry a homologous mutation found in the long-lived daf-2 mutant of Caenorhabditis elegans, showed enhanced resistance to oxidative stress cooperatively modulated by sex hormones and dietary signals (Baba et al., (2005. We herein investigated the lifespan of IR mutant mice to evaluate the biological significance of insulin signaling in mice. Under normoxia, mutant male mice had a lifespan comparable to that of wild-type male mice. IR mutant female mice also showed a lifespan similar to that of wild-type female mice, in spite of the fact that the IR mutant female mice acquired more resistance to oxidative stress than IR mutant male mice. On the other hand, IR mutant male and female mice both showed insulin resistance with hyperinsulinemia, but they did not develop hyperglycemia throughout their entire lifespan. These data indicate that the IR mutation does not impact the lifespan in mice, thus suggesting that insulin signaling might have a limited effect on the lifespan of mice.

  10. Sorghum Brown Midrib Mutants, Tools to Improve Biomass for Biofuels

    Science.gov (United States)

    To improve sorghum for cellulosic bioenergy uses, brown midrib mutants are being investigated for their ability to increase the conversion efficiency of biomass. brown midrib 6 and 12 (bmr6 and 12) mutants affect monolignol biosynthesis resulting in reduced lignin content and altered lignin composi...

  11. Unfolding intermediates of the mutant His-107-Tyr of human ...

    Indian Academy of Sciences (India)

    Srabani Taraphder

    Abstract. The mutant His-107-Tyr of human carbonic anhydrase II (HCA II) is highly unstable and has long been linked to a misfolding disease known as carbonic anhydrase deficiency syndrome (CADS). High temperature unfolding trajectories of the mutant are obtained from classical molecular dynamics simulations.

  12. Vaccines to Breast Cancer Based on p53 Mutants

    National Research Council Canada - National Science Library

    Ertl, Hildegund

    1997-01-01

    The aim of this proposal is to test vaccines expressing mouse mutant or wild-type p53 for induction of protective immunity against challenge with tumor cell lines expressing either mutant or high levels of wild-type p53...

  13. Characterization of human glucocerebrosidase from different mutant alleles

    NARCIS (Netherlands)

    Ohashi, T.; Hong, C. M.; Weiler, S.; Tomich, J. M.; Aerts, J. M.; Tager, J. M.; Barranger, J. A.

    1991-01-01

    Human cDNA was mutagenized to duplicate six naturally occurring mutations in the gene for glucocere-brosidase. The mutant genes were expressed in NIH 3T3 cells. The abnormal human enzymes were purified by immunoaffinity chromatography and characterized. The Asn370----Ser mutant protein differed from

  14. Mutants of Pseudomonas putida affected in poly-3-hydroxyalkanoate synthesis

    NARCIS (Netherlands)

    Ren, Q; Kessler, B; van der Leij, F; Witholt, B.

    The generation and characterization of Pseudomonas putida KT2442 mutants affected in poly-3-hydroxyalkanoate (PHA) synthesis are reported. The mutants from P. putida KT2442 carrying several copies of the PHA-polymerase-encoding gene (phaC) were isolated via N-methyl-N'-nitro-N-nitrosoguanidine

  15. Characteristics of mutant lines of sweet potato flour

    International Nuclear Information System (INIS)

    Aryanti

    2012-01-01

    Research on mutation induction of sweet potato Sari variety has been conducted. Flour mutant lines were obtained from selection of M1V5 tubers irradiated by gamma rays at the dose of 10 Gy. Flour was made by peeling of tubers, then dried, blended and sieved. The quality test of flour have been done by measuring degree of whiteness, proximate, amylose contents, water content, soluble water, swelling power, and flour characteristics. The result of this work showed that flour of C6.26.13 mutant line had higher protein content than the parent plant with concentration of 3.62 % and its amylose content was also higher than the other mutant lines. The soluble water value of mutant lines were significant different compared to the parent plant from 1.82 to 2.25 % and swelling power from 4.28 to 5.55 %. The flour granule of the mutant line was different compared to the parent plant. (author)

  16. Misfolded opsin mutants display elevated β-sheet structure.

    Science.gov (United States)

