Sample records for silencing method suitable

  1. A systemic gene silencing method suitable for high throughput, reverse genetic analyses of gene function in fern gametophytes

    Directory of Open Access Journals (Sweden)

    Tanurdzic Milos


    Full Text Available Abstract Background Ceratopteris richardii is a useful experimental system for studying gametophyte development and sexual reproduction in plants. However, few tools for cloning mutant genes or disrupting gene function exist for this species. The feasibility of systemic gene silencing as a reverse genetics tool was examined in this study. Results Several DNA constructs targeting a Ceratopteris protoporphyrin IX magnesium chelatase (CrChlI gene that is required for chlorophyll biosynthesis were each introduced into young gametophytes by biolistic delivery. Their transient expression in individual cells resulted in a colorless cell phenotype that affected most cells of the mature gametophyte, including the meristem and gametangia. The colorless phenotype was associated with a 7-fold decrease in the abundance of the endogenous transcript. While a construct designed to promote the transient expression of a CrChlI double stranded, potentially hairpin-forming RNA was found to be the most efficient in systemically silencing the endogenous gene, a plasmid containing the CrChlI cDNA insert alone was sufficient to induce silencing. Bombarded, colorless hermaphroditic gametophytes produced colorless embryos following self-fertilization, demonstrating that the silencing signal could be transmitted through gametogenesis and fertilization. Bombardment of young gametophytes with constructs targeting the Ceratopteris filamentous temperature sensitive (CrFtsZ and uroporphyrin dehydrogenase (CrUrod genes also produced the expected mutant phenotypes. Conclusion A method that induces the systemic silencing of target genes in the Ceratopteris gametophyte is described. It provides a simple, inexpensive and rapid means to test the functions of genes involved in gametophyte development, especially those involved in cellular processes common to all plants.

  2. Recent patents in RNA silencing in plants: constructs, methods and applications in plant biotechnology. (United States)

    López-Gomollón, Sara; Dalmay, Tamas


    RNA silencing is a recently discovered mechanism to regulate gene expression at transcriptional and posttranscriptional levels. It is based on the recognition and methylation of target genes or cleavage of target mRNAs by small RNA molecules, with length varying from 21 to 24 nucleotides. RNA silencing plays an important role modulating most of the important cell processes, such as growth, development or stress response. During the past few years, diverse strategies have been applied to exploit RNA silencing as a tool to create plants with enhanced economical properties or able to cope with pathogens or abiotic stress. This review describes the most important patents related to RNA silencing in plants, which disclose vectors designed to induce RNA silencing by hairpin RNAs, amplicons or virus-based plasmids, methods for detection and quantification of silencing as well as general uses in plant biotechnology.

  3. Hearing Silence: Toward a Mixed-Method Approach for Studying Genres' Exclusionary Potential (United States)

    Randazzo, Chalice


    Traditional Rhetorical Genre Study (RGS) methods are not well adapted to study exclusion because excluded information and people are typically absent from the genre, and some excluded information is simply unrelated to the genre because of genre conventions or social context. Within genre-based silences, how can scholars differentiate between an…

  4. A comparison between analytic and numerical methods for modelling automotive dissipative silencers with mean flow (United States)

    Kirby, R.


    Identifying an appropriate method for modelling automotive dissipative silencers normally requires one to choose between analytic and numerical methods. It is common in the literature to justify the choice of an analytic method based on the assumption that equivalent numerical techniques are more computationally expensive. The validity of this assumption is investigated here, and the relative speed and accuracy of two analytic methods are compared to two numerical methods for a uniform dissipative silencer that contains a bulk reacting porous material separated from a mean gas flow by a perforated pipe. The numerical methods are developed here with a view to speeding up transmission loss computation, and are based on a mode matching scheme and a hybrid finite element method. The results presented demonstrate excellent agreement between the analytic and numerical models provided a sufficient number of propagating acoustic modes are retained. However, the numerical mode matching method is shown to be the fastest method, significantly outperforming an equivalent analytic technique. Moreover, the hybrid finite element method is demonstrated to be as fast as the analytic technique. Accordingly, both numerical techniques deliver fast and accurate predictions and are capable of outperforming equivalent analytic methods for automotive dissipative silencers.

  5. Selection of suitable NDT methods for building inspection (United States)

    Pauzi Ismail, Mohamad


    Construction of modern structures requires good quality concrete with adequate strength and durability. Several accidents occurred in the civil constructions and were reported in the media. Such accidents were due to poor workmanship and lack of systematic monitoring during the constructions. In addition, water leaking and cracking in residential houses was commonly reported too. Based on these facts, monitoring the quality of concrete in structures is becoming more and more important subject. This paper describes major Non-destructive Testing (NDT) methods for evaluating structural integrity of concrete building. Some interesting findings during actual NDT inspections on site are presented. The NDT methods used are explained, compared and discussed. The suitable methods are suggested as minimum NDT methods to cover parameters required in the inspection.

  6. Determining the most suitable method of otolith preparation for ...

    African Journals Online (AJOL)

    This study compares sectioned and whole lapillus and asterisci otoliths as suitable structures for ageing tigerfish, Hydrocynus vittatus. Fifty tigerfish were collected from the Pongolapoort Dam, KwaZulu-Natal, South Africa. Growth zone counts on sectioned lapilli showed the greatest percentage agreement (52%) and highest ...

  7. Nanoscale array structures suitable for surface enhanced raman scattering and methods related thereto (United States)

    Bond, Tiziana C.; Miles, Robin; Davidson, James C.; Liu, Gang Logan


    Methods for fabricating nanoscale array structures suitable for surface enhanced Raman scattering, structures thus obtained, and methods to characterize the nanoscale array structures suitable for surface enhanced Raman scattering. Nanoscale array structures may comprise nanotrees, nanorecesses and tapered nanopillars.

  8. Are deterministic methods suitable for short term reserve planning?

    International Nuclear Information System (INIS)

    Voorspools, Kris R.; D'haeseleer, William D.


    Although deterministic methods for establishing minutes reserve (such as the N-1 reserve or the percentage reserve) ignore the stochastic nature of reliability issues, they are commonly used in energy modelling as well as in practical applications. In order to check the validity of such methods, two test procedures are developed. The first checks if the N-1 reserve is a logical fixed value for minutes reserve. The second test procedure investigates whether deterministic methods can realise a stable reliability that is independent of demand. In both evaluations, the loss-of-load expectation is used as the objective stochastic criterion. The first test shows no particular reason to choose the largest unit as minutes reserve. The expected jump in reliability, resulting in low reliability for reserve margins lower than the largest unit and high reliability above, is not observed. The second test shows that both the N-1 reserve and the percentage reserve methods do not provide a stable reliability level that is independent of power demand. For the N-1 reserve, the reliability increases with decreasing maximum demand. For the percentage reserve, the reliability decreases with decreasing demand. The answer to the question raised in the title, therefore, has to be that the probability based methods are to be preferred over the deterministic methods

  9. Comparison of protein extraction methods suitable for proteomics ...

    African Journals Online (AJOL)



    Jul 27, 2011 ... An efficient protein extraction method is a prerequisite for successful implementation of proteomics. In this study, seedling roots of Jerusalem artichoke were treated with the concentration of 250 mM. NaCl for 36 h. Subsequently, six different protocols of protein extraction were applied to seedling roots.

  10. Comparison of protein extraction methods suitable for proteomics ...

    African Journals Online (AJOL)

    An efficient protein extraction method is a prerequisite for successful implementation of proteomics. In this study, seedling roots of Jerusalem artichoke were treated with the concentration of 250 mM NaCl for 36 h. Subsequently, six different protocols of protein extraction were applied to seedling roots of Jerusalem artichoke ...

  11. Silence multiple

    DEFF Research Database (Denmark)

    Søndergaard, Katia Dupret

    The article highlights the importance of silences in the processes of innovation in organizations, and the claim is that silence and the absence of talk distribute authority, responsibility and decisions. The act of silencing is conceptualised as a central “configurating actor”. Using an Actor......-Network Theoretical approach to organization studies silence is conceptualised as both a means and an effect of innovative efforts. It is a way of ordering practices. Thus silencing is thought of as a central potential change agent both in composing a kind of specific organizational collectivity and in composing new...... working practices more generally. In line with the approach to destabilise the mundane, invisible and taken-for-granted aspects of innovative efforts in organisations (crucial for ANT and foucauldian post-structuralism more broadly), this article suggests to non-silence the silence and make...

  12. A simple method suitable to study de novo root organogenesis

    Directory of Open Access Journals (Sweden)

    Xiaodong eChen


    Full Text Available De novo root organogenesis is the process in which adventitious roots regenerate from detached or wounded plant tissues or organs. In tissue culture, appropriate types and concentrations of plant hormones in the medium are critical for inducing adventitious roots. However, in natural conditions, regeneration from detached organs is likely to rely on endogenous hormones. To investigate the actions of endogenous hormones and the molecular mechanisms guiding de novo root organogenesis, we developed a simple method to imitate natural conditions for adventitious root formation by culturing Arabidopsis thaliana leaf explants on B5 medium without additive hormones. Here we show that the ability of the leaf explants to regenerate roots depends on the age of the leaf and on certain nutrients in the medium. Based on these observations, we provide examples of how this method can be used in different situations, and how it can be optimized. This simple method could be used to investigate the effects of various physiological and molecular changes on the regeneration of adventitious roots. It is also useful for tracing cell lineage during the regeneration process by differential interference contrast observation of -glucuronidase staining, and by live imaging of proteins labeled with fluorescent tags.

  13. Suitability of voltage stability study methods for real-time assessment

    DEFF Research Database (Denmark)

    Perez, Angel; Jóhannsson, Hjörtur; Vancraeyveld, Pieter


    This paper analyzes the suitability of existing methods for long-term voltage stability assessment for real-time operation. An overview of the relevant methods is followed with a comparison that takes into account the accuracy, computational efficiency and characteristics when used for security...... assessment. The results enable an evaluation of the run time of each method with respect to the number of inputs. Furthermore, the results assist in identifying which of the methods is most suitable for realtime operation in future power system with production based on fluctuating energy sources....

  14. Method: low-cost delivery of the cotton leaf crumple virus-induced gene silencing system

    Directory of Open Access Journals (Sweden)

    Tuttle John


    Full Text Available Abstract Background We previously developed a virus-induced gene silencing (VIGS vector for cotton from the bipartite geminivirusCotton leaf crumple virus (CLCrV. The original CLCrV VIGS vector was designed for biolistic delivery by a gene gun. This prerequisite limited the use of the system to labs with access to biolistic equipment. Here we describe the adaptation of this system for delivery by Agrobacterium (Agrobacterium tumefaciens. We also describe the construction of two low-cost particle inflow guns. Results The biolistic CLCrV vector was transferred into two Agrobacterium binary plasmids. Agroinoculation of the binary plasmids into cotton resulted in silencing and GFP expression comparable to the biolistic vector. Two homemade low-cost gene guns were used to successfully inoculate cotton (G. hirsutum and N. benthamiana with either the CLCrV VIGS vector or the Tomato golden mosaic virus (TGMV VIGS vector respectively. Conclusions These innovations extend the versatility of CLCrV-based VIGS for analyzing gene function in cotton. The two low-cost gene guns make VIGS experiments affordable for both research and teaching labs by providing a working alternative to expensive commercial gene guns.

  15. Estimating Rooftop Suitability for PV: A Review of Methods, Patents, and Validation Techniques

    Energy Technology Data Exchange (ETDEWEB)

    Melius, J. [National Renewable Energy Lab. (NREL), Golden, CO (United States); Margolis, R. [National Renewable Energy Lab. (NREL), Golden, CO (United States); Ong, S. [National Renewable Energy Lab. (NREL), Golden, CO (United States)


    A number of methods have been developed using remote sensing data to estimate rooftop area suitable for the installation of photovoltaics (PV) at various geospatial resolutions. This report reviews the literature and patents on methods for estimating rooftop-area appropriate for PV, including constant-value methods, manual selection methods, and GIS-based methods. This report also presents NREL's proposed method for estimating suitable rooftop area for PV using Light Detection and Ranging (LiDAR) data in conjunction with a GIS model to predict areas with appropriate slope, orientation, and sunlight. NREL's method is validated against solar installation data from New Jersey, Colorado, and California to compare modeled results to actual on-the-ground measurements.

  16. Increasing protocol suitability for clinical trials in sub-Saharan Africa: a mixed methods study. (United States)

    Vischer, Nerina; Pfeiffer, Constanze; Kealy, Jennifer; Burri, Christian


    The trial protocol is the most important document for clinical trials and describes not only the design and methodology of a study, but also all practical aspects. The suitability of the protocol has a direct impact on the execution and results of the trial. However, suitability is rarely addressed in trial practice and research. The aim of our study was to investigate protocol suitability and to identify suitability-enhancing measures for trials in sub-Saharan Africa. We used an exploratory mixed methods design. First, we interviewed 36 trial staff at different organisational levels in Ghana, Burkina Faso and Senegal. Second, we conducted an online survey among trial staff in sub-Saharan Africa to investigate trial protocol suitability based on the main themes distilled from the interviews. Protocol suitability surfaced as a prominent topic in interviews with trial staff, critiqued for its lack of clarity, implementability and adaptation to trial participants as well as to the workforce and infrastructure available. Both qualitative and quantitative investigations identified local site staff involvement in protocol development as the most helpful mean of increasing protocol suitability. Careful assessment of the local context, capacity and cultures, and ensuring that staff understand the protocol were also cited as helpful measures. Our data suggests that protocol suitability can be increased by discussing and reviewing the protocol with trial staff in advance. Involving operationally experienced staff would be most useful. For multicentre trials, we suggest that at least one trial staff member from each of the sites with the highest expected recruitment rates be involved in developing the protocol. Carefully assessing the context prior to study start is indispensable to ensuring protocol suitability and should particularly focus on the workforce and infrastructure available, as well as the needs and availability of trial participants. To allow for protocol

  17. Comparison of Four Weighting Methods in Fuzzy-based Land Suitability to Predict Wheat Yield

    Directory of Open Access Journals (Sweden)

    Fatemeh Rahmati


    Full Text Available Introduction: Land suitability evaluation is a process to examine the degree of land fitness for specific utilization and also makes it possible to estimate land productivity potential. In 1976, FAO provided a general framework for land suitability classification. It has not been proposed a specific method to perform this classification in the framework. In later years, a collection of methods was presented based on the FAO framework. In parametric method, different land suitability aspects are defined as completely discrete groups and are separated from each other by distinguished and consistent ranges. Therefore, land units that have moderate suitability can only choose one of the characteristics of predefined classes of land suitability. Fuzzy logic is an extension of Boolean logic by LotfiZadeh in 1965 based on the mathematical theory of fuzzy sets, which is a generalization of the classical set theory. By introducing the notion of degree in the verification of a condition, fuzzy method enables a condition to be in a state other than true or false, as well as provides a very valuable flexibility for reasoning, which makes it possible to take into account inaccuracies and uncertainties. One advantage of fuzzy logic in order to formalize human reasoning is that the rules are set in natural language. In evaluation method based on fuzzy logic, the weights are used for land characteristics. The objective of this study was to compare four methods of weight calculation in the fuzzy logic to predict the yield of wheat in the study area covering 1500 ha in Kian town in Shahrekord (Chahrmahal and Bakhtiari province, Iran. Materials and Methods: In such investigations, climatic factors, and soil physical and chemical characteristics are studied. This investigation involves several studies including a lab study, and qualitative and quantitative land suitability evaluation with fuzzy logic for wheat. Factors affecting the wheat production consist of

  18. Suitability of leaching test methods for fly ash and slag: A review

    Directory of Open Access Journals (Sweden)

    Manoj Kumar Tiwari


    Full Text Available Fly ash and slag leachate pollution can be of great environmental concern due to generation of these wastes in huge quantities from their respective industrial units, mainly coal-based thermal power plants and iron and steel plants. For simulation of natural leaching in laboratory, various leaching methods are available, but selection of a method that can exactly simulate the real-life scenario for accurate estimation of various pollutants is challenging; particularly, the heavy metals present and impact due to reuse or disposal of these wastes. For choosing the most suitable leaching method according to specific situation, one must primarily consider the chemical and physical properties of wastes, the composition of the source, age of waste disposal, and the climatic conditions of the disposal area. Since these factors may not be specified, a variety of leaching methods with relevant equipment have been proposed by researchers; that are based on their required information to particular conditions in absence of a prescribed protocol and non standardization of equipment. The present review is an attempt to investigate the suitable leaching method for coal fly ash and slag.

  19. Suitability of various DNA extraction methods for a traditional Chinese paocai system. (United States)

    Zhao, Nan; Cai, Jialiang; Zhang, Chuchu; Guo, Zhuang; Lu, Wenwei; Yang, Bo; Tian, Feng-Wei; Liu, Xiao-Ming; Zhang, Hao; Chen, Wei


    Traditional paocai brine (PB), which is continuously propagated by back-slopping and contains various species of lactic acid bacteria (LAB), is critical for the flavor of paocai. Culture-independent approaches are commonly used to investigate the microbial communities of fermented food. To evaluate the influence of different DNA (DNA) extraction methods on estimates of bacterial community profiles from 4 PBs, the lysis efficiency, DNA yield, purity and denaturing gradient gel electrophoresis (DGGE) profiles of V3 region of a 16S ribosomal ribonucleic acid gene were acquired. The cell lysis pattern of SDS + beads and Lysing matrix E+ beads (methods 3 and 4) showed higher cell lysis efficiency than SDS and SDS + Lysozyme (methods 1 and 2) in all PBs. SDS + beads obtained the largest DNA yield of the 4 methods. Moreover, methods 3 and 4 resulted in higher H' values and generated more global bacteria profiles than other methods. Overall, our results demonstrate that the properties of PB significantly affect the efficiency of DNA extraction methods. Methods 3 and 4 were both suitable for DNA extraction from PB. Method 3 is more economic, simple and rapid than method 4 for large-scale studies of the bacterial profiles of PB.

  20. Identification of suitable genes contributes to lung adenocarcinoma clustering by multiple meta-analysis methods. (United States)

    Yang, Ze-Hui; Zheng, Rui; Gao, Yuan; Zhang, Qiang


    With the widespread application of high-throughput technology, numerous meta-analysis methods have been proposed for differential expression profiling across multiple studies. We identified the suitable differentially expressed (DE) genes that contributed to lung adenocarcinoma (ADC) clustering based on seven popular multiple meta-analysis methods. Seven microarray expression profiles of ADC and normal controls were extracted from the ArrayExpress database. The Bioconductor was used to perform the data preliminary preprocessing. Then, DE genes across multiple studies were identified. Hierarchical clustering was applied to compare the classification performance for microarray data samples. The classification efficiency was compared based on accuracy, sensitivity and specificity. Across seven datasets, 573 ADC cases and 222 normal controls were collected. After filtering out unexpressed and noninformative genes, 3688 genes were remained for further analysis. The classification efficiency analysis showed that DE genes identified by sum of ranks method separated ADC from normal controls with the best accuracy, sensitivity and specificity of 0.953, 0.969 and 0.932, respectively. The gene set with the highest classification accuracy mainly participated in the regulation of response to external stimulus (P = 7.97E-04), cyclic nucleotide-mediated signaling (P = 0.01), regulation of cell morphogenesis (P = 0.01) and regulation of cell proliferation (P = 0.01). Evaluation of DE genes identified by different meta-analysis methods in classification efficiency provided a new perspective to the choice of the suitable method in a given application. Varying meta-analysis methods always present varying abilities, so synthetic consideration should be taken when providing meta-analysis methods for particular research. © 2015 John Wiley & Sons Ltd.

  1. A simple method of genomic DNA extraction suitable for analysis of bulk fungal strains. (United States)

    Zhang, Y J; Zhang, S; Liu, X Z; Wen, H A; Wang, M


    A simple and rapid method (designated thermolysis) for extracting genomic DNA from bulk fungal strains was described. In the thermolysis method, a few mycelia or yeast cells were first rinsed with pure water to remove potential PCR inhibitors and then incubated in a lysis buffer at 85 degrees C to break down cell walls and membranes. This method was used to extract genomic DNA from large numbers of fungal strains (more than 92 species, 35 genera of three phyla) isolated from different sections of natural Ophiocordyceps sinensis specimens. Regions of interest from high as well as single-copy number genes were successfully amplified from the extracted DNA samples. The DNA samples obtained by this method can be stored at -20 degrees C for over 1 year. The method was effective, easy and fast and allowed batch DNA extraction from multiple fungal isolates. Use of the thermolysis method will allow researchers to obtain DNA from fungi quickly for use in molecular assays. This method requires only minute quantities of starting material and is suitable for diverse fungal species.

  2. Selection of Suitable Biological Method for the Synthesis of Silver Nanoparticles

    Directory of Open Access Journals (Sweden)

    Muhammad Ashraf Sabri


    Full Text Available This paper aims to present a brief overview of different biosynthesis routes of silver nanoparticles (NPs, their applications and influence of the method used on the size and morphology of these nanoparticles. A detailed and comprehensive study of available biological methods, also referred to as a bottom-up approach, as well as techniques reported, have been provided with an eye for details and comparison between the techniques involving fungi, bacteria, algae and plant extracts. Plant-derived bioreduc‐ tants such as leaf, stem or root extracts of various plants are seen as suitable solutions to green synthesis of silver NPs, implementing an easy, non-toxic, clean and environmen‐ tally friendly approach. Furthermore, reports on the antimicrobial activities with the zone of inhibition for various pathogens have also been included.

  3. The measurement of surface roughness to determine the suitability of different methods for stone cleaning

    International Nuclear Information System (INIS)

    Vazquez-Calvo, Carmen; Alvarez de Buergo, Monica; Fort, Rafael; Varas-Muriel, Maria Jose


    The roughness of stone surface was measured, before and after bead blasting-based cleaning methods, to select the most efficient one to be used in masonry and stonework of specific areas of the Cathedral of Segovia (Spain). These types of cleaning methods can, besides the removal of soiling and surface deposits, leave a rougher surface, which would mean higher and more rapid water retention and deposit accumulation due to a specific surface increase, therefore accelerating stone decay. Or, in contrast, the cleaning method can be so aggressive that it can smooth the surface by reducing its roughness, a fact that usually corresponds to excessive material removal—soot and deposits–-but also part of the stone substrate. Roughness results were complemented with scanning electron microscopy observations and analyses and colour measurements. Finally, it was possible to select the best cleaning method among the six that were analysed, for different areas and different stone materials. Therefore, this study confirms the measurement of surface roughness as a reliable test to determine the suitability of stone cleaning methods; it is a non-destructive technique, portable and friendly to use, which can help us to rapidly assess—together with other techniques—the efficacy and aggressiveness of the stone cleaning method. (paper)

  4. Comparing methods suitable for monitoring marine mammals in low visibility conditions during seismic surveys. (United States)

    Verfuss, Ursula K; Gillespie, Douglas; Gordon, Jonathan; Marques, Tiago A; Miller, Brianne; Plunkett, Rachael; Theriault, James A; Tollit, Dominic J; Zitterbart, Daniel P; Hubert, Philippe; Thomas, Len


    Loud sound emitted during offshore industrial activities can impact marine mammals. Regulations typically prescribe marine mammal monitoring before and/or during these activities to implement mitigation measures that minimise potential acoustic impacts. Using seismic surveys under low visibility conditions as a case study, we review which monitoring methods are suitable and compare their relative strengths and weaknesses. Passive acoustic monitoring has been implemented as either a complementary or alternative method to visual monitoring in low visibility conditions. Other methods such as RADAR, active sonar and thermal infrared have also been tested, but are rarely recommended by regulatory bodies. The efficiency of the monitoring method(s) will depend on the animal behaviour and environmental conditions, however, using a combination of complementary systems generally improves the overall detection performance. We recommend that the performance of monitoring systems, over a range of conditions, is explored in a modelling framework for a variety of species. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  5. Selection strategy for the most Suitable CEC method for clay barrier characterisation

    International Nuclear Information System (INIS)

    Dohrmann, R.; Kaufhold, S.


    competition to barium ions in smectite interlayers. The present study aims at proposing a selection strategy for CEC methods including exchangeable cation determination of different clay materials typically used in radioactive waste disposal studies. 1) If the clay/bentonite contains calcium carbonates but no gypsum several methods are available, all of them are based on a saturation of the exchange solution with calcite before the exchange experiment starts: AgTU-calcite, CoHex-calcite, and Cu-trien5xcalcite. If dolomite is present the initial calcite saturation minimizes dolomite dissolution as well (proved for AgTU-calcite). 2) If a bentonite contains carbonates and gypsum the only known successful method for determination of all exchangeable cations (including calcium) and the CEC is a combination of two separate results: i) calcite saturation of exchange solution (e.g. Cu-trien5xcalcite) and ii) quantification of gypsum with suitable mineralogical methods. Result i) is free of error caused by calcite dissolution; however it is still wrong because it contains significant amounts of Ca 2+ from gypsum dissolution. After proving that gypsum was completely dissolved during the exchange experiment result ii) was used to subtract the theoretical Ca 2+ portion of gypsum from result i). Gypsum saturation of exchange solutions was tested as not suitable owing to the high solubility of gypsum which results in high initial Ca 2+ concentrations. 3) If no such soluble minerals are present and if the pore water contains no excess electrolyte then usually any accepted CEC method can be used, however, the results should always be checked for plausibility (e.g. using mineralogical composition of the sample). Any accepted method means that methods with known systematic errors (hydrophobic interaction, dependency on layer charge density) are excluded. (authors)

  6. Is the molecular statics method suitable for the study of nanomaterials? A study case of nanowires

    International Nuclear Information System (INIS)

    Chang, I-L; Chen, Y-C


    Both molecular statics and molecular dynamics methods were employed to study the mechanical properties of copper nanowires. The size effect on both elastic and plastic properties of square cross-sectional nanowire was examined and compared systematically using two molecular approaches. It was found consistently from both molecular methods that the elastic and plastic properties of nanowires depend on the lateral size of nanowires. As the lateral size of nanowires decreases, the values of Young's modulus decrease and dislocation nucleation stresses increase. However, it was shown that the dislocation nucleation stress would be significantly influenced by the axial periodic length of the nanowire model using the molecular statics method while molecular dynamics simulations at two distinct temperatures (0.01 and 300 K) did not show the same dependence. It was concluded that molecular statics as an energy minimization numerical scheme is quite insensitive to the instability of atomic structure especially without thermal fluctuation and might not be a suitable tool for studying the behaviour of nanomaterials beyond the elastic limit

  7. An efficient method to enhance gene silencing by using precursor microRNA designed small hairpin RNAs. (United States)

    Shan, Zhixin; Lin, Qiuxiong; Deng, Chunyu; Li, Xiaohong; Huang, Wei; Tan, Honghong; Fu, Yongheng; Yang, Min; Yu, Xi-Yong


    Gene silencing can be mediated by small interfering RNA (siRNA) and microRNA (miRNA). To investigate the potential application of using a precursor microRNA (pre-miRNA) backbone for gene silencing, we studied the inhibition efficiency of exogenous GFP and endogenous GAPDH by conventional shRNA- and pre-miRNA-designed hairpins, respectively. In this study, the conventional shRNA-, pre-miRNA-30-, and pre-miRNA-155-designed hairpins targeting either GFP or GAPDH were transfected into the HEK293 cells that were mediated by the pSilencer-4.1-neo vector, which carries a modified RNA polymerase II-type CMV promoter. Comparisons with conventional GFP shRNA showed that GFP levels were reduced markedly by pre-miRNA-30- and pre-miRNA-155-designed GFP shRNAs by fluorescence microscopy. The consistent results from semi-quantitative RT-PCR and Western blot analysis revealed that pre-miRNA-30- and pre-miRNA-155-designed GFP shRNAs could suppress GFP expression significantly. As for endogenous GAPDH, the results from semi-quantitative RT-PCR and Western blot analysis showed that pre-miRNA-30- and pre-miRNA-155-designed GAPDH shRNAs could suppress GAPDH expression even more efficiently than conventional GAPDH shRNA. Together, this study confirmed the efficiency of gene silencing mediated by pre-miRNA-30- and pre-miRNA-155-designed shRNAs, demonstrating that pre-miRNA-designed hairpins are a good strategy for gene silencing.

  8. Potential Suitable Methods for Measuring the Effects of Animal-Assisted Activities and Therapy: a Review

    Directory of Open Access Journals (Sweden)

    Machová K.


    Full Text Available Dogs are companions of humans since time beyond memory and their use in rehabilitation processes is increasingly frequent. Dogs can be used for animal-assisted activities (AAA as well as animal-assisted therapy (AAT. The effects, however, have not been fully demonstrated, reasons for this including difficulties in providing evidence of positive action. According to previous studies, there is a decrease in heart and respiratory rate, blood pressure reduction, more positive mood as well as warming of muscles, which promotes relaxation of spastic areas. Regulation of hormone levels might also occur as a result of AAA/AAT. Indeed, increases in oxytocin levels and decreases of cortisol have been found. However, a unified methodology for clear measuring the entire impact of AAA/AAT on patients is missing. This survey evaluated different methods for measuring the effects of AAA/AAT, with results showing that the most suitable ones, selected on the basis of the effect of rehabilitation, comprise thermography, spectral analysis of heart rate, electromyography (EMG, polyelectromyography (PEMG, and blood sampling. Because AAT making use of dogs has not yet been recognized as an official method of treatment, it is very important to find out objective means to evidence its beneficial effects. Highlights:

  9. Optimization of suitable ethanol blend ratio for motorcycle engine using response surface method. (United States)

    Chen, Yu-Liang; Chen, Suming; Tsai, Jin-Ming; Tsai, Chao-Yin; Fang, Hsin-Hsiung; Yang, I-Chang; Liu, Sen-Yuan


    In view of energy shortage and air pollution, ethanol-gasoline blended fuel used for motorcycle engine was studied in this work. The emissions of carbon monoxide (CO), nitrogen oxides (NO(X)) and engine performance of a 125 cc four-stroke motorcycle engine with original carburetor using ethanol-gasoline fuels were investigated. The model of three-variable Box Behnken design (BBD) was used for experimental design, the ethanol blend ratios were prepared at 0, 10, 20 vol%; the speeds of motorcycle were selected as 30, 45, 60 km/h; and the throttle positions were set at 30, 60, 90 %. Both engine performance and air pollutant emissions were then analyzed by response surface method (RSM) to yield optimum operation parameters for tolerable pollutant emissions and maximum engine performance. The RSM optimization analysis indicated that the most suitable ethanol-gasoline blended ratio was found at the range of 3.92-4.12 vol% to yield a comparable fuel conversion efficiency, while considerable reductions of exhaust pollutant emissions of CO (-29 %) and NO(X) (-12 %) when compared to pure gasoline fuel. This study demonstrated low ethanol-gasoline blended fuels could be used in motorcycle carburetor engines without any modification to keep engine power while reducing exhaust pollutants.

  10. Selection of suitable propagation method for consistent plantlets production in Stevia rebaudiana (Bertoni) (United States)

    Khalil, Shahid Akbar; Zamir, Roshan; Ahmad, Nisar


    Stevia rebaudiana (Bert.) is an emerging sugar alternative and anti-diabetic plant in Pakistan. That is why people did not know the exact time of propagation. The main objective of the present study was to establish feasible propagation methods for healthy biomass production. In the present study, seed germination, stem cuttings and micropropagation were investigated for higher productivity. Fresh seeds showed better germination (25.51–40%) but lost viability after a few days of storage. In order to improve the germination percentage, seeds were irradiated with 2.5, 5.0, 7.5 and 10 Gy gamma doses. But gamma irradiation did not show any significant change in seed germination. A great variation in survival of stem cutting was observed in each month of 2012. October and November were found the most suitable months for stem cutting survival (60%). In order to enhance survival, stem cuttings were also dipped in different plant growth regulators (PGRs) solution. Only indole butyric acid (IBA; 1000 ppm) treated cutting showed a higher survival (33%) than control (11.1%). Furthermore, simple and feasible indirect regeneration system was established from leaf explants. Best callus induction (84.6%) was observed on MS-medium augmented with 6-benzyladenine (BA) and 2,4-dichlorophenoxyacetic acid (2,4-D; 2.0 mg l−1). For the first time, we obtained the highest number of shoots (106) on a medium containing BA (1.5 mg l−1) and gibberellic acid (GA3; 0.5 mg l−1). Plantlets were successfully acclimatized in plastic pots. The current results preferred micropropagation (85%) over seed germination (25.51–40%) and stem cutting (60%). PMID:25473365

  11. Antisense gene silencing

    DEFF Research Database (Denmark)

    Nielsen, Troels T; Nielsen, Jørgen E


    Since the first reports that double-stranded RNAs can efficiently silence gene expression in C. elegans, the technology of RNA interference (RNAi) has been intensively exploited as an experimental tool to study gene function. With the subsequent discovery that RNAi could also be applied...... to mammalian cells, the technology of RNAi expanded from being a valuable experimental tool to being an applicable method for gene-specific therapeutic regulation, and much effort has been put into further refinement of the technique. This review will focus on how RNAi has developed over the years and how...

  12. Are single-well "push-pull" tests suitable tracer methods for aquifer characterization? (United States)

    Hebig, Klaus; Zeilfelder, Sarah; Ito, Narimitsu; Machida, Isao; Scheytt, Traugott; Marui, Atsunao


    Recently, investigations were conducted for geological and hydrogeological characterisation of the sedimentary coastal basin of Horonobe (Hokkaido, Japan). Coastal areas are typical geological settings in Japan, which are less tectonically active than the mountain ranges. In Asia, and especially in Japan, these areas are often densely populated. Therefore, it is important to investigate the behaviour of solutes in such unconsolidated aquifers. In such settings sometimes only single boreholes or groundwater monitoring wells are available for aquifer testing for various reasons, e.g. depths of more than 100 m below ground level and slow groundwater velocities due to density driven flow. A standard tracer test with several involved groundwater monitoring wells is generally very difficult or even not possible at these depths. One of the most important questions in our project was how we can obtain information about chemical and hydraulic properties in such aquifers. Is it possible to characterize solute transport behaviour parameters with only one available groundwater monitoring well or borehole? A so-called "push-pull" test may be one suitable method for aquifer testing with only one available access point. In a push-pull test a known amount of several solutes including a conservative tracer is injected into the aquifer ("push") and afterwards extracted ("pull"). The measured breakthrough curve during the pumping back phase can then be analysed. This method has already been used previously with various aims, also in the recent project (e.g. Hebig et al. 2011, Zeilfelder et al. 2012). However, different test setups produced different tracer breakthrough curves. As no systematic evaluation of this aquifer tracer test method was done so far, nothing is known about its repeatability. Does the injection and extraction rate influence the shape of the breakthrough curve? Which role plays the often applied "chaser", which is used to push the test solution out from the

  13. A simple method of DNA extraction from coffee seeds suitable for ...

    African Journals Online (AJOL)

    Quality of DNA was confirmed by digestion using EcoRI, HindIII and PstI restriction endonucleases and complete digestion was observed. PCR with random decamer primers and consensus primers of mitochondria and chloroplast DNA and PCR-RFLP revealed the suitability of the DNA for PCR based marker techniques ...

  14. Comparing habitat suitability and connectivity modeling methods for conserving pronghorn migrations.

    Directory of Open Access Journals (Sweden)

    Erin E Poor

    Full Text Available Terrestrial long-distance migrations are declining globally: in North America, nearly 75% have been lost. Yet there has been limited research comparing habitat suitability and connectivity models to identify migration corridors across increasingly fragmented landscapes. Here we use pronghorn (Antilocapra americana migrations in prairie habitat to compare two types of models that identify habitat suitability: maximum entropy (Maxent and expert-based (Analytic Hierarchy Process. We used distance to wells, distance to water, NDVI, land cover, distance to roads, terrain shape and fence presence to parameterize the models. We then used the output of these models as cost surfaces to compare two common connectivity models, least-cost modeling (LCM and circuit theory. Using pronghorn movement data from spring and fall migrations, we identified potential migration corridors by combining each habitat suitability model with each connectivity model. The best performing model combination was Maxent with LCM corridors across both seasons. Maxent out-performed expert-based habitat suitability models for both spring and fall migrations. However, expert-based corridors can perform relatively well and are a cost-effective alternative if species location data are unavailable. Corridors created using LCM out-performed circuit theory, as measured by the number of pronghorn GPS locations present within the corridors. We suggest the use of a tiered approach using different corridor widths for prioritizing conservation and mitigation actions, such as fence removal or conservation easements.

  15. Comparing habitat suitability and connectivity modeling methods for conserving pronghorn migrations. (United States)

    Poor, Erin E; Loucks, Colby; Jakes, Andrew; Urban, Dean L


    Terrestrial long-distance migrations are declining globally: in North America, nearly 75% have been lost. Yet there has been limited research comparing habitat suitability and connectivity models to identify migration corridors across increasingly fragmented landscapes. Here we use pronghorn (Antilocapra americana) migrations in prairie habitat to compare two types of models that identify habitat suitability: maximum entropy (Maxent) and expert-based (Analytic Hierarchy Process). We used distance to wells, distance to water, NDVI, land cover, distance to roads, terrain shape and fence presence to parameterize the models. We then used the output of these models as cost surfaces to compare two common connectivity models, least-cost modeling (LCM) and circuit theory. Using pronghorn movement data from spring and fall migrations, we identified potential migration corridors by combining each habitat suitability model with each connectivity model. The best performing model combination was Maxent with LCM corridors across both seasons. Maxent out-performed expert-based habitat suitability models for both spring and fall migrations. However, expert-based corridors can perform relatively well and are a cost-effective alternative if species location data are unavailable. Corridors created using LCM out-performed circuit theory, as measured by the number of pronghorn GPS locations present within the corridors. We suggest the use of a tiered approach using different corridor widths for prioritizing conservation and mitigation actions, such as fence removal or conservation easements.

  16. Assessing stationary laboratory test methods for underground mining vehicles to determine their suitability in replicating real-world emissions

    CSIR Research Space (South Africa)

    Wattrus, MC


    Full Text Available adequately and that equipment is operating within acceptable parameters. An investigation into the required equipment and in-field test methods was conducted as the basis of this study. Stationary in-field vehicle emission tests were reproduced in a... controlled engine test cell to establish the repeatability and suitability of these methods. These stationary tests were compared against real-world data recorded during a load-haul- dump (LHD) vehicle underground operational cycle in order to devise...

  17. Method for assessment of the technical potential of the plant agricultural waste suitable for energy purposes in Bulgaria

    International Nuclear Information System (INIS)

    Georgiev, V.


    A method for assessment of technical potential of quantitatively important plant agriculture waste; straw from wheat and barley; corn stalks; sunflower stalks and heads; tobacco stalks; orchard prunings and vineyard prunings suitable for energy purposes for Bulgarian conditions is developed. Data for assessment of the technical potential for 2002 using this method are presented. A comparison between technical potential of mentioned wastes, final energy consumption in agriculture and all branches of Bulgarian economy for 2002 is made. (author)

  18. A Two-Point Newton Method Suitable for Nonconvergent Cases and with Super-Quadratic Convergence

    Directory of Open Access Journals (Sweden)

    Ababu Teklemariam Tiruneh


    Full Text Available An iterative formula based on Newton’s method alone is presented for the iterative solutions of equations that ensures convergence in cases where the traditional Newton Method may fail to converge to the desired root. In addition, the method has super-quadratic convergence of order 2.414 (i.e., . Newton method is said to fail in certain cases leading to oscillation, divergence to increasingly large number, or offshooting away to another root further from the desired domain or offshooting to an invalid domain where the function may not be defined. In addition when the derivative at the iteration point is zero, Newton method stalls. In most of these cases, hybrids of several methods such as Newton, bisection, and secant methods are suggested as substitute methods and Newton method is essentially blended with other methods or altogether abandoned. This paper argues that a solution is still possible in most of these cases by the application of Newton method alone without resorting to other methods and with the same computational effort (two functional evaluations per iteration like the traditional Newton method. In addition, the proposed modified formula based on Newton method has better convergence characteristics than the traditional Newton method.

  19. Evaluation of Deterministic and Complex Analytical Hierarchy Process Methods for Agricultural Land Suitability Analysis in a Changing Climate

    Directory of Open Access Journals (Sweden)

    Harmen Romeijn


    Full Text Available Land suitability analysis is employed to evaluate the appropriateness of land for a particular purpose whilst integrating both qualitative and quantitative inputs, which can be continuous in nature. However, in agricultural modelling there is often a disregard of this contiguous aspect. Therefore, some parametric procedures for suitability analysis compartmentalise units into defined membership classes. This imposition of crisp boundaries neglects the continuous formations found throughout nature and overlooks differences and inherent uncertainties found in the modelling. This research will compare two approaches to suitability analysis over three differing methods. The primary approach will use an Analytical Hierarchy Process (AHP, while the other approach will use a Fuzzy AHP over two methods; Fitted Fuzzy AHP and Nested Fuzzy AHP. Secondary to this, each method will be assessed into how it behaves in a climate change scenario to understand and highlight the role of uncertainties in model conceptualisation and structure. Outputs and comparisons between each method, in relation to area, proportion of membership classes and spatial representation, showed that fuzzy modelling techniques detailed a more robust and continuous output. In particular the Nested Fuzzy AHP was concluded to be more pertinent, as it incorporated complex modelling techniques, as well as the initial AHP framework. Through this comparison and assessment of model behaviour, an evaluation of each methods predictive capacity and relevance for decision-making purposes in agricultural applications is gained.

  20. Choosing Learning Methods Suitable for Teaching and Learning in Computer Science (United States)

    Taylor, Estelle; Breed, Marnus; Hauman, Ilette; Homann, Armando


    Our aim is to determine which teaching methods students in Computer Science and Information Systems prefer. There are in total 5 different paradigms (behaviorism, cognitivism, constructivism, design-based and humanism) with 32 models between them. Each model is unique and states different learning methods. Recommendations are made on methods that…

  1. New method suitable for measuring chromaticity and photometric quantity of laser displays (United States)

    Hieda, Keisuke; Maruyama, Tomoyuki; Takesako, Tomohiro; Narusawa, Fumio


    A new measurement method for laser displays is proposed to solve the problems caused by the extremely narrow spectra of lasers. This method is called the discrete centroid wavelength method, and is capable of simultaneously measuring the centroid wavelength and radiometric quantity of blue, green, and red lasers. First, its superiority to the tristimulus-value direct-reading method was verified by numerical analysis. Subsequently, the high accuracy of our method was confirmed by performing actual measurements of a uniform-luminance screen generated by RGB lasers.

  2. A simple method for plasma total vitamin C analysis suitable for routine clinical laboratory use. (United States)

    Robitaille, Line; Hoffer, L John


    In-hospital hypovitaminosis C is highly prevalent but almost completely unrecognized. Medical awareness of this potentially important disorder is hindered by the inability of most hospital laboratories to determine plasma vitamin C concentrations. The availability of a simple, reliable method for analyzing plasma vitamin C could increase opportunities for routine plasma vitamin C analysis in clinical medicine. Plasma vitamin C can be analyzed by high performance liquid chromatography (HPLC) with electrochemical (EC) or ultraviolet (UV) light detection. We modified existing UV-HPLC methods for plasma total vitamin C analysis (the sum of ascorbic and dehydroascorbic acid) to develop a simple, constant-low-pH sample reduction procedure followed by isocratic reverse-phase HPLC separation using a purely aqueous low-pH non-buffered mobile phase. Although EC-HPLC is widely recommended over UV-HPLC for plasma total vitamin C analysis, the two methods have never been directly compared. We formally compared the simplified UV-HPLC method with EC-HPLC in 80 consecutive clinical samples. The simplified UV-HPLC method was less expensive, easier to set up, required fewer reagents and no pH adjustments, and demonstrated greater sample stability than many existing methods for plasma vitamin C analysis. When compared with the gold-standard EC-HPLC method in 80 consecutive clinical samples exhibiting a wide range of plasma vitamin C concentrations, it performed equivalently. The easy set up, simplicity and sensitivity of the plasma vitamin C analysis method described here could make it practical in a normally equipped hospital laboratory. Unlike any prior UV-HPLC method for plasma total vitamin C analysis, it was rigorously compared with the gold-standard EC-HPLC method and performed equivalently. Adoption of this method could increase the availability of plasma vitamin C analysis in clinical medicine.

  3. Critical assessment of suitable methods used for determination of antibacterial properties at photocatalytic surfaces

    Energy Technology Data Exchange (ETDEWEB)

    Krysa, Josef, E-mail: [Institute of Chemical Technology Prague, Department of Water Technology and Environmental Engineering, Technicka 5, CZ-166 28 Prague (Czech Republic); Musilova, Eva [Institute of Chemical Technology Prague, Department of Water Technology and Environmental Engineering, Technicka 5, CZ-166 28 Prague (Czech Republic); Zita, Jiri [Institute of Chemical Technology Prague, Department of Inorganic Technology, Technicka 5, CZ-166 28 Prague (Czech Republic)


    Highlights: {yields} Methods for antibacterial activity of irradiated TiO{sub 2} films were critically assessed. {yields} As test bacteria gram negative E. coli and gram positive E. faecalis were employed. {yields} The ISO glass adhesion method is more appropriate than method with bacteria suspension. {yields} Some improvements of the ISO method were suggested. - Abstract: This work describes the development of methods necessary for antibacterial effect evaluation on irradiated TiO{sub 2} layers. Two methods using bacteria suspensions and the glass adhesion method (based on ISO 27447:2009(E)) were critically assessed and compared. As test bacteria gram negative Escherichia coli and gram positive Enterococcus faecalis were employed. The method using 50 cm{sup 3} of bacteria suspension is convenient for testing layers with strong antibacterial effect (prepared from powder photocatalysts). For the evaluation of the antibacterial effect of sol gel layers, the glass adhesion method based on the ISO is more appropriate than the method with 3 cm{sup 3} of bacteria suspension. The reason is that the later does not allow a distinction between the inhibition effect of TiO{sub 2} and UV light itself. Some improvements of the ISO method were suggested, namely the use of gelatinous pills (CCM) of bacteria, using saline solution instead of nutrient broth for bacteria suspension preparation and the application of selective media for bacteria cultivation. Decreasing the light intensity from 0.6 mW cm{sup -2} to 0.2 mW cm{sup -2} (fulfilling the requirements of the ISO) results in almost negligible effect of UV light itself, thus enabling proper testing of the antibacterial properties of TiO{sub 2} thin films.

  4. Critical assessment of suitable methods used for determination of antibacterial properties at photocatalytic surfaces

    International Nuclear Information System (INIS)

    Krysa, Josef; Musilova, Eva; Zita, Jiri


    Highlights: → Methods for antibacterial activity of irradiated TiO 2 films were critically assessed. → As test bacteria gram negative E. coli and gram positive E. faecalis were employed. → The ISO glass adhesion method is more appropriate than method with bacteria suspension. → Some improvements of the ISO method were suggested. - Abstract: This work describes the development of methods necessary for antibacterial effect evaluation on irradiated TiO 2 layers. Two methods using bacteria suspensions and the glass adhesion method (based on ISO 27447:2009(E)) were critically assessed and compared. As test bacteria gram negative Escherichia coli and gram positive Enterococcus faecalis were employed. The method using 50 cm 3 of bacteria suspension is convenient for testing layers with strong antibacterial effect (prepared from powder photocatalysts). For the evaluation of the antibacterial effect of sol gel layers, the glass adhesion method based on the ISO is more appropriate than the method with 3 cm 3 of bacteria suspension. The reason is that the later does not allow a distinction between the inhibition effect of TiO 2 and UV light itself. Some improvements of the ISO method were suggested, namely the use of gelatinous pills (CCM) of bacteria, using saline solution instead of nutrient broth for bacteria suspension preparation and the application of selective media for bacteria cultivation. Decreasing the light intensity from 0.6 mW cm -2 to 0.2 mW cm -2 (fulfilling the requirements of the ISO) results in almost negligible effect of UV light itself, thus enabling proper testing of the antibacterial properties of TiO 2 thin films.

  5. Comparison of Suitability of the Most Common Ancient DNA Quantification Methods. (United States)

    Brzobohatá, Kristýna; Drozdová, Eva; Smutný, Jiří; Zeman, Tomáš; Beňuš, Radoslav


    Ancient DNA (aDNA) extracted from historical bones is damaged and fragmented into short segments, present in low quantity, and usually copurified with microbial DNA. A wide range of DNA quantification methods are available. The aim of this study was to compare the five most common DNA quantification methods for aDNA. Quantification methods were tested on DNA extracted from skeletal material originating from an early medieval burial site. The tested methods included ultraviolet (UV) absorbance, real-time quantitative polymerase chain reaction (qPCR) based on SYBR ® green detection, real-time qPCR based on a forensic kit, quantification via fluorescent dyes bonded to DNA, and fragmentary analysis. Differences between groups were tested using a paired t-test. Methods that measure total DNA present in the sample (NanoDrop ™ UV spectrophotometer and Qubit ® fluorometer) showed the highest concentrations. Methods based on real-time qPCR underestimated the quantity of aDNA. The most accurate method of aDNA quantification was fragmentary analysis, which also allows DNA quantification of the desired length and is not affected by PCR inhibitors. Methods based on the quantification of the total amount of DNA in samples are unsuitable for ancient samples as they overestimate the amount of DNA presumably due to the presence of microbial DNA. Real-time qPCR methods give undervalued results due to DNA damage and the presence of PCR inhibitors. DNA quantification methods based on fragment analysis show not only the quantity of DNA but also fragment length.

  6. RAPD-PCR – still a suitable Method for Genetically Underexplored Species?

    Directory of Open Access Journals (Sweden)

    Konstanze Ursula Behrmann


    Full Text Available Saithe (Pollachius virens is a commercially important fish species; the annual catch quota in the Northeast Atlantic exceeds 100.000 t. Despite that saithe is underexplored from a fish population genetically view. Because saithe is a highly migratory species, which undergoes a long larval drift, the population structure of saithe within the Northeast Atlantic is not fully understood. Models used as a basis for the management plan are based on tagging studies, which have been carried out in the 1960th. But still there are doubts regarding the numbers of stocks living in the Northeast Atlantic. Migration routes are affected by salmon farming, growing steadily from the 1990th. In the last years a hyperstability of the saithe stock in the North Sea had been detected underlining the need to have a closer look on the saithe stocks in the Northeast Atlantic. Random amplified polymorphic DNA (RAPD - PCR is a DNA fingerprinting technique often used in species identification and population genetic research for species, whose genome has not been sequenced very extensive as being the case for most of the food fishes. We applied RAPD-PCR in a study of saithe populations from the North Atlantic. The suitability of RAPD-PCR was improved by optimisations for enhanced reproducibility. The “classical” protocol for RAPD-PCR was modified by increasing the annealing temperature and shortening the time of annealing, providing a much better reproducibility. Thus, RAPD-PCR was found to be a straightforward and low-cost way, compared to other population genetic tools, to get a first insight into the population structure of less sequenced fish species within a very short time, being useful for preliminary studies or laboratories without large capacities for DNA sequencing.

  7. A Rapid and Economical Method for Efficient DNA Extraction from Diverse Soils Suitable for Metagenomic Applications. (United States)

    Devi, Selvaraju Gayathri; Fathima, Anwar Aliya; Radha, Sudhakar; Arunraj, Rex; Curtis, Wayne R; Ramya, Mohandass


    A rapid, cost effective method of metagenomic DNA extraction from soil is a useful tool for environmental microbiology. The present work describes an improved method of DNA extraction namely "powdered glass method" from diverse soils. The method involves the use of sterile glass powder for cell lysis followed by addition of 1% powdered activated charcoal (PAC) as purifying agent to remove humic substances. The method yielded substantial DNA (5.87 ± 0.04 μg/g of soil) with high purity (A260/280: 1.76 ± 0.05) and reduced humic substances (A340: 0.047 ± 0.03). The quality of the extracted DNA was compared against five different methods based on 16S rDNA PCR amplification, BamHI digestion and validated using quantitative PCR. The digested DNA was used for a metagenomic library construction with the transformation efficiency of 4 X 106 CFU mL-1. Besides providing rapid, efficient and economical extraction of metgenomic DNA from diverse soils, this method's applicability is also demonstrated for cultivated organisms (Gram positive B. subtilis NRRL-B-201, Gram negative E. coli MTCC40, and a microalgae C. sorokiniana UTEX#1666).

  8. A Rapid and Economical Method for Efficient DNA Extraction from Diverse Soils Suitable for Metagenomic Applications.

    Directory of Open Access Journals (Sweden)

    Selvaraju Gayathri Devi

    Full Text Available A rapid, cost effective method of metagenomic DNA extraction from soil is a useful tool for environmental microbiology. The present work describes an improved method of DNA extraction namely "powdered glass method" from diverse soils. The method involves the use of sterile glass powder for cell lysis followed by addition of 1% powdered activated charcoal (PAC as purifying agent to remove humic substances. The method yielded substantial DNA (5.87 ± 0.04 μg/g of soil with high purity (A260/280: 1.76 ± 0.05 and reduced humic substances (A340: 0.047 ± 0.03. The quality of the extracted DNA was compared against five different methods based on 16S rDNA PCR amplification, BamHI digestion and validated using quantitative PCR. The digested DNA was used for a metagenomic library construction with the transformation efficiency of 4 X 106 CFU mL-1. Besides providing rapid, efficient and economical extraction of metgenomic DNA from diverse soils, this method's applicability is also demonstrated for cultivated organisms (Gram positive B. subtilis NRRL-B-201, Gram negative E. coli MTCC40, and a microalgae C. sorokiniana UTEX#1666.

  9. My Day of Silence. (United States)

    Brown, Scott C.


    A heterosexual doctoral student discusses his experiences when he tries to take part in a day of silence to help combat homophobia and heterosexism. His vow of silence teaches him that he will never fully understand the experience of a person who has been historically, socially, and legally silent. (Author/MKA)

  10. Suitability of thermoluminescence, chemiluminescence, ESR and viscosity measurements as detection method for the irradiation of medicinal herbs

    International Nuclear Information System (INIS)

    Schuettler, C.; Gebhardt, G.; Stock, A.; Helle, N.; Boegl, K.W.


    Chemiluminescence, electron spin resonance, thermoluminescence and viscosity measurements have been investigated for their suitability as detection method for the irradiation of the medicinal herbs anise seeds (anisi fructus), valerian roots (valerianae radix), redberry leaves (uvae ursi folium), birch leaves (betulae folium), greek hay seeds (foenugraeci semen), cayenne pepper (capsici fructus acer), black-aldertee bark (frangulae cortex), fennel fruits (feoniculi fructus), rose hip shells (cynosbati fructus), coltsfoot (farfarae folium), acorus roots (calami rhizoma), chamomile flowers (matricariae flos), caraway (carvi fructus), lavender flowers (lavandulae flos), linseed (lini semen), lime tree flowers (tiliae flos), St. Mary's thistle fruit (cardui mariae herba), lemon balm (melissae folium), java tea (orthosiphonis folium), peppermint (menthae piperitae folium), sage leaves (salviae folium), scouring rush (equiseti herba), senna leaves (sennae folium), plantain herbs (plantaginis lanceolata herba), thyme herbs (thymi herba), juniper berries (juniperi fructus), hawthorne herbs (crataegi folium), wheat starch (amylum tritici) and wormwood (absinthii herba). Depending on the herbs, the methods used were more or less suitable. Chemiluminescence measurements showed the smallest differences between untreated and irradiated samples whereas thermoluminescence measurements on isolated minerals from the vegetable drugs gave better results. In some herbs radiation-specific radicals could be identified by ESR spectroscopy. Viscosity measurement is suitable for some herbs as fast and inexpensive method for screening. (orig.) [de

  11. Test methods for determining the suitability of metal alloys for use in oxygen-enriched environments (United States)

    Stoltzfus, Joel M.; Gunaji, Mohan V.


    Materials are more flammable in oxygen rich environments than in air. When the structural elements of a system containing oxygen ignite and burn, the results are often catastrophic, causing loss of equipment and perhaps even human lives. Therefore, selection of the proper metallic and non-metallic materials for use in oxygen systems is extremely important. While test methods for the selection of non-metallic materials have been available for years, test methods for the selection of alloys have not been available until recently. Presented here are several test methods that were developed recently at NASA's White Sands Test Facility (WSTF) to study the ignition and combustion of alloys, including the supersonic and subsonic speed particle impact tests, the frictional heating and coefficient of friction tests, and the promoted combustion test. These test methods are available for commercial use.

  12. Modelling a suitable location for Urban Solid Waste Management using AHP method and GIS -A geospatial approach and MCDM Model (United States)

    Iqbal, M.; Islam, A.; Hossain, A.; Mustaque, S.


    Multi-Criteria Decision Making(MCDM) is advanced analytical method to evaluate appropriate result or decision from multiple criterion environment. Present time in advanced research, MCDM technique is progressive analytical process to evaluate a logical decision from various conflict. In addition, Present day Geospatial approach (e.g. Remote sensing and GIS) also another advanced technical approach in a research to collect, process and analyze various spatial data at a time. GIS and Remote sensing together with the MCDM technique could be the best platform to solve a complex decision making process. These two latest process combined very effectively used in site selection for solid waste management in urban policy. The most popular MCDM technique is Weighted Linear Method (WLC) where Analytical Hierarchy Process (AHP) is another popular and consistent techniques used in worldwide as dependable decision making. Consequently, the main objective of this study is improving a AHP model as MCDM technique with Geographic Information System (GIS) to select a suitable landfill site for urban solid waste management. Here AHP technique used as a MCDM tool to select the best suitable landfill location for urban solid waste management. To protect the urban environment in a sustainable way municipal waste needs an appropriate landfill site considering environmental, geological, social and technical aspect of the region. A MCDM model generate from five class related which related to environmental, geological, social and technical using AHP method and input the result set in GIS for final model location for urban solid waste management. The final suitable location comes out that 12.2% of the area corresponds to 22.89 km2 considering the total study area. In this study, Keraniganj sub-district of Dhaka district in Bangladesh is consider as study area which is densely populated city currently undergoes an unmanaged waste management system especially the suitable landfill sites for

  13. Evaluation of DNA Extraction Methods Suitable for PCR-based Detection and Genotyping of Clostridium botulinum

    DEFF Research Database (Denmark)

    Auricchio, Bruna; Anniballi, Fabrizio; Fiore, Alfonsina


    Sufficient quality and quantity of extracted DNA is critical to detecting and performing genotyping of Clostridium botulinum by means of PCR-based methods. An ideal extraction method has to optimize DNA yield, minimize DNA degradation, allow multiple samples to be extracted, and be efficient...... in terms of cost, time, labor, and supplies. Eleven botulinum toxin–producing clostridia strains and 25 samples (10 food, 13 clinical, and 2 environmental samples) naturally contaminated with botulinum toxin–producing clostridia were used to compare 4 DNA extraction procedures: Chelex® 100 matrix, Phenol......-Cloroform-Isoamyl alcohol, NucliSENS® magnetic extraction kit, and DNeasy® Blood & Tissue kit. Integrity, purity, and amount of amplifiable DNA were evaluated. The results show that the DNeasy® Blood & Tissue kit is the best extraction method evaluated because it provided the most pure, intact, and amplifiable DNA. However...

  14. Suitability of analytical methods to measure solubility for the purpose of nanoregulation. (United States)

    Tantra, Ratna; Bouwmeester, Hans; Bolea, Eduardo; Rey-Castro, Carlos; David, Calin A; Dogné, Jean-Michel; Jarman, John; Laborda, Francisco; Laloy, Julie; Robinson, Kenneth N; Undas, Anna K; van der Zande, Meike


    Solubility is an important physicochemical parameter in nanoregulation. If nanomaterial is completely soluble, then from a risk assessment point of view, its disposal can be treated much in the same way as "ordinary" chemicals, which will simplify testing and characterisation regimes. This review assesses potential techniques for the measurement of nanomaterial solubility and evaluates the performance against a set of analytical criteria (based on satisfying the requirements as governed by the cosmetic regulation as well as the need to quantify the concentration of free (hydrated) ions). Our findings show that no universal method exists. A complementary approach is thus recommended, to comprise an atomic spectrometry-based method in conjunction with an electrochemical (or colorimetric) method. This article shows that although some techniques are more commonly used than others, a huge research gap remains, related with the need to ensure data reliability.

  15. Optimization and Comparative Analysis of Plant Organellar DNA Enrichment Methods Suitable for Next-generation Sequencing. (United States)

    Miller, Marisa E; Liberatore, Katie L; Kianian, Shahryar F


    Plant organellar genomes contain large, repetitive elements that may undergo pairing or recombination to form complex structures and/or sub-genomic fragments. Organellar genomes also exist in admixtures within a given cell or tissue type (heteroplasmy), and an abundance of subtypes may change throughout development or when under stress (sub-stoichiometric shifting). Next-generation sequencing (NGS) technologies are required to obtain deeper understanding of organellar genome structure and function. Traditional sequencing studies use several methods to obtain organellar DNA: (1) If a large amount of starting tissue is used, it is homogenized and subjected to differential centrifugation and/or gradient purification. (2) If a smaller amount of tissue is used (i.e., if seeds, material, or space is limited), the same process is performed as in (1), followed by whole-genome amplification to obtain sufficient DNA. (3) Bioinformatics analysis can be used to sequence the total genomic DNA and to parse out organellar reads. All these methods have inherent challenges and tradeoffs. In (1), it may be difficult to obtain such a large amount of starting tissue; in (2), whole-genome amplification could introduce a sequencing bias; and in (3), homology between nuclear and organellar genomes could interfere with assembly and analysis. In plants with large nuclear genomes, it is advantageous to enrich for organellar DNA to reduce sequencing costs and sequence complexity for bioinformatics analyses. Here, we compare a traditional differential centrifugation method with a fourth method, an adapted CpG-methyl pulldown approach, to separate the total genomic DNA into nuclear and organellar fractions. Both methods yield sufficient DNA for NGS, DNA that is highly enriched for organellar sequences, albeit at different ratios in mitochondria and chloroplasts. We present the optimization of these methods for wheat leaf tissue and discuss major advantages and disadvantages of each approach in

  16. Is a QRA-based method suitable to guide Land Use Planning decisions in India?

    NARCIS (Netherlands)

    Sengupta, A.; Bandyopadhyay, D.; Van Westen, C. J.; Van Der Veen, A.; Nowakowski, T.


    In this paper, we have applied a quantitative risk assessment method to map cumulative risk levels arising from a number of hazardous installations located in Haldia, one of the industrial towns of India. The methodology consists of the following steps: frequency and consequences assessment of the

  17. A New Method to Implement Resampled Uniform PWM Suitable for Distributed Control of Modular Multilevel Converters

    DEFF Research Database (Denmark)

    Huang, Shaojun; Mathe, Laszlo; Teodorescu, Remus


    the proper switching instances needed for the resampling modulation technique. The software implementation of the proposed phase shifted PWM (PS-PWM) method, and its application in a distributed control system for MMC, are fully discussed in this paper. Simulation and experiment results show...... that the proposed solution can realize the resampled uniform PWM and provide high effective sampling frequency and low time delay, which is critical for the distributed control of MMC....

  18. A Suitable Method for DNA Extraction from Bones for Forensic Applications: A Case Study

    Directory of Open Access Journals (Sweden)

    Aqeela S. Abuidrees


    Full Text Available Human identification techniques are constantly developing. Before the discovery of DNA, anthropology accompanied with odontology was the most applicable technique for human identification. With the new era of molecular biology and the revolution of DNA and PCR techniques, DNA profiling has become the core of the human forensic identification process. Different types of samples can be exploited in forensic DNA analysis. In some extreme cases, bone samples are the only accessible samples of DNA due to the bad conditions of putrefaction or degradation of other biological materials and tissues. Therefore, an appropriate method should be determined to yield a full and clean profile. A case study is presented here in order to identify human remains and conclude the most appropriate method of DNA extraction from human remains. In addition, this study looks at the best part of the skeletal remains to be considered in the extraction of DNA for the purposes of identification. A suspect admitted that he buried his aborted son six months ago. The remains were recovered and DNA analysis was performed in order to determine any genetic link of the remains to the suspect and the female who delivered the baby. Two extraction methods were compared, the standard organic (phenol:chloroform:isoamyl alcohol and automated extraction using magnetic beads coated with silica (Qiagen EZ1 Advanced XL. Two bone parts, femur and clavicle, were also compared in terms of DNA yield. The efficiency of the two methods of DNA extraction from bones is illustrated quantitatively and qualitatively. Paternity testing was performed and the suspect was excluded from being the alleged father.

  19. Proposed Suitable Methods to Detect Transient Regime Switching to Improve Power Quality with Wavelet Transform

    Directory of Open Access Journals (Sweden)

    Javad Safaee Kuchaksaraee


    Full Text Available The increasing consumption of electrical energy and the use of non-linear loads that create transient regime states in distribution networks is increasing day by day. This is the only reason due to which the analysis of power quality for energy sustainability in power networks has become more important. Transients are often created by energy injection through switching or lightning and make changes in voltage and nominal current. Sudden increase or decrease in voltage or current makes characteristics of the transient regime. This paper shed some lights on the capacitor bank switching, which is one of the main causes for oscillatory transient regime states in the distribution network, using wavelet transform. The identification of the switching current of capacitor bank and the internal fault current of the transformer to prevent the unnecessary outage of the differential relay, it propose a new smart method. The accurate performance of this method is shown by simulation in EMTP and MATLAB (matrix laboratory software.

  20. Single-Case Research Methods: History and Suitability for a Psychological Science in Need of Alternatives. (United States)

    Hurtado-Parrado, Camilo; López-López, Wilson


    This paper presents a historical and conceptual analysis of a group of research strategies known as the Single-Case Methods (SCMs). First, we present an overview of the SCMs, their history, and their major proponents. We will argue that the philosophical roots of SCMs can be found in the ideas of authors who recognized the importance of understanding both the generality and individuality of psychological functioning. Second, we will discuss the influence that the natural sciences' attitude toward measurement and experimentation has had on SCMs. Although this influence can be traced back to the early days of experimental psychology, during which incipient forms of SCMs appeared, SCMs reached full development during the subsequent advent of Behavior Analysis (BA). Third, we will show that despite the success of SCMs in BA and other (mainly applied) disciplines, these designs are currently not prominent in psychology. More importantly, they have been neglected as a possible alternative to one of the mainstream approaches in psychology, the Null Hypothesis Significance Testing (NHST), despite serious controversies about the limitations of this prevailing method. Our thesis throughout this section will be that SCMs should be considered as an alternative to NHST because many of the recommendations for improving the use of significance testing (Wilkinson & the TFSI, 1999) are main characteristics of SCMs. The paper finishes with a discussion of a number of the possible reasons why SCMs have been neglected.

  1. Developing suitable methods for effective characterization of electrical properties of root segments (United States)

    Ehosioke, Solomon; Phalempin, Maxime; Garré, Sarah; Kemna, Andreas; Huisman, Sander; Javaux, Mathieu; Nguyen, Frédéric


    The root system represents the hidden half of the plant which plays a key role in food production and therefore needs to be well understood. Root system characterization has been a great challenge because the roots are buried in the soil. This coupled with the subsurface heterogeneity and the transient nature of the biogeochemical processes that occur in the root zone makes it difficult to access and monitor the root system over time. The traditional method of point sampling (root excavation, monoliths, minirhizotron etc.) for root investigation does not account for the transient nature and spatial variability of the root zone, and it often disturbs the natural system under investigation. The quest to overcome these challenges has led to an increase in the application of geophysical methods. Recent studies have shown a correlation between bulk electrical resistivity and root mass density, but an understanding of the contribution of the individual segments of the root system to that bulk signal is still missing. This study is an attempt to understand the electrical properties of roots at the segment scale (1-5cm) for more effective characterization of electrical signal of the full root architecture. The target plants were grown in three different media (pot soil, hydroponics and a mixture of sand, perlite and vermiculite). Resistance measurements were carried out on a single segment of each study plant using a voltmeter while the diameter was measured using a digital calliper. The axial resistance was calculated using the measured resistance and the geometric parameters. This procedure was repeated for each plant replica over a period of 75 days which enabled us to study the effects of age, growth media, diameter and length on the electrical response of the root segments of the selected plants. The growth medium was found to have a significant effect on the root electrical response, while the effect of root diameter on their electrical response was found to vary

  2. PID Testing Method Suitable for Process Control of Solar Cells Mass Production

    Directory of Open Access Journals (Sweden)

    Xianfang Gou


    Full Text Available Voltage bias of several hundred volts which are applied between solar cells and module frames may lead to significant power losses, so-called potential-induced degradation (PID, in normal photovoltaic (PV installations system. Modules and minimodules are used to conduct PID test of solar cells. The test procedure is time consuming and of high cost, which cannot be used as process monitoring method during solar cells fabrication. In this paper, three kinds of test including minimodule, Rsh, and V-Q test are conducted on solar cells or wafers with SiNx of different refractive index. All comparisons between test results of Rsh, V-Q, and minimodule tests have shown equal results. It is shown that Rsh test can be used as quality inspection of solar cells and V-Q test of coated wafer can be used as process control of solar cells.

  3. Development of a unit suitable for corrosion monitoring in district heating systems. Experiences with the LOCOR-cell test method

    DEFF Research Database (Denmark)

    Andersen, Asbjørn; Hilbert, Lisbeth Rischel


    A by-pass unit suitable for placement of a number of different probes for corrosion monitoring has been designed. Also measurements of water parameters are allowed in a side stream from the unit. The project is a part of the Nordic Innovation Fund project KORMOF. The by-pass unit has been installed...... in 6 pressurised circulating heating systems and in one cooling system. 7 different corrosion monitoring methods have been used to study corrosion rates and types in dependency of water chemistry. This paper describes the design of the by-pass unit including water analysis methods. It also describes...... other used corrosion monitoring methods in the project are described elsewhere (3) and (4). For future district heating corrosion monitoring the by-pass unit can be recommended for permanent installation and the two methods high sensitive ER-technique (Metricorr) and the LOCOR-Cell㤠(FORCE Technology...

  4. Personalized gene silencing therapeutics for Huntington disease. (United States)

    Kay, C; Skotte, N H; Southwell, A L; Hayden, M R


    Gene silencing offers a novel therapeutic strategy for dominant genetic disorders. In specific diseases, selective silencing of only one copy of a gene may be advantageous over non-selective silencing of both copies. Huntington disease (HD) is an autosomal dominant disorder caused by an expanded CAG trinucleotide repeat in the Huntingtin gene (HTT). Silencing both expanded and normal copies of HTT may be therapeutically beneficial, but preservation of normal HTT expression is preferred. Allele-specific methods can selectively silence the mutant HTT transcript by targeting either the expanded CAG repeat or single nucleotide polymorphisms (SNPs) in linkage disequilibrium with the expansion. Both approaches require personalized treatment strategies based on patient genotypes. We compare the prospect of safe treatment of HD by CAG- and SNP-specific silencing approaches and review HD population genetics used to guide target identification in the patient population. Clinical implementation of allele-specific HTT silencing faces challenges common to personalized genetic medicine, requiring novel solutions from clinical scientists and regulatory authorities. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  5. Laboratory assessment of novel oral anticoagulants: method suitability and variability between coagulation laboratories. (United States)

    Helin, Tuukka A; Pakkanen, Anja; Lassila, Riitta; Joutsi-Korhonen, Lotta


    Laboratory tests to assess novel oral anticoagulants (NOACs) are under evaluation. Routine monitoring is unnecessary, but under special circumstances bioactivity assessment becomes crucial. We analyzed the effects of NOACs on coagulation tests and the availability of specific assays at different laboratories. Plasma samples spiked with dabigatran (Dabi; 120 and 300 μg/L) or rivaroxaban (Riva; 60, 146, and 305 μg/L) were sent to 115 and 38 European laboratories, respectively. International normalized ratio (INR) and activated partial thromboplastin time (APTT) were analyzed for all samples; thrombin time (TT) was analyzed specifically for Dabi and calibrated anti-activated factor X (anti-Xa) activity for Riva. We compared the results with patient samples. Results of Dabi samples were reported by 73 laboratories (13 INR and 9 APTT reagents) and Riva samples by 22 laboratories (5 INR and 4 APTT reagents). Both NOACs increased INR values; the increase was modest, albeit larger, for Dabi, with higher CV, especially with Quick (vs Owren) methods. Both NOACs dose-dependently prolonged the APTT. Again, the prolongation and CVs were larger for Dabi. The INR and APTT results varied reagent-dependently (P laboratories, respectively. The screening tests INR and APTT are suboptimal in assessing NOACs, having high reagent dependence and low sensitivity and specificity. They may provide information, if laboratories recognize their limitations. The variation will likely increase and the sensitivity differ in clinical samples. Specific assays measure NOACs accurately; however, few laboratories applied them. © 2013 American Association for Clinical Chemistry.

  6. An incident flux expansion transport theory method suitable for coupling to diffusion theory methods in hexagonal geometry

    International Nuclear Information System (INIS)

    Hayward, Robert M.; Rahnema, Farzad; Zhang, Dingkang


    Highlights: ► A new hybrid stochastic–deterministic transport theory method to couple with diffusion theory. ► The method is implemented in 2D hexagonal geometry. ► The new method produces excellent results when compared with Monte Carlo reference solutions. ► The method is fast, solving all test cases in less than 12 s. - Abstract: A new hybrid stochastic–deterministic transport theory method, which is designed to couple with diffusion theory, is presented. The new method is an extension of the incident flux response expansion method, and it combines the speed of diffusion theory with the accuracy of transport theory. With ease of use in mind, the new method is derived in such a way that it can be implemented with only minimal modifications to an existing diffusion theory method. A new angular expansion, which is necessary for the diffusion theory coupling, is developed in 2D and 3D. The method is implemented in 2D hexagonal geometry, and an HTTR benchmark problem is used to test its accuracy in a standalone configuration. It is found that the new method produces excellent results (with average relative error in partial current less than 0.033%) when compared with Monte Carlo reference solutions. Furthermore, the method is fast, solving all test cases in less than 12 s

  7. Silencing criticism in Mexico

    Directory of Open Access Journals (Sweden)

    Ximena Suárez


    Full Text Available Journalists and human rights defenders in Mexico are being attacked in an attempt to silence their criticism. Many are forced to flee or risk being assassinated. The consequences are both personal and of wider social significance.

  8. Ombuds’ corner: Employee silence

    CERN Multimedia

    Vincent Vuillemin


    Although around a hundred cases a year are reported to the Ombuds, several issues may still not be disclosed due to employee silence*. The deliberate withholding of concerns, escalating misunderstandings or genuine conflicts can impede the global process of learning and development of a better respectful organizational workplace environment, and prevent the detection and correction of acts violating the CERN Code of Conduct.   For the employee him/herself, such silence can lead to feelings of anger, resentment, helplessness and humiliation. These feelings will inevitably contaminate personal and interpersonal relations, and poison creativity and effectiveness. Employee silence can be explained by many factors; sometimes it is connected to organizational forces. In their published paper*, authors Michael Knoll and Rolf van Dick found four forms of employee silence. People may stay silent if they feel that their opinion is neither welcomed nor valued by their management. They have gi...

  9. The Opinion of Latvian Teachers About the Most Suitable Teaching Methods and Possibilities to Make Lessons Interesting

    Directory of Open Access Journals (Sweden)

    Porozovs Juris


    Full Text Available Student’s learning motivation and learning outcomes depend on the ability of the teacher to interest students, the chosen teaching methods and proficiency to manage the learning process. Teacher who can successfully choose teaching content, material resources and different teaching methods is able to cause interest about his subject. Teacher who is interested in teaching process himself can help students to learn a subject. The aim of the study was to find out the Latvian teachers' opinion about the disturbing factors affecting the achievement of high learning outcomes for students, the most suitable teaching methods and the possibilities to make lessons interesting. The survey of teachers of different schools in Latvia regarding their students’ learning motivation was carried out. The data from 482 teachers’ answers were analysed. The results of the research showed that the majority of surveyed teachers consider that learning motivation of Latvia students has decreased during the last few years. Teachers point out several factors, which do not allow students to reach high learning outcomes, the most important of them are: the inability of students to link career and success with learning and lack of life goals; inability of students to concentrate attention during lessons; indisposition of students to do homework. The teachers consider laboratory works, discussions, project works, group works and teacher’s presentations and narrative of new material as the most suitable teaching methods. Teachers consider that the most important personality trait of teachers and learning motivation for students to improve their skills is the teacher’s ability to teach the subject in an interesting way. Teachers believe that lessons can be made interesting if modern technologies and teaching methods are used during lessons and when the teacher is able to connect the subject with real life.


    Directory of Open Access Journals (Sweden)

    Kenza SLIMANI


    Full Text Available The city of Batna, one of the Algerian cities, is the most exposed to flood risks, which are highly active and constitute a threat to the residents and the infrastructure. To better understand the vulnerability of this phenomenon, an approach of cartography inspired by the method of “suitability modeling” is used. This method allows us to determine the potential vulnerability levels of the study zone; therefore, we need to consider several criteria: socioeconomic factors like density of population, land use, and also physical criteria such as permeability, flow direction, and the like. Each of these various criteria was built on a database, which constituting an important element for building a model that helps us determine the vulnerable zones by using various GIS applications. The applied approach that depends on how the criteria are suitable give us 4 different levels of vulnerability in Batna, which are as follows: high vulnerability, 13% of the city’s total surface, average vulnerability 5%, low vulnerability 0.7%, and null vulnerability 82%.

  11. Suitable Method of Overloading for Fast Primary Frequency Control from Offshore Wind Power Plants in Multi-Terminal DC Grid

    DEFF Research Database (Denmark)

    Sakamuri, Jayachandra N.; Hansen, Anca Daniela; Altin, Müfit


    considering the operation of the WT at below rated wind speed. Moreover, the impact of release of overload on the dynamics of the wind turbine, therefore on the associated AC and DC grids are studied in this paper. Finally, the suitable overloading method is proposed based on the simulation and experimental...... results. The time domain simulations for fast primary frequency control are performed on an OWPP connected through a 3-terminal DC grid using DIgSILENT PowerFactory. The experiments are performed on OWPP model integrated to a laboratory scale 3-terminal DC grid test set up. Based on the simulations...... and experimental results, overloading method which considers the variation of WT output power during the overload provides better performance during and after release of the overload....

  12. Comparison of DNA extraction methods from small samples of newborn screening cards suitable for retrospective perinatal viral research. (United States)

    McMichael, Gai L; Highet, Amanda R; Gibson, Catherine S; Goldwater, Paul N; O'Callaghan, Michael E; Alvino, Emily R; MacLennan, Alastair H


    Reliable detection of viral DNA in stored newborn screening cards (NSC) would give important insight into possible silent infection during pregnancy and around birth. We sought a DNA extraction method with sufficient sensitivity to detect low copy numbers of viral DNA from small punch samples of NSC. Blank NSC were spotted with seronegative EDTA-blood and seropositive EBV EDTA-blood. DNA was extracted with commercial and noncommercial DNA extraction methods and quantified on a spectrofluorometer using a PicoGreen dsDNA quantification kit. Serial dilutions of purified viral DNA controls determined the sensitivity of the amplification protocol, and seropositive EBV EDTA-blood amplified by nested PCR (nPCR) validated the DNA extraction methods. There were considerable differences between the commercial and noncommercial DNA extraction methods (P=0.014; P=0.016). Commercial kits compared favorably, but the QIamp DNA micro kit with an added forensic filter step was marginally more sensitive. The mean DNA yield from this method was 3 ng/μl. The limit of detection was 10 viral genome copies in a 50-μl reaction. EBV nPCR detection in neat and 1:10 diluted DNA extracts could be replicated reliably. We conclude that the QIamp Micro DNA extraction method with the added forensic spin-filter step was suitable for retrospective DNA viral assays from NSC.

  13. Investigation of e-Linac tube construction method and implementation suitable method for IPM e-Linac

    Directory of Open Access Journals (Sweden)

    F ghasemi


    Full Text Available The goal of electron linear accelerator project in institute for research in fundamental sciences(IPM is to build its components as many as they can in Iran. This accelerator is a traveling wave type. Investigations show that there are various techniques in forming and connecting the accelerating tube cavities. The shrinking method applied for constructing the accelerating tube is selected based on the one applied for Stanford University’s Mark III accelerator. With success in building an 8-cavity test tube and finding the problems of the method, the construction of the final accelerating tube with 24 cavities has been accomplished. The results show that the obtained frequency of 2996.5 MHz and quality factor of 11200, satisfy the design desired values.

  14. An assessment of the long term suitability of present and proposed methods for the management of uranium mill tailings

    International Nuclear Information System (INIS)


    Proposals for safe, long-term containment of conventional tailings include 1) storage under water, 2) storage in active, abandoned or specially created underground mines and, 3) storage in open pits, with subsequent flooding or covering with overburden. The underwater proposal can meet most of the requirements of long term containment; however, extensive study of existing tailings deposits in deep water locations will be needed. Underground mines cannot provide sufficient storage capacity, since the tailings bulk during mill operation can occupy twice the volume of the original ore. It is possible to reduce the hazard by reducing the radium and thorium content of the tailings. Proposals for such an undertaking include ore beneficiation with rejection of the relatively innocuous fraction, radium-thorium removal in the mill, and significant changes in both ore processing and treatment of tailings. It is concluded that surface-stored tailings are vulnerable over the long term to dispersion by leaching and water erosion, and that access to a tailings site cannot be prevented, while only a major climatic or seismic event could disturb tailings stored in suitable underwater or underground mine sites. The criteria for determining suitability for each method, however, will need to be identified, tested, and accepted through the normal process of modeling, pilot plant evaluation, monitoring and evaluation. (author)

  15. A novel method for embedding neonatal murine calvaria in methyl methacrylate suitable for visualizing mineralization, cellular and structural detail. (United States)

    Horn, D A; Garrett, I R


    The study of undecalcified bone by histological methods is essential in the field of bone research. Culturing skeletal tissues such as neonatal murine calvaria provides a reliable bridge between assessment of bone formation in vitro and anabolic activity in vivo and contains most of the essential elements of bone for studying bone formation. Neonatal calvarial assay, supported by histological methods, is used to study the anabolic effects of a wide variety of factors and compounds on bone tissue. To optimize visualization and histomorphometric measurements using neonatal calvaria, we developed a method that provides high quality tissue sections suitable for routine and histochemical staining. Undecalcified neonatal mouse calvaria were processed and embedded using a low temperature methyl methacrylate procedure. Various staining methods were performed on deplastisized and floated sections to examine mineralization and to identify cells. The Von Kossa stain counterstained with a modified H & E yielded precise images of unmineralized bone including mineralization sites, and distinct osteoblasts and osteoclasts. Toluidine blue, Ladewig's trichrome, tartrate-resistant acid phosphatase, Goldner, H & E and Villanueva stains also were tested on the undecalcified neonatal calvaria sections.

  16. Memories Persist in Silence

    Directory of Open Access Journals (Sweden)

    Sandra Patricia Arenas Grisales


    Full Text Available This article exposes the hypothesis that memory artifacts, created to commemorate the victims of armed conflict in Colombia, are an expression of the underground memories and a way of political action in the midst of war. We analyze three cases of creations of memory artifacts in Medellín, Colombia, as forms of suffering, perceiving and resisting the power of armed groups in Medellín. The silence, inherent in these objects, should not be treated as an absence of language, but as another form of expression of memory. Silence is a tactic used to overcome losses and reset everyday life in contexts of protracted violence.

  17. Determining the most suitable areas for artificial groundwater recharge via an integrated PROMETHEE II-AHP method in GIS environment (case study: Garabaygan Basin, Iran). (United States)

    Nasiri, Hossein; Boloorani, Ali Darvishi; Sabokbar, Hassan Ali Faraji; Jafari, Hamid Reza; Hamzeh, Mohamad; Rafii, Yusef


    Flood spreading is a suitable strategy for controlling and benefiting from floods. Selecting suitable areas for flood spreading and directing the floodwater into permeable formations are amongst the most effective strategies in flood spreading projects. Having combined geographic information systems (GIS) and multi-criteria decision analysis approaches, the present study sought to locate the most suitable areas for flood spreading operation in the Garabaygan Basin of Iran. To this end, the data layers relating to the eight effective factors were prepared in GIS environment. This stage was followed by elimination of the exclusionary areas for flood spreading while determining the potentially suitable ones. Having closely examined the potentially suitable areas using the Preference Ranking Organization Method for Enrichment Evaluations (PROMETHEE) II and analytic hierarchy process (AHP) methods, the land suitability map for flood spreading was produced. The PROMETHEE II and AHP were used for ranking all the alternatives and weighting the criteria involved, respectively. The results of the study showed that most suitable areas for the artificial groundwater recharge are located in Quaternary Q(g) and Q(gsc) geologic units and in geomorphological units of pediment and Alluvial fans with slopes not exceeding 3%. Furthermore, significant correspondence between the produced map and the control areas, where the flood spreading projects were successfully performed, provided further evidence for the acceptable efficiency of the integrated PROMETHEE II-AHP method in locating suitable flood spreading areas.

  18. On the suitability of fast and frugal heuristics for designing values clarification methods in patient decision aids: a critical analysis. (United States)

    Pieterse, Arwen H; de Vries, Marieke


    Increasingly, patient decision aids and values clarification methods (VCMs) are being developed to support patients in making preference-sensitive health-care decisions. Many VCMs encourage extensive deliberation about options, without solid theoretical or empirical evidence showing that deliberation is advantageous. Research suggests that simple, fast and frugal heuristic decision strategies sometimes result in better judgments and decisions. Durand et al. have developed two fast and frugal heuristic-based VCMs. To critically analyse the suitability of the 'take the best' (TTB) and 'tallying' fast and frugal heuristics in the context of patient decision making. Analysis of the structural similarities between the environments in which the TTB and tallying heuristics have been proven successful and the context of patient decision making and of the potential of these heuristic decision processes to support patient decision making. The specific nature of patient preference-sensitive decision making does not seem to resemble environments in which the TTB and tallying heuristics have proven successful. Encouraging patients to consider less rather than more relevant information potentially even deteriorates their values clarification process. Values clarification methods promoting the use of more intuitive decision strategies may sometimes be more effective. Nevertheless, we strongly recommend further theoretical thinking about the expected value of such heuristics and of other more intuitive decision strategies in this context, as well as empirical assessments of the mechanisms by which inducing such decision strategies may impact the quality and outcome of values clarification. © 2011 John Wiley & Sons Ltd.

  19. Suitability of the infrared spectroscopy and the rheological method for distinguishing traditional cheese from industrial Turoš cheese

    Directory of Open Access Journals (Sweden)

    Kristijan Valkaj


    Full Text Available The aim of this paper was to determine suitability of the infrared spectroscopy and the rheological method for distinguishing traditional cheese from industrial Turoš cheese. Turoš cheese belongs to the group of sour, dried, cone shaped cheeses with added salt and red dry pepper. Cheeses were sampled from 15 family farms and from market (industrial cheeses from five different batches. The rheological parameters of analysed cheeses were in accordance with the chemical composition of the same samples. Infrared spectroscopy of cheeses show good relation with the chemical composition and it has been proved to be a fast and effective method when compared to textural and standard chemical analysis for monitoring the standard procedure of production of sour, dried cheeses such as Turoš cheese. The extensive variability of all the parameters was a result of unbalanced production of Turoš cheese among family farms. Industrial production of Turoš cheese demonstrates more uniformity in relation to traditional on-farm cheese production.

  20. The Fruit of Silence (United States)

    Nelson, Marilyn


    This presentation explores how contemplative practices, especially those anchored in an active listening to silence, are integrated into creative writing courses. It pays particular attention to a course taught at the United States Military Academy at West Point and to a course on the poetry of war and peace taught at the University of…

  1. Silence of the Genes

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education; Volume 12; Issue 4. Silence of the Genes - 2006 Nobel Prize in Physiology or Medicine. Utpal Nath Saumitra Das. General Article Volume 12 Issue 4 April 2007 pp 6-18. Fulltext. Click here to view fulltext PDF. Permanent link:

  2. Silence of the Genes

    Indian Academy of Sciences (India)


    The human genome codes for ~35,000 genes and all these genes are not expressed in every cell. The time and site of gene expression is very precisely regulated. In any cell, only. Silence of the Genes. 2006 Nobel Prize in Physiology or Medicine. Utpal Nath and Saumitra Das. Keywords. RNA interference, siRNA,.

  3. Evaluating methods to establish habitat suitability criteria: A case study in the upper Delaware River Basin, USA (United States)

    Galbraith, Heather S.; Blakeslee, Carrie J.; Cole, Jeffrey C.; Talbert, Colin; Maloney, Kelly O.


    Defining habitat suitability criteria (HSC) of aquatic biota can be a key component to environmental flow science. HSC can be developed through numerous methods; however, few studies have evaluated the consistency of HSC developed by different methodologies. We directly compared HSC for depth and velocity developed by the Delphi method (expert opinion) and by two primary literature meta-analyses (literature-derived range and interquartile range) to assess whether these independent methods produce analogous criteria for multiple species (rainbow trout, brown trout, American shad, and shallow fast guild) and life stages. We further evaluated how these two independently developed HSC affect calculations of habitat availability under three alternative reservoir management scenarios in the upper Delaware River at a mesohabitat (main channel, stream margins, and flood plain), reach, and basin scale. In general, literature-derived HSC fell within the range of the Delphi HSC, with highest congruence for velocity habitat. Habitat area predicted using the Delphi HSC fell between the habitat area predicted using two literature-derived HSC, both at the basin and the site scale. Predicted habitat increased in shallow regions (stream margins and flood plain) using literature-derived HSC while Delphi-derived HSC predicted increased channel habitat. HSC generally favoured the same reservoir management scenario; however, no favoured reservoir management scenario was the most common outcome when applying the literature range HSC. The differences found in this study lend insight into how different methodologies can shape HSC and their consequences for predicted habitat and water management decisions. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.