    Miller, Lisa M; Gragg, Megan; Kim, Tae Gyun; Park, Paul S-H

    2015-10-07

    Mutations in rhodopsin can cause misfolding and aggregation of the receptor, which leads to retinitis pigmentosa, a progressive retinal degenerative disease. The structure adopted by misfolded opsin mutants and the associated cell toxicity is poorly understood. Förster resonance energy transfer (FRET) and Fourier transform infrared (FTIR) microspectroscopy were utilized to probe within cells the structures formed by G188R and P23H opsins, which are misfolding mutants that cause autosomal dominant retinitis pigmentosa. Both mutants formed aggregates in the endoplasmic reticulum and exhibited altered secondary structure with elevated β-sheet and reduced α-helical content. The newly formed β-sheet structure may facilitate the aggregation of misfolded opsin mutants. The effects observed for the mutants were unrelated to retention of opsin molecules in the endoplasmic reticulum itself. Copyright © 2015 Federation of European Biochemical Societies. All rights reserved.

  17. Photosynthetic and nitrogen fixation capability in several soybean mutant lines

    International Nuclear Information System (INIS)

    Gandanegara, S.; Hendratno, K.

    1987-01-01

    Photosynthetic and nitrogen fixation capability in several soybean mutant lines. A greenhouse experiment has been carried out to study photosynthetic and nitrogen fixation capability of five mutant lines and two soybean varieties. An amount of 330 uCi of 14 CO 2 was fed to the plants including of the non-fixing reference crop (Chippewa non-nodulating isoline). Nitrogen fixation measurements was carried out using 15 N isotope dilution technique according to A-value concept. Results showed that beside variety/mutant lines, plant growth also has important role in photosynthetic and N fixing capability. Better growth and a higher photosynthetic capability in Orba, mutant lines nos. 63 and 65 resulted in a greater amount of N 2 fixed (mg N/plant) than other mutant lines. (author). 12 refs.; 5 figs

  18. The agronomic characters of a high protein rice mutant

    International Nuclear Information System (INIS)

    Harn, C.; Won, J.L.; Choi, K.T.

    1975-01-01

    Mutant lines (M 5 -M 9 ) of macro-phenotypic traits from several varieties were screened for the protein content. Mutant 398 (M 9 ) is one of the high protein mutants selected from Hokwang. Three years' tests revealed that it has a high protein line under any condition of cultivation. Except for early maturity and short culmness, other agronomic and yield characters were similar to the original variety. There was no difference between the mutant 398 and its mother variety in grain shape and weight, and also the size and protein content of the embryo. The high protein content of the mutant is attributable to the increase of protein in the endosperm. About 150 normal-looking or a few days-earlier-maturing selections were made from Jinheung variety in the M 3 and screened for protein. Promising lines in terms of the plant type, yield and protein were obtained. (author)

  19. Interesting Interest Points

    DEFF Research Database (Denmark)

    Aanæs, Henrik; Dahl, Anders Lindbjerg; Pedersen, Kim Steenstrup

    2012-01-01

    Not all interest points are equally interesting. The most valuable interest points lead to optimal performance of the computer vision method in which they are employed. But a measure of this kind will be dependent on the chosen vision application. We propose a more general performance measure bas...

  20. Poisson branching point processes

    International Nuclear Information System (INIS)

    Matsuo, K.; Teich, M.C.; Saleh, B.E.A.

    1984-01-01

    We investigate the statistical properties of a special branching point process. The initial process is assumed to be a homogeneous Poisson point process (HPP). The initiating events at each branching stage are carried forward to the following stage. In addition, each initiating event independently contributes a nonstationary Poisson point process (whose rate is a specified function) located at that point. The additional contributions from all points of a given stage constitute a doubly stochastic Poisson point process (DSPP) whose rate is a filtered version of the initiating point process at that stage. The process studied is a generalization of a Poisson branching process in which random time delays are permitted in the generation of events. Particular attention is given to the limit in which the number of branching stages is infinite while the average number of added events per event of the previous stage is infinitesimal. In the special case when the branching is instantaneous this limit of continuous branching corresponds to the well-known Yule--Furry process with an initial Poisson population. The Poisson branching point process provides a useful description for many problems in various scientific disciplines, such as the behavior of electron multipliers, neutron chain reactions, and cosmic ray showers