  4. A rapid method for selecting suitable animal species for studying pathogen interactions with plasma protein ligands in vivo. (United States)

    Naudin, Clément; Schumski, Ariane; Salo-Ahen, Outi M H; Herwald, Heiko; Smeds, Emanuel


    Species tropism constitutes a serious problem for developing relevant animal models of infection. Human pathogens can express virulence factors that show specific selectivity to human proteins, while their affinity for orthologs from other species can vary significantly. Suitable animal species must be used to analyse whether virulence factors are potential targets for drug development. We developed an assay that rapidly predicts applicable animal species for studying virulence factors binding plasma proteins. We used two well-characterized Staphylococcus aureus proteins, SSL7 and Efb, to develop an ELISA-based inhibition assay using plasma from different animal species. The interaction between SSL7 and human C5 and the binding of Efb to human fibrinogen and human C3 was studied. Affinity experiments and Western blot analyses were used to validate the assay. Human, monkey and cat plasma interfered with binding of SSL7 to human C5. Binding of Efb to human fibrinogen was blocked in human, monkey, gerbil and pig plasma, while human, monkey, gerbil, rabbit, cat and guinea pig plasma inhibited the binding of Efb to human C3. These results emphasize the importance of choosing correct animal models, and thus, our approach is a rapid and cost-effective method that can be used to prevent unnecessary animal experiments. © 2017 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

  5. Selection of the most suitable method for the extraction of DNA from foods and feeds for species identification

    Directory of Open Access Journals (Sweden)

    Michaela Nesvadbová


    Full Text Available High quality and purity of DNA isolated from food and feed is essential for species identification and has unpredictable influences an effect of analysis. In this study, the efficiency of eight different methods for DNA isolation was investigated. For DNA extraction, the raw chicken meet, ham, sausages, tinned lunch meat, pate, tinned feeds for dogs, complete granulated feeds for dogs and chicken flour were used. Kits of several different producers, i.e.: NucleoSpin Food (Marchery-Nagel, Wizard Genomic DNA Purification Kit (Promega, Invisorb Spin Food Kit I (Invitek, Wizard SV Genomic DNA Purification System (Promega, JetQuick Tissue DNA Spin Kit (Genomed, RNA Blue (Top-Bio, JetQuick Blood & Cell Culture Kit (Genomed, QIAamp DNA Mini Kit and QIAamp DNA Blood Mini Kit (Qiagen were employed in the study. Gel agarose electrophoresis for primary verification of DNA quality was performed. The isolates were subsequently assessed for quantity and quality using by spectrophotometer Nanodrop 2000 (Thermo Scientific. To verify of template usability and quality of isolated DNA, the polymerase chain reaction (PCR was used.Differences between isolated DNA from tinned products and meat, ham, sausage, granulated dog feed and chicken flour were found. In tinned food and feed, the DNA was more degraded, DNA content and DNA purity was lower and also PCR amplification was the most difficult. Overall DNA yield and quality have important influence on PCR products amplification. The best results were obtained with NucleoSpin Food and JetQuick Tissue DNA Spin Kit. DNA extracted by these methods proved highest yields, purity and template quality in all foods and feeds and the results of PCR analysis are excellent reproducible. Analyses showed that results depended on different food or feed using and dif­fe­rent isolation system.The results of this work will be utilized to choose the suitable isolating kit for educational course, which is designed for students and

  6. Biopharmaceutics permeability classification of lorcaserin, a selective 5-hydroxytryptamine 2C agonist: method suitability and permeability class membership. (United States)

    Chen, Chuan; Ma, Michael G; Fullenwider, Cody L; Chen, Weichao G; Sadeque, Abu J M


    The objectives of the study were (1) to demonstrate that a Caco-2 cell-based permeability assay, developed in our laboratory, is suitable to identify the permeability classification according to the US Food and Drug Administration Biopharmaceutics Classification System guidance, and (2) to use the validated Caco-2 method to determine permeability class membership of lorcaserin. Lorcaserin, marketed in United States as Belviq, is a selective human 5-hydroxytryptamine 2C agonist used for weight management. First, the permeability of twenty commercially available drugs was determined in the apical-to-basolateral direction at a final concentration of 10 μM, with the pH of transporter buffer in the apical and basolateral compartments being 6.8 and 7.4, respectively. A rank-order relationship between in vitro permeability results and the extent of human intestinal absorption for the drugs tested was observed. Second, the apparent permeability coefficient values of lorcaserin at 2, 20, and 200 μM and apical pH values of 6.8 and 7.4 in the apical-to-basolateral direction were determined using the validated method and found to be comparable to those of the high-permeability internal standard metoprolol. Lorcaserin permeability across Caco-2 cell monolayers was not dependent on the variation of apical pH. Furthermore, lorcaserin was not a substrate for efflux transporters such as P-glycoprotein. In conclusion, using the validated Caco-2 permeability assay, it was shown that lorcaserin is a highly permeable compound.

  7. Radioimmunoassay method suitable for analysis of salivary cortisol in patients with breast cancer in treatment by teletherapy - a preliminary study

    International Nuclear Information System (INIS)

    Souza, Kellen Verena S.; Santos, Jane Lima dos


    Determination of salivary cortisol using radioimmunoassay and the relationship between stress levels were investigated before and after of the psychological intervention of 21 patients during teletherapy treatment of breast cancer. The benefits of psychological intervention during cancer treatment has been observed in some studies and evidence show that the patient's emotional state directly influences adherence to treatment, good prognosis and under stress. Thus, in present study 21 patients aged 50-70 years, females in the treatment of breast cancer at the Radiotherapy Service of Santa Casa de Misericordia in Belo Horizonte, were divided randomly into two groups: control (without psychological intervention) and Group TCC (with psychological intervention). Salivary cortisol concentrations are reportedly about 5.10% of those in serum, the levels has been used as an objective marker of hypothalamic-pituitary-adrenal (HPA) axis activity and avoided the confounding effects of venepuncture stress on cortisol levels. The assessment of cortisol in saliva has proven a valid and reliable measure of unbound steroid in the circulation. The salivary cortisol measured by radioimmunoassay in control patients and TCC showed no significant differences. The radioimmunoassay method show be suitable for analysis of salivary cortisol because allows measure low detection limits and have high sensibility. The values of TCC group and Control group were higher than reference values, except for one patient of the control group. At the beginning of treatment, both groups were homogeneous for Teletherapy. Both the TCC group as the control group had . 80% of participants in phase with stress. It was observed in the TCC group after radiotherapy showing a reversal 60% of participants without stress. The data show that cognitive-behavioral therapy has benefits for the control of psychological stress and enhance relevance of the psychologist to provide reduction or stress control during

  8. Radioimmunoassay method suitable for analysis of salivary cortisol in patients with breast cancer in treatment by teletherapy - a preliminary study

    Energy Technology Data Exchange (ETDEWEB)

    Souza, Kellen Verena S. [Departamento de Engenharia Nuclear. Universidade Federal de Minas Gerais, Pampulha, MG (Brazil); Santos, Jane Lima dos [Departamento de Ciencias Biologicas, Universidade Estadual de Santa Cruz, Ilheus, BA (Brazil)


    Determination of salivary cortisol using radioimmunoassay and the relationship between stress levels were investigated before and after of the psychological intervention of 21 patients during teletherapy treatment of breast cancer. The benefits of psychological intervention during cancer treatment has been observed in some studies and evidence show that the patient's emotional state directly influences adherence to treatment, good prognosis and under stress. Thus, in present study 21 patients aged 50-70 years, females in the treatment of breast cancer at the Radiotherapy Service of Santa Casa de Misericordia in Belo Horizonte, were divided randomly into two groups: control (without psychological intervention) and Group TCC (with psychological intervention). Salivary cortisol concentrations are reportedly about 5.10% of those in serum, the levels has been used as an objective marker of hypothalamic-pituitary-adrenal (HPA) axis activity and avoided the confounding effects of venepuncture stress on cortisol levels. The assessment of cortisol in saliva has proven a valid and reliable measure of unbound steroid in the circulation. The salivary cortisol measured by radioimmunoassay in control patients and TCC showed no significant differences. The radioimmunoassay method show be suitable for analysis of salivary cortisol because allows measure low detection limits and have high sensibility. The values of TCC group and Control group were higher than reference values, except for one patient of the control group. At the beginning of treatment, both groups were homogeneous for Teletherapy. Both the TCC group as the control group had . 80% of participants in phase with stress. It was observed in the TCC group after radiotherapy showing a reversal 60% of participants without stress. The data show that cognitive-behavioral therapy has benefits for the control of psychological stress and enhance relevance of the psychologist to provide reduction or stress control during

  9. Selection of Suitable DNA Extraction Methods for Genetically Modified Maize 3272, and Development and Evaluation of an Event-Specific Quantitative PCR Method for 3272. (United States)

    Takabatake, Reona; Masubuchi, Tomoko; Futo, Satoshi; Minegishi, Yasutaka; Noguchi, Akio; Kondo, Kazunari; Teshima, Reiko; Kurashima, Takeyo; Mano, Junichi; Kitta, Kazumi


    A novel real-time PCR-based analytical method was developed for the event-specific quantification of a genetically modified (GM) maize, 3272. We first attempted to obtain genome DNA from this maize using a DNeasy Plant Maxi kit and a DNeasy Plant Mini kit, which have been widely utilized in our previous studies, but DNA extraction yields from 3272 were markedly lower than those from non-GM maize seeds. However, lowering of DNA extraction yields was not observed with GM quicker or Genomic-tip 20/G. We chose GM quicker for evaluation of the quantitative method. We prepared a standard plasmid for 3272 quantification. The conversion factor (Cf), which is required to calculate the amount of a genetically modified organism (GMO), was experimentally determined for two real-time PCR instruments, the Applied Biosystems 7900HT (the ABI 7900) and the Applied Biosystems 7500 (the ABI7500). The determined Cf values were 0.60 and 0.59 for the ABI 7900 and the ABI 7500, respectively. To evaluate the developed method, a blind test was conducted as part of an interlaboratory study. The trueness and precision were evaluated as the bias and reproducibility of the relative standard deviation (RSDr). The determined values were similar to those in our previous validation studies. The limit of quantitation for the method was estimated to be 0.5% or less, and we concluded that the developed method would be suitable and practical for detection and quantification of 3272.

  10. Ceramic material suitable for repair of a space vehicle component in a microgravity and vacuum environment, method of making same, and method of repairing a space vehicle component (United States)

    Riedell, James A. (Inventor); Easler, Timothy E. (Inventor)


    A precursor of a ceramic adhesive suitable for use in a vacuum, thermal, and microgravity environment. The precursor of the ceramic adhesive includes a silicon-based, preceramic polymer and at least one ceramic powder selected from the group consisting of aluminum oxide, aluminum nitride, boron carbide, boron oxide, boron nitride, hafnium boride, hafnium carbide, hafnium oxide, lithium aluminate, molybdenum silicide, niobium carbide, niobium nitride, silicon boride, silicon carbide, silicon oxide, silicon nitride, tin oxide, tantalum boride, tantalum carbide, tantalum oxide, tantalum nitride, titanium boride, titanium carbide, titanium oxide, titanium nitride, yttrium oxide, zirconium diboride, zirconium carbide, zirconium oxide, and zirconium silicate. Methods of forming the ceramic adhesive and of repairing a substrate in a vacuum and microgravity environment are also disclosed, as is a substrate repaired with the ceramic adhesive.

  11. Memories Persist in Silence


    Sandra Patricia Arenas Grisales


    This article exposes the hypothesis that memory artifacts, created to commemorate the victims of armed conflict in Colombia, are an expression of the underground memories and a way of political action in the midst of war. We analyze three cases of creations of memory artifacts in Medellín, Colombia, as forms of suffering, perceiving and resisting the power of armed groups in Medellín. The silence, inherent in these objects, should not be treated as an absence of language, but as another form ...

  12. Voice and silence in organizations

    Directory of Open Access Journals (Sweden)

    Moaşa, H.


    Full Text Available Unlike previous research on voice and silence, this article breaksthe distance between the two and declines to treat them as opposites. Voice and silence are interrelated and intertwined strategic forms ofcommunication which presuppose each other in such a way that the absence of one would minimize completely the other’s presence. Social actors are not voice, or silence. Social actors can have voice or silence, they can do both because they operate at multiple levels and deal with multiple issues at different moments in time.

  13. Wheat germ agglutinin staining as a suitable method for detection and quantification of fibrosis in cardiac tissue after myocardial infarction

    Directory of Open Access Journals (Sweden)

    B. Emde


    Full Text Available The quantification of fibrotic tissue is an important task in the analysis of cardiac remodeling. The use of established fibrosis staining techniques is limited on frozen cardiac tissue sections due to a reduced color contrast compared to paraffin embedded sections. We therefore used FITC-labeled wheat germ agglutinin (WGA, which marks fibrotic tissue in comparable quality as the established picrosirius red (SR staining, for the staining of post myocardial infarction scar tissue. The fibrosis amount was quantified in a histogram-based approach using the non-commercial image processing program ImageJ. Our results clearly demonstrate that WGA-FITC is a suitable marker for cardiac fibrosis in frozen tissue sections. In combination with the histogram-based analysis, this new quantification approach is i easy and fast to perform; ii suitable for raw frozen tissue sections; and iii allows the use of additional antibodies in co-immunostaining. 

  14. "Listening Silence" and Its Discursive Effects (United States)

    Applebaum, Barbara


    While researchers have studied how white silence protects white innocence and white ignorance, in this essay Barbara Applebaum explores a form of white silence that she refers to as "listening silence" in which silence protects white innocence but does not necessarily promote resistance to learning. White listening silence can appear to…

  15. Bodies, Spaces, Voices, Silences

    Directory of Open Access Journals (Sweden)

    Donatella Mazzoleni


    Full Text Available A good architecture should not only allow functional, formal and technical quality for urban spaces, but also let the voice of the city be perceived, listened, enjoyed. Every city has got its specific sound identity, or “ISO” (R. O. Benenzon, made up of a complex texture of background noises and fluctuation of sound figures emerging and disappearing in a game of continuous fadings. For instance, the ISO of Naples is characterized by a spread need of hearing the sound return of one’s/others voices, by a hate of silence. Cities may fall ill: illness from noise, within super-crowded neighbourhoods, or illness from silence, in the forced isolation of peripheries. The proposal of an urban music therapy denotes an unpublished and innovative enlarged interdisciplinary research path, where architecture, music, medicine, psychology, communication science may converge, in order to work for rebalancing spaces and relation life of the urban collectivity, through the care of body and sound dimensions.

  16. Implementation of USP antibody standard for system suitability in capillary electrophoresis sodium dodecyl sulfate (CE-SDS) for release and stability methods. (United States)

    Esterman, Abbie L; Katiyar, Amit; Krishnamurthy, Girija


    Capillary electrophoresis sodium dodecyl sulfate (CE-SDS) is widely used for purity analysis of monoclonal antibody therapeutics for release and stability to demonstrate product consistency and shelf life during the manufacturing and life cycle of the product. CE-SDS method development is focused on exploring the method capability to provide the information about the product purity and product related degradants (fragmentation, aggregation etc.). In order to establish the functionality of the instrumentation, software, and sample preparation; system suitability criteria need to be defined for analytical methods using a well characterized reference standard run under the same protocol and analysis as the test articles. Typically the reference standard is produced using a manufacturing process representative of the clinical material. The qualification, control, and maintenance of in-house reference standards are established through rigorous quality and regulatory guidelines. The U.S. Pharmacopeia (USP) has developed a monoclonal IgG System Suitability Reference Standard to be utilized for assessment of system suitability in CE-SDS methods. In this communication, we evaluate the system suitability acceptance criteria performance of the USP IgG standard using two methods, the recommended USP protocol provided in monograph and a molecule specific Bristol-Myers Squibb (BMS) CE-SDS method. The results from USP IgG standard were compared with two in-house monoclonal antibody reference standards. The data suggest that the USP CE-SDS method may not be suitable for CE-SDS analysis for release and stability of monoclonal antibody therapeutics due to the high level of method induced partial reduction observed for all molecules tested. This high level of fragmentation observed utilizing the USP method will result in reporting lower purity levels, which will impact the overall quality assessment of the molecule. The system suitability criteria recommended by the USP method can be

  17. Organizational Silence in Sports Employees (United States)

    Bastug, Gulsum; Pala, Adem; Yilmaz, Taner; Duyan, Mehdi; Gunel, Ilker


    Organizational silence can be defined as a way of behaviour belonging to men and women employees in the organization exhibited without reflecting their feelings, ideas, concerns and suggestions related with their workplaces, works for which they are responsible or other activities of the organization. In the period of organizational silence,…

  18. Silence, an Eye of Knowledge (United States)

    Aghamohammadi, Mehdi


    One of the conspicuous features of the twentieth-century West was silence. This idea could be supported by examining reflections of Ludwig Wittgenstein, Fritz Mauthner, John Cage, Samuel Beckett, Ihab Hassan, Franz Kafka, Wassily Kandinsky, Jean-Paul Sartre, Virginia Woolf, Wolfgang Iser, Jacques Derrida, and Pierre Macherey. To me, silence is not…

  19. Fast Swinnex Filtration (FSF): A fast and robust sampling and extraction method suitable for metabolomics analysis of cultures grown in complex media

    DEFF Research Database (Denmark)

    McCloskey, Douglas; Utrilla, Jose; Naviaux, Robert K.


    , we develop a fast-filtration method using pressuredriven Swinnex filters. We show that the method is fast enough to provide an accurate snapshot of intracellular metabolism, reduces matrix interference from the media to improve the number of compounds that can be detected, and is applicable...... of a suitable method for anaerobic cultures grown in complex media. Given that a vast majority of bacteria are facultative or obligate anaerobes that grow to low biomass density and need to be cultured in complex media, a suitable sampling and extraction strategy for anaerobic cultures is needed. In this work...... to anaerobic and aerobic liquid cultures grown in a variety of culturing systems. Furthermore, we apply the fast filtration method to investigate differences in the absolute intracellular metabolite levels of anaerobic cultures grown in minimal and complex media....

  20. A suitable method to detect potential fraud of bringing Malayan box turtle (Cuora amboinensis) meat into the food chain. (United States)

    Ali, Md Eaqub; Asing; Hamid, Sharifah Bee Abd; Razzak, Md Abdur; Rashid, Nur Raifana Abd; Al Amin, Md; Mustafa, Shuhaimi


    Malayan box turtle (Cuora amboinensis) has been a wildlife-protected vulnerable turtle species in Malaysia since 2005. However, because of its purported usage in traditional medicine, tonic foods and feeds, clandestine black market trade is rampant. Several polymerase chain reaction (PCR) assays for the taxonomic detection and classification of turtle species have been proposed. These assays are based on long-length target amplicons which are assumed to break down under compromised states and, hence, might not be suitable for the forensic tracing and tracking of turtle trafficking. For the first time this paper develops a very short-amplicon-length PCR assay (120 bp) for the detection of Malayan box turtle meat in raw, processed and mixed matrices, and experimental evidence is produced that such an assay is not only more stable and reliable but also more sensitive than those previously published. We checked the assay specificity against 20 different species and no cross-species detection was observed. The possibility of any false-negative detection was eliminated by a universal endogenous control for eukaryotes. The assay detection limit was 0.0001 ng of box turtle DNA from pure meat and 0.01% turtle meat in binary and ternary admixtures and commercial meatballs. Superior target stability and sensitivity under extreme treatments of boiling, autoclaving and microwave cooking suggested that this newly developed assay would be suitable for any forensic and/or archaeological identification of Malayan box turtle species, even in severely degraded specimens. Further, in silico studies indicated that the assay has the potential to be used as a universal probe for the detection of nine Cuora species, all of which are critically endangered.

  1. Bioinformatics tools for achieving better gene silencing in plants. (United States)

    Ahmed, Firoz; Dai, Xinbin; Zhao, Patrick Xuechun


    RNA interference (RNAi) is one of the most popular and effective molecular technologies for knocking down the expression of an individual gene of interest in living organisms. Yet the technology still faces the major issue of nonspecific gene silencing, which can compromise gene functional characterization and the interpretation of phenotypes associated with individual gene knockdown. Designing an effective and target-specific small interfering RNA (siRNA) for induction of RNAi is therefore the major challenge in RNAi-based gene silencing. A 'good' siRNA molecule must possess three key features: (a) the ability to specifically silence an individual gene of interest, (b) little or no effect on the expressions of unintended siRNA gene targets (off-target genes), and (c) no cell toxicity. Although several siRNA design and analysis algorithms have been developed, only a few of them are specifically focused on gene silencing in plants. Furthermore, current algorithms lack a comprehensive consideration of siRNA specificity, efficacy, and nontoxicity in siRNA design, mainly due to lack of integration of all known rules that govern different steps in the RNAi pathway. In this review, we first describe popular RNAi methods that have been used for gene silencing in plants and their serious limitations regarding gene-silencing potency and specificity. We then present novel, rationale-based strategies in combination with computational and experimental approaches to induce potent, specific, and nontoxic gene silencing in plants.

  2. Experimental Evaluation of Suitability of Selected Multi-Criteria Decision-Making Methods for Large-Scale Agent-Based Simulations. (United States)

    Tučník, Petr; Bureš, Vladimír


    Multi-criteria decision-making (MCDM) can be formally implemented by various methods. This study compares suitability of four selected MCDM methods, namely WPM, TOPSIS, VIKOR, and PROMETHEE, for future applications in agent-based computational economic (ACE) models of larger scale (i.e., over 10 000 agents in one geographical region). These four MCDM methods were selected according to their appropriateness for computational processing in ACE applications. Tests of the selected methods were conducted on four hardware configurations. For each method, 100 tests were performed, which represented one testing iteration. With four testing iterations conducted on each hardware setting and separated testing of all configurations with the-server parameter de/activated, altogether, 12800 data points were collected and consequently analyzed. An illustrational decision-making scenario was used which allows the mutual comparison of all of the selected decision making methods. Our test results suggest that although all methods are convenient and can be used in practice, the VIKOR method accomplished the tests with the best results and thus can be recommended as the most suitable for simulations of large-scale agent-based models.

  3. Assessing future suitability of tree species under climate change by multiple methods: a case study in southern Germany

    Directory of Open Access Journals (Sweden)

    Helge Walentowski


    Full Text Available We compared results derived using three different approaches to assess the suitability of common tree species on the Franconian Plateau in southern Germany under projected warmer and drier climate conditions in the period 2061-2080. The study area is currently a relatively warm and dry region of Germany. We calculated species distribution models (SDMs using information on species’ climate envelopes to predict regional species spectra under 63 different climate change scenarios. We complemented this with fine-scale ecological niche analysis using data from 51 vegetation surveys in seven forest reserves in the study area, and tree-ring analysis (TRA from local populations of five tree species to quantify their sensitivity to climatic extreme years. The SDMs showed that predicted future climate change in the region remains within the climate envelope of certain species (e.g. Quercus petraea, whilst for e.g. Fagus sylvatica, future climate conditions in one third of the scenarios are too warm and dry. This was confirmed by the TRA: sensitivity to drought periods is lower for Q. petraea than for F. sylvatica. The niche analysis shows that the local ecological niches of Quercus robur and Fraxinus excelsior are mainly characterized by soils providing favorable water supply than by climate, and Pinus sylvestris (planted is strongly influenced by light availability. The best adapted species for a warmer and potentially drier climate in the study region are Acer campestre, Sorbus torminalis, S. aria, Ulmus minor, and Tilia platyphyllos, which should therefore play a more prominent role in future climate-resilient mixed forest ecosystems.

  4. Preparation of the new certified reference material Virginia Tobacco Leaves (CTA-VTL-2) and development of suitable methods for checking the homogeneity

    International Nuclear Information System (INIS)

    Dybczynski, R.; Polkowska-Motrenko, H.; Samczynski, Z.; Szopa, Z.


    The aim of this project in the long run has been reparation of a new biological reference material: Tobacco leaves of the 'Virginia' type and its certification for the content of possibly great number of trace elements. Further aims have been: development of the suitable methods for checking the homogeneity with the special emphasis on homogeneity of small samples and the critical analysis of the performance of various analytical techniques

  5. A Rapid Method for the Extraction and Analysis of Carotenoids and Other Hydrophobic Substances Suitable for Systems Biology Studies with Photosynthetic Bacteria

    Directory of Open Access Journals (Sweden)

    Oliver Sawodny


    Full Text Available A simple, rapid, and inexpensive extraction method for carotenoids and other non-polar compounds present in phototrophic bacteria has been developed. The method, which has been extensively tested on the phototrophic purple non-sulphur bacterium Rhodospirillum rubrum, is suitable for extracting large numbers of samples, which is common in systems biology studies, and yields material suitable for subsequent analysis using HPLC and mass spectroscopy. The procedure is particularly suitable for carotenoids and other terpenoids, including quinones, bacteriochlorophyll a and bacteriopheophytin a, and is also useful for the analysis of polar phospholipids. The extraction procedure requires only a single step extraction with a hexane/methanol/water mixture, followed by HPLC using a Spherisorb C18 column, with a mobile phase consisting of acetone-water and a non-linear gradient of 50%–100% acetone. The method was employed for examining the carotenoid composition observed during microaerophilic growth of R. rubrum strains, and was able to determine 18 carotenoids, 4 isoprenoid-quinones, bacteriochlorophyll a and bacteriopheophytin a as well as four different phosphatidylglycerol species of different acyl chain compositions. The analytical procedure was used to examine the dynamics of carotenoid biosynthesis in the major and minor pathways operating simultaneously in a carotenoid biosynthesis mutant of R. rubrum.

  6. On the suitability of fast and frugal heuristics for designing values clarification methods in patient decision aids : A critical analysis

    NARCIS (Netherlands)

    Pieterse, A.H.; de Vries, M.


    Background  Increasingly, patient decision aids and values clarification methods (VCMs) are being developed to support patients in making preference-sensitive health-care decisions. Many VCMs encourage extensive deliberation about options, without solid theoretical or empirical evidence showing that

  7. Silence, an Eye of Knowledge

    Directory of Open Access Journals (Sweden)

    Mehdi Aghamohammadi


    Full Text Available One of the conspicuous features of the twentieth-century West was silence. This idea could be supported by examining reflections of Ludwig Wittgenstein, Fritz Mauthner, John Cage, Samuel Beckett, Ihab Hassan, Franz Kafka, Wassily Kandinsky, Jean-Paul Sartre, Virginia Woolf, Wolfgang Iser, Jacques Derrida, and Pierre Macherey. To me, silence is not a mere theory, but rather a phenomenon from which we can get practical benefits. I believe silence is an eye, eye of knowledge. We can broaden our knowledge of the world through silence. To convey the idea that silence is an eye, I have concocted the word slence, where  has replaced the letter i and stands for the eye. This means knowledge can enable us to see, thereby acquiring knowledge of, what used to be invisible, and accordingly unknowable. In other words, through silence, we can achieve a certain type of literacy. I substantiate this claim by exploring the Horus myth, Ojo de Dios, John Cage’s 4' 33", the nature of Expressionist paintings, Hinduism, thoughts of Hermes Trismegistus and Ibn al-Arabi, and practices of Mohammad, the prophet of Islam.

  8. A multi-criteria, ecosystem-service value method used to assess catchment suitability for potential wetland reconstruction in Denmark

    DEFF Research Database (Denmark)

    Odgaard, Mette Vestergaard; Turner, Katrine Grace; Bøcher, Peder Klith


    Wetlands provide a range of ecosystem services such as drought resistance, flood resistance, nutrient deposition, biodiversity, etc. This study presents a new multi-criteria, ecosystems service value-driven method to drive the optimal placement of restored wetlands in terms of maximizing selected...... actually hold a presently reconstructed wetland. This indicates a prior placement with almost no consideration of maximizing ecosystem services benefits. We recommend a systematic approach, such as the ecosystem service value-driven method demonstrated in the present case study, to target more services...... placements of wetlands. Furthermore, scenario testing and hotspot analysis were combined to provide future recommendations for optimal placements of wetlands. The scenarios investigated were Climate Adaptation and Protection of Aquatic Environment, Land-Based Economy, and Rich Nature. Based...

  9. Method for the production of a solid waste form suitable for ultimate disposal of tritium-bearing water

    International Nuclear Information System (INIS)

    Kunz, W.; Gramatte, W.; Schmitt, R.E.; Pohl, U.


    A method for the production of a cement product for the final storage of tritium-containing waters is described, in which activated bentonite capable of swelling is dispersively mixed with the tritium-containing water and the suspension obtained is subjected to a swelling process. The expanded suspension is dispersively mixed with the cement. The set product has a water content of more than 75% by weight. (orig.) [de

  10. Comparing two remote video survey methods for spatial predictions of the distribution and environmental niche suitability of demersal fishes. (United States)

    Galaiduk, Ronen; Radford, Ben T; Wilson, Shaun K; Harvey, Euan S


    Information on habitat associations from survey data, combined with spatial modelling, allow the development of more refined species distribution modelling which may identify areas of high conservation/fisheries value and consequentially improve conservation efforts. Generalised additive models were used to model the probability of occurrence of six focal species after surveys that utilised two remote underwater video sampling methods (i.e. baited and towed video). Models developed for the towed video method had consistently better predictive performance for all but one study species although only three models had a good to fair fit, and the rest were poor fits, highlighting the challenges associated with modelling habitat associations of marine species in highly homogenous, low relief environments. Models based on baited video dataset regularly included large-scale measures of structural complexity, suggesting fish attraction to a single focus point by bait. Conversely, models based on the towed video data often incorporated small-scale measures of habitat complexity and were more likely to reflect true species-habitat relationships. The cost associated with use of the towed video systems for surveying low-relief seascapes was also relatively low providing additional support for considering this method for marine spatial ecological modelling.

  11. Novel extraction method of genomic DNA suitable for long-fragment amplification from small amounts of milk. (United States)

    Liu, Y F; Gao, J L; Yang, Y F; Ku, T; Zan, L S


    Isolation of genomic DNA is a prerequisite for assessment of milk quality. As a source of genomic DNA, milk somatic cells from milking ruminants are practical, animal friendly, and cost-effective sources. Extracting DNA from milk can avoid the stress response caused by blood and tissue sampling of cows. In this study, we optimized a novel DNA extraction method for amplifying long (>1,000 bp) DNA fragments and used it to evaluate the isolation of DNA from small amounts of milk. The techniques used for the separation of milk somatic cell were explored and combined with a sodium dodecyl sulfate (SDS)-phenol method for optimizing DNA extraction from milk. Spectrophotometry was used to determine the concentration and purity of the extracted DNA. Gel electrophoresis and DNA amplification technologies were used for to determine DNA size and quality. The DNA of 112 cows was obtained from milk (samples of 13 ± 1 mL) and the corresponding optical density ratios at 260:280 nm were between 1.65 and 1.75. Concentrations were between 12 and 45 μg/μL and DNA size and quality were acceptable. The specific PCR amplification of 1,019- and 729-bp bovine DNA fragments was successfully carried out. This novel method can be used as a practical, fast, and economical mean for long genomic DNA extraction from a small amount of milk. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  12. Suitability of monitoring methods for the optimisation of Radiological Protection in the case of internal exposure through inhalation

    International Nuclear Information System (INIS)

    Degrange, J.P.; Gibert, B.; Basire, D.


    The radiological protection system recommended by the International Commission for Radiological Protection (ICRP) for justified practices relied pn the limitation and optimisation principles. The monitoring of internal exposure is most often based on the periodic assessment of individual exposure in order to essentially insure the simple compliance with the annual dose limits. Optimisation of protection implies a realistic, sensitive and analytical assessment of individual and collective exposures in order to allow the indentification of the main sources of exposure (main sources of contamination, most exposed operators, work activities contributing the most to the exposure) and the selection of the optimal protection options. Therefore the monitoring methods must allow the realistic assessment of individual dose levels far lower than annual limits together with measurements as frequent as possible. The aim of this presentation is to discuss the ability of various monitoring methods (collective and individual air sampling, in vivo and in vitro bioassays) to fulfil those needs. This discussion is illustrated by the particular case of the internal exposure to natural uranium compounds through inhalation. Firstly, the sensitivity and the degree to which each monitoring method is realistic are quantified and discussed on the basis of the application of the new ICRP dosimetric model, and their analytical capability for the optimisation of radiological protection is then indicated. Secondly, a case study is presented which shows the capability of individual air sampling techniques to analyse the exposure of the workers and the inadequacy of static air sampling to accurately estimate the exposures when contamination varies significantly over time and space in the workstations. As far as exposure to natural uranium compounds through inhalation is concerned, the study for assessing the sensitivity, analytic ability and accuracy of the different measuring systems shows that

  13. A standardized scoring method for the copy of cube test, developed to be suitable for use in psychiatric populations

    Directory of Open Access Journals (Sweden)

    Terzoglou Vassiliki A


    Full Text Available Abstract Background Although the 'copy of cube test', a version of which is included in the Short Test of Mental Status (STMS, has existed for years, little has been done to standardize it in detail. The aim of the current study was to develop a novel and detailed standardized method of administration and scoring this test. Methods The study sample included 93 healthy control subjects (53 women and 40 men aged 35.87 ± 12.62 and 127 patients suffering from schizophrenia (54 women and 73 men aged 34.07 ± 9.83 years. The psychometric assessment included the Positive and Negative Symptoms Scale (PANSS the Young Mania Rating Scale (YMRS, and the Montgomery-Åsberg Depression Rating Scale (MADRS. Results A scoring method was developed based on the frequencies of responses of healthy controls. Cronbach's α was equal to 0.75 and inter-rater reliability was 0.90. Three indices and five subscales of the Standardized Copy of the Cube Test (SCCT were eventually developed. They included the Deficit Index (DcI, which includes the Missing Elements (ME Mirror Image (M subscales, the Deformation Index (DfI which includes the Deformation (D and the Rotation (R subscales and the Closing-In Index (CiI. Discussion The SCCT seems to be a reliable, valid and sensitive to change instrument for the testing of psychiatric patients. The great advantage of this instrument is the fact that it only requires paper and a pencil, and is this easily administered and brief. Further research is necessary to test its usefulness as a neuropsychological test.

  14. Development of a suitable test method for evaluating the toxicity of contaminated soils to earthworms in Canada

    Energy Technology Data Exchange (ETDEWEB)

    Stephenson, G.L. [Univ. of Guelph, Ontario (Canada); Scroggins, R. [Environment Canada, Gloucester, Ontario (Canada). Method Development and Application Section


    Environment Canada has embarked on a five year program to develop, standardize, and validate a battery of soil toxicity tests which can be used to assess the relative toxicity of contaminants in soils to terrestrial organisms. These tests must be applicable to soil conditions typically found in Canadian environments and the test species must be representative of the species of soil invertebrates or plants inhabiting soil ecosystems in Canada. One of the toxicity tests being developed is designed to assess the toxicity of contaminated soils to earthworms. Five of the potential test species belong to the Lumbricidae family and include the Canadian worm (Allobophora calignosa/Aporrectodea tuberculate), the European bark worm (Dendrodtilus rubidus (rubida)), the pink soil worm (Eisenia rosea), the red marsh worm (Lumbricus rubellus), and the Canadian night crawler or dew worm (Lumbricus terrestris). The sixth species, the white pot worm (Enchytraeus albidus), belongs to the Enchytraeidae family. Further assessment reduced the number of representative species to three. Most earthworm test methods have been developed to assess the toxicity of chemically-spiked artificial soils to Eisenia fetida or E. andrei. Test methods have also been developed to assess the relative toxicity of contaminated soils from hazardous waste sites. Comparative acute toxicity data for three species of earthworm exposed to a hydrocarbon contamination will be presented. Comparative toxicity data for the same three species of earthworm will also be presented using test procedures and conditions that have been modified to accommodate biological differences among the species of earthworm. Recommendations regarding test design, methods, and conditions optimal for each test species will be summarized and discussed with respect to the precision of test results.

  15. Suitability of Secondary PEEK Telescopic Crowns on Zirconia Primary Crowns: The Influence of Fabrication Method and Taper

    Directory of Open Access Journals (Sweden)

    Susanne Merk


    Full Text Available This study investigates the retention load (RL between ZrO2 primary crowns and secondary polyetheretherketone (PEEK crowns made by different fabrication methods with three different tapers. Standardized primary ZrO2 crowns were fabricated with three different tapers: 0°, 1°, and 2° (n = 10/group. Ten secondary crowns were fabricated (i milled from breCam BioHPP blanks (PM; (ii pressed from industrially fabricated PEEK pellets (PP (BioHPP Pellet; or (iii pressed from granular PEEK (PG (BioHPP Granulat. One calibrated operator adjusted all crowns. In total, the RL of 90 secondary crowns were measured in pull-off tests at 50 mm/min, and each specimen was tested 20 times. Two- and one-way ANOVAs followed by a Scheffé’s post-hoc test were used for data analysis (p < 0.05. Within crowns with a 0° taper, the PP group showed significantly higher retention load values compared with the other groups. Among the 1° taper, the PM group presented significantly lower retention loads than the PP group. However, the pressing type had no impact on the results. Within the 2° taper, the fabrication method had no influence on the RL. Within the PM group, the 2° taper showed significantly higher retention load compared with the 1° taper. The taper with 0° was in the same range value as the 1° and 2° tapers. No impact of the taper on the retention value was observed between the PP groups. Within the PG groups, the 0° taper presented significantly lower RL than the 1° taper, whereas the 2° taper showed no differences. The fabrication method of the secondary PEEK crowns and taper angles showed no consistent effect within all tested groups.

  16. Phorbol myristate acetate stimulated NBT test: a simple method suitable for antenatal diagnosis of chronic granulomatous disease. (United States)

    Levinsky, R J; Harvey, B A; Rodeck, C H; Soothill, J F


    When endotoxin was compared with phorbol myristate acetate (PMA) for stimulation of phagocytes in the nitroblue tetrazolium (NBT) test, both methods discriminated between affected patients with X-linked chronic granulomatous disease (CGD) and controls, but only the PMA NBT test distinguished female carriers of CGD. Endotoxin provided no stimulation of normal fetal blood whereas PMA was an effective stimulator. Our results indicate the superiority of the PMA NBT test for diagnosis of patients and carriers of CGD and should allow accurate antenatal diagnosis of the disease. Images Fig. 3 PMID:6360441

  17. A Method for Teaching the Modeling of Manikins Suitable for Third-Person 3-D Virtual Worlds and Games

    Directory of Open Access Journals (Sweden)

    Nick V. Flor


    Full Text Available Virtual Worlds have the potential to transform the way people learn, work, and play. With the emerging fields of service science and design science, professors and students at universities are in a unique position to lead the research and development of innovative and value-adding virtual worlds. However, a key barrier in the development of virtual worlds—especially for business, technical, and non-artistic students—is the ability to model human figures in 3-D for use as avatars and automated characters in virtual worlds. There are no articles in either research or teaching journals which describe methods that non-artists can use to create 3-D human figures. This paper presents a repeatable and flexible method I have taught successfully to both artists and business students, which allows them to quickly model human-like figures (manikins that are sufficient for prototype purposes and that allows students and researchers alike to explore the development of new kinds of virtual worlds.

  18. The ''in vivo'' distribution of carbon 11 labeled-nicotine in animals. A method suitable for use in man

    International Nuclear Information System (INIS)

    Maziere, M.; Berger, G.; Plummer, D.; Comar, D.; Masse, R.


    A method is described to label nicotine with carbon 11. A hundred millicuries can be obtained, in 45 minutes, with a high specific activity. This labeling of nicotine has allowed an ''in vivo'' study of the distribution of this very toxic drug in animals. Five minutes after injection in rabbits or monkeys, it was shown with a gamma camera or with a positron camera that the radioactivity was very rapidly distributed throughout the tissues especially in brain, lungs and kidneys. 11 C-nicotine readily penetrates the blood-brain barrier and the brain radioactivity decreases very sharply with time. The eyes however retained activity, possibly in the retina. Unfortunately the monkey is not the ideal subject for 11 C-nicotine brain study because: the brain is small, considering the resolution of the cameras and the cerebral lobes are also quite overlaped in this animal; Japanese authors have shown that compared with dogs the nicotine brain uptake is lower, due to the high affinity of nicotine for skeletal muscle which occupies approximately forty to fifty % of the body weight of the monkey. Also in monkeys, the nicotine destruction is faster than in dogs because there is a higher enzyme nicotine metabolizing activity in the liver of this animal. The differences observed between various animals studies using nicotine indicate that we should not draw any firm conclusions about the behaviour of this drug in humans. In order to do so, examinations must be conducted in man and the method described in spite of its limitations provides a means for such a study

  19. The most suitable techiniques and methods to identify high alumina cement and based portland cement in concretes

    Directory of Open Access Journals (Sweden)

    Blanco, M. T.


    Full Text Available Instrumental techniques are indicated and the most adequated methodologies for determining the nature of the binder in concretes are explained. These methods are: a Determination of the Silicic Moduli through chemical analysis of the sample. This test reveáis very different valúes between cement portland based concrete and high alumina cement based concretes. b X-ray diffraction. It is considered as the best method. In the present paper the main diffraction Unes corresponding to the components of hydrated portland cement and high alumina cement, are given. As complementary methods having as limited use, DTA and TG are studied. The most significative changes of the hydrated phases of both cements when heated are shown. A valoration of the Oxine test is also done as well as a description of using the characteristic colour of the high alumina cement as a practical indicator for determining its presence in concretes. Finally, comments on the significance of the presence of different CaCO3 polymorhisms in portland cement concrete and high alumina cement concrétete are also done.

    Se indican las técnicas y se explican las metodologías más adecuadas para determinar la naturaleza del conglomerante aluminoso en hormigones. Estos métodos son: a Determinación del Módulo Silícico, a través de Análisis Químico de la muestra problema, al considerar la acusada diferencia de aquel valor en hormigones de cemento portland, respecto a hormigones de cemento aluminoso. b Difracción de Rayos X. Se considera la técnica idónea. Se dan en el presente trabajo las principales líneas de difracción de los compuestos característicos de los cementos hidratados, portland y aluminoso. Como métodos complementarios y de utilidad limitada se estudia el ATD y el TG. Se proponen las temperaturas más significativas de los compuestos hidratados de aquellos cementos, a través de lo cual se puede deducir la presencia de uno u otro

  20. Suitability of thermoluminescence, chemiluminescence, ESR and viscosity measurements as detection method for the irradiation of medicinal herbs; Eignung von Thermolumineszenz-, Chemilumineszenz-, ESR- und Viskositaetsmessungen zur Identifizierung strahlenbehandelter Arzneidrogen

    Energy Technology Data Exchange (ETDEWEB)

    Schuettler, C.; Gebhardt, G.; Stock, A.; Helle, N.; Boegl, K.W.


    Chemiluminescence, electron spin resonance, thermoluminescence and viscosity measurements have been investigated for their suitability as detection method for the irradiation of the medicinal herbs anise seeds (anisi fructus), valerian roots (valerianae radix), redberry leaves (uvae ursi folium), birch leaves (betulae folium), greek hay seeds (foenugraeci semen), cayenne pepper (capsici fructus acer), black-aldertee bark (frangulae cortex), fennel fruits (feoniculi fructus), rose hip shells (cynosbati fructus), coltsfoot (farfarae folium), acorus roots (calami rhizoma), chamomile flowers (matricariae flos), caraway (carvi fructus), lavender flowers (lavandulae flos), linseed (lini semen), lime tree flowers (tiliae flos), St. Mary`s thistle fruit (cardui mariae herba), lemon balm (melissae folium), java tea (orthosiphonis folium), peppermint (menthae piperitae folium), sage leaves (salviae folium), scouring rush (equiseti herba), senna leaves (sennae folium), plantain herbs (plantaginis lanceolata herba), thyme herbs (thymi herba), juniper berries (juniperi fructus), hawthorne herbs (crataegi folium), wheat starch (amylum tritici) and wormwood (absinthii herba). Depending on the herbs, the methods used were more or less suitable. Chemiluminescence measurements showed the smallest differences between untreated and irradiated samples whereas thermoluminescence measurements on isolated minerals from the vegetable drugs gave better results. In some herbs radiation-specific radicals could be identified by ESR spectroscopy. Viscosity measurement is suitable for some herbs as fast and inexpensive method for screening. (orig.) [Deutsch] Die Eignung von Chemilumineszenz-, ESR-, Thermolumineszenz- und Viskositaetsmessungen als Nachweismethode fuer die Behandlung von Arzneidrogen mit ionisierenden Strahlen wurde an Anis, Baldrianwurzel, Baerentraubenblaettern, Birkenblaettern, Bockshornsamen, Cayennepfeffer, Faulbaumrinde, Fenchel, Hagebuttenschalen, Huflattichblaettern

  1. Suitability of an ion trap GC/MS/MS method for routine analysis of PCDD/Fs in fishery products and by-products

    Energy Technology Data Exchange (ETDEWEB)

    Sobrado, C.; Quintela, C.; Blanco, L.; Gonzalez, J.C.; Vieites, J.M. [ANFACO-CECOPESCA, Vigo (Spain); Cabaleiro, S. [CETGA, Riveira (Spain)


    Fishery products and by-products constitute an important vector of dioxins towards humans due to bioacumulation and biomagnification, so fish processing industry is highly interested in the development of analytical procedures cheaper and faster than the the standard HRGCHRMS. The aim of this study was to demonstrate the suitability of a method of determination of the 17 congeners of dioxins and furans in fishery products and by-products, using a high resolution gas chromatography connected to an ion trap tandem mass spectrometer (HRGCMS/ MS) based on the patern of fragmentation of the congeners by MS/MS and quantification by isotope dilution.

  2. Most suitable mother wavelet for the analysis of fractal properties of stride interval time series via the average wavelet coefficient method. (United States)

    Zhang, Zhenwei; VanSwearingen, Jessie; Brach, Jennifer S; Perera, Subashan; Sejdić, Ervin


    Human gait is a complex interaction of many nonlinear systems and stride intervals exhibiting self-similarity over long time scales that can be modeled as a fractal process. The scaling exponent represents the fractal degree and can be interpreted as a "biomarker" of relative diseases. The previous study showed that the average wavelet method provides the most accurate results to estimate this scaling exponent when applied to stride interval time series. The purpose of this paper is to determine the most suitable mother wavelet for the average wavelet method. This paper presents a comparative numerical analysis of 16 mother wavelets using simulated and real fractal signals. Simulated fractal signals were generated under varying signal lengths and scaling exponents that indicate a range of physiologically conceivable fractal signals. The five candidates were chosen due to their good performance on the mean square error test for both short and long signals. Next, we comparatively analyzed these five mother wavelets for physiologically relevant stride time series lengths. Our analysis showed that the symlet 2 mother wavelet provides a low mean square error and low variance for long time intervals and relatively low errors for short signal lengths. It can be considered as the most suitable mother function without the burden of considering the signal length. Copyright © 2016 Elsevier Ltd. All rights reserved.

  3. Technical advances in trigger-induced RNA interference gene silencing in the parasite Entamoeba histolytica. (United States)

    Khalil, Mohamed I; Foda, Bardees M; Suresh, Susmitha; Singh, Upinder


    Entamoeba histolytica has a robust endogenous RNA interference (RNAi) pathway. There are abundant 27 nucleotide (nt) anti-sense small RNAs (AS sRNAs) that target genes for silencing and the genome encodes many genes involved in the RNAi pathway such as Argonaute proteins. Importantly, an E. histolytica gene with numerous AS sRNAs can function as a "trigger" to induce silencing of a gene that is fused to the trigger. Thus, the amebic RNAi pathway regulates gene expression relevant to amebic biology and has additionally been harnessed as a tool for genetic manipulation. In this study we have further improved the trigger-induced gene silencing method. We demonstrate that rather than using the full-length gene, a short portion of the coding region fused to a trigger is sufficient to induce silencing; the first 537 bp of the E. histolytica rhomboid gene (EhROM1) fused in-frame to the trigger was sufficient to silence EhROM1. We also demonstrated that the trigger method could silence two amebic genes concomitantly; fusion of the coding regions of EhROM1 and transcription factor, EhMyb, in-frame to a trigger gene resulted in both genes being silenced. Alternatively, two genes can be silenced sequentially: EhROM1-silenced parasites with no drug selection plasmid were transfected with trigger-EhMyb, resulting in parasites with both EhROM1 and EhMyb silenced. With all approaches tested, the trigger-mediated silencing was substantive and silencing was maintained despite loss of the G418 selectable marker. All gene silencing was associated with generation of AS sRNAs to the silenced gene. We tested the reversibility of the trigger system using inhibitors of histone modifications but found that the silencing was highly stable. This work represents a technical advance in the trigger gene silencing method in E. histolytica. Approaches that readily silence multiple genes add significantly to the genetic toolkit available to the ameba research community. Copyright © 2016

  4. Method for estimating critical properties of heavy compounds suitable for cubic equations of state and its application to the prediction of vapor pressures

    DEFF Research Database (Denmark)

    Kontogeorgis, Georgios; Ioannis, Smirlis; Iakovos, Yakoumis


    Cubic equations of state (EoS) are often used for correlating and predicting phase equilibria. Before extending any EoS to mixtures, reliable vapor-pressure prediction is essential. This requires experimental, if possible, critical temperatures T-c, pressures P-c, and acentric factor omega...... or extensive pure-compound vapor-pressure data which, for heavy and/or complex compounds, are often not available. This work presents a method for estimating T-c, P-c, and omega values for heavy compounds (typically with MW > 130) suitable for vapor-pressure calculations with generalized cubic Eo......S. The proposed scheme employs a recent group-contribution method (Constantinou et al. Fluid Phase Equilib. 1995, 103 (1), 11) for estimating the acentric factor. The two critical properties are estimated via a generalized correlation for the ratio T-c/P-c (with the van der Waals surface area) and the cubic Eo...

  5. Communicative Silences: Forms and Functions (United States)

    Bruneau, Thomas J.


    The nature of silence is discussed as an imposition of mind, as an interdependent signification ground for speech signs, as a relationship to mental time (as opposed to artificial time), and as it relates to sensation, perception and metaphorical movement. (Author)

  6. Silence in the creative process

    Directory of Open Access Journals (Sweden)

    Francesco Wilhelm


    Full Text Available A research about silence as the moment where creative thinking takes place, as an art object and as a way of interpreting artworks, is the main issue of the graduation thesis named Il silenzio nel processo creativo (2014. Here is proposed an offprint, which summarizes its fundamental steps.

  7. The silence of the archive

    CERN Document Server

    Thomas, David; Thomas, David


    This new book will provide a ground breaking discussion of a major but little considered issue - the silence of the archive: why archives, sometimes seen as the repositories of truth, often fail to satisfy users because they do not contain information which they expect to find.

  8. Breaching cultural silence: enhancing resilience among Ugandan ...

    African Journals Online (AJOL)

    Cultural silence is frequently the outcome of deep-seated taboos regarding adults talking to children about sex and death. This paper examines the impact of cultural silence on the resilience of children orphaned by AIDS in Uganda. Cultural silence is often linked with denial. This article explores the complexities of cultural ...

  9. Proceedings of the Workshop on Justifying the Suitability of Nuclear Licensee Organisational Structure, Resources and Competencies - Methods, Approaches and Good Practices

    International Nuclear Information System (INIS)


    The nuclear industry is currently facing a range of organisational challenges. The nuclear renaissance is resulting in renewed interest in new reactor build programmes; existing plants are being modernised; ageing plants and an ageing workforce are being replaced. The industry is developing new models of working in a competitive, and increasingly global market which has seen increased use of contractors and organisational change taking place at an unparalleled rate. It is clear that the way in which nuclear licensees' organisations are structured and resourced has a potential impact on nuclear safety. For example, nuclear safety may be challenged if organisational structures create uncertainty concerning authority and responsibilities or if nuclear safety functions are not adequately resourced. Inasmuch as this is so, then it is reasonable to expect both licensees and regulatory bodies to seek assurance that licensee organisations are suitable to manage nuclear safety and discharge the responsibilities associated with operating as a nuclear licensee. Although licensees should have the authority to organise their plant activities in different ways, they should also be able to demonstrate that they understand the potential impact that these activities may have on plant safety. They should be able to show how their organisations are designed to carry out these activities safely and effectively, and to verify that the nuclear safety functions are being delivered as expected. There is a growing interest from some nuclear regulatory bodies, as well as licensees, in methods and approaches that can be used to ensure that the licensee organisations are well structured and have sufficient resources and competencies to manage safety. To address these and other nuclear plant organisational safety-related issues a NEA/CSNI workshop was held in Uppsala (Sweden) hosted by the Swedish Radiation Safety Authority with support from the European Union's Joint Research Centre

  10. Friction coefficient measurements of silencers on specialized duct tract

    Directory of Open Access Journals (Sweden)

    Sehnalek Stanislav


    Full Text Available This article describes test methods on air duct track in Laboratory of Environmental Engineering. It focuses on measurement of silencer parameter like is pressure loss coeffcient. Firstly, the paper describe the measurement apparatus with description of calculation method by standard ISO 7235 and energy equation. Then the paper presents three ways how to accomplish measurement because such way is not covered by procedure in standard. Then follows the evaluation of results of measurements on three types of silencer designed for HVAC applications. The article is concluded with discussion over measured data with outline for further research.

  11. Comparison of Eleven Methods for Genomic DNA Extraction Suitable for Large-Scale Whole-Genome Genotyping and Long-Term DNA Banking Using Blood Samples (United States)

    Psifidi, Androniki; Dovas, Chrysostomos I.; Bramis, Georgios; Lazou, Thomai; Russel, Claire L.; Arsenos, Georgios; Banos, Georgios


    Over the recent years, next generation sequencing and microarray technologies have revolutionized scientific research with their applications to high-throughput analysis of biological systems. Isolation of high quantities of pure, intact, double stranded, highly concentrated, not contaminated genomic DNA is prerequisite for successful and reliable large scale genotyping analysis. High quantities of pure DNA are also required for the creation of DNA-banks. In the present study, eleven different DNA extraction procedures, including phenol-chloroform, silica and magnetic beads based extractions, were examined to ascertain their relative effectiveness for extracting DNA from ovine blood samples. The quality and quantity of the differentially extracted DNA was subsequently assessed by spectrophotometric measurements, Qubit measurements, real-time PCR amplifications and gel electrophoresis. Processing time, intensity of labor and cost for each method were also evaluated. Results revealed significant differences among the eleven procedures and only four of the methods yielded satisfactory outputs. These four methods, comprising three modified silica based commercial kits (Modified Blood, Modified Tissue, Modified Dx kits) and an in-house developed magnetic beads based protocol, were most appropriate for extracting high quality and quantity DNA suitable for large-scale microarray genotyping and also for long-term DNA storage as demonstrated by their successful application to 600 individuals. PMID:25635817

  12. Silence of the Genes

    Indian Academy of Sciences (India)


    tary to the endogenous sense mRNA produced by the normal gene. The antisense strand binds to the sense strand and blocks protein synthesis. This method of gene inhibition was termed antisense technology. The antisense technology soon became a. RNA interference. (RNAi) is a novel mechanism for controlling gene.

  13. Virus-induced gene silencing and Agrobacterium tumefaciens-mediated transient expression in Nicotiana tabacum

    NARCIS (Netherlands)

    Zhang, Z.; Thomma, B.P.H.J.


    Virus-induced gene silencing (VIGS) is a rapid method for transient silencing of plant genes. In this chapter, we describe the methodology for Tobacco rattle virus (TRV)-based VIGS in Nicotiana tabacum. In combination with subsequent co-expression of the tomato immune receptor Ve1 and the

  14. Testing the suitability of different high-performance liquid chromatographic methods to determine aflatoxin M1 in a soft fresh Italian cheese. (United States)

    Cattaneo, T M P; Marinoni, L; Barzaghi, S; Cremonesi, K; Monti, L


    Aflatoxin M1 (AFM1) is a toxic undesirable compound in milk. AFM1 affinity for caseins causes a concentration effect during milk process for dairy transformation. In spite of this, no official method of analysis, nor maximum tolerance level for aflatoxin M1 in cheese have been established. Thus, the aim of this work was to test the suitability of different HPLC methods for the AFM1 quantification in soft cheese samples at three different contamination levels (low, medium and high, at respectively nearly 30, 100 and 250 ng/kg). Nine participants were selected among Italian laboratories accredited by the Italian accreditation body (ACCREDIA) for HPLC toxin analysis. They were asked to analyze samples applying the method routinely used. The different applied methods were compared, and precision and accuracy parameters were evaluated. The main differences among HPLC procedures were registered at the level of extraction step. The use of an enzymatic digestion for the extraction of the toxin from cheese seemed to be particularly advantageous and the use of immunoaffinity columns seemed to be determinant for the improvement of sensitivity at low contamination levels. In general, the applied methods well discriminated the 3 levels of contamination, even though they performed better at the medium and high concentration levels (100 and 250 ng/kg) than at the low one (30 ng/kg). In fact relative standard deviation for reproducibility at low level was higher (60.1%) than the same value at medium and high levels (22.8% and 28.9%, respectively). Copyright © 2011 Elsevier B.V. All rights reserved.

  15. Transcriptional Silencing of Retroviral Vectors

    DEFF Research Database (Denmark)

    Lund, Anders Henrik; Duch, M.; Pedersen, F.S.


    Although retroviral vector systems have been found to efficiently transduce a variety of cell types in vitro, the use of vectors based on murine leukemia virus in preclinical models of somatic gene therapy has led to the identification of transcriptional silencing in vivo as an important problem....... Extinction of long-term vector expression has been observed after implantation of transduced hematopoietic cells as well as fibroblasts, myoblasts and hepatocytes. Here we review the influence of vector structure, integration site and cell type on transcriptional silencing. While down-regulation of proviral...... transcription is known from a number of cellular and animal models, major insight has been gained from studies in the germ line and embryonal cells of the mouse. Key elements for the transfer and expression of retroviral vectors, such as the viral transcriptional enhancer and the binding site for the t...

  16. Assessment of a novel multi-array normalization method based on spike-in control probes suitable for microRNA datasets with global decreases in expression. (United States)

    Sewer, Alain; Gubian, Sylvain; Kogel, Ulrike; Veljkovic, Emilija; Han, Wanjiang; Hengstermann, Arnd; Peitsch, Manuel C; Hoeng, Julia


    suitable for reliable downstream analysis. The multi-array miRNA raw data normalization method was implemented in an R software package called ExiMiR and deposited in the Bioconductor repository.

  17. Posttranscriptional gene silencing in nuclei (United States)

    Hoffer, Paul; Ivashuta, Sergey; Pontes, Olga; Vitins, Alexa; Pikaard, Craig; Mroczka, Andrew; Wagner, Nicholas; Voelker, Toni


    In plants, small interfering RNAs (siRNAs) with sequence homology to transcribed regions of genes can guide the sequence-specific degradation of corresponding mRNAs, leading to posttranscriptional gene silencing (PTGS). The current consensus is that siRNA-mediated PTGS occurs primarily in the cytoplasm where target mRNAs are localized and translated into proteins. However, expression of an inverted-repeat double-stranded RNA corresponding to the soybean FAD2-1A desaturase intron is sufficient to silence FAD2-1, implicating nuclear precursor mRNA (pre-mRNA) rather than cytosolic mRNA as the target of PTGS. Silencing FAD2-1 using intronic or 3′-UTR sequences does not affect transcription rates of the target genes but results in the strong reduction of target transcript levels in the nucleus. Moreover, siRNAs corresponding to pre-mRNA–specific sequences accumulate in the nucleus. In Arabidopsis, we find that two enzymes involved in PTGS, Dicer-like 4 and RNA-dependent RNA polymerase 6, are localized in the nucleus. Collectively, these results demonstrate that siRNA-directed RNA degradation can take place in the nucleus, suggesting the need for a more complex view of the subcellular compartmentation of PTGS in plants. PMID:21173264


    Directory of Open Access Journals (Sweden)

    Ruslan Wirosoedarmo


    Full Text Available Land suitability assessment was needed to plan productive and sustainable land use. The aim of this research was to de- liver an informative data about land suitability of corn using spatial analysis model. The research was conducted from July 2007 to January 2008, located in Blitar district. The land survey and analysis covered several parameters needed for suitability assessment for corn such as temperature, rainfall, soil drainage, soil texture, pH, effective depth of soil, organic- C, soil content, Cation Exchange Capacity of clay, slope, altitude, and erosion endangered. The land suitability classes were classified into four classes, those were very suitable, suitable, marginally suitable, and not suitable. The data were finally analyzed using Map Calculator in Arc View GIS Software.The results showed that there were three classes of land suitability for corn i.e. very suitable, suitable and marginally suitable which accounted for 85 %, 10 %, and 5 % of 150.96 hectare areas in Blitar. The suitable class mainly located in the northern of Blitar district, while the marginally suitable class mostly located in more than 1200 meters height above sea level covered 10, 117, and 52 hectares area of Wlingi, Gandusari, and Doko county area respectively. ABSTRAK Evaluasi kesesuaian lahan diperlukan untuk perencanaan penggunaan lahan yang produktif dan lestari. Tujuan peneli- tian adalah untuk menyajikan data dan informasi tentang evaluasi kesesuaian lahan bagi tanaman jagung menggunakan model analisa spasial. Penelitian dilaksanakan mulai bulan Juli 2007 sampai bulan Januari 2008. Lokasi penelitian dilaksanakan di Blitar, Jawa Timur. Data yang diperlukan meliputi data spasial berupa peta kontur dan peta jenis tanah skala 1:25000 serta data atribut berupa data klimatologi wilayah dan hasil pengamatan lapangan terhadap sifat fisik, morfologi dan kimia tanah. Parameter kesesuaian untuk tanaman jagung yang ditetapkan meliputi temperatur, curah hujan

  19. MGMT gene silencing and benefit from temozolomide in glioblastoma

    NARCIS (Netherlands)

    Hegi, ME; Diserens, A; Gorlia, T; Hamou, M; de Tribolet, N; Weller, M; Kros, JM; Hainfellner, JA; Mason, W; Mariani, L; Bromberg, JEC; Hau, P; Mirimanoff, RO; Cairncross, JG; Janzer, RC; Stupp, R


    BACKGROUND: Epigenetic silencing of the MGMT (O(sup 6)-methylguanine-DNA methyltransferase) DNA-repair gene by promoter methylation compromises DNA repair and has been associated with longer survival in patients with glioblastoma who receive alkylating agents. METHODS: We tested the relationship

  20. 5-Azacytidine mediated reactivation of silenced transgenes in potato (Solanum tuberosum) at the whole plant level. (United States)

    Tyč, Dimitrij; Nocarová, Eva; Sikorová, Lenka; Fischer, Lukáš


    Transient 5-azacytidine treatment of leaf explants from potato plants with transcriptionally silenced transgenes allows de novo regeneration of plants with restored transgene expression at the whole plant level. Transgenes introduced into plant genomes frequently become silenced either at the transcriptional or the posttranscriptional level. Transcriptional silencing is usually associated with DNA methylation in the promoter region. Treatments with inhibitors of maintenance DNA methylation were previously shown to allow reactivation of transcriptionally silenced transgenes in single cells or tissues, but not at the whole plant level. Here we analyzed the effect of DNA methylation inhibitor 5-azacytidine (AzaC) on the expression of two silenced reporter genes encoding green fluorescent protein (GFP) and neomycin phosphotransferase (NPTII) in potato plants. Whereas no obvious reactivation was observed in AzaC-treated stem cuttings, transient treatment of leaf segments with 10 μM AzaC and subsequent de novo regeneration of shoots on the selective medium with kanamycin resulted in the production of whole plants with clearly reactivated expression of previously silenced transgenes. Reactivation of nptII expression was accompanied by a decrease in cytosine methylation in the promoter region of the gene. Using the plants with reactivated GFP expression, we found that re-silencing of this transgene can be accidentally triggered by de novo regeneration. Thus, testing the incidence of transgene silencing during de novo regeneration could be a suitable procedure for negative selection of transgenic lines (insertion events) which have an inclination to be silenced. Based on our analysis of non-specific inhibitory effects of AzaC on growth of potato shoots in vitro, we estimated that AzaC half-life in the culture media is approximately 2 days.

  1. [E. M. Jellinek's silenced and silencing transgenerational story]. (United States)

    Kelemen, Gábor; Márk, Mónika


    Jellinek is a kind of archetypal character for future generations in the field of addiction studies. His implosion in the arena of alcoholism around the age of 50 was an unexpected challenge to medical science. We know very little about his own role models giving an intellectual and moral compass to his pragmatic creativity. More than 30 years has passed since Jellinek's death when an American sociologist Ron Roizen started unearthing his silent story. Roizen discerned that there are a lot of unsaid and muted issues in his personal Hungarian past. Our paper, based on the authors' research in Hungarian archives and other sources reveals that not just Jellinek's personal but his transgenerational narrative has been not-yet-said. This silenced and silencing history appears an unfinished business of acculturation of the family, which started prior to four generations. Authors have been concluding that the issue of religious conversion is a critical point in the process of acculturation. They examine the counter move of loyalty to family values and driving force of assimilation making their story unspeakable.

  2. A simple and reliable anion-exchange resin method for sulfate extraction and purification suitable for multiple O- and S-isotope measurements. (United States)

    Le Gendre, Erwann; Martin, Erwan; Villemant, Benoit; Cartigny, Pierre; Assayag, Nelly


    The O- and S-isotope compositions of sulfates can be used as key tracers of the fate and sink of sulfate in both terrestrial and extra-terrestrial environments. However, their application remains limited in those geological systems where sulfate occurs in low concentrations. Here we present a simple and reliable method to extract, purify and concentrate sulfate from natural samples. The method allows us to take into account the separation of nitrate, which is known to be an issue in O-isotope analysis. The separation and concentration of sulfate from other anions in any aqueous solution are performed within a few hours via anion-exchange resin. The possible O- (δ 18 O and Δ 17 O) and S- (δ 34 S, Δ 33 S and Δ 36 S) isotope exchanges, fractionations and/or contaminations are for the first time monitored during the whole procedure using initial O- and S-mass-dependent and mass-independent sulfate solutions. After elution in HCl, pure sulfate is fully retrieved and precipitated into BaSO 4 , which is suitable for O- and S-isotopic measurements using established techniques. The analysis of retrieved barite presents no variation within 2σ uncertainties: ±0.5‰ and ±0.1‰ in O- (δ 18 O, Δ 17 O) and ±0.2‰, ±0.02‰ and ±0.09‰ in S- (δ 34 S, Δ 33 S and Δ 36 S) isotope ratios, respectively. This study shows that the resin method for sulfate extraction and purification, in addition to being cheap, simple and quick, is applicable for the measurements of all O- and S-isotopic ratios in sulfates (including the Δ 17 O, Δ 33 S and Δ 36 S values). Therefore, this method can be easily used for a high range of natural samples in which sulfate occurs in low concentration including aerosols, ice cores, sediments, volcanic deposits, (paleo)soils and rainwater, and thus it can be a key to our understanding of the sulfur cycle on Earth. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  3. Silence (United States)

    Cogswell, J.


    On the occasion of the International Year of Astronomy, I was commissioned to create a mural for the University of Michigan Department of Astronomy, responding to an array of scientific images based on astronomical research, with special focus on the work of University of Michigan astronomers carried out within the building. My paper illustrates the development of this and several subsequent projects, explaining the implications for my artistic practice of entering into this conversation with astronomers and their work.

  4. "The Silence Itself Is Enough of a Statement": The Day of Silence and LGBTQ Awareness Raising (United States)

    Woolley, Susan W.


    This ethnographic study of a high school gay-straight alliance club examines unintended consequences of silence during the Day of Silence, a day of action aimed at addressing anti-LGBTQ bias in schools. While this strategy calls for students to engage in intentional silences to raise awareness of anti-LGBTQ bias, it does not necessarily lead…

  5. Application of a GIS Multi-Criteria Decision Analysis for the Identification of Intrinsic Suitable Sites in Costa Rica for the Application of Managed Aquifer Recharge (MAR through Spreading Methods

    Directory of Open Access Journals (Sweden)

    José Pablo Bonilla Valverde


    Full Text Available Costa Rica’s annual mean precipitation is above 3300 mm, but this precipitation is not evenly distributed in time or space, producing clear differentiated wet and dry seasons in most of the country. Droughts are also common phenomena which greatly affect the availability of water resources. Managed aquifer recharge (MAR schemes are being taken into consideration to enhance the underground water storage capacity of the country. The present study constitutes the first assessment for the identification of suitable sites for the implementation of MAR technology spreading methods (SM in Costa Rica. The suitable sites are identified by means of a geographic information system multi-criteria decision analysis (GIS-MCDA based on four criteria: hydrogeological aptitude, terrain slope, top soil texture and drainage network density. Four steps are performed in order to identify these sites: problem definition, screening for suitable areas, suitability mapping, and sensitivity analysis. The suitability map was divided in two zones after the screening: suitable and unsuitable, the first zone was further divided in five classes according to the weighted linear combination (WLC ranking. The results indicate that 61% of the country is suitable for conducting SM. This map is a tool for future implementation of MAR techniques in the country.

  6. Optical silencing of C. elegans cells with arch proton pump.

    Directory of Open Access Journals (Sweden)

    Ayako Okazaki

    Full Text Available BACKGROUND: Optogenetic techniques using light-driven ion channels or ion pumps for controlling excitable cells have greatly facilitated the investigation of nervous systems in vivo. A model organism, C. elegans, with its small transparent body and well-characterized neural circuits, is especially suitable for optogenetic analyses. METHODOLOGY/PRINCIPAL FINDINGS: We describe the application of archaerhodopsin-3 (Arch, a recently reported optical neuronal silencer, to C. elegans. Arch::GFP expressed either in all neurons or body wall muscles of the entire body by means of transgenes were localized, at least partially, to the cell membrane without adverse effects, and caused locomotory paralysis of worms when illuminated by green light (550 nm. Pan-neuronal expression of Arch endowed worms with quick and sustained responsiveness to such light. Worms reliably responded to repeated periods of illumination and non-illumination, and remained paralyzed under continuous illumination for 30 seconds. Worms expressing Arch in different subsets of motor neurons exhibited distinct defects in the locomotory behavior under green light: selective silencing of A-type motor neurons affected backward movement while silencing of B-type motor neurons affected forward movement more severely. Our experiments using a heat-shock-mediated induction system also indicate that Arch becomes fully functional only 12 hours after induction and remains functional for more than 24 hour. CONCLUSIONS/SGNIFICANCE: Arch can be used for silencing neurons and muscles, and may be a useful alternative to currently widely used halorhodopsin (NpHR in optogenetic studies of C. elegans.

  7. Baseline Suitability Analysis (United States)


    recreation organizations that receive funding through profits earned by the organization). An applicant can be deemed suitable for federal...of previous bad conduct, such as driving while intoxicated, possessing or using marijuana , or experiencing indebtedness, do not automatically

  8. Epigenetic Silencing and Resistance to Imatinib Mesylate in CML

    National Research Council Canada - National Science Library

    Issa, Jean-Pierre


    ...). In this project, we are exploring the hypothesis that epigenetic silencing associated with promoter DNA methylation mediates resistance in selected cases, and that reversal of silencing by decitabine...

  9. Epigenetic Silencing and Resistance to Imatinib Mesylate in CML

    National Research Council Canada - National Science Library

    Issa, Jean-Pierre


    ...). In this project, we are exploring the hypothesis that epigenetic silencing associated with promoter DNA methylation mediates resistance in selected cases, and that reversal of silencing by decitabine...

  10. Epigenetic Silencing and Resistance to Imatinib Mesylate in CML

    National Research Council Canada - National Science Library

    Issa, Jean-Pierre


    ...). In this project we are exploring the hypothesis that epigenetic silencing associated with promoter DNA methylation mediates resistance in selected cases and that reversal of silencing by decitabine...

  11. The gifts of silence and solitude. (United States)

    Schmidt Bunkers, Sandra


    In this column the author describes the importance of finding silence and solitude amid the noise and technology present today in the teaching-learning academy. Three gifts of silence and solitude are identified: the gift of comforting aloneness, the gift of vision for new horizons, and the gift of a sense of freedom. A humanbecoming perspective is used to explore the implications of these gifts. This column introduces a column by Diana Vander Woude describing her teaching-learning experience in leadership focusing on silence and solitude.

  12. Bacterial Cellular Engineering by Genome Editing and Gene Silencing

    Directory of Open Access Journals (Sweden)

    Nobutaka Nakashima


    Full Text Available Genome editing is an important technology for bacterial cellular engineering, which is commonly conducted by homologous recombination-based procedures, including gene knockout (disruption, knock-in (insertion, and allelic exchange. In addition, some new recombination-independent approaches have emerged that utilize catalytic RNAs, artificial nucleases, nucleic acid analogs, and peptide nucleic acids. Apart from these methods, which directly modify the genomic structure, an alternative approach is to conditionally modify the gene expression profile at the posttranscriptional level without altering the genomes. This is performed by expressing antisense RNAs to knock down (silence target mRNAs in vivo. This review describes the features and recent advances on methods used in genomic engineering and silencing technologies that are advantageously used for bacterial cellular engineering.

  13. Long Double-Stranded Multiplex siRNAs for Dual Genes Silencing (United States)

    Peng, Wei; Chen, Jianxin; Qin, Yinchao; Yang, Zhenjun


    Simultaneous suppression of multiple oncogenes is an attractive strategy to treat cancers. Herein we present a series of long double-stranded multiplex small interfering RNAs (multi-siRNAs) that is suitable for dual genes silencing through a sequence-specific RNA interference process without inducing significant immune responses. A gap feature structurally designed in either of the nucleotide strands of the multi-siRNAs was proved to be essential toward silencing target genes and avoiding immune responses. Furthermore, the silencing effect of multi-siRNAs against SURVIVIN and BCL-2 genes was shown to be effective and resulted in up-regulation of caspase-3 related apoptosis and, in turn, inhibition of bladder cancer cell proliferation. Our observation suggested that the rationally designed multi-siRNAs would have great potential for therapeutic siRNA design. PMID:23656495

  14. Small RNA-Mediated Epigenetic Myostatin Silencing

    Directory of Open Access Journals (Sweden)

    Thomas C Roberts


    Full Text Available Myostatin (Mstn is a secreted growth factor that negatively regulates muscle mass and is therefore a potential pharmacological target for the treatment of muscle wasting disorders such as Duchenne muscular dystrophy. Here we describe a novel Mstn blockade approach in which small interfering RNAs (siRNAs complementary to a promoter-associated transcript induce transcriptional gene silencing (TGS in two differentiated mouse muscle cell lines. Silencing is sensitive to treatment with the histone deacetylase inhibitor trichostatin A, and the silent state chromatin mark H3K9me2 is enriched at the Mstn promoter following siRNA transfection, suggesting epigenetic remodeling underlies the silencing effect. These observations suggest that long-term epigenetic silencing may be feasible for Mstn and that TGS is a promising novel therapeutic strategy for the treatment of muscle wasting disorders.

  15. Virus-induced gene silencing and Agrobacterium tumefaciens-mediated transient expression in Nicotiana tabacum. (United States)

    Zhang, Zhao; Thomma, Bart P H J


    Virus-induced gene silencing (VIGS) is a rapid method for transient silencing of plant genes. In this chapter, we describe the methodology for Tobacco rattle virus (TRV)-based VIGS in Nicotiana tabacum. In combination with subsequent co-expression of the tomato immune receptor Ve1 and the corresponding Verticillium effector Ave1 through Agrobacterium tumefaciens-mediated transient transformation (agroinfiltration), we established a rapid system for assessing the requirement of candidate plant genes for Ve1-mediated immune signaling.

  16. Artificial MiRNA Knockdown of Platelet Glycoprotein lbα: A Tool for Platelet Gene Silencing.

    Directory of Open Access Journals (Sweden)

    Tim Thijs

    Full Text Available In recent years, candidate genes and proteins implicated in platelet function have been identified by various genomic approaches. To elucidate their exact role, we aimed to develop a method to apply miRNA interference in platelet progenitor cells by using GPIbα as a proof-of-concept target protein. After in silico and in vitro screening of siRNAs targeting GPIbα (siGPIBAs, we developed artificial miRNAs (miGPIBAs, which were tested in CHO cells stably expressing GPIb-IX complex and megakaryoblastic DAMI cells. Introduction of siGPIBAs in CHO GPIb-IX cells resulted in 44 to 75% and up to 80% knockdown of GPIbα expression using single or combined siRNAs, respectively. Conversion of siGPIBAs to miGPIBAs resulted in reduced silencing efficiency, which could however be circumvented by tandem integration of two hairpins targeting different regions of GPIBA mRNA where 72% GPIbα knockdown was achieved. CHO GPIb-IX cells transfected with the miGPIBA construct displayed a significant decrease in their ability to aggregate characterized by lower aggregate numbers and size compared to control CHO GPIb-IX cells. More importantly, we successfully silenced GPIbα in differentiating megakaryoblastic DAMI cells that exhibited morphological changes associated with actin organization. In conclusion, we here report the successful use of miRNA technology to silence a platelet protein in megakaryoblastic cells and demonstrate its usefulness in functional assays. Hence, we believe that artificial miRNAs are suitable tools to unravel the role of a protein of interest in stem cells, megakaryocytes and platelets, thereby expanding their application to novel fields of basic and translational research.

  17. Titration and hysteresis in epigenetic chromatin silencing (United States)

    Dayarian, Adel; Sengupta, Anirvan M.


    Epigenetic mechanisms of silencing via heritable chromatin modifications play a major role in gene regulation and cell fate specification. We consider a model of epigenetic chromatin silencing in budding yeast and study the bifurcation diagram and characterize the bistable and the monostable regimes. The main focus of this paper is to examine how the perturbations altering the activity of histone modifying enzymes affect the epigenetic states. We analyze the implications of having the total number of silencing proteins, given by the sum of proteins bound to the nucleosomes and the ones available in the ambient, to be constant. This constraint couples different regions of chromatin through the shared reservoir of ambient silencing proteins. We show that the response of the system to perturbations depends dramatically on the titration effect caused by the above constraint. In particular, for a certain range of overall abundance of silencing proteins, the hysteresis loop changes qualitatively with certain jump replaced by continuous merger of different states. In addition, we find a nonmonotonic dependence of gene expression on the rate of histone deacetylation activity of Sir2. We discuss how these qualitative predictions of our model could be compared with experimental studies of the yeast system under anti-silencing drugs.

  18. Titration and hysteresis in epigenetic chromatin silencing

    International Nuclear Information System (INIS)

    Dayarian, Adel; Sengupta, Anirvan M


    Epigenetic mechanisms of silencing via heritable chromatin modifications play a major role in gene regulation and cell fate specification. We consider a model of epigenetic chromatin silencing in budding yeast and study the bifurcation diagram and characterize the bistable and the monostable regimes. The main focus of this paper is to examine how the perturbations altering the activity of histone modifying enzymes affect the epigenetic states. We analyze the implications of having the total number of silencing proteins, given by the sum of proteins bound to the nucleosomes and the ones available in the ambient, to be constant. This constraint couples different regions of chromatin through the shared reservoir of ambient silencing proteins. We show that the response of the system to perturbations depends dramatically on the titration effect caused by the above constraint. In particular, for a certain range of overall abundance of silencing proteins, the hysteresis loop changes qualitatively with certain jump replaced by continuous merger of different states. In addition, we find a nonmonotonic dependence of gene expression on the rate of histone deacetylation activity of Sir2. We discuss how these qualitative predictions of our model could be compared with experimental studies of the yeast system under anti-silencing drugs. (paper)

  19. Potential of Metal Fibre Felts as Passive Absorbers in Absorption Silencers

    Directory of Open Access Journals (Sweden)

    Björn Hinze


    Full Text Available The growing noise exposure of residents, due to a rising number of flights, causes significant impacts on physical health. Therefore it is necessary to reduce the noise emission of aircrafts. During take-off, the noise generated by the jet engines is dominating. One way to lower the noise emission of jet engines is to build an absorption silencer by using porous liners. Because of the high thermic and corrosive attacks as well as high fatigue loads, conventional absorbers cannot be used. A promising material is sintered metal fibre felts. This study investigates the suitability of metal fibre felts for the use as absorption material in silencers. The influences of pore morphology, absorption coefficient, determined with perpendicular sound incidence, as well as geometric parameters of the silencer to the damping are identified. To characterise the material, the parameters fibre diameter, porosity and thickness are determined using three-dimensional computer tomography images. The damping potential of absorption silencers is measured using an impedance tube, which was modified for transmission measurements. The essential parameter to describe the acoustic characteristics of porous materials is the flow resistivity. It depends on the size, shape and number of open pores in the material. Finally a connection between pore morphology, flow resistivity of the metal fibre felts and damping potential of the absorption silencer is given.

  20. Virus-induced gene silencing in detached tomatoes and biochemical effects of phytoene desaturase gene silencing

    NARCIS (Netherlands)

    Romero, I.; Tikunov, Y.M.; Bovy, A.G.


    Virus-induced gene silencing (VIGS) is a technology that has rapidly emerged for gene function studies in plants. Many advances have been made in applying this technique in an increasing number of crops. Recently, VIGS has been successfully used to silence genes in tomato fruit through

  1. Surface functionalisation of PLGA nanoparticles for gene silencing

    DEFF Research Database (Denmark)

    Andersen, Morten Østergaard; Lichawska, Agata; Arpanaei, Ayyoob


    . In addition, particles containing cetylated-PEI achieved 64% silencing of TNFα in J774.1 cells. This rapid method for surface modification of PLGA nanoparticles promotes its application for alternative cetylated functional derivatives as a strategy to control specific biological properties of nanoparticles.......This work presents a method for decorating the surface of poly (lactide-co-glycolide) (PLGA) nanoparticles with polyethyleneimine (PEI) utilising a cetyl derivative to improve surface functionalisation and siRNA delivery. Sub-micron particles were produced by an emulsion-diffusion method using...

  2. Climate and suitable clothing. (United States)

    Tamas, R


    Climate may be described as refined weather The meteorologist notes the day-to-day variations of the weather components, extracts the principal frequencies which then describe the seasons, and once the average values have been derived over long periods of time, there emerges the concept of climate. Advances in preventive and curative medicine are such that man is left virtually with only his two oldest enemies to combat: his fellow men and the environment and he will be able to give better attention to the former if the latter is neutralised by the supply of suitable clothing designed to keep him comfortable.

  3. A novel multiplex RT-qPCR method based on dual-labelled probes suitable for typing all known genotypes of viral haemorrhagic septicaemia virus

    DEFF Research Database (Denmark)

    Vázquez, D.; López-Vázquez, C.; Skall, Helle Frank


    , resulting in a correct detection and typing of all strains. The analytical sensitivity was evaluated in a comparative assay with titration in cell culture, observing that both methods provided similar limits of detection. The proposed method can be a powerful tool for epidemiological analysis of VHSV...

  4. Spatiotemporal flicker detector model of motion silencing. (United States)

    Choi, Lark Kwon; Bovik, Alan C; Cormack, Lawrence K


    Motion can impair the perception of other visual changes. Suchow and Alvarez (2011a, Current Biology, 21, 140-143) recently demonstrated a striking 'motion silencing' illusion, in which the salient changes among a group of objects' luminances (or colors, etc) appear to cease in the presence of large, coherent object motion. To understand why the visual system might be insensitive to changes in object luminances ('flicker') in the presence of object motion, we constructed similar stimuli and did a systematic spectral analysis of them. We conducted human psychophysical experiments to examine motion silencing as a function of stimulus velocity, flicker frequency, and spacing; and we created a simple filter-based model as a working hypothesis of motion silencing. From the results, we found that the threshold of silencing occurs when the log frequency of object replacement is roughly one quarter of the log flicker frequency (the mean slope is approximately 0.27). The dependence of silencing on object spacing may be explained as a phenomenon of temporal sampling of the stimuli by the visual system. Our proposed model successfully captures the psychophysical data over a wide range of velocities and flicker frequencies.

  5. Stress Degradation Behavior of Atorvastatin Calcium and Development of a Suitable Stability-Indicating LC Method for the Determination of Atorvastatin, its Related Impurities, and its Degradation Products. (United States)

    Vukkum, Pallavi; Moses Babu, J; Muralikrishna, R


    A rapid, reversed-phase liquid chromatographic method was developed for the quantitative determination of Atorvastatin calcium, its related substances (12 impurities), and degradation impurities in bulk drugs. The chromatographic separation was achieved on a Zorbax Bonus-RP column by employing a gradient elution with water-acetonitrile-trifluoroacetic acid as the mobile phase in a shorter run time of 25 min. The flow rate was 1.0 mL/min and the detection wavelength was 245 nm. The drug substance was subjected to stress studies such as hydrolysis, oxidation, photolysis, and thermal degradation, and considerable degradation was observed in acidic hydrolysis, oxidative, thermal, and photolytic stress conditions. The formed degradation products were reported and were well-resolved from the Atorvastatin and its related substances. The stressed samples were quantified against a qualified reference standard and the mass balance was found to be close to 99.5% (w/w) when the response of the degradant was considered to be equal to the analyte (i.e. Atorvastatin), which demonstrates the stability-indicating capability of the method. The method was validated in agreement with ICH requirements. The method developed here was single and shorter (25 min method for the determination of all 12 related impurities of Atorvastatin and its degradation products), with clearly better resolution and higher sensitivity than the European (85 min method for the determination of six impurities) and United States pharmacopeia (115 min and 55 min, two different methods for the determination of six related substances).

  6. Organizational Silence, from Roots to Solutions: A Case Study in Iran Petroleum Industry

    Directory of Open Access Journals (Sweden)

    Mehdi Afkhami Ardakani


    Full Text Available Organizational silence is defined as the lack of effective interactions among staff and it stands opposite to the concept of organizational voice. In the present research, the purpose is to measure the silence behavior among the Research Institute of Petroleum Industry (RIPI staff before and after the implementation of a comprehensive suggestion system. A suggestion system is an internal structure easily accessed by all the staff to state their suggestions in a pre-structured format. The roots of silence behavior are studied based on a deep literature review to find out possible solutions to improve organizational voice. To conduct the research, a self-structured questionnaire has been developed and distributed among all the staff. A quasi-experimental methodology has been adopted to compare pretest and post-test results of silence status before and after implementing the suggestion system. The results show that the silence behavior has been meaningfully reduced. This is based on a simple t-test performed by SPSS software, where there is a meaningful difference between the silence status of pre-test and post-test. In other words, a suggestion system could be a communication opportunity to encourage staff to provide suggestions and to cooperate for promoting the organization, which will finally reduce the organization silence. A major gap within the studies of Iranian scholars about organizational silence is the failure to introduce effective solutions to reduce it. However, this research is innovative in the sense that it fills the mentioned gap. This research shows that large scale organizations like RIPI need to consider methods like suggestion systems to break bureaucratic obstacles so that their staff can easily find open routes to share their ideas and suggestions in a prestructured format. This cooperating will lead to mutual benefits for both parts, since suggestions could be used to enhance organizational structure and performance and

  7. An Investigation of Acoustic Attenuation Performance of Silencers with Mean Flow Based on Three-Dimensional Numerical Simulation

    Directory of Open Access Journals (Sweden)

    Wei Fan


    Full Text Available Transmission loss (TL is often used to evaluate the acoustic attenuation performance of a silencer. In this work, a three-dimensional (3D finite element method (FEM is employed to calculate the TL of some representative silencers, namely, circular expansion chamber silencer and straight-through perforated pipe silencer. In order to account for the effect of mean flow that exists inside the silencer, the 3D FEM is used in conjunction with the Computational Fluid Dynamics (CFD simulation of the flow field. More concretely, the 3D mean flow field is computed by firstly using CFD, and then the obtained mean flow data are imported to an acoustic solution undertaken using FEM. The data transfer between the two steps is accomplished by mesh mapping. The results presented demonstrate good agreement between present TL predictions and previously published experimental and numerical works. Also, the details of the flow inside the silencers may be studied. Furthermore, the effect of mean flow velocity on acoustic attenuation performance of the silencers is investigated. It is concluded that for the studied silencers, in general, increasing flow velocity increases the TL and decreases the resonance peaks.

  8. Silence as a Response to Everyday Violence

    DEFF Research Database (Denmark)

    Gammeltoft, Tine


    Across the world, existing research indicates that many women respond with silence to marital abuse. This article offers an ethnographic investigation of the social and psychic forces behind Vietnamese women’s silencing of violence and a theoretical exploration of how the psychoanalytic concept...... of fantasy—understood as unconscious or subconscious mental processes—may contribute to the analysis of everyday violence and psychic distress. Distinguishing between what I term deliberate and subconscious silence, I explore the role that fantasy plays when Vietnamese women silently endure intimate partner...... violence. Closer ethnographic attention to the fantasy-constructions that sustain day-to-day lives can, I argue, strengthen the capacity of anthropology to comprehend how systems of everyday violence are upheld and rendered socially invisible....

  9. Granulation of core particles suitable for film coating by agitation fluidized bed I. Optimum formulation for core particles and development of a novel friability test method. (United States)

    Hamashita, Tomohiro; Nakagawa, Yasuo; Aketo, Takao; Watano, Satoru


    To prepare powdered medicines without bitter taste, film coating is required to cover the surface of core particles. In this study, effect of formulation and operating conditions of agitation fluidized bed on the core particle properties was investigated. In order to prevent breakage of the core particles during coating process, which sometimes causes variation of drug dissolution rate, addition of maltose syrup powder during the formulation process of the core particles was investigated. Also, a method for friability test in which the core particles were subjected to strong impact was proposed to evaluate strength of the core particles. The friability of the core particles determined by this test method correlated well with the actual friability of the particles during the coating process. Based on this result, we confirmed this novel friability test method could predict the core particle endurance during the coating process.

  10. Influence of the Determination Methods of K and C Parameters on the Ability of Weibull Distribution to Suitably Estimate Wind Potential and Electric Energy

    Directory of Open Access Journals (Sweden)

    Ruben M. Mouangue


    Full Text Available The modeling of the wind speed distribution is of great importance for the assessment of wind energy potential and the performance of wind energy conversion system. In this paper, the choice of two determination methods of Weibull parameters shows theirs influences on the Weibull distribution performances. Because of important calm winds on the site of Ngaoundere airport, we characterize the wind potential using the approach of Weibull distribution with parameters which are determined by the modified maximum likelihood method. This approach is compared to the Weibull distribution with parameters which are determined by the maximum likelihood method and the hybrid distribution which is recommended for wind potential assessment of sites having nonzero probability of calm. Using data provided by the ASECNA Weather Service (Agency for the Safety of Air Navigation in Africa and Madagascar, we evaluate the goodness of fit of the various fitted distributions to the wind speed data using the Q – Q plots, the Pearson’s coefficient of correlation, the mean wind speed, the mean square error, the energy density and its relative error. It appears from the results that the accuracy of the Weibull distribution with parameters which are determined by the modified maximum likelihood method is higher than others. Then, this approach is used to estimate the monthly and annual energy productions of the site of the Ngaoundere airport. The most energy contribution is made in March with 255.7 MWh. It also appears from the results that a wind turbine generator installed on this particular site could not work for at least a half of the time because of higher frequency of calm. For this kind of sites, the modified maximum likelihood method proposed by Seguro and Lambert in 2000 is one of the best methods which can be used to determinate the Weibull parameters.

  11. A new method suitable for calculating accurately wetting temperature over a wide range of conditions: Based on the adaptation of continuation algorithm to classical DFT (United States)

    Zhou, Shiqi


    A new scheme is put forward to determine the wetting temperature (Tw) by utilizing the adaptation of arc-length continuation algorithm to classical density functional theory (DFT) used originally by Frink and Salinger, and its advantages are summarized into four points: (i) the new scheme is applicable whether the wetting occurs near a planar or a non-planar surface, whereas a zero contact angle method is considered only applicable to a perfectly flat solid surface, as demonstrated previously and in this work, and essentially not fit for non-planar surface. (ii) The new scheme is devoid of an uncertainty, which plagues a pre-wetting extrapolation method and originates from an unattainability of the infinitely thick film in the theoretical calculation. (iii) The new scheme can be similarly and easily applied to extreme instances characterized by lower temperatures and/or higher surface attraction force field, which, however, can not be dealt with by the pre-wetting extrapolation method because of the pre-wetting transition being mixed with many layering transitions and the difficulty in differentiating varieties of the surface phase transitions. (iv) The new scheme still works in instance wherein the wetting transition occurs close to the bulk critical temperature; however, this case completely can not be managed by the pre-wetting extrapolation method because near the bulk critical temperature the pre-wetting region is extremely narrow, and no enough pre-wetting data are available for use of the extrapolation procedure.

  12. Active compressor engine silencer reduces exhaust noise

    International Nuclear Information System (INIS)

    Denenberg, J.N.; Miller, S.K.; Jay, M.A.


    An active industrial silencer on a compressor engine at a Tenneco Gas station has reduced low-frequency 'rumbling' noise by 8 dB during trials while lowering backpressure about 90$. This 8 dB reduction of the piston firing frequency corresponds to a more than 80% decrease in emitted acoustic power. The silencing unit, installed on one of six engines at the station near Eden, N.Y., continues in operation. Based on the results, the manufacturer is identifying additional compressor sites for further tests. This paper reviews this project

  13. On the acoustic performance of rectangular splitter silencers in the presence of mean flow (United States)

    Kirby, Ray; Amott, Ken; Williams, Paul T.; Duan, Wenbo


    Dissipative splitter silencers are often used to reduce the noise emitted in ventilation and gas turbine systems. It is well known that the acoustic performance of a splitter silencer changes under the influence of the convective effects of a mean gas flow and so in this article a theoretical model is developed to include the effects of mean flow. The theoretical model is based on a hybrid finite element method which enables the inclusion of bull nose fairings and a perforated screen separating the mean gas flow from a bulk reacting porous material. Predictions are compared against experimental measurements obtained both with and without mean flow. Good agreement between prediction and measurement is generally observed in the absence of mean flow, although it is seen that for silencers with a low percentage open area the silencer insertion loss is over predicted at higher frequencies. When mean flow is present, problems with the experimental methodology are observed at relatively modest mean flow velocities, and so comparison between prediction and experiment is limited to relatively low face velocities. However, experiment and theory both show that the insertion loss reduces at low frequencies when mean flow is in the direction of sound propagation, and at high frequencies the influence of mean flow is generally much smaller. Following additional theoretical investigations it is concluded that the influence of mean flow on splitter silencer performance should be accounted for at low frequencies when silencer airway velocities are greater than about 20 m/s; however, at higher frequencies one may generally neglect the effect of mean flow, even at higher velocities. Predictions obtained using the hybrid method are also compared to a simplified point collocation approach and it is demonstrated that the computationally efficient point collocation method may be used to investigate the effects of mean flow in a splitter silencer without loss of accuracy.

  14. “The Silence Itself is Enough of a Statement”: Unintended Consequences of Silence as an Awareness-Raising Strategy


    Woolley, Susan W.


    The strategy of silence for voice, as seen in the Day of Silence, deploys silence in order to draw attention to the ways in which an individual or group has been silenced and to establish possibilities for voice. The Day of Silence is a nation-wide day of action aimed at addressing anti-LGBTQ bias and harassment in schools. This ethnographic study of a high school gay-straight alliance (GSA) club examines the unintended consequences of silence as an awareness-raising strategy during events ...

  15. An easy, rapid, and cost-effective method for DNA extraction from various lichen taxa and specimens suitable for analysis of fungal and algal strains. (United States)

    Park, Sook-Young; Jang, Seol-Hwa; Oh, Soon-Ok; Kim, Jung A; Hur, Jae-Seoun


    Lichen studies, including biodiversity, phylogenetic relationships, and conservation concerns require definitive species identification, however many lichens can be challenging to identify at the species level. Molecular techniques have shown efficacy in discriminating among lichen taxa, however, obtaining genomic DNA from herbarium and fresh lichen thalli by conventional methods has been difficult, because lichens contain high proteins, polysaccharides, and other complex compounds in their cell walls. Here we report a rapid, easy, and inexpensive protocol for extracting PCR-quality DNA from various lichen species. This method involves the following two steps: first, cell breakage using a beadbeater; and second, extraction, isolation, and precipitation of genomic DNA. The procedure requires approximately 10 mg of lichen thalli and can be completed within 20 min. The obtained DNAs were of sufficient quality and quantity to amplify the internal transcribed spacer region from the fungal and algal lichen components, as well as to sequence the amplified products. In addition, 26 different lichen taxa were tested, resulting in successful PCR products. The results of this study validated the experimental protocols, and clearly demonstrated the efficacy and value of our KCl extraction method applied in the fungal and algal samples.

  16. Phenotyping of VIGS-mediated gene silencing in rice using a vector derived from a DNA virus. (United States)

    Kant, Ravi; Dasgupta, Indranil


    Target genes in rice can be optimally silenced if inserted in antisense or hairpin orientation in the RTBV-derived VIGS vector and plants grown at 28 °C and 80% humidity after inoculation. Virus induced gene silencing (VIGS) is a method used to transiently silence genes in dicot as well as monocot plants. For the important monocot species rice, the Rice tungro bacilliform virus (RTBV)-derived VIGS system (RTBV-VIGS), which uses agroinoculation to initiate silencing, has not been standardized for optimal use. Here, using RTBV-VIGS, three sets of conditions were tested to achieve optimal silencing of the rice marker gene phytoene desaturase (pds). The effect of orientation of the insert in the RTBV-VIGS plasmid (sense, antisense and hairpin) on the silencing of the target gene was then evaluated using rice magnesium chelatase subunit H (chlH). Finally, the rice Xa21 gene, conferring resistance against bacterial leaf blight disease (BLB) was silenced using RTBV-VIGS system. In each case, real-time PCR-based assessment indicated approximately 40-80% fall in the accumulation levels of the transcripts of pds, chlH and Xa21. In the case of pds, the appearance of white streaks in the emerging leaves, and for chlH, chlorophyll levels and F v /F m ratio were assessed as phenotypes for silencing. For Xa21, the resistance levels to BLB were assessed by measuring the lesion length and the percent diseased areas of leaves, following challenge inoculation with Xanthomonas oryzae. In each case, the RTBV-MVIGS system gave rise to a discernible phenotype indicating the silencing of the respective target gene using condition III (temperature 28 °C, humidity 80% and 1 mM MES and 20 µM acetosyringone in secondary agrobacterium culture), which revealed the robustness of this gene silencing system for rice.

  17. Suitability of quality control materials for prostate-specific antigen (PSA) measurement: inter-method variability of common tumor marker control materials. (United States)

    Vucetic, Zivjena; Dnistrian, Ann; Nilsson, Olle; Lilja, Hans G; Plebani, Mario


    Quality control materials with minimal inter-assay differences and clinically relevant proportions of different molecular forms of the analyte are needed to optimize intra- and inter-laboratory accuracy and precision. We assessed if clinically relevant total prostate-specific antigen (tPSA) levels were present in seven commercially available Multi Constituent Tumor Marker Controls (MC-TMC). Further, we determined the concentration of free PSA (fPSA) and calculated the percentage of free PSA (%fPSA) in all materials. Finally, we determined variability of TMC materials across several commonly used PSA platforms. All MC-TMC materials contained at least one concentration of tPSA in normal and pathologic range. Control materials varied in the amount of fPSA and %fPSA, with most controls consisting of fPSA only and only one MC-TMC containing medically relevant levels of around 35% fPSA. Only a minority of MC-TMC materials showed minimal variability across four PSA methods while the majority of PSA controls showed wide inter-method differences. Use of many commercially available controls for PSA could lead to biased PSA measurements because they contain medically irrelevant proportions of fPSA and show significant variation among different PSA assay platforms.

  18. Surprised by Bird, Bard, and Bach: Language, Silence, and Transcendence. (United States)

    Suhor, Charles


    Argues the importance of the relationships among silence and literature, the arts, and other experiences that point toward transcendence. Suggests that English teachers can expand the repertoire of classroom activities and teaching techniques that make use of silence. (KEH)

  19. Histone Methylation and Epigenetic Silencing in Breast Cancer

    National Research Council Canada - National Science Library

    Simon, Jeffrey A; Lange, Carol A


    .... EZH2 is a histone methyltransferase which modifies lysine-27 of histone H3 an epigenetic mark which is generally linked to gene silencing and is implicated in tumor suppressor silencing during breast cancer progression...

  20. A marker based kinematic method of identifying initial contact during gait suitable for use in real-time visual feedback applications. (United States)

    De Asha, A R; Robinson, M A; Barton, G J


    A gait cycle is typically defined as being from heel strike or initial contact (IC) to the next ipsilateral IC using kinetic data. When these data are not available other methods of event definition are required. An algorithm based upon sagittal plane kinematics of the hip, which defines IC at contralateral peak hip extension (PHE) is presented. Kinematic and kinetic data were recorded while 10 unimpaired participants each completed a minimum of 25 overground gait cycles. The accuracy of 551 IC events was evaluated by comparing the agreement of PHE to other kinematic and kinetic algorithms. The mean temporal difference in IC between the PHE algorithm and a kinetic algorithm was +0.0006±0.008 s. The 95% Limits of Agreement was ±0.018 s. This new PHE algorithm provides simple to implement and accurate gait events for use when kinetic data are not available. Copyright © 2012 Elsevier B.V. All rights reserved.

  1. Development and validation of an X-ray stress measurement device using an image plate suitable for the cosα method

    International Nuclear Information System (INIS)

    Maruyama, Yoichi; Sasaki, Toshihiko; Miyazaki, Toshiyuki


    We have developed a novel residual stress measurement instrument based on the X-ray diffraction technique. An image plate was used as the detector to sample the full diffraction ring. The image plate and the scanning unit were built into the one system. Compared with the traditional X-ray stress measurement systems, the physical size and weight of the instrument were significantly reduced. The instrument also has the capability to measure the distance from the specimen to the image plate. This capability contributes to improve the accuracy and simplicity of the stress measurement. The developed instrument was focused on measuring a ferrite sample and the acquired diffraction ring was analyzed using the cosα method. In the present work, we evaluated the accuracy of the measurement data from the instrument using a ferrite powder compared with strain gauge results obtained from four-point bending measurements. (author)

  2. Suitability of near-infrared methods for the determination of moisture in a freeze-dried injection product containing different amounts of the active ingredient. (United States)

    Last, I R; Prebble, K A


    A near-infrared reflectance (NIR) method for determination of moisture in an experimental freeze-dried injection product was developed and validated. NIR spectra were collected through the bases of unopened product vials using a horizontal instrument accessory, before generating primary reference data on the same individual vials by Karl-Fischer titration. Data were collected for product containing different concentrations of the active ingredient in the same matrix. NIR calibrations were developed with second derivative spectral data using regression facilities within the NIR software, and validated using independent test sets. An assessment is given of the applicability of moisture calibrations developed on product at one active ingredient level to the prediction of moisture contents in the product containing a different concentration of active ingredient.

  3. The impact of organizational culture on employees’ organizational silence In Shiraz University of Medical Sciences

    Directory of Open Access Journals (Sweden)

    Ebrahim Parcham


    Full Text Available Introduction: Organizational Culture is one of the most important factors that can change the climate of silence. The main aim of this research was to investigate the influence of organizational culture on employees’ organizational silence in Shiraz University of Medical Sciences. Method: This research was a descriptive-correlation one. The target population was chosen from 1900 staff of the University of Medical sciences and Health Care headquarter in Shiraz. Thus 311 employees were selected using the Krejcie and Morgan sampling table. The instrument used in this research was Denison (2006 organizational culture questionnaire and Dimitris Buratas and Maria Vacula (2007 organizational culture. Cornbrash’s alpha method was used to calculate the reliability. The Item analysis and expert consensus were applied to calculate the validity of instruments. All gathered data analyzed with PLS software. Results: The results showed that the four dimensions of organizational culture include organizational involvement, organizational adaptability, organizational concistency and organizational mission was moderate and the mean scores obtained for each factor were 2.85, 2.82, 2.94 and 2.93 respectively. Structural equation model showed Organizational culture has a significant positive impact on organizational silence (β=0.68; P<.001. Conclusion: Based on the results and impact of organizational culture on organizational silence that is positive and significant; The organization further efforts to strengthen various aspects of organizational culture, especially the employees’ involvement in decision making; Employees can better express their opinions and thus reduced their organizational silence. In other words strengthening corporate culture is combined with the reduction of organizational silence. Medical organizations can establish appropriate reward system for creative ideas and suggestions to encourage people express their ideas As a result, reduced

  4. Silence in the second language classroom

    CERN Document Server

    King, J


    Why are second language learners in Japan's universities so silent? This book investigates the perplexing but intriguing phenomenon of classroom silence and draws on ideas from psychology, sociolinguistics and anthropology to offer a unique insight into the reasons why some learners are either unable or unwilling to speak in a foreign language.

  5. Mainstream Television, Adolescent Homosexuality, and Significant Silence. (United States)

    Kielwasser, Alfred P.; Wolf, Michelle A.


    Argues that the symbolic annihilation of gay and lesbian youth exhibited by network television contributes to a dysfunctional isolation supported by the mutually reinforcing invisibility of homosexual adolescents on television and in the real world. Suggests that the spiral of silence also partially accounts for the inefficacy of oppositional…

  6. From talking to silence: a confessional journey


    Heddon, D.; Howells, A.


    This article provides an account of work created by Adrian Howells over the duration of his AHRC Creative Fellowship. Howells provides a creator's perspective; Heddon, a spectator's perspective. Both chart the shift from confessional performances anchored around speech, to the use of silence as a way to engage intimate connections between performer and audience.

  7. Development of a driving method suitable for ultrahigh-speed shooting in a 2M-fps 300k-pixel single-chip color camera (United States)

    Yonai, J.; Arai, T.; Hayashida, T.; Ohtake, H.; Namiki, J.; Yoshida, T.; Etoh, T. Goji


    We have developed an ultrahigh-speed CCD camera that can capture instantaneous phenomena not visible to the human eye and impossible to capture with a regular video camera. The ultrahigh-speed CCD was specially constructed so that the CCD memory between the photodiode and the vertical transfer path of each pixel can store 144 frames each. For every one-frame shot, the electric charges generated from the photodiodes are transferred in one step to the memory of all the parallel pixels, making ultrahigh-speed shooting possible. Earlier, we experimentally manufactured a 1M-fps ultrahigh-speed camera and tested it for broadcasting applications. Through those tests, we learned that there are cases that require shooting speeds (frame rate) of more than 1M fps; hence we aimed to develop a new ultrahigh-speed camera that will enable much faster shooting speeds than what is currently possible. Since shooting at speeds of more than 200,000 fps results in decreased image quality and abrupt heating of the image sensor and drive circuit board, faster speeds cannot be achieved merely by increasing the drive frequency. We therefore had to improve the image sensor wiring layout and the driving method to develop a new 2M-fps, 300k-pixel ultrahigh-speed single-chip color camera for broadcasting purposes.

  8. Virus-induced gene silencing (VIGS) in Nicotiana benthamiana and tomato. (United States)

    Velásquez, André C; Chakravarthy, Suma; Martin, Gregory B


    RNA interference (RNAi) is a highly specific gene-silencing phenomenon triggered by dsRNA. This silencing mechanism uses two major classes of RNA regulators: microRNAs, which are produced from non-protein coding genes and short interfering RNAs (siRNAs). Plants use RNAi to control transposons and to exert tight control over developmental processes such as flower organ formation and leaf development. Plants also use RNAi to defend themselves against infection by viruses. Consequently, many viruses have evolved suppressors of gene silencing to allow their successful colonization of their host. Virus-induced gene silencing (VIGS) is a method that takes advantage of the plant RNAi-mediated antiviral defense mechanism. In plants infected with unmodified viruses the mechanism is specifically targeted against the viral genome. However, with virus vectors carrying sequences derived from host genes, the process can be additionally targeted against the corresponding host mRNAs. VIGS has been adapted for high-throughput functional genomics in plants by using the plant pathogen Agrobacterium tumefaciens to deliver, via its Ti plasmid, a recombinant virus carrying the entire or part of the gene sequence targeted for silencing. Systemic virus spread and the endogenous plant RNAi machinery take care of the rest. dsRNAs corresponding to the target gene are produced and then cleaved by the ribonuclease Dicer into siRNAs of 21 to 24 nucleotides in length. These siRNAs ultimately guide the RNA-induced silencing complex (RISC) to degrade the target transcript. Different vectors have been employed in VIGS and one of the most frequently used is based on tobacco rattle virus (TRV). TRV is a bipartite virus and, as such, two different A. tumefaciens strains are used for VIGS. One carries pTRV1, which encodes the replication and movement viral functions while the other, pTRV2, harbors the coat protein and the sequence used for VIGS. Inoculation of Nicotiana benthamiana and tomato seedlings

  9. Suitability of imaging methods (X-ray, CT, MRI) in the diagnostics of Ewing’s sarcoma in children – analysis of own material (United States)

    Kuleta-Bosak, Elżbieta; Kluczewska, Ewa; Machnik-Broncel, Joanna; Madziara, Wojciech; Ciupińska-Kajor, Monika; Sojka, Dorota; Rogala, Wojciech; Juszczyk, Jan; Wilk, Robert


    Summary Background: Ewing sarcoma is a malignant, small round cell bone tumor, presenting predominantly in children and adolescents. Ewing sarcoma may develop in every bone; diaphyses of long bones, ribs and flat bones are the main locations. Local and systemic clinical symptoms are nonspecific - pain, swelling, fever or ill-being. The aim of the study was to assess the role of radiography, computed tomography and magnetic resonance imaging in the analysis of bone lesions in children and young adults with Ewing sarcoma. Material/Methods: Twenty-seven patients, aged between 1 year and 10 months, and 17 years and 2 months, with histologically verified Ewing sarcoma of the bone, referred to the Radiological Department of University Hospital No 6., John Paul II Upper Silesian Centre for Child Health Katowice, in the period from 1996 to 2007, were included in the study.Plain radiography was performed in every child, CT in 20 and MRI in 12 individuals. Tumour location, extension of the tumour, soft tissue mass, and periosteal reaction were taken into consideration in the evaluation of the lesion. In some cases, pathological features of the MRI and CT were compared. The prevalence of some radiological features was compared to the literature data. Results: The most common site of tumor was: ribs (6 children), femoral bone (6 children), pelvis (4 children) and tibia (3 children). In 2 children, a primary tumor was diagnosed in the spine (multifocal in 1 child). X-rays revealed: periosteal reaction in 17 children (63%), soft tissue involvement in 19 children (70%), permeative component in 16 children (59%), and sclerotic component in 5 children (19%). In 10 children (37%), periosteal reaction was not detected. The examination revealed: soft tissue calcifications in 7 cases (26%), a well-delineated focus of destruction within bones in 3 children (11%), cortical thickening in 4 children (15%), cortical destruction in 4 children (15%), saucerisation in 3 children (11%), bone

  10. An Improved Brome mosaic virus Silencing Vector: Greater Insert Stability and More Extensive VIGS1[OPEN (United States)


    Virus-induced gene silencing (VIGS) is used extensively for gene function studies in plants. VIGS is inexpensive and rapid compared with silencing conducted through stable transformation, but many virus-silencing vectors, especially in grasses, induce only transient silencing phenotypes. A major reason for transient phenotypes is the instability of the foreign gene fragment (insert) in the vector during VIGS. Here, we report the development of a Brome mosaic virus (BMV)-based vector that better maintains inserts through modification of the original BMV vector RNA sequence. Modification of the BMV RNA3 sequence yielded a vector, BMVCP5, that better maintained phytoene desaturase and heat shock protein70-1 (HSP70-1) inserts in Nicotiana benthamiana and maize (Zea mays). Longer maintenance of inserts was correlated with greater target gene silencing and more extensive visible silencing phenotypes displaying greater tissue penetration and involving more leaves. The modified vector accumulated similarly to the original vector in N. benthamiana after agroinfiltration, thus maintaining a high titer of virus in this intermediate host used to produce virus inoculum for grass hosts. For HSP70, silencing one family member led to a large increase in the expression of another family member, an increase likely related to the target gene knockdown and not a general effect of virus infection. The cause of the increased insert stability in the modified vector is discussed in relationship to its recombination and accumulation potential. The modified vector will improve functional genomic studies in grasses, and the conceptual methods used to improve the vector may be applied to other VIGS vectors. PMID:29127260

  11. An ImprovedBrome mosaic virusSilencing Vector: Greater Insert Stability and More Extensive VIGS. (United States)

    Ding, Xin Shun; Mannas, Stephen W; Bishop, Bethany A; Rao, Xiaolan; Lecoultre, Mitchell; Kwon, Soonil; Nelson, Richard S


    Virus-induced gene silencing (VIGS) is used extensively for gene function studies in plants. VIGS is inexpensive and rapid compared with silencing conducted through stable transformation, but many virus-silencing vectors, especially in grasses, induce only transient silencing phenotypes. A major reason for transient phenotypes is the instability of the foreign gene fragment (insert) in the vector during VIGS. Here, we report the development of a Brome mosaic virus (BMV)-based vector that better maintains inserts through modification of the original BMV vector RNA sequence. Modification of the BMV RNA3 sequence yielded a vector, BMVCP5, that better maintained phytoene desaturase and heat shock protein70-1 ( HSP70-1 ) inserts in Nicotiana benthamiana and maize ( Zea mays ). Longer maintenance of inserts was correlated with greater target gene silencing and more extensive visible silencing phenotypes displaying greater tissue penetration and involving more leaves. The modified vector accumulated similarly to the original vector in N. benthamiana after agroinfiltration, thus maintaining a high titer of virus in this intermediate host used to produce virus inoculum for grass hosts. For HSP70 , silencing one family member led to a large increase in the expression of another family member, an increase likely related to the target gene knockdown and not a general effect of virus infection. The cause of the increased insert stability in the modified vector is discussed in relationship to its recombination and accumulation potential. The modified vector will improve functional genomic studies in grasses, and the conceptual methods used to improve the vector may be applied to other VIGS vectors. © 2018 American Society of Plant Biologists. All Rights Reserved.

  12. Development of RNA Interference Trigger-Mediated Gene Silencing in Entamoeba invadens. (United States)

    Suresh, Susmitha; Ehrenkaufer, Gretchen; Zhang, Hanbang; Singh, Upinder


    Entamoeba histolytica, a protozoan parasite, is an important human pathogen and a leading parasitic cause of death. The organism has two life cycle stages, trophozoites, which are responsible for tissue invasion, and cysts, which are involved in pathogen transmission. Entamoeba invadens is the model system to study Entamoeba developmental biology, as high-grade regulated encystation and excystation are readily achievable. However, the lack of gene-silencing tools in E. invadens has limited the molecular studies that can be performed. Using the endogenous RNA interference (RNAi) pathway in Entamoeba, we developed an RNAi-based trigger gene-silencing approach inE. invadens We demonstrate that a gene's coding region that has abundant antisense small RNAs (sRNAs) can trigger silencing of a gene that is fused to it. The trigger fusion leads to the generation of abundant antisense sRNAs that map to the target gene, with silencing occurring independently of trigger location at the 5' or 3' end of a gene. Gene silencing is stably maintained during development, including encystation and excystation. We have used this approach to successfully silence two E. invadens genes: a putative rhomboid protease gene and a SHAQKY family Myb gene. The Myb gene is upregulated during oxidative stress and development, and its downregulation led, as predicted, to decreased viability under oxidative stress and decreased cyst formation. Thus, the RNAi trigger silencing method can be used to successfully investigate the molecular functions of genes inE. invadens Dissection of the molecular basis of Entamoeba stage conversion is now possible, representing an important technical advance for the system. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  13. Detailed characterization of the posttranscriptional gene-silencing-related small RNA in a GUS gene-silenced tobacco

    NARCIS (Netherlands)

    Hutvágner, G.; Mlynarova, L.; Nap, J.P.H.


    Posttranscriptional gene-silencing phenomena such as cosuppression and RNA interference are associated with the occurrence of small, about 21-23 nt short RNA species homologous to the silenced gene. We here show that the small RNA present in silenced transgenic plants can easily be detected in total

  14. The Relationship between Organizational Climate and the Organizational Silence of Administrative Staff in Education Department (United States)

    Pozveh, Asghar Zamani; Karimi, Fariba


    The aim of the present study was to determine the relationship between organizational climate and the organizational silence of administrative staff in Education Department in Isfahan. The research method was descriptive and correlational-type method. The study population was administrative staff of Education Department in Isfahan during the…

  15. Identification of process suitable diluent

    Energy Technology Data Exchange (ETDEWEB)

    Dean R. Peterman


    The Sigma Team for Minor Actinide Separation (STMAS) was formed within the USDOE Fuel Cycle Research and Development (FCRD) program in order to develop more efficient methods for the separation of americium and other minor actinides (MA) from used nuclear fuel. The development of processes for MA separations is driven by the potential benefits; reduced long-term radiotoxicty of waste placed in a geologic repository, reduced timeframe of waste storage, reduced repository heat load, the possibility of increased repository capacity, and increased utilization of energy potential of used nuclear fuel. The research conducted within the STMAS framework is focused upon the realization of significant simplifications to aqueous recycle processes proposed for MA separations. This report describes the research efforts focused upon the identification of a process suitable diluent for a flowsheet concept for the separation of MA which is based upon the dithiophosphinic acid (DPAH) extractants previously developed at the Idaho National Laboratory (INL).

  16. Lactate dehydrogenase A silencing in IDH mutant gliomas. (United States)

    Chesnelong, Charles; Chaumeil, Myriam M; Blough, Michael D; Al-Najjar, Mohammad; Stechishin, Owen D; Chan, Jennifer A; Pieper, Russell O; Ronen, Sabrina M; Weiss, Samuel; Luchman, H Artee; Cairncross, J Gregory


    Mutations of the isocitrate dehydrogenase 1 and 2 gene (IDH1/2) were initially thought to enhance cancer cell survival and proliferation by promoting the Warburg effect. However, recent experimental data have shown that production of 2-hydroxyglutarate by IDH mutant cells promotes hypoxia-inducible factor (HIF)1α degradation and, by doing so, may have unexpected metabolic effects. We used human glioma tissues and derived brain tumor stem cells (BTSCs) to study the expression of HIF1α target genes in IDH mutant ((mt)) and IDH wild-type ((wt)) tumors. Focusing thereafter on the major glycolytic enzyme, lactate dehydrogenase A (LDHA), we used standard molecular methods and pyrosequencing-based DNA methylation analysis to identify mechanisms by which LDHA expression was regulated in human gliomas. We found that HIF1α-responsive genes, including many essential for glycolysis (SLC2A1, PDK1, LDHA, SLC16A3), were underexpressed in IDH(mt) gliomas and/or derived BTSCs. We then demonstrated that LDHA was silenced in IDH(mt) derived BTSCs, including those that did not retain the mutant IDH1 allele (mIDH(wt)), matched BTSC xenografts, and parental glioma tissues. Silencing of LDHA was associated with increased methylation of the LDHA promoter, as was ectopic expression of mutant IDH1 in immortalized human astrocytes. Furthermore, in a search of The Cancer Genome Atlas, we found low expression and high methylation of LDHA in IDH(mt) glioblastomas. To our knowledge, this is the first demonstration of downregulation of LDHA in cancer. Although unexpected findings, silencing of LDHA and downregulation of several other glycolysis essential genes raise the intriguing possibility that IDH(mt) gliomas have limited glycolytic capacity, which may contribute to their slow growth and better prognosis.

  17. Suitability of tracers

    International Nuclear Information System (INIS)

    Klotz, D.


    Hydrological tracer techniques are a means of making statements on the direction and speed of underground water. One of the simpler tasks is to find out whether there is hydrological communication between two given points. This requires a determination of the direction of flow, which places less exacting demands on the properties of the tracer than does the task of determining the flow velocity of underground water. Tracer methods can serve to infer from flow velocity the distance (flow) velocity, which is defined as the ratio between the distance between two points located in flow direction and the actual time it takes water to flow from one to the other [de

  18. Break the silence: Let's talk about AIDS. (United States)


    The organizers of the Confederation of East and Central Africa Football Associations¿ (CECAFA) Youth Tournament in Kenya will organize a campaign on AIDS prevention. ¿Break the Silence: Let¿s Talk About AIDS¿ is the campaign slogan, which will be used before, during, and after the 2-week football tournament. The campaign will involve an estimated 300 players and coaches and millions of fans who will attend, watch television, or listen to radios. AIDS prevention efforts linked to the tournament include: radio and television AIDS prevention spots featuring famous football players; campaign orientation and training in AIDS outreach and education for coaches and team managers; telephone hotlines for HIV/AIDS counseling through public phone booths and health information booths; AIDS information caravans to support community awareness rallies about AIDS prevention; and a minute of silence at the beginning of all matches to commemorate soccer heroes and other loved ones who have died of AIDS.

  19. Homology-dependent Gene Silencing in Paramecium (United States)

    Ruiz, Françoise; Vayssié, Laurence; Klotz, Catherine; Sperling, Linda; Madeddu, Luisa


    Microinjection at high copy number of plasmids containing only the coding region of a gene into the Paramecium somatic macronucleus led to a marked reduction in the expression of the corresponding endogenous gene(s). The silencing effect, which is stably maintained throughout vegetative growth, has been observed for all Paramecium genes examined so far: a single-copy gene (ND7), as well as members of multigene families (centrin genes and trichocyst matrix protein genes) in which all closely related paralogous genes appeared to be affected. This phenomenon may be related to posttranscriptional gene silencing in transgenic plants and quelling in Neurospora and allows the efficient creation of specific mutant phenotypes thus providing a potentially powerful tool to study gene function in Paramecium. For the two multigene families that encode proteins that coassemble to build up complex subcellular structures the analysis presented herein provides the first experimental evidence that the members of these gene families are not functionally redundant. PMID:9529389

  20. Virus-induced gene silencing as a tool for comparative functional studies in Thalictrum.

    Directory of Open Access Journals (Sweden)

    Verónica S Di Stilio

    Full Text Available Perennial woodland herbs in the genus Thalictrum exhibit high diversity of floral morphology, including four breeding and two pollination systems. Their phylogenetic position, in the early-diverging eudicots, makes them especially suitable for exploring the evolution of floral traits and the fate of gene paralogs that may have shaped the radiation of the eudicots. A current limitation in evolution of plant development studies is the lack of genetic tools for conducting functional assays in key taxa spanning the angiosperm phylogeny. We first show that virus-induced gene silencing (VIGS of a PHYTOENE DESATURASE ortholog (TdPDS can be achieved in Thalictrum dioicum with an efficiency of 42% and a survival rate of 97%, using tobacco rattle virus (TRV vectors. The photobleached leaf phenotype of silenced plants significantly correlates with the down-regulation of endogenous TdPDS (P<0.05, as compared to controls. Floral silencing of PDS was achieved in the faster flowering spring ephemeral T. thalictroides. In its close relative, T. clavatum, silencing of the floral MADS box gene AGAMOUS (AG resulted in strong homeotic conversions of floral organs. In conclusion, we set forth our optimized protocol for VIGS by vacuum-infiltration of Thalictrum seedlings or dormant tubers as a reference for the research community. The three species reported here span the range of floral morphologies and pollination syndromes present in Thalictrum. The evidence presented on floral silencing of orthologs of the marker gene PDS and the floral homeotic gene AG will enable a comparative approach to the study of the evolution of flower development in this group.

  1. On the suitability of an allometric proxy for nondestructive estimation of average leaf dry weight in eelgrass shoots I: sensitivity analysis and examination of the influences of data quality, analysis method, and sample size on precision. (United States)

    Echavarría-Heras, Héctor; Leal-Ramírez, Cecilia; Villa-Diharce, Enrique; Cazarez-Castro, Nohe


    The effects of current anthropogenic influences on eelgrass (Zostera marina) meadows are noticeable. Eelgrass ecological services grant important benefits for mankind. Preservation of eelgrass meadows include several transplantation methods. Evaluation of establishing success relies on the estimation of standing stock and productivity. Average leaf biomass in shoots is a fundamental component of standing stock. Existing methods of leaf biomass measurement are destructive and time consuming. These assessments could alter shoot density in developing transplants. Allometric methods offer convenient indirect assessments of individual leaf biomass. Aggregation of single leaf projections produce surrogates for average leaf biomass in shoots. Involved parameters are time invariant, then derived proxies yield simplified nondestructive approximations. On spite of time invariance local factors induce relative variability of parameter estimates. This influences accuracy of surrogates. And factors like analysis method, sample size and data quality also impact precision. Besides, scaling projections are sensitive to parameter fluctuation. Thus the suitability of the addressed allometric approximations requires clarification. The considered proxies produced accurate indirect assessments of observed values. Only parameter estimates fitted from raw data using nonlinear regression, produced robust approximations. Data quality influenced sensitivity and sample size for an optimal precision. Allometric surrogates of average leaf biomass in eelgrass shoots offer convenient nondestructive assessments. But analysis method and sample size can influence accuracy in a direct manner. Standardized routines for data quality are crucial on granting cost-effectiveness of the method.

  2. Gas turbine exhaust system silencing design

    International Nuclear Information System (INIS)

    Ozgur, D.


    Gas turbines are the preferred prime mover in many applications because of their high efficiency, fuel flexibility, and low environmental impact. A typical mid-size machine might have a power rating of 80 MW, a flow of about 1000 kg/hr, and an exhaust temperature of over 500C. The most powerful single source of noise is generally the exhaust, which may generate over a kilowatt of acoustic energy. This paper reports that there are two important ways in which exhaust systems can radiate noise. The first is through the discharge of the exhaust duct, with the exhaust gas. Because of the large quantity of hot gas, the duct exit is always oriented vertically; it may be fairly high in the air in order to promote dispersion of the exhaust plume. This source is almost always attenuated by means of a silencer located somewhere in the ductwork. The second source of noise is often called breakout; it is the radiation of exhaust noise through the walls of the ducting. Breakout is most important for those sections of the exhaust duct which lie upstream of the silencer, where sound levels inside the ducting are highest. Both exhaust duct exit noise and breakout noise can be calculated from the sound power level of the gas turbine exhaust and the sound transmission loss (TL) of the silencer and ducting

  3. Using Machine Learning for Land Suitability Classification ...

    African Journals Online (AJOL)

    Artificial intelligence and machine learning methods can be used to automate the land suitability classification. Multiple Classifier System (MCS) or ensemble methods are rapidly growing and receiving a lot of attention and proved to be more accurate and robust than an excellent single classifier in many fields. In this study ...

  4. Using Machine Learning for Land Suitability Classification

    African Journals Online (AJOL)


    Abstract. Artificial intelligence and machine learning methods can be used to automate the land suitability classification. Multiple Classifier System (MCS) or ensemble methods are rapidly growing and receiving a lot of attention and proved to be more accurate and robust than an excellent single classifier in many fields.

  5. The development and application of a multiple gene co-silencing system using endogenous URA3 as a reporter gene in Ganoderma lucidum.

    Directory of Open Access Journals (Sweden)

    Dashuai Mu

    Full Text Available Ganoderma lucidum is one of the most important medicinal mushrooms; however, molecular genetics research on this species has been limited due to a lack of reliable reverse genetic tools. In this study, the endogenous orotidine 5'-monophosphate decarboxylase gene (URA3 was cloned as a silencing reporter, and four gene-silencing methods using hairpin, sense, antisense, and dual promoter constructs, were introduced into G. lucidum through a simple electroporation procedure. A comparison and evaluation of silencing efficiency demonstrated that all of the four methods differentially suppressed the expression of URA3. Our data unequivocally indicate that the dual promoter silencing vector yields the highest rate of URA3 silencing compared with other vectors (up to 81.9%. To highlight the advantages of the dual promoter system, we constructed a co-silencing system based on the dual promoter method and succeeded in co-silencing URA3 and laccase in G. lucidum. The reduction of the mRNA levels of the two genes were correlated. Thus, the screening efficiency for RNAi knockdown of multiple genes may be improved by the co-silencing of an endogenous reporter gene. The molecular tools developed in this study should facilitate the isolation of genes and the characterization of the functions of multiple genes in this pharmaceutically important species, and these tools should be highly useful for the study of other basidiomycetes.

  6. [Preparation of a gel-forming ointment base applicable to the recovery stage of bedsore and clinical evaluation of a treatment method with different ointment bases suitable to each stage of bedsore]. (United States)

    Shigeyama, Masato


    An ointment base for better treatment of bedsores was developed to improve the release rate of minocycline hydrochloride (MH) and the water absorption capacity using various types of hydrophobic to hydrophilic ointment bases. The effect of purified lanolin (PL) on the release behavior of MH from a hydrophilic ointment (HO) base was the primary focus. It was found that the drug release rate from the ointment base was modified according to the method of preparation of the ointment base and the type of cyclodextrins admixed. The physicochemical properties, such as viscosity, elution volume, and water absorption, of the ointment base were also modified by those factors. To develop an ointment formulation suitable for the recovery stages of bedsores, including the proliferation period of granulation and the formative period of epidermis, the physicochemical properties of Macrogol ointment containing various hydrophilic polymers, which have gel-forming ability, were tested. A novel ointment base suitable for the treatment in the recovery stage of bedsores was developed using hydrophobically modified hydroxypropyl methylcellulose (HM-HPMC). Considerably sustained release of MH (T50 of 170 at a min) was attained with a macrogol ointment mixed with HM-HPMC and Carbopol formulation ratio of 3:7. We clinically evaluated the effectiveness of bedsore treatment by applying different ointment bases to patients with different stages of bedsores.

  7. Identifying areas suitable for wine tourism through the use of multi-criteria and geographic information system: the method and its application in the countryside around Mount Etna (Sicily

    Directory of Open Access Journals (Sweden)

    Lara Riguccio


    Full Text Available Vineyards are among the crops that shape quality landscapes. Many places in the world are famous for their unique wine landscapes which play an important role in the development of tourism in the rural areas. Among these, the wine landscape surrounding mount Etna (Sicily emerges due to its undisputed value, as it is an important component of the territory recognised as a World Heritage Site by UNESCO. This work was conducted with that in mind, in order to identify the most suitable areas for wine tourism on the slopes of our volcano. The method used assigns a great importance to the quality of the landscape, an indispensable resource for encouraging wine tourism, and considers it to be of equal importance with the production of the wines themselves. The present work uses multi-criteria analysis in combination with geographic information system (GIS. Numerous indicators describing local resources were weighed and spatialized. The GIS analysis allowed for the development of various intermediate maps, which allowed to draw up the final suitability map for wine tourism, identifying areas larger than those of the actual vineyards. The value of these areas and the quality of their landscapes are closely connected to the production of the wines in the zone. It could be the target for specific plans and projects aimed at using the available resources, to develop wine tourism in rural areas. Although the study only covers a limited geographical area, the methodology used has general validity and could be used in other contexts.

  8. Optimized cDNA libraries for virus-induced gene silencing (VIGS using tobacco rattle virus

    Directory of Open Access Journals (Sweden)

    Page Jonathan E


    Full Text Available Abstract Background Virus-induced gene silencing (VIGS has emerged as a method for performing rapid loss-of-function experiments in plants. Despite its expanding use, the effect of host gene insert length and other properties on silencing efficiency have not been systematically tested. In this study, we probed the optimal properties of cDNA fragments of the phytoene desaturase (PDS gene for efficient VIGS in Nicotiana benthamiana using tobacco rattle virus (TRV. Results NbPDS inserts of between 192 bp and 1304 bp led to efficient silencing as determined by analysis of leaf chlorophyll a levels. The region of the NbPDS cDNA used for silencing had a small effect on silencing efficiency with 5' and 3' located inserts performing more poorly than those from the middle. Silencing efficiency was reduced by the inclusion of a 24 bp poly(A or poly(G homopolymeric region. We developed a method for constructing cDNA libraries for use as a source of VIGS-ready constructs. Library construction involved the synthesis of cDNA on a solid phase support, digestion with RsaI to yield short cDNA fragments lacking poly(A tails and suppression subtractive hybridization to enrich for differentially expressed transcripts. We constructed two cDNA libraries from methyl-jasmonate treated N. benthamiana roots and obtained 2948 ESTs. Thirty percent of the cDNA inserts were 401–500 bp in length and 99.5% lacked poly(A tails. To test the efficiency of constructs derived from the VIGS-cDNA libraries, we silenced the nicotine biosynthetic enzyme, putrescine N-methyltransferase (PMT, with ten different VIGS-NbPMT constructs ranging from 122 bp to 517 bp. Leaf nicotine levels were reduced by more than 90% in all plants infected with the NbPMT constructs. Conclusion Based on the silencing of NbPDS and NbPMT, we suggest the following design guidelines for constructs in TRV vectors: (1 Insert lengths should be in the range of ~200 bp to ~1300 bp, (2 they should be positioned in

  9. Epigenetic silencing of serine protease HTRA1 drives polyploidy

    International Nuclear Information System (INIS)

    Schmidt, Nina; Irle, Inga; Ripkens, Kamilla; Lux, Vanda; Nelles, Jasmin; Johannes, Christian; Parry, Lee; Greenow, Kirsty; Amir, Sarah; Campioni, Mara; Baldi, Alfonso; Oka, Chio; Kawaichi, Masashi; Clarke, Alan R.; Ehrmann, Michael


    Increased numbers and improperly positioned centrosomes, aneuploidy or polyploidy, and chromosomal instability are frequently observed characteristics of cancer cells. While some aspects of these events and the checkpoint mechanisms are well studied, not all players have yet been identified. As the role of proteases other than the proteasome in tumorigenesis is an insufficiently addressed question, we investigated the epigenetic control of the widely conserved protease HTRA1 and the phenotypes of deregulation. Mouse embryonal fibroblasts and HCT116 and SW480 cells were used to study the mechanism of epigenetic silencing of HTRA1. In addition, using cell biological and genetic methods, the phenotypes of downregulation of HTRA1 expression were investigated. HTRA1 is epigenetically silenced in HCT116 colon carcinoma cells via the epigenetic adaptor protein MBD2. On the cellular level, HTRA1 depletion causes multiple phenotypes including acceleration of cell growth, centrosome amplification and polyploidy in SW480 colon adenocarcinoma cells as well as in primary mouse embryonic fibroblasts (MEFs). Downregulation of HTRA1 causes a number of phenotypes that are hallmarks of cancer cells suggesting that the methylation state of the HtrA1 promoter may be used as a biomarker for tumour cells or cells at risk of transformation. The online version of this article (doi:10.1186/s12885-016-2425-8) contains supplementary material, which is available to authorized users

  10. Development of male sterility by silencing Bcp1 gene of Arabidopsis ...

    African Journals Online (AJOL)

    The development of male sterility is one of the most important steps in hybrid seed production. Several methods for the abortion of pollens based on conventional as well as genetic engineering are reported for the various crop species. Here we have investigated the use of RNA interference (RNAi) technology to silence a ...

  11. Graft-accelerated virus-induced gene silencing facilitates functional genomics in rose flowers. (United States)

    Yan, Huijun; Shi, Shaochuan; Ma, Nan; Cao, Xiaoqian; Zhang, Hao; Qiu, Xianqin; Wang, Qigang; Jian, Hongying; Zhou, Ningning; Zhang, Zhao; Tang, Kaixue


    Rose has emerged as a model ornamental plant for studies of flower development, senescence, and morphology, as well as the metabolism of floral fragrances and colors. Virus-induced gene silencing (VIGS) has long been used in functional genomics studies of rose by vacuum infiltration of cuttings or seedlings with an Agrobacterium suspension carrying TRV-derived vectors. However, VIGS in rose flowers remains a challenge because of its low efficiency and long time to establish silencing. Here we present a novel and rapid VIGS method that can be used to analyze gene function in rose, called 'graft-accelerated VIGS', where axillary sprouts are cut from the rose plant and vacuum infiltrated with Agrobacterium. The inoculated scions are then grafted back onto the plants to flower and silencing phenotypes can be observed within 5 weeks, post-infiltration. Using this new method, we successfully silenced expression of the RhDFR1, RhAG, and RhNUDX1 in rose flowers, and affected their color, petal number, as well as fragrance, respectively. This grafting method will facilitate high-throughput functional analysis of genes in rose flowers. Importantly, it may also be applied to other woody species that are not currently amenable to VIGS by conventional leaf or plantlet/seedling infiltration methods. © 2017 Institute of Botany, Chinese Academy of Sciences.

  12. An Eastern Learning Paradox: Paradoxes in Two Korean Mathematics Teachers' Pedagogy of Silence in the Classroom (United States)

    Lee, Kyeonghwa; Sriraman, Bharath


    Eastern philosophies of education such as Confucianism and Taosim advocate the use of silence in the teacher-pupil tradition of pedagogy. We investigate contemporary classrooms in Korea, and study whether teachers in Korea today incorporate this method implicitly or explicitly in their classrooms. Empirical data in the form of video-taped…

  13. Sinus lifting before Le Fort I maxillary osteotomy: a suitable method for oral rehabilitation of edentulous patients with skelettal class-III conditions: review of the literature and report of a case

    Directory of Open Access Journals (Sweden)

    Meyer Ulrich


    Full Text Available Abstract Background Functional rehabilitation of patients afflicted with severe mandibular and maxillary alveolar atrophy might be challenging especially in malformed patients. Methods Treatment planning using sinus lifting and implant placement before Le Fort I maxillary osteotomy in a patient with severe mandibular and posterior maxillary alveolar atrophy and skelettal class-III conditions due to cleft palate are described. Results A full functional and esthetic rehabilitation of the patient was achieved by a stepwise surgical approach performed through sinus lifting as the primary approach followed by implant placement and subsequent Le Fort I maxillary osteotomy to correct the maxillo-mandibular relation. Conclusion Stabilisation of the maxillary complex by a sinus lifting procedure in combination with computer aided implant placement as preorthodontic planning procedure before Le Fort I maxillary osteotomy seems to be suitable in order to allow ideal oral rehabilitation especially in malformed patients.

  14. Rock suitability classification RSC 2012

    Energy Technology Data Exchange (ETDEWEB)

    McEwen, T. (ed.) [McEwen Consulting, Leicester (United Kingdom); Kapyaho, A. [Geological Survey of Finland, Espoo (Finland); Hella, P. [Saanio and Riekkola, Helsinki (Finland); Aro, S.; Kosunen, P.; Mattila, J.; Pere, T.


    This report presents Posiva's Rock Suitability Classification (RSC) system, developed for locating suitable rock volumes for repository design and construction. The RSC system comprises both the revised rock suitability criteria and the procedure for the suitability classification during the construction of the repository. The aim of the classification is to avoid such features of the host rock that may be detrimental to the favourable conditions within the repository, either initially or in the long term. This report also discusses the implications of applying the RSC system for the fulfilment of the regulatory requirements concerning the host rock as a natural barrier and the site's overall suitability for hosting a final repository of spent nuclear fuel.

  15. Solid waste dumping site suitability analysis using geographic ...

    African Journals Online (AJOL)

    The maps were prepared by overlay and suitability analysis of geographic information system (GIS), remote sensing techniques and multi criteria analysis methods. The final suitability map was prepared by overlay analyses on Arc map and leveled as high, moderate, less suitable, and unsuitable regions of the study area ...

  16. Veiled Word(s) – Sacred Silence

    DEFF Research Database (Denmark)

    Isar, Nicoletta


    This paper defines the Byzantine altar as a sacred space of participative performance in the mystery of the Eucharist – a paradigm of “timeless re-enactment of the Christian drama.” Reading a series of Byzantine ekphraseis, the paper addresses such concepts as inner vision (theoria), silent...... or secret prayer, and divine silence, which are at the very centre of the Byzantine altar. The main focus is to investigate the liminal nature of the Mystery, manifested through concealing-revealing devices, which are thresholds in the liturgical participation of the Byzantine subject. Fear and secrecy...

  17. Validation of RNAi Silencing Efficiency Using Gene Array Data shows 18.5% Failure Rate across 429 Independent Experiments

    Directory of Open Access Journals (Sweden)

    Gyöngyi Munkácsy


    Full Text Available No independent cross-validation of success rate for studies utilizing small interfering RNA (siRNA for gene silencing has been completed before. To assess the influence of experimental parameters like cell line, transfection technique, validation method, and type of control, we have to validate these in a large set of studies. We utilized gene chip data published for siRNA experiments to assess success rate and to compare methods used in these experiments. We searched NCBI GEO for samples with whole transcriptome analysis before and after gene silencing and evaluated the efficiency for the target and off-target genes using the array-based expression data. Wilcoxon signed-rank test was used to assess silencing efficacy and Kruskal–Wallis tests and Spearman rank correlation were used to evaluate study parameters. All together 1,643 samples representing 429 experiments published in 207 studies were evaluated. The fold change (FC of down-regulation of the target gene was above 0.7 in 18.5% and was above 0.5 in 38.7% of experiments. Silencing efficiency was lowest in MCF7 and highest in SW480 cells (FC = 0.59 and FC = 0.30, respectively, P = 9.3E−06. Studies utilizing Western blot for validation performed better than those with quantitative polymerase chain reaction (qPCR or microarray (FC = 0.43, FC = 0.47, and FC = 0.55, respectively, P = 2.8E−04. There was no correlation between type of control, transfection method, publication year, and silencing efficiency. Although gene silencing is a robust feature successfully cross-validated in the majority of experiments, efficiency remained insufficient in a significant proportion of studies. Selection of cell line model and validation method had the highest influence on silencing proficiency.

  18. A Resonant Silence: For an Ethnographic History of Abortion

    Directory of Open Access Journals (Sweden)

    Flávia de Mattos Motta


    Full Text Available By using methods of the fields of History and Anthropology, this study investigated the practice of abortion in southern Brazil in the first half of the twentieth century. The examination of primary sources and interviews with elderly women intended to unveil a huge silence around this practice, which leaves very few historical sources to be investigated, and hides those memories. This text presents data collected by this survey and analyzes the diversity of representations over abortion and the ambiguities that this theme causes on the subjects involved. The practice of abortion must be understood as inserted into a context of power. The subjects operate from multiple sources of knowledge and within a morality network composed of contradictory elements.

  19. Hearing the Silence: Children’s Voices on Violence

    Directory of Open Access Journals (Sweden)

    Kathryn S. Collins


    Full Text Available Each year more than five million children in the United States are exposed to traumatizing events in their communities. This paper presents a discussion of multiple areas in violence and victimization research that needs to be continued as well as provides suggestions on how to un-silence child survivors through bridging the gaps between research and practice. It reviews the overarching problem of violence in the U.S. at the domestic and global levels and the effects of victimization. Suggestions on how to study possible mediators and moderators of victimization and individual and family adjustment, including: 1 The ecological perspective; 2 Court process and verdict; and 3Utilization of victim services, are explored. Finally, a rationale and examples of combining qualitative and quantitative methods in future research that uses children’s attributions of violence as a mediator are presented.

  20. Prognostic significance of aberrantly silenced ANPEP expression in prostate cancer

    DEFF Research Database (Denmark)

    Sørensen, Karina Dalsgaard; Abildgaard, Mette Opstrup; Haldrup, Christa


    Background:Novel biomarkers for prostate cancer (PC) are urgently needed. This study investigates the expression, epigenetic regulation, and prognostic potential of ANPEP in PC.Methods:Aminopeptidase N (APN; encoded by ANPEP) expression was analysed by immunohistochemistry using tissue microarrays...... in three hypermethylated prostate cell lines, suggesting epigenetic silencing. Negative APN immunoreactivity was significantly associated with short RFS and short CSS in the RP and CT cohort, respectively, independently of routine clinicopathological predictors. Combining APN with a known angiogenesis...... marker (vascular endothelial growth factor or microvessel density) improved risk prediction significantly in both cohorts.Conclusion:Our results suggest negative APN immunoreactivity as a new independent adverse prognostic factor for patients with clinically localised PC and, furthermore, that epigenetic...

  1. Projective synchronization based on suitable separation

    International Nuclear Information System (INIS)

    Li Guohui; Xiong Chuan; Sun Xiaonan


    A new approach for constructing a projective-synchronized chaotic slave system is proposed in this paper. This method is based on suitable separation by decomposing the system as the linear part and the nonlinear one. From matrix measure theory, some simple but efficient criteria are derived for projective synchronization of chaotic system. Numerical simulations for the Lorenz system show that this control method works very well

  2. Collaborative silence and some unguided prerogative utterances of ...

    African Journals Online (AJOL)

    This essay attempts to demystify the silence and non-prosecution syndrome surrounding unguided prerogative utterances of Nigerian leaders as a contemporary culture, which has continued to threaten the unity and stability of the country. In doing this, the silence and withdrawal of key Nigerian agencies, such as; the ...

  3. Echoes of Silence: Empathy and Making Connections through Writing Process (United States)

    Freedman, Joel M.


    On April 25, 2008, students on college and public school campuses collectively committed to a vow of silence commemorating an event known as the National Day of Silence. This student-generated, nationwide action theatrically "speaks out" in solidarity with lesbian, gay, bisexual, and transgender (LGBT) people who for one reason or another fear…

  4. The Educators' Guide to the Day of Silence (United States)

    Gay, Lesbian and Straight Education Network (GLSEN), 2010


    The Day of Silence is the largest single student-led action towards creating safer schools for all, regardless of sexual orientation, gender identity or gender expression. From the first-ever Day of Silence at the University of Virginia in 1996, to the organizing efforts in over 8,000 middle schools, high schools, colleges and universities across…

  5. Reflections on the Silencing the Self Scale and Its Origins (United States)

    Jack, Dana Crowley


    In this article, the author reflects on the Silencing the Self Scale (STSS) and blends her personal and professional thoughts about self-silencing, gender, and depression. For her, the despair of depression deeply involves questions of value and meaning, culture and freedom. The STSS grew from listening to depressed women's voices. From them, the…

  6. Antiviral RNA silencing viral counter defense in plants

    NARCIS (Netherlands)

    Bucher, E.C.


    The research described in this thesis centres around the mechanism of RNA silencing in relation to virus-host interaction, an area of increasing importance. It shows how this recently disclosed mechanism can be used to produce virus-resistant plants. Based on the activity of the RNA silencing

  7. Language and silence: persoonlike herinneringe aan Adam Small ...

    African Journals Online (AJOL)


    En hy was self nooit kategories of dogmaties in sy denke of uitsprake nie. Die titel van my voordrag is “Language and silence: persoonlike herinneringe aan. 'n digter-intellektueel”. Dis Adam wat my as student voorgestel het aan George Steiner se “Language and Silence: Essays on Language, Literature and the Inhumane.

  8. Strategic Silence as a Tool of Political Communication: A Reflection ...

    African Journals Online (AJOL)

    The paper, therefore, recommended that politicians should be cautious in the use of strategic silence as a means of political communication in view of the nature of politics which calls for self- expression in the use of persuasion, clarification and conviction to win public support and sympathy. Maintaining silence when one is ...

  9. No-Big-Silence teeb klubituuri / Urmas Hännile

    Index Scriptorium Estoniae

    Hännile, Urmas


    Rockansamblist No-Big-Silence ja dark-popansamblist Sinine, nende kontsertttuurist mööda Eestimaad, tutvustamisel bändide uued albumid (No-Big-Silence "Starstealer" ja Sinine "Butterflies"), Pärnus on kontsert 24. oktoobril klubis Sugar

  10. The splicing machinery promotes RNA-directed DNA methylation and transcriptional silencing in Arabidopsis (United States)

    Zhang, Cui-Jun; Zhou, Jin-Xing; Liu, Jun; Ma, Ze-Yang; Zhang, Su-Wei; Dou, Kun; Huang, Huan-Wei; Cai, Tao; Liu, Renyi; Zhu, Jian-Kang; He, Xin-Jian


    DNA methylation in transposons and other DNA repeats is conserved in plants as well as in animals. In Arabidopsis thaliana, an RNA-directed DNA methylation (RdDM) pathway directs de novo DNA methylation. We performed a forward genetic screen for suppressors of the DNA demethylase mutant ros1 and identified a novel Zinc-finger and OCRE domain-containing Protein 1 (ZOP1) that promotes Pol IV-dependent siRNA accumulation, DNA methylation, and transcriptional silencing. Whole-genome methods disclosed the genome-wide effects of zop1 on Pol IV-dependent siRNA accumulation and DNA methylation, suggesting that ZOP1 has both RdDM-dependent and -independent roles in transcriptional silencing. We demonstrated that ZOP1 is a pre-mRNA splicing factor that associates with several typical components of the splicing machinery as well as with Pol II. Immunofluorescence assay revealed that ZOP1 overlaps with Cajal body and is partially colocalized with NRPE1 and DRM2. Moreover, we found that the other development-defective splicing mutants tested including mac3a3b, mos4, mos12 and mos14 show defects in RdDM and transcriptional silencing. We propose that the splicing machinery rather than specific splicing factors is involved in promoting RdDM and transcriptional silencing. PMID:23524848

  11. Chinanteco children’s silences in different classroom situations

    Directory of Open Access Journals (Sweden)

    Valeria Rebolledo Angulo


    Full Text Available This article analyzes, from an ethnographic perspective and a sociocultural framework, the construction of silences in the interaction between students and teachers in a multilingual classroom situation in an indigenous community in méxico. the analysis reveals how the silence of the chinanteco speaking children when asked to answer certain questions in class is not always due to their failure to understand spoken and written spanish that is used in class. their silences are responses taking different meanings in specific situations. the silence of the children can be a way of resisting, a way of hiding, and, sometimes, their voices are silenced.

  12. The RNA silencing pathway: the bits and pieces that matter.

    Directory of Open Access Journals (Sweden)

    Marian A C Groenenboom


    Full Text Available Cellular pathways are generally proposed on the basis of available experimental knowledge. The proposed pathways, however, may be inadequate to describe the phenomena they are supposed to explain. For instance, by means of concise mathematical models we are able to reveal shortcomings in the current description of the pathway of RNA silencing. The silencing pathway operates by cleaving siRNAs from dsRNA. siRNAs can associate with RISC, leading to the degradation of the target mRNA. We propose and analyze a few small extensions to the pathway: a siRNA degrading RNase, primed amplification of aberrant RNA pieces, and cooperation between aberrant RNA to trigger amplification. These extensions allow for a consistent explanation for various types of silencing phenomena, such as virus induced silencing, transgene and transposon induced silencing, and avoidance of self-reactivity, as well as for differences found between species groups.

  13. Dynamics of Chromatin Silencing at Telomeres: Deterministic and Stochastic Aspects (United States)

    Apratim, Manjul; Dayarian, Adel; Sontag, Eduardo; Sengupta, Anirvan


    Epigenetic silencing modifications of are often associated with well-defined domains. We study potential mechanisms of formation of boundary of silenced regions. We specially focus on the possibility that some telomeric silencing boundaries are formed in a self-organized manner, as opposed to being defined by specific boundary elements. In particular, we examine systems where a titration-induced feedback can stabilize the boundary of the silenced region. A consequence of having multiple such boundaries is large stochastic cell-to-cell variation of boundary locations. We proceed to make an argument about the nature of the fall-off of the average silencing protein occupancy, coming from such variability, and test the predictions against HA-Sir3 ChIP-seq data from experiments performed on yeast.

  14. Scaled signal intensity of uterine fibroids based on T2-weighted MR images: a potential objective method to determine the suitability for magnetic resonance-guided focused ultrasound surgery of uterine fibroids

    Energy Technology Data Exchange (ETDEWEB)

    Park, Hyun [CHA University, Comprehensive Gynecologic Cancer Center, CHA Bundang Medical Center, Gyunggi-do (Korea, Republic of); Yoon, Sang-Wook [CHA University, Department of Diagnostic Radiology, CHA Bundang Medical Center, Sungnam-si, Gyunggi-do (Korea, Republic of); Sokolov, Amit [InSightec Ltd., Haifa (Israel)


    Magnetic Resonance-guided Focused Ultrasound Surgery (MRgFUS) is a non-invasive method to treat uterine fibroids. To help determine the patient suitability for MRgFUS, we propose a new objective measure: the scaled signal intensity (SSI) of uterine fibroids in T2 weighted MR images (T2WI). Forty three uterine fibroids in 40 premenopausal women were included in this retrospective study. SSI of each fibroid was measured from the screening T2WI by standardizing its mean signal intensity to a 0-100 scale, using reference intensities of rectus abdominis muscle (0) and subcutaneous fat (100). Correlation between the SSI and the non-perfused volume (NPV) ratio (a measure for treatment success) was calculated. Pre-treatment SSI showed a significant inverse-correlation with post treatment NPV ratio (p < 0.05). When dichotomizing NPV ratio at 45 %, the optimal cut off value of the SSI was found to be 16.0. A fibroid with SSI value 16.0 or less can be expected to have optimal responses. The SSI of uterine fibroids in T2WI can be suggested as an objective parameter to help in patient selection for MRgFUS. (orig.)

  15. Rewiring of neuronal networks during synaptic silencing. (United States)

    Wrosch, Jana Katharina; Einem, Vicky von; Breininger, Katharina; Dahlmanns, Marc; Maier, Andreas; Kornhuber, Johannes; Groemer, Teja Wolfgang


    Analyzing the connectivity of neuronal networks, based on functional brain imaging data, has yielded new insight into brain circuitry, bringing functional and effective networks into the focus of interest for understanding complex neurological and psychiatric disorders. However, the analysis of network changes, based on the activity of individual neurons, is hindered by the lack of suitable meaningful and reproducible methodologies. Here, we used calcium imaging, statistical spike time analysis and a powerful classification model to reconstruct effective networks of primary rat hippocampal neurons in vitro. This method enables the calculation of network parameters, such as propagation probability, path length, and clustering behavior through the measurement of synaptic activity at the single-cell level, thus providing a fuller understanding of how changes at single synapses translate to an entire population of neurons. We demonstrate that our methodology can detect the known effects of drug-induced neuronal inactivity and can be used to investigate the extensive rewiring processes affecting population-wide connectivity patterns after periods of induced neuronal inactivity.

  16. Post-translational selective intracellular silencing of acetylated proteins with de novo selected intrabodies. (United States)

    Chirichella, Michele; Lisi, Simonetta; Fantini, Marco; Goracci, Martina; Calvello, Mariantonietta; Brandi, Rossella; Arisi, Ivan; D'Onofrio, Mara; Di Primio, Cristina; Cattaneo, Antonino


    The ability to selectively interfere with post-translationally modified proteins would have many biological and therapeutic applications. However, post-translational modifications cannot be selectively targeted by nucleic-acid-based interference approaches. Here we describe post-translational intracellular silencing antibody technology (PISA), a method for selecting intrabodies against post-translationally modified proteins. We demonstrate our method by generating intrabodies against native acetylated proteins and showing functional interference in living cells.

  17. Development of Agrobacterium-mediated virus-induced gene silencing and performance evaluation of four marker genes in Gossypium barbadense.

    Directory of Open Access Journals (Sweden)

    Jinhuan Pang

    Full Text Available Gossypiumbarbadense is a cultivated cotton species and possesses many desirable traits, including high fiber quality and resistance to pathogens, especially Verticilliumdahliae (a devastating pathogen of Gossypium hirsutum, the main cultivated species. These elite traits are difficult to be introduced into G. hirsutum through classical breeding methods. In addition, genetic transformation of G. barbadense has not been successfully performed. It is therefore important to develop methods for evaluating the function and molecular mechanism of genes in G. barbadense. In this study, we had successfully introduced a virus-induced gene silencing (VIGS system into three cultivars of G. barbadense by inserting marker genes into the tobacco rattle virus (TRV vector. After we optimized the VIGS conditions, including light intensity, photoperiod, seedling age and Agrobacterium strain, 100% of plants agroinfiltrated with the GaPDS silencing vector showed white colored leaves. Three other marker genes, GaCLA1, GaANS and GaANR, were employed to further test this VIGS system in G. barbadense. The transcript levels of the endogenous genes in the silenced plants were reduced by more than 99% compared to control plants; these plants presented phenotypic symptoms 2 weeks after inoculation. We introduced a fusing sequence fragment of GaPDS and GaANR gene silencing vectors into a single plant, which resulted in both photobleaching and brownish coloration. The extent of silencing in plants agroinfiltrated with fusing two-gene-silencing vector was consistent with plants harboring a single gene silencing vector. The development of this VIGS system should promote analysis of gene function in G. barbadense, and help to contribute desirable traits for breeding of G. barbadense and G. hirsutum.

  18. The Curious Silence of the Dog and Paul of Tarsus; Revisiting The Argument from Silence

    Directory of Open Access Journals (Sweden)

    Michael Gary Duncan


    Full Text Available In this essay I propose an interpretative and explanatory structure for the so-called argumentum ex silento, or argument from silence (henceforth referred to as the AFS. To this end, I explore two examples, namely, Sherlock Holmes’s oft-quoted notice of the “curious incident of the dog in the night-time” from Arthur Conan Doyle’s short story “Silver Blaze,” and the historical question of Paul of Tarsus’s silence on biographical details of the historical Jesus. Through these cases, I conclude that the AFS serves as a dialogical topos best evaluated and understood through the perceived authority of the arguer and the willingness of the audience to accept that authority, due to the “curious” nature of the negative evidence that the argument employed.

  19. The ethics of silence: Does conflict of interest explain employee silence? (United States)

    Anderson, James


    Employee silence constitutes a significant threat to organizational success. This article argues that silence is a by-product of a structural Conflict of Interest (COI) between employees and their employers. This argument turns on the claim, also defended here, that employees are in a privileged position vis-à-vis knowledge of their work and that leaders-whether they recognize it or not-are dependent on their employees for reliable information about the work they are doing. Employee voice, therefore, is an organizational necessity. It is also a moral achievement as it involves risking one's personal interests for the sake of the organization. Leaders must take steps to mitigate COI and encourage employee voice; this article provides several strategies for doing exactly that.

  20. Viral counterdefense on RNA silencing : analysis of RNA silencing suppressors from arthropod-borne negative strand RNA plant viruses

    NARCIS (Netherlands)

    Schnettler, E.


    This thesis describes that RNA silencing suppressor (RSS) proteins encoded by negative-stranded RNA plant viruses are able to interfere with different RNA silencing pathways in a variety of organisms by interacting with double stranded (ds)RNA molecules. These RSS proteins are able to counteract the

  1. Virus-induced gene silencing in diverse maize lines using the Brome Mosaic virus-based silencing vector (United States)

    Virus-induced gene silencing (VIGS) is a widely used tool for gene function studies in many plant species, though its use in monocots has been limited. Using a Brome mosaic virus (BMV) vector designed to silence the maize phytoene desaturase gene, a genetically diverse set of maize inbred lines was ...

  2. Environmental Suitability for Leasing Property (United States)

    This is a Memorandum of Understanding between the U.S. Environmental Protection Agency and the U.S. Department of Defense, regarding the procedures to determine environmental suitability for leasing property available as a result of a base closure or realignment.

  3. Readability and Suitability of COPD Consumer Information

    Directory of Open Access Journals (Sweden)

    Kathryn Fullmann


    Full Text Available Background. Information leaflets have been shown to positively or negatively impact adherence, depending on their content. The objective of this study was to perform an appraisal of the consumer information provided in COPD inhaler monographs. Methods. COPD inhalers were identified from the Health Canada Drug Product Database. Medication information and instructions for inhaler use were analyzed for readability by seven formulas, with an acceptability threshold of grades 6–8. Three researchers rated suitability using a modified Suitability Assessment of Materials (SAM tool and assessed leaflets for explicit warnings. Results. Twenty-six inhalers with a COPD indication were evaluated. Medication information sections were rated as “difficult to read” or “hard,” and 85% (22/26 had a reading level above grade 8. The instructions for inhaler use were rated as “easy” or “fairly easy” to read and 63% (16/26 met the threshold by all formulas. While all leaflets achieved superior suitability ratings, extreme warnings included risk of premature death (n=12, risks of serious injury (n=26, serious interactions (n=26, and statements that convey a serious consequence to therapy (n=26. Conclusion. While COPD information leaflets in Canada performed well in terms of readability and suitability, overemphasis on side effects, warnings, and precautions may contribute to patient fear and nonadherence.

  4. Transcriptional silencing of multiple genes in trophozoites of Entamoeba histolytica.

    Directory of Open Access Journals (Sweden)

    Rivka Bracha


    Full Text Available In a previous work we described the transcriptional silencing of the amoebapore A (AP-A gene (Ehap-a of Entamoeba histolytica strain HM-1:IMSS. The silencing occurred following transfection with a plasmid containing a 5' upstream region (473 bp of Ehap-a that included a truncated segment (140 bp of a short interspersed nuclear element (SINE1. Silencing remained in effect even after removal of the plasmid (clone G3. Neither short interfering RNA nor methylated DNA were detected, but the chromatin domain of Ehap-a in the gene-silenced trophozoites was modified. Two other similar genes (Ehap-b and one encoding a Saposin-like protein, SAPLIP 1 also became silenced. In the present work we demonstrate the silencing of a second gene of choice, one that encodes the light subunit of the Gal/GalNAc inhibitable lectin (Ehlgl1 and the other, the cysteine proteinase 5 (EhCP-5. This silencing occurred in G3 trophozoites transfected with a plasmid in which the 473 bp 5' upstream Ehap-a fragment was directly ligated to the second gene. Transcriptional silencing occurred in both the transgene and the chromosomal gene. SINE1 sequences were essential, as was a direct connection between the Ehap-a upstream region and the beginning of the open reading frame of the second gene. Gene silencing did not occur in strain HM-1:IMSS with any of these plasmid constructs. The trophozoites with two silenced genes were virulence-attenuated as were those of clone G3. In addition, trophozoites not expressing Lgl1 and AP-A proteins had a significantly reduced ability to cap the Gal/GalNAc-lectin to the uroid region when incubated with antibodies against the heavy (170 kDa subunit of the lectin. Lysates of trophozoites lacking cysteine proteinase 5 and AP-A proteins had 30% less cysteine proteinase activity than those of HM-1:IMSS strain or the G3 clone. Silencing of other genes in G3 amoebae could provide a model to study their various functions. In addition, double gene-silenced

  5. Habitat Suitability Index Models: Osprey (United States)

    Vana-Miller, Sandra L.


    A review and synthesis of existing information were used to develop a Habitat Suitability Index (HSI) model for the osprey (Pandion haliaetus). The model consolidates habitat use information into a framework appropriate for field application, and is scaled to produce an index between 0.0 (unsuitable habitat) to 1.0 (optimum habitat). HSI models are designed to be used with Habitat Evaluation Procedures previously developed by the U.S. Fish and Wildlife Service.

  6. Utility of P19 Gene-Silencing Suppressor for High Level Expression of Recombinant Human Therapeutic Proteins in Plant Cells

    Directory of Open Access Journals (Sweden)

    Maryam Zangi


    Full Text Available Background: The potential of plants, as a safe and eukaryotic system, is considered in the production of recombinant therapeutic human protein today; but the expression level of heterologous proteins is limited by the post-transcriptional gene silencing (PTGS response in this new technology. The use of viral suppressors of gene silencing can prevent PTGS and improve transient expression level of foreign proteins. In this study, we investigated the effect of p19 silencing suppressor on recombinant human nerve growth factor expression in Nicotiana benthamiana. Materials and Methods: The p19 coding region was inserted in the pCAMBIA using NcoI and BstEII recognition sites. Also, the cloned synthesized recombinant human NGF (rhNGF fragment was cloned directly into PVX vector by ClaI and SalI restriction enzymes. The co-agroinfiltration of rhNGF with p19 viral suppressor of gene silencing was evaluated by dot-blot and SDS-PAGE. The amount of expressed rhNGF protein was calculated by AlphaEaseFC software. Results: Co-agroinfiltration of hNGF with P19 suppressor showed about forty-fold increase (8% total soluble protein (TSP when compared to the absence of P19 suppressor (0.2%TSP. Conclusion: The results presented here confirmed that the use of P19 gene silencing suppressor derived from tomato bushy stunt virus (TBSV could efficiently increase the transient expression of recombinant proteins in Nicotiana benthamiana manifold.

  7. Use of a Virus Gene Silencing Vector for Maize Functional Genomics Research. (United States)

    Zhou, Tao; Liu, Xuedong; Fan, Zaifeng


    Virus-induced gene silencing (VIGS) is a genetic technology that exploits the RNA-mediated defense against virus. The method has great potential for plant reverse genetics as it could knock down gene expression in a rapid way, which is triggered by a replicating viral genome engineered to carry a fragment of host gene to be silenced. A number of efficient VIGS vectors are available for dicots, such as for model plant Nicotiana benthamiana; however, only a few of VIGS vectors for monocotyledonous cereal crops. Here, we describe the method for the use of a newly developed VIGS vector based on a maize-infecting Cucumber mosaic virus (CMV) strain ZMBJ-CMV for maize. The RNA2 of ZMBJ-CMV was modified as a vector pCMV201-2b N81 having multiple cloning sites for the insert of 100-300 bp fragment of target gene. Using a method of vascular puncture inoculation of maize seeds with crude sap prepared from Agrobacterium-infiltrated N. benthamiana leaves, silencing of target genes could be obtained in 4 weeks.

  8. Oriana: the voice and the silences

    Directory of Open Access Journals (Sweden)

    Mercedes Rosúa


    Full Text Available Commentators did their best to bury the late Oriana Fallaci’s ideas and books written since the 11 S under tons of praises to her previous works, and under deep silence or global discrediting her denunciation of Islamic fundamentalism and of terrorist blackmail and all its forms of getting a grip on Europe. Obituary notices avoided carefully the analysis of facts and data brought forward by Oriana in her three last books: The rage and the pride. The power of the reason and Oriana Fallaci’s inter- view with herself. The Apocalypse. But the italian journalist, who past away on 15th Sep- tember 2006, denounced substantiated facts, true dangers and a real collusion of wes- tern sectors-venal, corrupt, fainthearted and ignorant-with the strategy of penetration and manipulation of public opinion of an islamic movement who is the enemy of democracies, Civil Rights, freedom and the western systems of open society.Key words: Oriana Fallaci, Islamic fundamentalism, Defence freedom, Denun- ciation fear, Europe ́s surrender, Terrorism, Western censorship. 

  9. Breaking the Silence: Feminism and Post humanism

    Directory of Open Access Journals (Sweden)

    Debika Saha


    Full Text Available Postmodernism starts its journey by challenging the master narratives of metaphysics and philosophy. In this journey the narrative get replaced either by an emancipation from narratives that claim to possess eternal truth or by a turn to a kind of informative technology. But both the ways keep at their centre the process of “denaturing” or disassociating something of its natural qualities. This postmodernist turn has now taken a new form. The choice is not limited between the natural body and the culturally constructed body, but between different areas of bodily reconstruction bearing different social and cultural implications. As Spivak in her famous writing raises the question «Can the subaltern speak?» In fact, Spivak narrates how the marginalized, the less privileged depend upon the benevolence of the more privileged. To engage with the other means not only to talk, listen, learn or know about her but also to identify with her. There must be some ways for open dialogues that can diminish the effects of cultural dominance. The imperial silence should be broken.

  10. Evidence for gene silencing by endogenous DNA methylation (United States)

    Holliday, Robin; Ho, Thu


    Transformed cells can spontaneously silence genes by de novo methylation, and it is generally assumed that this is due to DNA methyltransferase activity. We have tested the alternative hypothesis that gene silencing could be due to the uptake of 5-methyl-dCMP into DNA, via the di- and triphosphonucleotides. 5-Methyl-dCMP would be present in cells from the ongoing repair of DNA. We have isolated a strain of Chinese hamster ovary (CHO) cells, designated HAM−, which spontaneously silences two tested genes at a very high frequency. We have shown that this strain incorporates 5-[3H]methyldeoxycytidine into 5-methylcytosine and thymine in DNA. It also has low 5-methyl-dCMP deaminase activity. Another HAM+ strain has high deaminase activity and a very low frequency of gene silencing. The starting strain, CHO K1, has a phenotype intermediate between HAM− and HAM+. PMID:9671746

  11. Mutations in Ran system affected telomere silencing in Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    Hayashi, Naoyuki; Kobayashi, Masahiko; Shimizu, Hiroko; Yamamoto, Ken-ichi; Murakami, Seishi; Nishimoto, Takeharu


    The Ran GTPase system regulates the direction and timing of several cellular events, such as nuclear-cytosolic transport, centrosome formation, and nuclear envelope assembly in telophase. To gain insight into the Ran system's involvement in chromatin formation, we investigated gene silencing at the telomere in several mutants of the budding yeast Saccharomyces cerevisiae, which had defects in genes involved in the Ran system. A mutation of the RanGAP gene, rna1-1, caused reduced silencing at the telomere, and partial disruption of the nuclear Ran binding factor, yrb2-Δ2, increased this silencing. The reduced telomere silencing in rna1-1 cells was suppressed by a high dosage of the SIR3 gene or the SIT4 gene. Furthermore, hyperphosphorylated Sir3 protein accumulated in the rna1-1 mutant. These results suggest that RanGAP is required for the heterochromatin structure at the telomere in budding yeast

  12. Locus-specific ribosomal RNA gene silencing in nucleolar dominance.

    Directory of Open Access Journals (Sweden)

    Michelle S Lewis


    Full Text Available The silencing of one parental set of rRNA genes in a genetic hybrid is an epigenetic phenomenon known as nucleolar dominance. We showed previously that silencing is restricted to the nucleolus organizer regions (NORs, the loci where rRNA genes are tandemly arrayed, and does not spread to or from neighboring protein-coding genes. One hypothesis is that nucleolar dominance is the net result of hundreds of silencing events acting one rRNA gene at a time. A prediction of this hypothesis is that rRNA gene silencing should occur independent of chromosomal location. An alternative hypothesis is that the regulatory unit in nucleolar dominance is the NOR, rather than each individual rRNA gene, in which case NOR localization may be essential for rRNA gene silencing. To test these alternative hypotheses, we examined the fates of rRNA transgenes integrated at ectopic locations. The transgenes were accurately transcribed in all independent transgenic Arabidopsis thaliana lines tested, indicating that NOR localization is not required for rRNA gene expression. Upon crossing the transgenic A. thaliana lines as ovule parents with A. lyrata to form F1 hybrids, a new system for the study of nucleolar dominance, the endogenous rRNA genes located within the A. thaliana NORs are silenced. However, rRNA transgenes escaped silencing in multiple independent hybrids. Collectively, our data suggest that rRNA gene activation can occur in a gene-autonomous fashion, independent of chromosomal location, whereas rRNA gene silencing in nucleolar dominance is locus-dependent.

  13. Highly efficient virus-induced gene silencing in apple and soybean by apple latent spherical virus vector and biolistic inoculation. (United States)

    Yamagishi, Noriko; Yoshikawa, Nobuyuki


    Virus-induced gene silencing (VIGS) is an effective tool for the analysis of the gene function in plants within a short time. However, in woody fruit tree like apple, some of Solanum crops, and soybean, it is generally difficult to inoculate virus vector by conventional inoculation methods. Here, we show efficient VIGS in apple and soybean by Apple latent spherical virus (ALSV) vector and biolistic inoculation. The plants inoculated with ALSV vectors by particle bombardment showed uniform silenced phenotypes of target genes within 2-3 weeks post inoculation.

  14. RNA silencing is resistant to low-temperature in grapevine.

    Directory of Open Access Journals (Sweden)

    Marjorie Romon

    Full Text Available RNA silencing is a natural defence mechanism against viruses in plants, and transgenes expressing viral RNA-derived sequences were previously shown to confer silencing-based enhanced resistance against the cognate virus in several species. However, RNA silencing was shown to dysfunction at low temperatures in several species, questioning the relevance of this strategy in perennial plants such as grapevines, which are often exposed to low temperatures during the winter season. Here, we show that inverted-repeat (IR constructs trigger a highly efficient silencing reaction in all somatic tissues in grapevines. Similarly to other plant species, IR-derived siRNAs trigger production of secondary transitive siRNAs. However, and in sharp contrast to other species tested to date where RNA silencing is hindered at low temperature, this process remained active in grapevine cultivated at 4°C. Consistently, siRNA levels remained steady in grapevines cultivated between 26°C and 4°C, whereas they are severely decreased in Arabidopsis grown at 15°C and almost undetectable at 4°C. Altogether, these results demonstrate that RNA silencing operates in grapevine in a conserved manner but is resistant to far lower temperatures than ever described in other species.

  15. 21 CFR 640.63 - Suitability of donor. (United States)


    ... the plasma; (9) Freedom from any disease, other than malaria, transmissible by blood transfusion... ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Source Plasma § 640.63 Suitability of donor. (a) Method of determining. The suitability of a donor for Source Plasma shall be determined by a qualified...

  16. 21 CFR 640.3 - Suitability of donor. (United States)


    ... ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Whole Blood § 640.3 Suitability of donor. (a) Method of determining. The suitability of a donor as a source of Whole Blood shall be determined by a... transmissible by blood transfusion, insofar as can be determined by history and examinations indicated above...

  17. Fast Silencing Reveals a Lost Role for Reciprocal Inhibition in Locomotion (United States)

    Moult, Peter R.; Cottrell, Glen A.; Li, Wen-Chang


    Summary Alternating contractions of antagonistic muscle groups during locomotion are generated by spinal “half-center” networks coupled in antiphase by reciprocal inhibition. It is widely thought that reciprocal inhibition only coordinates the activity of these muscles. We have devised two methods to rapidly and selectively silence neurons on just one side of Xenopus tadpole spinal cord and hindbrain, which generate swimming rhythms. Silencing activity on one side led to rapid cessation of activity on the other side. Analyses reveal that this resulted from the depression of reciprocal inhibition connecting the two sides. Although critical neurons in intact tadpoles are capable of pacemaker firing individually, an effect that could support motor rhythms without inhibition, the swimming network itself requires ∼23 min to regain rhythmic activity after blocking inhibition pharmacologically, implying some homeostatic changes. We conclude therefore that reciprocal inhibition is critical for the generation of normal locomotor rhythm. PMID:23312521

  18. Platinum Interference with siRNA Non-seed Regions Fine-Tunes Silencing Capacity

    DEFF Research Database (Denmark)

    Hedman, Hanna K; Kirpekar, Finn; Elmroth, Sofi K C


    , cisplatin and oxaliplatin, on siRNA's silencing capacity has been evaluated. More specifically, siRNAs targeting the 3' UTR region of Wnt-5a mRNA (NM_003352) were constructed, and the biologically active antisense RNA strand was pre-platinated. Platinum adducts were detected and characterized...... by a combination of gel electrophoresis and MALDI-MS techniques, and the silencing capacity was evaluated in cellular luciferase-expressing systems using HB2 cells. Data show that platination of the antisense strand of the siRNAs results in adducts with protection against hydrolytic cleavage in the proximity...... of the platination sites, i.e., with altered degradation patterns compared to native RNAs. The MALDI-MS method was successfully used to further identify and characterize platinated RNA, with the naturally occurring platinum isotopic patterns serving as sensitive fingerprints for metalated sites. Expression assays...

  19. The Enamovirus P0 protein is a silencing suppressor which inhibits local and systemic RNA silencing through AGO1 degradation

    Energy Technology Data Exchange (ETDEWEB)

    Fusaro, Adriana F. [University of Sydney, NSW 2006 (Australia); CSIRO Plant Industry, Canberra, P.O. Box 1600, ACT 2601 (Australia); Correa, Regis L. [CSIRO Plant Industry, Canberra, P.O. Box 1600, ACT 2601 (Australia); Depto. de Virologia, IMPPG, UFRJ, 21941-902 (Brazil); Nakasugi, Kenlee; Jackson, Craig [University of Sydney, NSW 2006 (Australia); Kawchuk, Lawrence [Research Centre, Agriculture and Agri-Food Canada, Lethbridge, AB T1J4B1 (Canada); Vaslin, Maite F.S. [Depto. de Virologia, IMPPG, UFRJ, 21941-902 (Brazil); Waterhouse, Peter M., E-mail: [University of Sydney, NSW 2006 (Australia); CSIRO Plant Industry, Canberra, P.O. Box 1600, ACT 2601 (Australia)


    The P0 protein of poleroviruses and P1 protein of sobemoviruses suppress the plant's RNA silencing machinery. Here we identified a silencing suppressor protein (SSP), P0{sup PE}, in the Enamovirus Pea enation mosaic virus-1 (PEMV-1) and showed that it and the P0s of poleroviruses Potato leaf roll virus and Cereal yellow dwarf virus have strong local and systemic SSP activity, while the P1 of Sobemovirus Southern bean mosaic virus supresses systemic silencing. The nuclear localized P0{sup PE} has no discernable sequence conservation with known SSPs, but proved to be a strong suppressor of local silencing and a moderate suppressor of systemic silencing. Like the P0s from poleroviruses, P0{sup PE} destabilizes AGO1 and this action is mediated by an F-box-like domain. Therefore, despite the lack of any sequence similarity, the poleroviral and enamoviral SSPs have a conserved mode of action upon the RNA silencing machinery.

  20. Breaking the Silence: A Phenomenological Exploration of Secondary Traumatic Stress in U.S. College Student Affairs Professionals (United States)

    Lynch, Robert Jason


    Breaking the Silence: A Phenomenological Exploration of Secondary Traumatic Stress in U.S. College Student Affairs Professionals is a qualitative-intensive mixed methods study using phenomenology and art-based research techniques to uncover the essence of secondary traumatic stress in U.S. college student affairs professionals. Researchers in the…

  1. Mind the gap: In-session silences are associated with client attachment insecurity, therapeutic alliance, and treatment outcome

    DEFF Research Database (Denmark)

    Daniel, Sarah Ingrid Franksdatter; Folke, Sofie; Lunn, Susanne


    Objective: The association between in-session silences and client attachment, therapeutic alliance, and treatment outcome was investigated in two treatments for bulimia nervosa. Method: 69 women and one man were randomized to two years of psychoanalytic psychotherapy (PPT) or 20 sessions of cogni...

  2. The Relationship between Organizational Trust and Organizational Silence with Job Satisfaction and Organizational Commitment of the Employees of University (United States)

    Fard, Parastoo Gashtasebi; Karimi, Fariba


    This study aimed to investigate the structural model between organizational trust and organizational silence with job satisfaction and organizational commitment of the employees of Islamic Azad University of Isfahan, (Khorasgan) branch. The study method is descriptive-correlation. The study population is the employees of Islamic Azad University of…

  3. Suitable footwear for enhanced safety

    CERN Document Server


    Safety shoes are the theme of a new safety campaign. Always remember that accidents can happen - even to your feet! When entering hazardous areas such as underground halls, work sites, experiment assembly sites, workshops etc., sandals, ordinary shoes or similar light footwear should not be worn. Whatever the risks to which you may be exposed, always think safety and wear suitable footwear, i.e. safety shoes, which have non-slip soles and steel reinforcements to protect your feet from being crushed, fractured or pierced. Is it serious, Doctor? "Some traumas resulting from foot-related accidents - open fractures for instance - can be quite serious," explains CERN Works Doctor Véronique Fassnacht. "But the most common injuries are sprained ankles sustained during simple falls caused by differences in floor-levels (e.g. false floors). Fractures, bruising, surface wounds or deep wounds caused by objects falling onto the top of the foot are also quite common." ...

  4. Method

    Directory of Open Access Journals (Sweden)

    Ling Fiona W.M.


    Full Text Available Rapid prototyping of microchannel gain lots of attention from researchers along with the rapid development of microfluidic technology. The conventional methods carried few disadvantages such as high cost, time consuming, required high operating pressure and temperature and involve expertise in operating the equipment. In this work, new method adapting xurography method is introduced to replace the conventional method of fabrication of microchannels. The novelty in this study is replacing the adhesion film with clear plastic film which was used to cut the design of the microchannel as the material is more suitable for fabricating more complex microchannel design. The microchannel was then mold using polymethyldisiloxane (PDMS and bonded with a clean glass to produce a close microchannel. The microchannel produced had a clean edge indicating good master mold was produced using the cutting plotter and the bonding between the PDMS and glass was good where no leakage was observed. The materials used in this method is cheap and the total time consumed is less than 5 hours where this method is suitable for rapid prototyping of microchannel.

  5. MicroRNA-208a Silencing Attenuates Doxorubicin Induced Myocyte Apoptosis and Cardiac Dysfunction

    Directory of Open Access Journals (Sweden)

    Hasahya Tony


    Full Text Available Aims. GATA4 depletion is a distinct mechanism by which doxorubicin leads to cardiomyocyte apoptosis, and preservation of GATA4 mitigates doxorubicin induced myocyte apoptosis and cardiac dysfunction. We investigated a novel approach of attenuating doxorubicin induced cardiac toxicity by silencing miR-208a, a heart specific microRNA known to target GATA4. Methods and Results. Eight-week-old female Balb/C mice were randomly assigned to sham, antagomir, and control groups. Antagomir group were pretreated with miR-208a antagomir 4 days before doxorubicin administration. At day 0, control and antagomir groups received 20 mg/kg of doxorubicin, while sham mice received phosphate buffered solution. Echocardiography was done at day 7, after which animals were sacrificed and hearts harvested and assessed for apoptosis and expression of miR-208a, GATA4, and BCL-2. Doxorubicin significantly upregulated miR-208a, downregulated GATA4, and increased myocyte apoptosis, with resulting decrease in cardiac function. In contrast, therapeutic silencing of miR-208a salvaged GATA4 and BCL-2 and decreased apoptosis, with improvement in cardiac function. Conclusion. Doxorubicin upregulates miR-208a and promotes cardiomyocyte apoptosis, while therapeutic silencing of miR-208a attenuates doxorubicin induced myocyte apoptosis with subsequent improvement in cardiac function. These novel results highlight the therapeutic potential of targeting miR-208a to prevent doxorubicin cardiotoxicity.

  6. An efficient virus-induced gene silencing vector for maize functional genomics research. (United States)

    Wang, Rong; Yang, Xinxin; Wang, Nian; Liu, Xuedong; Nelson, Richard S; Li, Weimin; Fan, Zaifeng; Zhou, Tao


    Maize is a major crop whose rich genetic diversity provides an advanced resource for genetic research. However, a tool for rapid transient gene function analysis in maize that may be utilized in most maize cultivars has been lacking, resulting in reliance on time-consuming stable transformation and mutation studies to obtain answers. We developed an efficient virus-induced gene silencing (VIGS) vector for maize based on a naturally maize-infecting cucumber mosaic virus (CMV) strain, ZMBJ-CMV. An infectious clone of ZMBJ-CMV was constructed, and a vascular puncture inoculation method utilizing Agrobacterium was optimized to improve its utility for CMV infection of maize. ZMBJ-CMV was then modified to function as a VIGS vector. The ZMBJ-CMV vector induced mild to moderate symptoms in many maize lines, making it useful for gene function studies in critically important maize cultivars, such as the sequenced reference inbred line B73. Using this CMV VIGS system, expression of two endogenous genes, ZmPDS and ZmIspH, was found to be decreased by 75% and 78%, respectively, compared with non-silenced tissue. Inserts with lengths of 100-300 bp produced the most complete transcriptional and visual silencing phenotypes. Moreover, genes related to autophagy, ZmATG3 and ZmATG8a, were also silenced, and it was found that they function in leaf starch degradation. These results indicate that our ZMBJ-CMV VIGS vector provides a tool for rapid and efficient gene function studies in maize. © 2016 The Authors The Plant Journal © 2016 John Wiley & Sons Ltd.

  7. Synthetic versions of firefly luciferase and Renilla luciferase reporter genes that resist transgene silencing in sugarcane (United States)


    Background Down-regulation or silencing of transgene expression can be a major hurdle to both molecular studies and biotechnology applications in many plant species. Sugarcane is particularly effective at silencing introduced transgenes, including reporter genes such as the firefly luciferase gene. Synthesizing transgene coding sequences optimized for usage in the host plant is one method of enhancing transgene expression and stability. Using specified design rules we have synthesised new coding sequences for both the firefly luciferase and Renilla luciferase reporter genes. We have tested these optimized versions for enhanced levels of luciferase activity and for increased steady state luciferase mRNA levels in sugarcane. Results The synthetic firefly luciferase (luc*) and Renilla luciferase (Renluc*) coding sequences have elevated G + C contents in line with sugarcane codon usage, but maintain 75% identity to the native firefly or Renilla luciferase nucleotide sequences and 100% identity to the protein coding sequences. Under the control of the maize pUbi promoter, the synthetic luc* and Renluc* genes yielded 60x and 15x higher luciferase activity respectively, over the native firefly and Renilla luciferase genes in transient assays on sugarcane suspension cell cultures. Using a novel transient assay in sugarcane suspension cells combining co-bombardment and qRT-PCR, we showed that synthetic luc* and Renluc* genes generate increased transcript levels compared to the native firefly and Renilla luciferase genes. In stable transgenic lines, the luc* transgene generated significantly higher levels of expression than the native firefly luciferase transgene. The fold difference in expression was highest in the youngest tissues. Conclusions We developed synthetic versions of both the firefly and Renilla luciferase reporter genes that resist transgene silencing in sugarcane. These transgenes will be particularly useful for evaluating the expression patterns conferred

  8. Anti-viral RNA silencing: do we look like plants ?

    Directory of Open Access Journals (Sweden)

    Lecellier Charles-Henri


    Full Text Available Abstract The anti-viral function of RNA silencing was first discovered in plants as a natural manifestation of the artificial 'co-suppression', which refers to the extinction of endogenous gene induced by homologous transgene. Because silencing components are conserved among most, if not all, eukaryotes, the question rapidly arose as to determine whether this process fulfils anti-viral functions in animals, such as insects and mammals. It appears that, whereas the anti-viral process seems to be similarly conserved from plants to insects, even in worms, RNA silencing does influence the replication of mammalian viruses but in a particular mode: micro(miRNAs, endogenous small RNAs naturally implicated in translational control, rather than virus-derived small interfering (siRNAs like in other organisms, are involved. In fact, these recent studies even suggest that RNA silencing may be beneficial for viral replication. Accordingly, several large DNA mammalian viruses have been shown to encode their own miRNAs. Here, we summarize the seminal studies that have implicated RNA silencing in viral infection and compare the different eukaryotic responses.

  9. Precedents and consequences of prosocial behaviors of voice and silence

    Directory of Open Access Journals (Sweden)

    Alicia Omar


    Full Text Available Prosocial behavior is that which that encourages solidarity and harmony in interpersonal relationships, and produce personal or collective benefits. Although early research on job prosociality was focused on the study of conventional behaviors such as help, courtesy and sportsmanship, the identification and operationalization of new dimensions is rapidly expanding this nomological network. Such is the case of prosocial voice and prosocial silence, recently introduced in the scientific literature. The aim of this study is to explore possible relationships between employee’s voice and employee’s silence, and their personality structure; and examine the role of interpersonal justice perceptions on such relationships. We worked with a sample of 316 employees- aged 37 years and holding a 4.2-year signority working at public and private companies in southern and central Rosario (Argentina. The subjects completed Colquitt’ Justice Organizational Scale, Eysenck Personality Questionnaire, and Van Dyne’s Prosocial Voice and Prosocial Silence Scales. Extraversion and neuroticism emerged as the strongest predictors of prosocial voice and prosocial silence, respectively. Interpersonal justice perceptions emerged as moderators of the ‘natural’ tendency of extraverted workers to engage in prosocial voice, and emotionally controlled workers to engage in prosocial silence. Such findings would indicate that the promotion of high levels of interpersonal justice on job contexts could help workers to engage in more prosocial behavior with positive effects for the organization. 

  10. Designing of highly effective complementary and mismatch siRNAs for silencing a gene. (United States)

    Ahmed, Firoz; Raghava, Gajendra P S


    In past, numerous methods have been developed for predicting efficacy of short interfering RNA (siRNA). However these methods have been developed for predicting efficacy of fully complementary siRNA against a gene. Best of author's knowledge no method has been developed for predicting efficacy of mismatch siRNA against a gene. In this study, a systematic attempt has been made to identify highly effective complementary as well as mismatch siRNAs for silencing a gene.Support vector machine (SVM) based models have been developed for predicting efficacy of siRNAs using composition, binary and hybrid pattern siRNAs. We achieved maximum correlation 0.67 between predicted and actual efficacy of siRNAs using hybrid model. All models were trained and tested on a dataset of 2182 siRNAs and performance was evaluated using five-fold cross validation techniques. The performance of our method desiRm is comparable to other well-known methods. In this study, first time attempt has been made to design mutant siRNAs (mismatch siRNAs). In this approach we mutated a given siRNA on all possible sites/positions with all possible nucleotides. Efficacy of each mutated siRNA is predicted using our method desiRm. It is well known from literature that mismatches between siRNA and target affects the silencing efficacy. Thus we have incorporated the rules derived from base mismatches experimental data to find out over all efficacy of mutated or mismatch siRNAs. Finally we developed a webserver, desiRm ( for designing highly effective siRNA for silencing a gene. This tool will be helpful to design siRNA to degrade disease isoform of heterozygous single nucleotide polymorphism gene without depleting the wild type protein.

  11. Optical silencing of C. elegans cells with light-driven proton pumps. (United States)

    Okazaki, Ayako; Takahashi, Megumi; Toyoda, Naoya; Takagi, Shin


    Recent development of optogenetic techniques, which utilize light-driven ion channels or ion pumps for controlling the activity of excitable cells, has greatly facilitated the investigation of nervous systems in vivo. A new generation of optical silencers includes outward-directed proton pumps, such as Arch, which have several advantages over currently widely used halorhodopsin (NpHR). These advantages include the resistance to inactivation during prolonged illumination and the ability to generate a larger optical current from low intensity light. C. elegans, with its small transparent body and well-characterized neural circuits, is especially suitable for optogenetic analyses. In this article, we will outline the practical aspects of using of Arch and other proton pumps as optogenetic tools in C. elegans. Copyright © 2014 Elsevier Inc. All rights reserved.


    Directory of Open Access Journals (Sweden)

    Fitnat Nazlı Sayğan


    Full Text Available In this study, the affective commitment that is oneof the components oforganizational commitment put forth by Allen and Meyer (Allen, Meyer 1996will be differentiated from the other commitment components. The importance ofcreating an emotional commitment to organizations will be examined and theorganizational factors needed to form organizational commitment will beinvestigated. Also, organizational silence is a situation that the company avoided.In the study, the reasons and the drawbacks of silence are focused on and thefactors that cause employees to remain silent are discussed.The aim of this study is intended to manifest the relationship of ‘organizationalsilence' with affective commitment’ which is one ofthe components of‘organizational commitment’ on the basis of literature. In this study, a negativecorrelation between affective commitment and organizational silence is suggested

  13. RNA silencing can explain chlorotic infection patterns on plant leaves

    Directory of Open Access Journals (Sweden)

    Hogeweg Paulien


    Full Text Available Abstract Background RNA silencing has been implicated in virus symptom development in plants. One common infection symptom in plants is the formation of chlorotic tissue in leaves. Chlorotic and healthy tissue co-occur on a single leaf and form patterns. It has been shown that virus levels in chlorotic tissue are high, while they are low in healthy tissue. Additionally, the presence of siRNAs is confined to the chlorotic spots and the boundaries between healthy and infected tissue. These results strongly indicate that the interaction between virus growth and RNA silencing plays a role in the formation of infection patterns on leaves. However, how RNA silencing leads to the intricate patterns is not known. Results Here we elucidate the mechanisms leading to infection patterns and the conditions which lead to the various patterns observed. We present a modeling approach in which we combine intra- and inter-cellular dynamics of RNA silencing and viral growth. We observe that, due to the spread of viruses and the RNA silencing response, parts of the tissue become infected while other parts remain healthy. As is observed in experiments high virus levels coincide with high levels of siRNAs, and siRNAs are also present in the boundaries between infected and healthy tissue. We study how single- and double-stranded cleavage by Dicer and amplification by RNA-dependent RNA polymerase can affect the patterns formed. Conclusion This work shows that RNA silencing and virus growth within a cell, and the local spread of virions and siRNAs between cells can explain the heterogeneous spread of virus in leaf tissue, and therewith the observed infection patterns in plants.

  14. Re-discovery of silence in pastoral care. (United States)

    Moriichi, Shuji


    Pastoral care within the greater American cultural milieu strongly appraises the use of spoken words and tends to undervalue silence as a powerful and creative element for mutual connection with others and with God. In this essay the author proposes a "counter-cultural" adjustment in the perception of silence and its implications to the practice of, and theological understanding of, pastoral care. The author bases his theses on his own cross-cultural learning experiences, including his training in Clinical Pastoral Education (CPE), in the re-vitalization of ancient Christian wisdom, and in his current ministry to nursing home residents.

  15. Uncertainty in Various Habitat Suitability Models and Its Impact on Habitat Suitability Estimates for Fish

    Directory of Open Access Journals (Sweden)

    Yu-Pin Lin


    Full Text Available Species distribution models (SDMs are extensively used to project habitat suitability of species in stream ecological studies. Owing to complex sources of uncertainty, such models may yield projections with varying degrees of uncertainty. To better understand projected spatial distributions and the variability between habitat suitability projections, this study uses five SDMs that are based on the outputs of a two-dimensional hydraulic model to project the suitability of habitats and to evaluate the degree of variability originating from both differing model types and the split-sample procedure. The habitat suitability index (HSI of each species is based on two stream flow variables, including current velocity (V, water depth (D, as well as the heterogeneity of these flow conditions as quantified by the information entropy of V and D. The six SDM approaches used to project fish abundance, as represented by HSI, included two stochastic models: the generalized linear model (GLM and the generalized additive model (GAM; as well as three machine learning models: the support vector machine (SVM, random forest (RF and the artificial neural network (ANN, and an ensemble model (where the latter is the average of the preceding five models. The target species Sicyopterus japonicas was found to prefer habitats with high current velocities. The relationship between mesohabitat diversity and fish abundance was indicated by the trends in information entropy and weighted usable area (WUA over the study area. This study proposes a method for quantifying habitat suitability, and for assessing the uncertainties in HSI and WUA that are introduced by the various SDMs and samples. This study also demonstrated both the merits of the ensemble modeling approach and the necessity of addressing model uncertainty.

  16. 45 CFR 12a.4 - Suitability determination. (United States)


    ... 45 Public Welfare 1 2010-10-01 2010-10-01 false Suitability determination. 12a.4 Section 12a.4... PROPERTY TO ASSIST THE HOMELESS § 12a.4 Suitability determination. (a) Suitability determination. Within 30... determination to the landholding agency. Properties that are under lease, contract, license, or agreement by...

  17. An Abundant Class of Non-coding DNA Can Prevent Stochastic Gene Silencing in the C. elegans Germline

    DEFF Research Database (Denmark)

    Frøkjær-Jensen, Christian; Jain, Nimit; Hansen, Loren


    Cells benefit from silencing foreign genetic elements but must simultaneously avoid inactivating endogenous genes. Although chromatin modifications and RNAs contribute to maintenance of silenced states, the establishment of silenced regions will inevitably reflect underlying DNA sequence and/or s...

  18. Investigating plasmodesmata genetics with virus-induced gene silencing and an agrobacterium-mediated GFP movement assay. (United States)

    Brunkard, Jacob O; Burch-Smith, Tessa M; Runkel, Anne M; Zambryski, Patricia


    Plasmodesmata (PD) are channels that connect the cytoplasm of adjacent plant cells, permitting intercellular transport and communication. PD function and formation are essential to plant growth and development, but we still know very little about the genetic pathways regulating PD transport. Here, we present a method for assaying changes in the rate of PD transport following genetic manipulation. Gene expression in leaves is modified by virus-induced gene silencing. Seven to ten days after infection with Tobacco rattle virus carrying a silencing trigger, the gene(s) of interest is silenced in newly arising leaves. In these new leaves, individual cells are then transformed with Agrobacterium to express GFP, and the rate of GFP diffusion via PD is measured. By measuring GFP diffusion both within the epidermis and between the epidermis and mesophyll, the assay can be used to study the effects of silencing a gene(s) on PD transport in general, or transport through secondary PD specifically. Plant biologists working in several fields will find this assay useful, since PD transport impacts plant physiology, development, and defense.

  19. Gene Overexpression and RNA Silencing Tools for the Genetic Manipulation of the S-(+)-Abscisic Acid Producing Ascomycete Botrytis cinerea. (United States)

    Ding, Zhong-Tao; Zhang, Zhi; Luo, Di; Zhou, Jin-Yan; Zhong, Juan; Yang, Jie; Xiao, Liang; Shu, Dan; Tan, Hong


    The phytopathogenic ascomycete Botrytis cinerea produces several secondary metabolites that have biotechnical significance and has been particularly used for S-(+)-abscisic acid production at the industrial scale. To manipulate the expression levels of specific secondary metabolite biosynthetic genes of B. cinerea with Agrobacterium tumefaciens-mediated transformation system, two expression vectors (pCBh1 and pCBg1 with different selection markers) and one RNA silencing vector, pCBSilent1, were developed with the In-Fusion assembly method. Both expression vectors were highly effective in constitutively expressing eGFP, and pCBSilent1 effectively silenced the eGFP gene in B. cinerea. Bcaba4, a gene suggested to participate in ABA biosynthesis in B. cinerea, was then targeted for gene overexpression and RNA silencing with these reverse genetic tools. The overexpression of bcaba4 dramatically induced ABA formation in the B. cinerea wild type strain Bc-6, and the gene silencing of bcaba4 significantly reduced ABA-production in an ABA-producing B. cinerea strain.

  20. HvCKX2 gene silencing by biolistic or Agrobacterium-mediated transformation in barley leads to different phenotypes. (United States)

    Zalewski, Wojciech; Orczyk, Wacław; Gasparis, Sebastian; Nadolska-Orczyk, Anna


    CKX genes encode cytokinin dehydrogenase enzymes (CKX), which metabolize cytokinins in plants and influence developmental processes. The genes are expressed in different tissues and organs during development; however, their exact role in barley is poorly understood. It has already been proven that RNA interference (RNAi)-based silencing of HvCKX1 decreased the CKX level, especially in those organs which showed the highest expression, i.e. developing kernels and roots, leading to higher plant productivity and higher mass of the roots [1]. The same type of RNAi construct was applied to silence HvCKX2 and analyze the function of the gene. Two cultivars of barley were transformed with the same silencing and selection cassettes by two different methods: biolistic and via Agrobacterium. The mean Agrobacterium-mediated transformation efficiency of Golden Promise was 3.47% (±2.82). The transcript level of HvCKX2 in segregating progeny of T(1) lines was decreased to 34%. The reduction of the transcript in Agrobacterium-derived plants resulted in decreased CKX activity in the developing and developed leaves as well as in 7 DAP (days after pollination) spikes. The final phenotypic effect was increased productivity of T(0) plants and T(1) lines. Higher productivity was the result of the higher number of seeds and higher grain yield. It was also correlated with the higher 1000 grain weight, increased (by 7.5%) height of the plants and higher (from 0.5 to 2) numbers of spikes. The transformation efficiency of Golden Promise after biolistic transformation was more than twice as low compared to Agrobacterium. The transcript level in segregating progeny of T(1) lines was decreased to 24%. Otherwise, the enzyme activity found in the leaves of the lines after biolistic transformation, especially in cv. Golden Promise, was very high, exceeding the relative level of the control lines. These unbalanced ratios of the transcript level and the activity of the CKX enzyme negatively

  1. Assessing the tobacco-rattle-virus-based vectors system as an efficient gene silencing technique in Datura stramonium (Solanaceae). (United States)

    Eftekhariyan Ghamsari, Mohammad Reza; Karimi, Farah; Mousavi Gargari, Seyed Latif; Hosseini Tafreshi, Seyed Ali; Salami, Seyed Alireza


    Datura stramonium is a well-known medicinal plant, which is important for its alkaloids. There are intrinsic limitations for the natural production of alkaloids in plants; metabolic engineering methods can be effectively used to conquer these limitations. In order for this the genes involved in corresponding pathways need to be studied. Virus-Induced Gene Silencing is known as a functional genomics technique to knock-down expression of endogenous genes. In this study, we silenced phytoene desaturase as a marker gene in D. stramonium in a heterologous and homologous manner by tobacco-rattle-virus-based VIGS vectors. Recombinant TRV vector containing pds gene from D. stramonium (pTRV2-Dspds) was constructed and injected into seedlings. The plants injected with pTRV2-Dspds showed photobleaching 2 weeks after infiltration. Spectrophotometric analysis demonstrated that the amount of chlorophylls and carotenoids in leaves of the bleached plants decreased considerably compared to that of the control plants. Semi-Quantitative RT-PCR results also confirmed that the expression of pds gene in the silenced plants was significantly reduced in comparison with the control plants. The results showed that the viral vector was able to influence the levels of total alkaloid content in D. stramonium. Our results illustrated that TRV-based VIGS vectors are able to induce effective and reliable functional gene silencing in D. stramonium as an alternative tool for studying the genes of interest in this plant, such as the targeted genes in tropane alkaloid biosynthetic pathway. The present work is the first report of establishing VIGS as an efficient method for transient silencing of any gene of interest in D. stramonium.

  2. Silence about encounters with dying among healthcare professionals in a society that ‘de-tabooises’ death

    Directory of Open Access Journals (Sweden)

    Ellen Ramvi


    Full Text Available Background: Empirical studies on healthcare personnel indicate that professionals’ experiences with dying and death become silenced and unutterable within the healthcare service. Aim: To explore and interpret silence about encounters with death and dying among healthcare professionals in Norway. Method: The method used was theoretical exploration, using a psychosocial approach. Findings: This analysis reveals complex interrelations and two-way dynamics between subject-worlds, sociocultural and societal worlds when it comes to dealing with death and dying at work. A performance culture saturates these worlds, and may be implicated in silencing death within the healthcare institutions of the Norwegian welfare state. Conclusions: This article suggests that silence about death and dying among healthcare professionals is indicative of crucial emerging and unresolved tensions in the neoliberal episteme, accompanied and reinforced by the ineluctable basic conditions of life and intrapsychic defence against threats towards the self. Implications for practice: Silence about death and dying presents a serious challenge for dying patients and next of kin. Healthcare professionals should be enabled to acknowledge their thoughts and emotions about death in order to be able to support and contain patients and next of kin Learning activities such as peer support and supervision can help the processing of difficult psychological content and allow for emotional aspects of professionals’ work to be acknowledged and thought about in a way that encourages reflective and sound practice Clinical managers should address whether performance pressures induce shameful feelings in staff, who may believe that by providing appropriate levels of care they are compromising productivity. Shame in turn, may undermine professionals’ emotional wellbeing and ability to continue to provide attuned and adequate care for dying patients Creative approaches to facilitate

  3. Small RNA binding is a common strategy to suppress RNA silencing by several viral suppressors (United States)

    Lakatos, Lóránt; Csorba, Tibor; Pantaleo, Vitantonio; Chapman, Elisabeth J; Carrington, James C; Liu, Yu-Ping; Dolja, Valerian V; Calvino, Lourdes Fernández; López-Moya, Juan José; Burgyán, József


    RNA silencing is an evolutionarily conserved system that functions as an antiviral mechanism in higher plants and insects. To counteract RNA silencing, viruses express silencing suppressors that interfere with both siRNA- and microRNA-guided silencing pathways. We used comparative in vitro and in vivo approaches to analyse the molecular mechanism of suppression by three well-studied silencing suppressors. We found that silencing suppressors p19, p21 and HC-Pro each inhibit the intermediate step of RNA silencing via binding to siRNAs, although the molecular features required for duplex siRNA binding differ among the three proteins. None of the suppressors affected the activity of preassembled RISC complexes. In contrast, each suppressor uniformly inhibited the siRNA-initiated RISC assembly pathway by preventing RNA silencing initiator complex formation. PMID:16724105

  4. Silencing Huntington's chorea: Is RNA Interference a Potential Cure?

    Directory of Open Access Journals (Sweden)

    Gerlinde A. Metz


    Full Text Available In 1872, George Huntington described Huntington's disease as characterized by motor, cognitive and psychiatric impairments. Huntington's disease is a dominant and autosomal mutation on chromosome 4 featuring the insertion of numerous CAG repeats. CAG codes for the amino acid, glutmanine that forms part of the Huntingtin protein (htt. Excess glutamine attachments make htt prone to accumulate in neurons. Three genes can be considered when developing therapies for Huntington's disease. They include targeting the symptoms of the disease, the progression of the disease and the cause of the disease. By using RNA interference (RNAi, the cause of the disease can be targeted. RNAi is a method that could potentially silence the formation of abnormal htt. This paper will discuss how RNAi could potentially cure Huntington's disease, by describing the genetic and proteinomic basis of Huntington's disease, the function of RNAi in Huntington's disease and the problems of benefits of RNAi. Preliminary work using RNAi in transgenic mice has shown a decrease in the behavioural expression of the mutant Huntington gene. There are several limitations associated with using RNAi as a gene therapy. For example, the effects of RNAi are short lived. A transposition system such as Sleeping Beauty can be used to increase the integration of the gene, however, for patients who currently have Huntington's disease, RNAi may potentially be used in combination with drugs or other treatments to target both symptoms and the underlying cause of Huntington's disease. This combination could eventually alleviate many painful symptoms associated with Huntington's disease and could even stop the progressive neurodegeneration of Huntington's disease. This review concludes that a substantial amount of new research is still necessary before RNAi is directly applicable to human patients with Huntington's disease.

  5. Suppressors of RNA silencing encoded by tomato leaf curl ...

    Indian Academy of Sciences (India)


    Jan 6, 2013 ... step in the RNA-silencing pathway that occurs after siRNA production (Zrachya et al. 2007). Geminiviruses (family Geminiviridae) are a diverse group of plant viruses with circular single-stranded DNA genomes that are composed of one or two components of 2700–. 3000 bp length which are encapsidated ...

  6. Organizational Justice As a Predictor of Organizational Silence (United States)

    Tan, Çetin


    In this study, relation between teachers' perception for organizational justice and their organizational silence was examined. Sample of this study consists of 300 teachers who work at elementary schools in Siirt. Relational Scanning model was utilized in performance of this study. In this study, Organizational Justice Scale and Organizational…

  7. For the Sake of Silence | Kearney | Current Writing: Text and ...

    African Journals Online (AJOL)

    The Order of Cistercians of the Strict Observance – better known as “Trappists” after their famous monastery, Notre Dame de la Trappe – is the strictest and most ascetic Order in the Catholic Church. In addition to vows of poverty, chastity and obedience taken by all religious, Trappists bind themselves to silence and stability ...

  8. Survivors' Voices Breaking the Silence: Owning the Past. (United States)

    Benton, Carol L.

    For the adult survivor of incest, the evolution from victim to survivor occurs in complex and creative ways. Often journal writing can be a means of exposing and exploring past abuse. The process of journal writing is special in that it allows the adult survivor to break the silence and shame imposed by the perpetrator while remaining essentially…

  9. Bureaucratic Constructions of Sexual Diversity: "Sensitive", "Controversial" and Silencing (United States)

    Ullman, Jacqueline; Ferfolja, Tania


    National research illustrates the high degree of discrimination that prevails against lesbian, gay, bisexual, transgender and queer (LGBTQ) students resulting in diminished educational outcomes, both academic and social. This phenomenon is influenced by the prevalence of whole-school silences around LGBTQ topics in many Australian schools. This…

  10. Identification of a maize chlorotic dwarf virus silencing suppressor protein. (United States)

    Stewart, Lucy R; Jarugula, Sridhar; Zhao, Yujing; Qu, Feng; Marty, DeeMarie


    Maize chlorotic dwarf virus (MCDV), a member of the genus Waikavirus, family Secoviridae, has a 11784 nt (+)ssRNA genome that encodes a 389kDa proteolytically processed polyprotein. We show that the N-terminal 78kDa polyprotein (R78) of MCDV acts as a suppressor of RNA silencing in a well-established assay system. We further demonstrate that R78 is cleaved by the viral 3C-like protease into 51 and 27kDa proteins (p51 and p27), and that p51 is responsible for silencing suppressor activity. Silencing suppressor activity of R78 is conserved in three divergent MCDV strains (MCDV-Severe, MCDV-M1, and MCDV-Tennessee), as well as the waikavirus Bellflower vein chlorosis virus, but was not detected for orthologous protein of Rice tungro spherical virus (RTSV-A) or the similarly-positioned protein from the sequivirus Parsnip yellow fleck virus (PYFV). This is the first identification of a virus suppressor of RNA silencing encoded by a waikavirus. Copyright © 2017. Published by Elsevier Inc.

  11. The effects of thinking in silence on creativity and innovation

    NARCIS (Netherlands)

    de Vet, A.J.


    This dissertation consists of three empirical studies on the effects of thinking in silence on creativity and innovation. In these studies I use a social psychology and cognitive psychology lens to study creativity and innovation at the individual and at the team level of analysis, using randomized

  12. RNAi-based silencing of genes encoding the vacuolar- ATPase ...

    African Journals Online (AJOL)

    RNAi-based silencing of genes encoding the vacuolar- ATPase subunits a and c in pink bollworm (Pectinophora gossypiella). Ahmed M. A. Mohammed. Abstract. RNA interference is a post- transcriptional gene regulation mechanism that is predominantly found in eukaryotic organisms. RNAi demonstrated a successful ...

  13. 47 CFR 80.304 - Watch requirement during silence periods. (United States)


    ....304 Section 80.304 Telecommunication FEDERAL COMMUNICATIONS COMMISSION (CONTINUED) SAFETY AND SPECIAL RADIO SERVICES STATIONS IN THE MARITIME SERVICES Safety Watch Requirements and Procedures Ship Station Safety Watches § 80.304 Watch requirement during silence periods. Each ship station operating on...

  14. Silence is Golden: The Lack of Direction on Compensation for ...

    African Journals Online (AJOL)

    Silence is Golden: The Lack of Direction on Compensation for Expropriation in the 2011 Green Paper on Land Reform. ... Frustrated at the slow pace of land reform, the ruling party is starting to indicate that the state will in future rely on its expropriation powers to acquire such land. Section 25 of the Constitution makes it ...

  15. Maternal Silences: Motherhood and Voluntary Childlessness in Contemporary Christianity

    Directory of Open Access Journals (Sweden)

    Dawn Llewellyn


    Full Text Available In Christianity, there is an ideology of motherhood that pervades scripture, ritual, and doctrine, yet there is an academic silence that means relatively little space has been given to motherhood and mothering, and even less to voluntary childlessness, from a faith perspective. By drawing on qualitative in-depth interviews with Christian women living in Britain, narrating their experiences of motherhood and voluntary childlessness, I suggest there are also lived maternal silences encountered by women in contemporary Christianity. There is a maternal expectation produced through church teaching, liturgy and culture that constructs women as ‘maternal bodies’ (Gatrell 2008; this silences and marginalises women from articulating their complex relationship with religion, motherhood, and childlessness in ways that challenge their spiritual development. However, this article also introduces the everyday and intentional tactics women employ to disrupt the maternal expectation, and hereby interrupt the maternal silence.

  16. Shame and Silence | Janz | South African Journal of Philosophy

    African Journals Online (AJOL)

    Samantha Vice's proposal on how to live in 'this strange place' of contemporary South Africa, includes an appeal to the concepts of shame and silence. In this paper, I use Emmanuel Levinas and Giorgio Agamben to move the discussion of shame from a moral to an existential question. The issue is not about how one ...

  17. Identification of a maize chlorotic dwarf virus silencing suppressor protein (United States)

    Maize chlorotic dwarf virus (MCDV), a member of the genus Waikavirus, family Secoviridae, has a 11784 nt (+)ssRNA genome that encodes a 389 kDa proteolytically processed polyprotein. We show that an N-terminal 78kDa polyprotein (R78) has silencing suppressor activity, that it is cleaved by the viral...

  18. Relationship between Organizational Mobbing and Silence Behavior among Teachers (United States)

    Hüsrevsahi, Selda Polat


    This study mainly aims to investigate the correlation between teachers' exposure to mobbing in their workplaces and their display of the act of silence. This study is based on a survey design where data from 312 teachers were collected and analyzed using correlation and regression analyses. Specifically, "The Structure and Dimensions of…

  19. Inhibition of tobacco axillary bud differentiation by silencing CUP ...

    African Journals Online (AJOL)



    Feb 23, 2012 ... exhibited some important phenotypes, such as leaf fusion, leaf curl and incomplete leaf. CUC3 in N. tabacum was silenced successfully by RNA interference (RNAi), which .... 0.1 mg NAA L-1 and 1.0 mg BAP L-1 was used as co-culture medium. Co-culture medium with hygromycin B 50 mg hygromycin.

  20. Studies of the silencing of Baculovirus DNA binding protein

    NARCIS (Netherlands)

    Quadt, I.; Lent, van J.W.M.; Knebel-Morsdorf, D.


    Baculovirus DNA binding protein (DBP) binds preferentially single-stranded DNA in vitro and colocalizes with viral DNA replication sites. Here, its putative role as viral replication factor has been addressed by RNA interference. Silencing of DBP in Autographa californica multiple

  1. Tacit Authorization : A Legal Solution for Administrative Silence

    NARCIS (Netherlands)

    Hoogstra, Nicole; de Graaf, K.J.


    This article discusses one of the current legal instruments to stimulate timely decision-making by administrative authorities, namely the ‘Lex silencio positivo’ or the ‘Silence is Consent’ rule. Tacit authorization prescribes that the license sought by the applicant will be granted automatically if

  2. Suppressors of RNA silencing encoded by tomato leaf curl ...

    Indian Academy of Sciences (India)

    In the present study suppressor function of ORF C1 of three betasatellites Tomato leaf curl Bangalore betasatellite ToLCBB-[IN:Hess:08], Cotton leaf curl Multan betasatellite CLCuMB–[IN:Sri:02] and Luffa leaf distortion betasatellite LuLDB-[IN:Lu:04] were examined. Agroinfiltration of GFP-silenced Nicotiana tabaccum cv.

  3. Mobile gene silencing in Arabidopsis is regulated by hydrogen peroxide

    Directory of Open Access Journals (Sweden)

    Dacheng Liang


    Full Text Available In plants and nematodes, RNAi can spread from cells from which it is initiated to other cells in the organism. The underlying mechanism controlling the mobility of RNAi signals is not known, especially in the case of plants. A genetic screen designed to recover plants impaired in the movement but not the production or effectiveness of the RNAi signal identified RCI3, which encodes a hydrogen peroxide (H2O2-producing type III peroxidase, as a key regulator of silencing mobility in Arabidopsis thaliana. Silencing initiated in the roots of rci3 plants failed to spread into leaf tissue or floral tissue. Application of exogenous H2O2 reinstated the spread in rci3 plants and accelerated it in wild-type plants. The addition of catalase or MnO2, which breaks down H2O2, slowed the spread of silencing in wild-type plants. We propose that endogenous H2O2, under the control of peroxidases, regulates the spread of gene silencing by altering plasmodesmata permeability through remodelling of local cell wall structure, and may play a role in regulating systemic viral defence.

  4. Is our knowledge of voice and silence in organizations growing?

    DEFF Research Database (Denmark)

    Knoll, Michael; Wegge, Jürgen; Unterrainer, Christine


    This article has three objectives. Firstly, we seek to demonstrate the relevance of voice and silence – that is, whether employees contribute or withhold information, ideas, views and/or concerns at work – for the sustainable development of individuals, organizations and societies. Our second obj...

  5. Silencing of HMGA2 promotes apoptosis and inhibits migration and ...

    Indian Academy of Sciences (India)


    Apr 5, 2016 ... Silencing of HMGA2 promotes apoptosis and inhibits migration and invasion of prostate cancer cells. ZHAN SHI, DING WU, RUN TANG, XIANG LI, RENFU CHEN, SONG XUE, CHENGJING ZHANG and XIAOQING SUN*. Department of Urology, The Affiliated Hospital of Xuzhou Medical College,. Xuzhou ...

  6. RNAi-based silencing of genes encoding the vacuolar- ATPase ...

    African Journals Online (AJOL)


    Nov 9, 2016 ... Spodoptera exigua larval development by silencing chitin synthase gene with RNA interference. Bull. Entomol. Res. 98:613-619. Dow JAT (1999). The Multifunctional Drosophila melanogaster V-. ATPase is encoded by a multigene family. J. Bioenerg. Biomembr. 31:75-83. Fire A, Xu SQ, Montgomery MK, ...

  7. Breaking the Silence Surrounding Mental Health on Campus (United States)

    Roper, Larry D.


    Mentally ill students are able to participate in higher education at unprecedented rates. While colleges and universities have been responsive to the therapeutic needs, we have failed to successfully create supportive campus climates. Campus leaders are challenged to demonstrate ethical leadership that breaks the silence and confronts the stigma…

  8. Suppressors of RNA silencing encoded by tomato leaf curl ...

    Indian Academy of Sciences (India)


    Jan 6, 2013 ... satellite DNA β which are predicted to function as silencing suppressors. In the present study suppressor function ... of plant viruses with circular single-stranded DNA genomes that are composed of one or two components of ..... 1 2 3 4 5 6 7 8 9 10 11 12 13 14 M. Figure 4. Southern blot analysis of DNA ...

  9. The code of silence: Revisiting South African police integrity | Ivkovic ...

    African Journals Online (AJOL)

    ... results provide further evidence of the presence of the code of silence covering various forms of police misconduct. At least one quarter of the respondents would protect a fellow officer who verbally abused citizens, covered up police driving under the influence (DUI) accident, accepted gratuities, or failed to react to graffiti.

  10. Strain Specific Factors Control Effector Gene Silencing in Phytophthora sojae.

    Directory of Open Access Journals (Sweden)

    Sirjana Devi Shrestha

    Full Text Available The Phytophthora sojae avirulence gene Avr3a encodes an effector that is capable of triggering immunity on soybean plants carrying the resistance gene Rps3a. P. sojae strains that express Avr3a are avirulent to Rps3a plants, while strains that do not are virulent. To study the inheritance of Avr3a expression and virulence towards Rps3a, genetic crosses and self-fertilizations were performed. A cross between P. sojae strains ACR10 X P7076 causes transgenerational gene silencing of Avr3a allele, and this effect is meiotically stable up to the F5 generation. However, test-crosses of F1 progeny (ACR10 X P7076 with strain P6497 result in the release of silencing of Avr3a. Expression of Avr3a in the progeny is variable and correlates with the phenotypic penetrance of the avirulence trait. The F1 progeny from a direct cross of P6497 X ACR10 segregate for inheritance for Avr3a expression, a result that could not be explained by parental imprinting or heterozygosity. Analysis of small RNA arising from the Avr3a gene sequence in the parental strains and hybrid progeny suggests that the presence of small RNA is necessary but not sufficient for gene silencing. Overall, we conclude that inheritance of the Avr3a gene silenced phenotype relies on factors that are variable among P. sojae strains.

  11. Marie Nimier, au cœur du silence

    Directory of Open Access Journals (Sweden)

    Joëlle Papillon


    Full Text Available Dans La Reine du silence, Marie Nimier se confronte à la figure de son père, l’écrivain Roger Nimier, mort lorsqu’elle avait cinq ans. Elle y montre le poids qui pèse sur l’enfant d’écrivain, mais aussi celui de l’héritage du secret familial et de l’injonction au silence. La difficulté de l’élaboration de son récit de filiation se révèle dans les constants recommencements et reformulations, qui constituent la marque de la tension angoissante entre l’obligation de dire et celle de taire. In La Reine du silence, Marie Nimier confronts her father’s memory – the writer Roger Nimier, who died when she was five years old. The novel describes the burden of being a writer’s child, along with that of inheriting family secrets and submitting to a code of silence. The difficulty of recounting her relationship with her late father is evidenced by the narrator’s numerous “false starts” and her constant rewritings. The hesitant nature of the narration captures an anguish born of two irreconcilable obligations : the need to put things into words and the pressure to remain silent.

  12. Assessment of RNAi-induced silencing in banana (Musa spp.). (United States)

    Dang, Tuong Vi T; Windelinckx, Saskia; Henry, Isabelle M; De Coninck, Barbara; Cammue, Bruno P A; Swennen, Rony; Remy, Serge


    In plants, RNA- based gene silencing mediated by small RNAs functions at the transcriptional or post-transcriptional level to negatively regulate target genes, repetitive sequences, viral RNAs and/or transposon elements. Post-transcriptional gene silencing (PTGS) or the RNA interference (RNAi) approach has been achieved in a wide range of plant species for inhibiting the expression of target genes by generating double-stranded RNA (dsRNA). However, to our knowledge, successful RNAi-application to knock-down endogenous genes has not been reported in the important staple food crop banana. Using embryogenic cell suspension (ECS) transformed with ß-glucuronidase (GUS) as a model system, we assessed silencing of gusAINT using three intron-spliced hairpin RNA (ihpRNA) constructs containing gusAINT sequences of 299-nt, 26-nt and 19-nt, respectively. Their silencing potential was analysed in 2 different experimental set-ups. In the first, Agrobacterium-mediated co-transformation of banana ECS with a gusAINT containing vector and an ihpRNA construct resulted in a significantly reduced GUS enzyme activity 6-8 days after co-cultivation with either the 299-nt and 19-nt ihpRNA vectors. In the second approach, these ihpRNA constructs were transferred to stable GUS-expressing ECS and their silencing potential was evaluated in the regenerated in vitro plants. In comparison to control plants, transgenic plants transformed with the 299-nt gusAINT targeting sequence showed a 4.5 fold down-regulated gusA mRNA expression level, while GUS enzyme activity was reduced by 9 fold. Histochemical staining of plant tissues confirmed these findings. Northern blotting used to detect the expression of siRNA in the 299-nt ihpRNA vector transgenic in vitro plants revealed a negative relationship between siRNA expression and GUS enzyme activity. In contrast, no reduction in GUS activity or GUS mRNA expression occurred in the regenerated lines transformed with either of the two gusAINT oligo target

  13. "I Don't Want to Hear That!": Legitimating Whiteness through Silence in Schools (United States)

    Castagno, Angelina E.


    In this article, I examine the ways in which silences around race contribute to the maintenance and legitimation of Whiteness. Drawing on ethnographic data from two demographically different schools, I highlight patterns of racially coded language, teacher silence, silencing students' race talk, and the conflating of culture with race, equality…

  14. Eloquent silences: A musical and lexical analysis of conversation between oncologists and their patients. (United States)

    Bartels, Josef; Rodenbach, Rachel; Ciesinski, Katherine; Gramling, Robert; Fiscella, Kevin; Epstein, Ronald


    Silences in doctor-patient communication can be "connectional" and communicative, in contrast to silences that indicate awkwardness or distraction. Musical and lexical analyses can identify and characterize connectional silences in consultations between oncologists and patients. Two medical students and a professor of voice screened all 1211 silences over 2s in length from 124 oncology office visits. We developed a "strength of connection" taxonomy and examined ten connectional silences for lexical and musical features including pitch, volume, and speaker turn-taking rhythm. We identified connectional silences with good reliability. Typical dialog rhythms surrounding connectional silences are characterized by relatively equal turn lengths and frequent short vocalizations. We found no pattern of volume and pitch variability around these silences. Connectional silences occurred in a wide variety of lexical contexts. Particular patterns of dialog rhythm mark connectional silences. Exploring structures of connectional silence extends our understanding of the audio-linguistic conditions that mark patient-clinician connection. Communicating with an awareness of pitch, rhythm, and silence - in addition to lexical content - can facilitate shared understanding and emotional connection. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  15. RNA-mediated gene silencing of superoxide dismutase (bcsod1) in Botrytis cinerea

    NARCIS (Netherlands)

    Patel, R.M.; Kan, van J.A.L.; Bailey, A.M.; Foster, G.D.


    Gene silencing is a powerful tool utilized for identification of gene function and analysis in plants, animals, and fungi. Here, we report the silencing of superoxide dismutase (bcsod1) in Botrytis cinerea through sense and antisense-mediated silencing mechanisms. Because superoxide dismutase (SOD)

  16. Simultaneous Silencing of Xylanase Genes in Botrytis cinerea

    Directory of Open Access Journals (Sweden)

    Néstor García


    Full Text Available The endo-β-1,4-xylanase BcXyn11A is one of several plant cell-wall degrading enzymes that the phytopathogenic fungus Botrytis cinerea secretes during interaction with its hosts. In addition to its enzymatic activity, this protein also acts as an elicitor of the defense response in plants and has been identified as a virulence factor. In the present work, other four endoxylanase coding genes (Bcxyn11B, Bcxyn11C, Bcxyn10A, and Bcxyn10B were identified in the B. cinerea genome and the expression of all five genes was analyzed by Q-RT- PCR in vitro and in planta. A cross-regulation between xylanase genes was identified analyzing their expression pattern in the ΔBcxyn11A mutant strain and a putative BcXyn11A-dependt induction of Bcxyn10B gene was found. In addition, multiple knockdown strains were obtained for the five endoxylanase genes by transformation of B. cinerea with a chimeric DNA construct composed of 50-nt sequences from the target genes. The silencing of each xylanase gene was analyzed in axenic cultures and during infection and the results showed that the efficiency of the multiple silencing depends on the growth conditions and on the cross-regulation between them. Although the simultaneous silencing of the five genes was observed by Q-RT-PCR when the silenced strains were grown on medium supplemented with tomato extract, the endoxylanase activity measured in the supernatants was reduced only by 40%. Unexpectedly, the silenced strains overexpressed the Bcxyn11A and Bcxyn11C genes during the infection of tomato leaves, making difficult the analysis of the role of the endo-β-1,4-xylanases in the virulence of the fungus.

  17. Increasing the amylose content of durum wheat through silencing of the SBEIIa genes

    Directory of Open Access Journals (Sweden)

    Masci Stefania


    Full Text Available Abstract Background High amylose starch has attracted particular interest because of its correlation with the amount of Resistant Starch (RS in food. RS plays a role similar to fibre with beneficial effects for human health, providing protection from several diseases such as colon cancer, diabetes, obesity, osteoporosis and cardiovascular diseases. Amylose content can be modified by a targeted manipulation of the starch biosynthetic pathway. In particular, the inactivation of the enzymes involved in amylopectin synthesis can lead to the increase of amylose content. In this work, genes encoding starch branching enzymes of class II (SBEIIa were silenced using the RNA interference (RNAi technique in two cultivars of durum wheat, using two different methods of transformation (biolistic and Agrobacterium. Expression of RNAi transcripts was targeted to the seed endosperm using a tissue-specific promoter. Results Amylose content was markedly increased in the durum wheat transgenic lines exhibiting SBEIIa gene silencing. Moreover the starch granules in these lines were deformed, possessing an irregular and deflated shape and being smaller than those present in the untransformed controls. Two novel granule bound proteins, identified by SDS-PAGE in SBEIIa RNAi lines, were investigated by mass spectrometry and shown to have strong homologies to the waxy proteins. RVA analysis showed new pasting properties associated with high amylose lines in comparison with untransformed controls. Finally, pleiotropic effects on other starch genes were found by semi-quantitative and Real-Time reverse transcription-polymerase chain reaction (RT-PCR. Conclusion We have found that the silencing of SBEIIa genes in durum wheat causes obvious alterations in granule morphology and starch composition, leading to high amylose wheat. Results obtained with two different methods of transformation and in two durum wheat cultivars were comparable.

  18. Expression of RNA-interference/antisense transgenes by the cognate promoters of target genes is a better gene-silencing strategy to study gene functions in rice. (United States)

    Li, Jing; Jiang, Dagang; Zhou, Hai; Li, Feng; Yang, Jiawei; Hong, Laifa; Fu, Xiao; Li, Zhibin; Liu, Zhenlan; Li, Jianming; Zhuang, Chuxiong


    Antisense and RNA interference (RNAi)-mediated gene silencing systems are powerful reverse genetic methods for studying gene function. Most RNAi and antisense experiments used constitutive promoters to drive the expression of RNAi/antisense transgenes; however, several reports showed that constitutive promoters were not expressed in all cell types in cereal plants, suggesting that the constitutive promoter systems are not effective for silencing gene expression in certain tissues/organs. To develop an alternative method that complements the constitutive promoter systems, we constructed RNAi and/or antisense transgenes for four rice genes using a constitutive promoter or a cognate promoter of a selected rice target gene and generated many independent transgenic lines. Genetic, molecular, and phenotypic analyses of these RNAi/antisense transgenic rice plants, in comparison to previously-reported transgenic lines that silenced similar genes, revealed that expression of the cognate promoter-driven RNAi/antisense transgenes resulted in novel growth/developmental defects that were not observed in transgenic lines expressing constitutive promoter-driven gene-silencing transgenes of the same target genes. Our results strongly suggested that expression of RNAi/antisense transgenes by cognate promoters of target genes is a better gene-silencing approach to discovery gene function in rice.

  19. Design And Construction Of A Die Casting Mould Suitable For ...

    African Journals Online (AJOL)

    Efforts have been made to group the various types of key that are most commonly used by the public at large. A suitable die casting method is used taking into account such limitations as the sizes of the keys, their serviceability, rate of production, as they affect the method of producing the unit price of the key blanks and the ...

  20. Efficient and nontoxic biological response carrier delivering TNF-α shRNA for gene silencing in a murine model of rheumatoid arthritis

    Directory of Open Access Journals (Sweden)

    Jialin Song


    Full Text Available Small interfering RNA (siRNA is an effective and specific method for silencing genes. However, an efficient and nontoxic carrier is needed to deliver the siRNA into the target cells. Tumor necrosis factor α (TNF-α plays a central role in the occurrence and progression of rheumatoid arthritis. In this study, we pre-synthetized a degradable cationic polymer (PDAPEI from 2,6-pyridinedicarboxaldehyde and low molecular weight polyethyleneimine (PEI, Mw=1.8 kDa as a gene vector for the delivery of TNF-α shRNA. The PDAPEI/pDNA complex showed a suitable particle size and stable zeta potential for transfection. In vitro study of the PDAPEI/pDNA complex revealed a lower cytotoxicity and higher transfection efficiency when transfecting TNF-α shRNA to macrophages by significantly down-regulating the expression of TNF-α. Moreover, the complex was extremely efficient in decreasing the severity of arthritis in mice with collagen-induced arthritis (CIA. PDAPEI delivered TNF-α shRNA has great potential in the treatment of rheumatoid arthritis.

  1. Silencing the SpMPK1, SpMPK2, and SpMPK3 Genes in Tomato Reduces Abscisic Acid—Mediated Drought Tolerance

    Directory of Open Access Journals (Sweden)

    Yan Liang


    Full Text Available Drought is a major threat to agriculture production worldwide. Mitogen-activated protein kinases (MAPKs play a pivotal role in sensing and converting stress signals into appropriate responses so that plants can adapt and survive. To examine the function of MAPKs in the drought tolerance of tomato plants, we silenced the SpMPK1, SpMPK2, and SpMPK3 genes in wild-type plants using the virus-induced gene silencing (VIGS method. The results indicate that silencing the individual genes or co-silencing SpMPK1, SpMPK2, and SpMPK3 reduced the drought tolerance of tomato plants by varying degrees. Co-silencing SpMPK1 and SpMPK2 impaired abscisic acid (ABA-induced and hydrogen peroxide (H2O2-induced stomatal closure and enhanced ABA-induced H2O2 production. Similar results were observed when silencing SpMPK3 alone, but not when SpMPK1 and SpMPK2 were individually silenced. These data suggest that the functions of SpMPK1 and SpMPK2 are redundant, and they overlap with that of SpMPK3 in drought stress signaling pathways. In addition, we found that SpMPK3 may regulate H2O2 levels by mediating the expression of CAT1. Hence, SpMPK1, SpMPK2, and SpMPK3 may play crucial roles in enhancing tomato plants’ drought tolerance by influencing stomatal activity and H2O2 production via the ABA-H2O2 pathway.

  2. Alligator, Alligator mississippiensis, habitat suitability index model (United States)

    Waddle, J. Hardin


    The 2012 Coastal Master Plan utilized Habitat Suitability Indices (HSIs) to evaluate potential project effects on wildlife species. Even though HSIs quantify habitat condition, which may not directly correlate to species abundance, they remain a practical and tractable way to assess changes in habitat quality from various restoration actions. As part of the legislatively mandated five year update to the 2012 plan, the wildlife habitat suitability indices were updated and revised using literature and existing field data where available. The outcome of these efforts resulted in improved, or in some cases entirely new suitability indices. This report describes the development of the habitat suitability indices for the American alligator, Alligator mississippiensis.

  3. Toward a science of silence: The consequences of leaving a memory unsaid

    DEFF Research Database (Denmark)

    Stone, Charles; Coman, Alin; Brown, Adam


    Silence about the past permeates acts of remembering, with marked mnemonic consequences. Mnemonic silence—the absence of expressing a memory—is public in nature and is embedded within communicative acts, such as conversations. As such, silence has the potential to affect both speakers—the source...... the silence is intentional or unintentional, and whether the silenced memory is related or unrelated to the memories emerging in a conversation. These factors appear to be critical when considering the mnemonic consequences. Moreover, the influence of silence on memory varies between speaker and listener...

  4. Sample Preparation and Extraction in Small Sample Volumes Suitable for Pediatric Clinical Studies: Challenges, Advances, and Experiences of a Bioanalytical HPLC-MS/MS Method Validation Using Enalapril and Enalaprilat (United States)

    Burckhardt, Bjoern B.; Laeer, Stephanie


    In USA and Europe, medicines agencies force the development of child-appropriate medications and intend to increase the availability of information on the pediatric use. This asks for bioanalytical methods which are able to deal with small sample volumes as the trial-related blood lost is very restricted in children. Broadly used HPLC-MS/MS, being able to cope with small volumes, is susceptible to matrix effects. The latter restrains the precise drug quantification through, for example, causing signal suppression. Sophisticated sample preparation and purification utilizing solid-phase extraction was applied to reduce and control matrix effects. A scale-up from vacuum manifold to positive pressure manifold was conducted to meet the demands of high-throughput within a clinical setting. Faced challenges, advances, and experiences in solid-phase extraction are exemplarily presented on the basis of the bioanalytical method development and validation of low-volume samples (50 μL serum). Enalapril, enalaprilat, and benazepril served as sample drugs. The applied sample preparation and extraction successfully reduced the absolute and relative matrix effect to comply with international guidelines. Recoveries ranged from 77 to 104% for enalapril and from 93 to 118% for enalaprilat. The bioanalytical method comprising sample extraction by solid-phase extraction was fully validated according to FDA and EMA bioanalytical guidelines and was used in a Phase I study in 24 volunteers. PMID:25873972

  5. Sample Preparation and Extraction in Small Sample Volumes Suitable for Pediatric Clinical Studies: Challenges, Advances, and Experiences of a Bioanalytical HPLC-MS/MS Method Validation Using Enalapril and Enalaprilat. (United States)

    Burckhardt, Bjoern B; Laeer, Stephanie


    In USA and Europe, medicines agencies force the development of child-appropriate medications and intend to increase the availability of information on the pediatric use. This asks for bioanalytical methods which are able to deal with small sample volumes as the trial-related blood lost is very restricted in children. Broadly used HPLC-MS/MS, being able to cope with small volumes, is susceptible to matrix effects. The latter restrains the precise drug quantification through, for example, causing signal suppression. Sophisticated sample preparation and purification utilizing solid-phase extraction was applied to reduce and control matrix effects. A scale-up from vacuum manifold to positive pressure manifold was conducted to meet the demands of high-throughput within a clinical setting. Faced challenges, advances, and experiences in solid-phase extraction are exemplarily presented on the basis of the bioanalytical method development and validation of low-volume samples (50 μL serum). Enalapril, enalaprilat, and benazepril served as sample drugs. The applied sample preparation and extraction successfully reduced the absolute and relative matrix effect to comply with international guidelines. Recoveries ranged from 77 to 104% for enalapril and from 93 to 118% for enalaprilat. The bioanalytical method comprising sample extraction by solid-phase extraction was fully validated according to FDA and EMA bioanalytical guidelines and was used in a Phase I study in 24 volunteers.


    African Journals Online (AJOL)


    hence the application of GIS and Remote Sensing to waste disposal Site Suitability analysis and route optimization for solid waste in Yola town. Different factors or criteria were considered in relation to solid waste disposal sites. In order to achieve the aim of this paper, the Boolean logic was used, where suitable areas were ...

  7. Three distinct suppressors of RNA silencing encoded by a 20-kb viral RNA genome (United States)

    Lu, Rui; Folimonov, Alexey; Shintaku, Michael; Li, Wan-Xiang; Falk, Bryce W.; Dawson, William O.; Ding, Shou-Wei


    Viral infection in both plant and invertebrate hosts requires a virus-encoded function to block the RNA silencing antiviral defense. Here, we report the identification and characterization of three distinct suppressors of RNA silencing encoded by the 20-kb plus-strand RNA genome of citrus tristeza virus (CTV). When introduced by genetic crosses into plants carrying a silencing transgene, both p20 and p23, but not coat protein (CP), restored expression of the transgene. Although none of the CTV proteins prevented DNA methylation of the transgene, export of the silencing signal (capable of mediating intercellular silencing spread) was detected only from the F1 plants expressing p23 and not from the CP- or p20-expressing F1 plants, demonstrating suppression of intercellular silencing by CP and p20 but not by p23. Thus, intracellular and intercellular silencing are each targeted by a CTV protein, whereas the third, p20, inhibits silencing at both levels. Notably, CP suppresses intercellular silencing without interfering with intracellular silencing. The novel property of CP suggests a mechanism distinct to p20 and all of the other viral suppressors known to interfere with intercellular silencing and that this class of viral suppressors may not be consistently identified by Agrobacterium coinfiltration because it also induces RNA silencing against the infiltrated suppressor transgene. Our analyses reveal a sophisticated viral counter-defense strategy that targets the silencing antiviral pathway at multiple steps and may be essential for protecting CTV with such a large RNA genome from antiviral silencing in the perennial tree host. RNA interference | citrus tristeza virus | virus synergy | antiviral immunity

  8. [An analysis of "silence" in public opinion about public works]. (United States)

    Fujii, Satoshi


    An Internet survey targeting at from 200 to 500 samples in every prefecture in Japan (n=15,316) was conducted in order to analyze the expression of attitudes or silence regarding public works. The data indicated those who had different attitudes from the perceived public attitude were likely to express their attitudes, those whose attitude was neutral were not likely to express it, and those with positive attitudes were likely to express more than those with negative attitudes. The tendency that those with positive attitudes were more likely to express them than those with negative attitudes decreased as the residential population increased. Those with positive attitudes in Tokyo kept "silence" regarding their attitudes, similar to those with neutral attitudes. Analysis regarding the attitudes in different prefectures indicated that attitudes toward public works were positively related to the yearly public stake in the prefecture.

  9. cDNA libraries for virus-induced gene silencing. (United States)

    Todd, Andrea T; Liu, Enwu; Page, Jonathan E


    Virus-induced gene silencing (VIGS) exploits endogenous plant antiviral defense mechanisms to posttranscriptionally silence the expression of targeted plant genes. VIGS is quick and relatively easy to perform and therefore serves as a powerful tool for high-throughput functional genomics in plants. Combined with the use of subtractive cDNA libraries for generating a collection of VIGS-ready cDNA inserts, VIGS can be utilized to screen a large number of genes to determine phenotypes resulting from the knockdown/knockout of gene function. Taking into account the optimal insert design for VIGS, we describe a methodology for producing VIGS-ready cDNA libraries enriched for inserts relevant to the biological process of interest.

  10. The great silence science and philosophy of Fermi's paradox

    CERN Document Server

    Cirkovic, Milan M


    The Great Silence explores the multifaceted problem named after the great Italian physicist Enrico Fermi and his legendary 1950 lunchtime question "Where is everybody?" In many respects, Fermi's paradox is the richest and the most challenging problem for the entire field of astrobiology and the Search for ExtraTerrestrial Intelligence (SETI) studies. This book shows how Fermi's paradox is intricately connected with many fields of learning, technology, arts, and even everyday life. It aims to establish the strongest possible version of the problem, to dispel many related confusions, obfuscations, and prejudices, as well as to offer a novel point of entry to the many solutions proposed in existing literature. Cirkovic argues that any evolutionary worldview cannot avoid resolving the Great Silence problem in one guise or another.

  11. Hush… : The Dangers of Silence in Academic Libraries

    Directory of Open Access Journals (Sweden)

    Jessica Schomberg


    Full Text Available In Brief This article critiques the idea that civility rhetoric decreases workplace bullying or discrimination. We use Critical Discourse Analysis (CDA to do a rhetorical analysis of a campus-wide civility campaign in contrast with literature about civility in libraries. To combat discrimination and bullying, we need to be attentive to systemic power dynamics and to rhetoric designed to enforce compliance and conformity. We conclude with recommendations about how to raise our voices instead of silencing our peers.

  12. Hypoxia Silences Retrotrapezoid Nucleus Respiratory Chemoreceptors via Alkalosis


    Basting, Tyler M.; Burke, Peter G.R.; Kanbar, Roy; Viar, Kenneth E.; Stornetta, Daniel S.; Stornetta, Ruth L.; Guyenet, Patrice G.


    In conscious mammals, hypoxia or hypercapnia stimulates breathing while theoretically exerting opposite effects on central respiratory chemoreceptors (CRCs). We tested this theory by examining how hypoxia and hypercapnia change the activity of the retrotrapezoid nucleus (RTN), a putative CRC and chemoreflex integrator. Archaerhodopsin-(Arch)-transduced RTN neurons were reversibly silenced by light in anesthetized rats. We bilaterally transduced RTN and nearby C1 neurons with Arch (PRSx8-ArchT...



    Berta Bekti Retnawati


    There are powerful forces in many organzations that cause widespread withholding of information about potential problems or issues by employess, this collective-level phenomenon as ‘organizational silence’. One significant effect of organizational silence relates to lack of organizational creativity.There are five major organizational factors that enhance creativity in a work environment: organizational climate, leadership style, organizational culture, resources and skills, the structure and...

  14. Developing Gene Silencing for the Study and Treatment of Dystonia (United States)


    that this is possible but using cells growing in a dish in the laboratory, not in living animals. In this project, we aim to answer several specific... important therapeutic implication of these experiments is that either the striatum is not the primary site of dysfunction in DYT1, or that the molecular...1 AWARD NUMBER: W81XWH-14-1-0282 TITLE: Developing Gene Silencing for the Study and Treatment of Dystonia PRINCIPAL INVESTIGATOR: Pedro

  15. Another Piece of the "Silence in PBL" Puzzle: Students' Explanations of Dominance and Quietness as Complementary Group Roles (United States)

    Skinner, Vicki J.; Braunack-Mayer, Annette; Winning, Tracey A.


    A problem-based learning (PBL) assumption is that silence is incompatible with collaborative learning. Although sociocultural studies have reinterpreted silence as collaborative, we must understand how silence occurs in PBL groups. This essay presents students' explanations of dominance, leadership, and silence as PBL group roles. An ethnographic…

  16. The Application of Coconut Fiber as Dissipative Silencer (United States)

    Madlan, M. A.; Ghazali, M. I.; Zaman, I.; Kasron, M. Z.; Ying, T. C.


    Heat ventilation air conditioning system (HVAC) is one of the ducting systems that broadly applied in the building. There are HVAC silencers in the market, however the sound absorptive material commonly used is mineral wool. In this research study, a sound absorptive material made of coconut fiber was tested to identify its performance as a potential replacement of green material for ducting silencer. The experiment was carried out in a testing apparatus that follows the BS EN ISO 11691:2009 standard. Different configurations of sound absorptive material and contents of coconut fiber were investigated in the study. The trend of insertion loss at 1/3 octave frequency was identified where at frequency below 3000Hz, the insertion loss of dissipative silencer is observed high at certain frequency with a very narrow range. At 3000Hz, the insertion loss of 4dB to 6dB is constant until 4000Hz and drops until 5000Hz before it increases again steadily up to 13dB at 10000Hz. A similar trend was observed for different configuration of sound absorptive material. Despite the configuration different, the outcome shows that the insertion loss is increasing with higher content of coconut fiber.

  17. Epigenetic silencing of tumor suppressor genes: Paradigms, puzzles, and potential. (United States)

    Kazanets, Anna; Shorstova, Tatiana; Hilmi, Khalid; Marques, Maud; Witcher, Michael


    Cancer constitutes a set of diseases with heterogeneous molecular pathologies. However, there are a number of universal aberrations common to all cancers, one of these being the epigenetic silencing of tumor suppressor genes (TSGs). The silencing of TSGs is thought to be an early, driving event in the oncogenic process. With this in consideration, great efforts have been made to develop small molecules aimed at the restoration of TSGs in order to limit tumor cell proliferation and survival. However, the molecular forces that drive the broad epigenetic reprogramming and transcriptional repression of these genes remain ill-defined. Undoubtedly, understanding the molecular underpinnings of transcriptionally silenced TSGs will aid us in our ability to reactivate these key anti-cancer targets. Here, we describe what we consider to be the five most logical molecular mechanisms that may account for this widely observed phenomenon: 1) ablation of transcription factor binding, 2) overexpression of DNA methyltransferases, 3) disruption of CTCF binding, 4) elevation of EZH2 activity, 5) aberrant expression of long non-coding RNAs. The strengths and weaknesses of each proposed mechanism is highlighted, followed by an overview of clinical efforts to target these processes. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

  18. Silencing Deafness: Displacing Disability in the Nineteenth Century

    Directory of Open Access Journals (Sweden)

    Esme Cleall


    Full Text Available This article traces the way in which the language of displacement and silence were used in nineteenth-century discussions of deafness and connects this tendency to the marginalised place deaf experience occupies historically. Throughout the nineteenth century, a period which saw the consolidation of ‘the deaf and dumb’ as a social category, the word ‘forgetting’ crept into numerous discussions of deafness by both deaf and hearing commentators. Some, such as the educationalist Alexander Graeme Bell, were overt in their desire to forget deafness, demanding disability was ‘bred out’ and deaf culture condemned to the forgotten past. Others used the term ambivalently and sometimes metaphorically discussing the deaf as ‘forgotten’ by society, and ‘children of silence’. Some even pleaded that people who were deaf were not forgotten. But, though varied, the use of the imagery of forgetting and silence to evoke deafness is recurrent, and may, therefore, be seen to reveal something about how deaf experience can be approached as a displacement where deafness was spatially and imaginatively marginalised. I argue that one of the consequences of the conceptual framing of deafness through the language of forgetting was actively to silence deafness and to neutralise the idea that disability should be marginal and could be forgotten.

  19. Epigenetic silencing of CYP24 in the tumor microenvironment (United States)

    Johnson, Candace S.; Chung, Ivy; Trump, Donald L.


    Calcitriol (1,25 dihydroxycholecalciferol) has significant antitumor activity in vitro and in vivo in a number of tumor model systems. We developed a system for isolation of fresh endothelial cells from tumors and Matrigel environments which demonstrate that CYP24, the catabolic enzyme involved in vitamin D signaling, is epigenetically silenced selectively in tumor-derived endothelial cells (TDEC). TDEC maintain phenotypic characteristics which are distinct from endothelial cells isolated from normal tissues and from Matrigel plugs (MDEC). In TDEC, calcitriol induces G0/G1 arrest, modulates p27 and p21, and induces apoptotic cell death and decreases P-Erk and P-Akt. In contrast, endothelial cells isolated from normal tissues and MDEC are unresponsive to calcitriol-mediated anti-proliferative effects despite intact signaling through the vitamin D receptor (VDR). In TDEC, which is sensitive to calcitriol, the CYP24 promoter is hypermethylated in two CpG island regions located at the 5′end; this hypermethylation may contribute to gene silencing of CYP24. The extent of methylation in these two regions is significantly less in MDEC. Lastly, treatment of TDEC with a DNA methyltransferase inhibitor restores calcitriol-mediated induction of CYP24 and resistance to calcitriol. These data suggest that epigenetic silencing of CYP24 modulates cellular responses to calcitriol. PMID:20304059

  20. 76 FR 82201 - General Site Suitability Criteria for Nuclear Power Stations (United States)


    ..., and 52 [NRC-2011-0297] General Site Suitability Criteria for Nuclear Power Stations AGENCY: Nuclear... Suitability Criteria for Nuclear Power Stations.'' This guide describes a method that the NRC staff considers acceptable to implement the site suitability requirements for nuclear power stations. DATES: Submit comments...

  1. Silencing of Target Chitinase Genes via Oral Delivery of dsRNA Caused Lethal Phenotypic Effects in Mythimna separata (Lepidoptera: Noctuidae). (United States)

    Cao, Budao; Bao, Wenhua; Wuriyanghan, Hada


    Mythimna separata walker (Lepidoptera: Noctuidae) is a polyphagous, migratory corn pest. Outbreak of M. separata has led to severe damage to corn production recently in China. RNAi (RNA interference) is a gene silencing technology applied both in model and non-model organisms, and it is especially useful for the latter in which the reverse genetic research tools are not available. RNAi approach was broadly investigated in many plant pathogens and was used for the generation of anti-pest transgenic plants. We are proposing to use this technology to silence M. separata endogenous genes, thereby, providing a biocontrol method for this insect. Feeding of dsRNAs for target Chitinase genes resulted in substantial decreases of their transcript levels in M. separata. Furthermore, silencing of target Chitinase genes led to phenotypic effects such as reduced body weight and increased mortality. Our study provided both reverse genetic research tool and potential control strategy for this insect species.

  2. Method for the appraisal and classification of the suitability for use of watercourses - Final report; Methodik zur Bewertung und Klassierung der Nutzungseignung von Fliessgewaesserstrecken. Grundlagen fuer die raeumliche Prioritaetensetzung bei der Wasserkraftnutzung und dem Schutz von Gewaessern - Schlussbericht

    Energy Technology Data Exchange (ETDEWEB)

    Wehse, H.


    This final report for the Swiss Federal Office of Energy (SFOE) discusses the fundamentals for setting priorities on the use of hydropower and the protection of streams and rivers. New general conditions for the realisation of hydropower projects in Switzerland resulting from the cost-covering remuneration scheme for renewable energy sources have led to an increasing number of new projects. The article discussed the conflicts of interest between power station projects and environmental protectionists and anglers. A method is proposed that is to help resolve such conflicts. The development of the proposed methodology is discussed and the results of a test application of the methodology are examined. The report is enhanced with a comprehensive appendix.

  3. Multi-gene epigenetic silencing of tumor suppressor genes in T-cell lymphoma cells; delayed expression of the p16 protein upon reversal of the silencing

    DEFF Research Database (Denmark)

    Nagasawa, T; Zhang, Q; Raghunath, P N


    )-expressing T-cell lymphomas. p16 gene was epigenetically silenced in all but one of the 10 malignant T-cell lines examined, p15 gene silenced in roughly half of the lines, and p14 was the least frequently affected. Extensive methylation of the p16 promoter was seen in six out of 10 cutaneous T-cell lymphoma...... promoter demethylation and required up to 3 weeks to occur, seemingly reflecting late activation of the p16 gene. These findings indicate that epigenetic silencing affects in T-cell malignancies, often simultaneously, several tumor suppressor genes that impact on key cell functions. The observed...... differential silencing of p16 and p14, and to a lesser degree p15 gene, indicates that the silencing is governed by precise, promoter region-specific mechanisms. The study provides also further rationale for treatment of at least some types of T-cell lymphomas with DNA methyltransferase inhibitors to target...

  4. Selection of suitable stainless steels for nuclear reprocessing plants: application of chemical and electrochemical testing methods to austenitic CrNi steel AISI type 304L in various chemical compositions

    International Nuclear Information System (INIS)

    Leistikow, S.; Kraft, R.


    DIN Standard Huey testing has been performed in boiling 14.4n nitric acid during 5-15 periods (240-720 h) for selection of appropriate nitric acid resistant materials for nuclear fuel reprocessing applications. The paper describes the testing process during which the intermediate and final results of metal loss by dissolution are directly transferred from the balance to the computer, stored and activated - besides material properties data - for documentation purposes. Further routine evaluation of these experiments includes metallography in cross-section and surface microscopy to look after uniform and local metal dissolution phenomena and their relationship to the bulk structure. A large variety of materials have been tested this way through the last years. It was shown how sensitively the chosen testing conditions are able to differ between materials of the same nominal composition AISI 304L/Material No. 1.4306 in different contents of residual elements. Especially, for the purest electroslag-molten steel (ESU) results of parameter studies concerning the influence of sensitization, cold deformation, grain size and sheet thickness (in respect to end grain attack) are given. Within an attempt to define faster methods of corrosion testing, e.g. to differ within a group of materials of similar composition, but different corrosion behaviour, electrochemical tests in heated nitric acid were performed under potentiostatic conditions. The necessary electrochemical equipment and the results of its application by potentiostatic tests on AISI 304L in above mentioned three chemical compositions at 1250 mV, 14n HNO 3 are presented. The evaluation by light and electron microscopy of the corroded surfaces, supported by measurements of current density, weight change, metallography and surface roughness, proved that within one hour a remarkable differentiation of the corrosion behaviour took place which can serve as a basis of materials preselection and to diminish the extent of

  5. Regional technical innovation suitability and economic growth in China

    NARCIS (Netherlands)

    Zhou, Qing; Zhu, Dan; Brouwers-Ren, Liqin


    This paper aims to use quantitative method to analyse the suitability of regional technical innovation and further compare its influence to regional economic growth in China. Based on literature review, the authors develop an evaluation index system containing four aspects (namely, innovators,

  6. Validation of standards suitable for genome size estimation of fungi. (United States)

    Talhinhas, Pedro; Tavares, Daniela; Ramos, Ana Paula; Gonçalves, Susana; Loureiro, João


    Genome size information is fundamental to genome sequencing and may also uncover genomic aspects of evolution. Flow Cytometry, the preferred method for genome size estimation, requires suitable standards. Here we validate Inonotus hispidus, Colletotrichum acutatum and Cenococcum geophilum (41, 68 and 203Mbp), as standards for fungal genome size estimation. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Selecting Suitable Drainage Pattern to Minimize Flooding in ...

    African Journals Online (AJOL)

    Water shed analysis is a geographic information system (GIS) based technique designed to model the way surface water flows on the earth surface. This was the method adopted to select suitable drainage pattern to minimized flood in some parts of sangere. The process of watershed computes the local direction of flow ...

  8. Inducible and titratable silencing of Caenorhabditis elegans neurons in vivo with histamine-gated chloride channels (United States)

    Pokala, Navin; Liu, Qiang; Gordus, Andrew; Bargmann, Cornelia I.


    Recent progress in neuroscience has been facilitated by tools for neuronal activation and inactivation that are orthogonal to endogenous signaling systems. We describe here a chemical-genetic approach for inducible silencing of Caenorhabditis elegans neurons in intact animals, using the histamine-gated chloride channel HisCl1 from Drosophila and exogenous histamine. Administering histamine to freely moving C. elegans that express HisCl1 transgenes in neurons leads to rapid and potent inhibition of neural activity within minutes, as assessed by behavior, functional calcium imaging, and electrophysiology of neurons expressing HisCl1. C. elegans does not use histamine as an endogenous neurotransmitter, and exogenous histamine has little apparent effect on wild-type C. elegans behavior. HisCl1-histamine silencing of sensory neurons, interneurons, and motor neurons leads to behavioral effects matching their known functions. In addition, the HisCl1-histamine system can be used to titrate the level of neural activity, revealing quantitative relationships between neural activity and behavioral output. We use these methods to dissect escape circuits, define interneurons that regulate locomotion speed (AVA, AIB) and escape-related omega turns (AIB), and demonstrate graded control of reversal length by AVA interneurons and DA/VA motor neurons. The histamine-HisCl1 system is effective, robust, compatible with standard behavioral assays, and easily combined with optogenetic tools, properties that should make it a useful addition to C. elegans neurotechnology. PMID:24550306

  9. Aflatoxin-free transgenic maize using host-induced gene silencing. (United States)

    Thakare, Dhiraj; Zhang, Jianwei; Wing, Rod A; Cotty, Peter J; Schmidt, Monica A


    Aflatoxins, toxic secondary metabolites produced by some Aspergillus species, are a universal agricultural economic problem and a critical health issue. Despite decades of control efforts, aflatoxin contamination is responsible for a global loss of millions of tons of crops each year. We show that host-induced gene silencing is an effective method for eliminating this toxin in transgenic maize. We transformed maize plants with a kernel-specific RNA interference (RNAi) gene cassette targeting the aflC gene, which encodes an enzyme in the Aspergillus aflatoxin biosynthetic pathway. After pathogen infection, aflatoxin could not be detected in kernels from these RNAi transgenic maize plants, while toxin loads reached thousands of parts per billion in nontransgenic control kernels. A comparison of transcripts in developing aflatoxin-free transgenic kernels with those from nontransgenic kernels showed no significant differences between these two groups. These results demonstrate that small interfering RNA molecules can be used to silence aflatoxin biosynthesis in maize, providing an attractive and precise engineering strategy that could also be extended to other crops to improve food security.

  10. Investigating Gene Function in Cereal Rust Fungi by Plant-Mediated Virus-Induced Gene Silencing. (United States)

    Panwar, Vinay; Bakkeren, Guus


    Cereal rust fungi are destructive pathogens, threatening grain production worldwide. Targeted breeding for resistance utilizing host resistance genes has been effective. However, breakdown of resistance occurs frequently and continued efforts are needed to understand how these fungi overcome resistance and to expand the range of available resistance genes. Whole genome sequencing, transcriptomic and proteomic studies followed by genome-wide computational and comparative analyses have identified large repertoire of genes in rust fungi among which are candidates predicted to code for pathogenicity and virulence factors. Some of these genes represent defence triggering avirulence effectors. However, functions of most genes still needs to be assessed to understand the biology of these obligate biotrophic pathogens. Since genetic manipulations such as gene deletion and genetic transformation are not yet feasible in rust fungi, performing functional gene studies is challenging. Recently, Host-induced gene silencing (HIGS) has emerged as a useful tool to characterize gene function in rust fungi while infecting and growing in host plants. We utilized Barley stripe mosaic virus-mediated virus induced gene silencing (BSMV-VIGS) to induce HIGS of candidate rust fungal genes in the wheat host to determine their role in plant-fungal interactions. Here, we describe the methods for using BSMV-VIGS in wheat for functional genomics study in cereal rust fungi.

  11. Delivery of siRNA silencing P-gp in peptide-functionalized nanoparticles causes efflux modulation at the blood-brain barrier

    DEFF Research Database (Denmark)

    Gomes, Maria João; Kennedy, Patrick J; Martins, Susana


    AIM: Explore the use of transferrin-receptor peptide-functionalized nanoparticles (NPs) targeting blood-brain barrier (BBB) as siRNA carriers to silence P-glycoprotein (P-gp). MATERIALS & METHODS: Permeability experiments were assessed through a developed BBB cell-based model; P-gp mRNA expressio...

  12. Suitable long tendon technologies and practices

    CSIR Research Space (South Africa)

    Altounyan, P


    Full Text Available effective long tendon support systems suitable in South African coalmines. Appropriate test procedures for the evaluation of the performance of long tendon systems were also determined and are detailed in this report....

  13. Proposal of an Algorithm to Synthesize Music Suitable for Dance (United States)

    Morioka, Hirofumi; Nakatani, Mie; Nishida, Shogo

    This paper proposes an algorithm for synthesizing music suitable for emotions in moving pictures. Our goal is to support multi-media content creation; web page design, animation films and so on. Here we adopt a human dance as a moving picture to examine the availability of our method. Because we think the dance image has high affinity with music. This algorithm is composed of three modules. The first is the module for computing emotions from an input dance image, the second is for computing emotions from music in the database and the last is for selecting music suitable for input dance via an interface of emotion.

  14. The Role Of Self - Efficacy Perception in Silencing Organization: A St udy On Accommodation Enterprises in Alanya

    Directory of Open Access Journals (Sweden)

    Engin Üngüren


    Full Text Available Qualified employees has an undeniable a place and importance in the labor-intensive tourism sector to achieve competitive advantage, improve service and product quality, carry out sustainable development. As well qualified employees increase the competitiveness of countries in the international market. Providing opportunities for using qualified employees’ full potential is an important method in obtaining competitive advantage of the company. In this context, the main aim of the research is to determine the impact of self-efficacy on organizational silence. Sub-objectives of this research is to determine employees’ ideas and thoughts which provide contribution to business, whether there is a difference among employees’ perceptions of organizational silence and perceptions of self efficacy according to employees’ demographic and occupational characteristics. The research data were obtained by the survey carried out in the accommodation businesses. On the basis of the quantitative paradigm, descriptive research model, comparative model, the relational model and the relational model approaches are used. The findings of the research pointed out that perception of self-efficacy had decisive influence on organizational silence. As a result of research was determined that employees, who have low self-efficacy, was in silent by not expressing ideas and thought and employees, who have high self-efficacy, did not hesitate to share their ideas and thoughts.

  15. Normalization of Overexpressed α-Synuclein Causing Parkinson's Disease By a Moderate Gene Silencing With RNA Interference

    Directory of Open Access Journals (Sweden)

    Masaki Takahashi


    Full Text Available The α-synuclein (SNCA gene is a responsible gene for Parkinson's disease (PD; and not only nucleotide variations but also overexpression of SNCA appears to be involved in the pathogenesis of PD. A specific inhibition against mutant SNCA genes carrying nucleotide variations may be feasible by a specific silencing such as an allele-specific RNA interference (RNAi; however, there is no method for restoring the SNCA overexpression to a normal level. Here, we show that an atypical RNAi using small interfering RNAs (siRNAs that confer a moderate level of gene silencing is capable of controlling overexpressed SNCA genes to return to a normal level; named “expression-control RNAi” (ExCont-RNAi. ExCont-RNAi exhibited little or no significant off-target effects in its treated PD patient's fibroblasts that carry SNCA triplication. To further assess the therapeutic effect of ExCont-RNAi, PD-model flies that carried the human SNCA gene underwent an ExCont-RNAi treatment. The treated PD-flies demonstrated a significant improvement in their motor function. Our current findings suggested that ExCont-RNAi might be capable of becoming a novel therapeutic procedure for PD with the SNCA overexpression, and that siRNAs conferring a moderate level of gene silencing to target genes, which have been abandoned as useless siRNAs so far, might be available for controlling abnormally expressed disease-causing genes without producing adverse effects.

  16. Cancelation and its simulation using Matlab according to active noise control case study of automotive noise silencer (United States)

    Alfisyahrin; Isranuri, I.


    Active Noise Control is a technique to overcome noisy with noise or sound countered with sound in scientific terminology i.e signal countered with signals. This technique can be used to dampen relevant noise in accordance with the wishes of the engineering task and reducing automotive muffler noise to a minimum. Objective of this study is to develop a Active Noise Control which should cancel the noise of automotive Exhaust (Silencer) through Signal Processing Simulation methods. Noise generator of Active Noise Control is to make the opponent signal amplitude and frequency of the automotive noise. The steps are: Firstly, the noise of automotive silencer was measured to characterize the automotive noise that its amplitude and frequency which intended to be expressed. The opposed sound which having similar character with the signal source should be generated by signal function. A comparison between the data which has been completed with simulation calculations Fourier transform field data is data that has been captured on the muffler (noise silencer) Toyota Kijang Capsule assembly 2009. MATLAB is used to simulate how the signal processing noise generated by exhaust (silencer) using FFT. This opponent is inverted phase signal from the signal source 180° conducted by Instruments of Signal Noise Generators. The process of noise cancelation examined through simulation using computer software simulation. The result is obtained that attenuation of sound (noise cancellation) has a difference of 33.7%. This value is obtained from the comparison of the value of the signal source and the signal value of the opponent. So it can be concluded that the noisy signal can be attenuated by 33.7%.

  17. Evaluation of methods to test chemicals suitability for umbilical applications

    Energy Technology Data Exchange (ETDEWEB)

    Allenson, S. J.; Lindeman, O. E.; Cenegy, L. M.


    Offshore deep-water projects are increasingly deploying chemicals to sub-sea wellheads through umbilical lines. There is no margin for error in umbilical chemical treatment programs since any flow blockage in a sub-sea line would result in a multi-million dollar problem. Chemicals for umbilical delivery must also meet strict requirements in their performance and especially their handling properties. Umbilical delivery must be effective at low concentrations in preventing corrosion, scale, hydrates, asphaltenes, paraffin and a host of other problems. Chemical transiting an umbilical can experience pressures as high as 15,000 psi and temperatures ranging from near 0 deg C to greater than 120 deg C. Since some umbilicals are as long as 80 km, a week or more can elapse from the time the chemical is injected at the platform until it reaches the sub-sea well. Therefore, the chemical must not only be stable under all temperature and pressure conditions that it may experience in the umbilical line, it must also be stable under these conditions for a long period of time. Since many umbilical lines actually terminate into sub-sea valves and connectors that are only a few hundred microns in diameter, it is critical that the injected chemical have a low viscosity at sub-sea temperatures and pressures and that it be completely free of particles. These issues present substantial challenges in formulating and manufacturing chemicals for umbilical applications that must be addressed prior to approval of a product for use. Each of these challenges was taken into consideration and a series of tests were developed to assure reliable chemical pump ability through an umbilical line. The tests developed included enhanced formulation stability tests under umbilical temperature and pressure conditions, NAS Class rating, extensive material compatibility testing to include all metals and elastomers that may be used in umbilical injection systems and comprehensive physical property testing. Once the chemicals passed all tests they were further evaluated in a full size operational umbilical unit. The test protocols developed are fully discussed in this paper. (Author) (tk)

  18. 10 CFR 963.16 - Postclosure suitability evaluation method. (United States)


    ... barriers, including those processes that would adversely affect natural barriers, (such as degradation of... the capability of the natural and engineered barriers important to isolating radioactive waste, taking... basis for the description of the capability of the natural and engineered barriers important to...

  19. Exon silencing by UAGG motifs in response to neuronal excitation.

    Directory of Open Access Journals (Sweden)

    Ping An


    Full Text Available Alternative pre-mRNA splicing plays fundamental roles in neurons by generating functional diversity in proteins associated with the communication and connectivity of the synapse. The CI cassette of the NMDA R1 receptor is one of a variety of exons that show an increase in exon skipping in response to cell excitation, but the molecular nature of this splicing responsiveness is not yet understood. Here we investigate the molecular basis for the induced changes in splicing of the CI cassette exon in primary rat cortical cultures in response to KCl-induced depolarization using an expression assay with a tight neuron-specific readout. In this system, exon silencing in response to neuronal excitation was mediated by multiple UAGG-type silencing motifs, and transfer of the motifs to a constitutive exon conferred a similar responsiveness by gain of function. Biochemical analysis of protein binding to UAGG motifs in extracts prepared from treated and mock-treated cortical cultures showed an increase in nuclear hnRNP A1-RNA binding activity in parallel with excitation. Evidence for the role of the NMDA receptor and calcium signaling in the induced splicing response was shown by the use of specific antagonists, as well as cell-permeable inhibitors of signaling pathways. Finally, a wider role for exon-skipping responsiveness is shown to involve additional exons with UAGG-related silencing motifs, and transcripts involved in synaptic functions. These results suggest that, at the post-transcriptional level, excitable exons such as the CI cassette may be involved in strategies by which neurons mount adaptive responses to hyperstimulation.

  20. Epigenetic silencing may aid evolution by gene duplication. (United States)

    Rodin, Sergei N; Riggs, Arthur D


    Gene duplication is commonly regarded as the main evolutionary path toward the gain of a new function. However, even with gene duplication, there is a loss-versus-gain dilemma: most newly born duplicates degrade to pseudogenes, since degenerative mutations are much more frequent than advantageous ones. Thus, something additional seems to be needed to shift the loss versus gain equilibrium toward functional divergence. We suggest that epigenetic silencing of duplicates might play this role in evolution. This study began when we noticed in a previous publication (Lynch M, Conery JS [2000] Science 291:1151-1155) that the frequency of functional young gene duplicates is higher in organisms that have cytosine methylation (H. sapiens, M. musculus, and A. thaliana) than in organisms that do not have methylated genomes (S. cerevisiae, D. melanogaster, and C. elegans). We find that genome data analysis confirms the likelihood of much more efficient functional divergence of gene duplicates in mammals and plants than in yeast, nematode, and fly. We have also extended the classic model of gene duplication, in which newly duplicated genes have exactly the same expression pattern, to the case when they are epigenetically silenced in a tissue- and/or developmental stage-complementary manner. This exposes each of the duplicates to negative selection, thus protecting from "pseudogenization." Our analysis indicates that this kind of silencing (i) enhances evolution of duplicated genes to new functions, particularly in small populations, (ii) is quite consistent with the subfunctionalization model when degenerative but complementary mutations affect different subfunctions of the gene, and (iii) furthermore, may actually cooperate with the DDC (duplication-degeneration-complementation) process.

  1. DNA methylation, genomic silencing, and links to nutrition and cancer. (United States)

    McCabe, Dale C; Caudill, Marie A


    DNA methylation is a heritable epigenetic feature that is associated with transcriptional silencing, X-chromosome inactivation, genetic imprinting, and genomic stability. The addition of the methyl group is catalyzed by a family of DNA methyltransferases whose co-substrates are DNA and S-adenosylmethionine, the latter being derived from the methionine cycle. Aberrant DNA methylation is linked to numerous pathologies, including cancer. The purpose of this review is to describe DNA methylation and its functions, to examine the relationship between dietary methyl insufficiency and DNA methylation, and to evaluate the associations between DNA methylation and cancer.

  2. Silent God in a Wordy World. Silence in Ignatian Spirituality


    García de Castro Valdés, S.J., José


    The Society of Jesus is an apostolic religious order involved in many different activities and missions. Jesuits live far away from monasteries and strict contemplative life. Nevertheless, one of the most well-known peculiarities of the Ignatian Spiritual Exercises is that a complete and absolute silence is required during the time of the retreat. Where and how should we place “silence” in the life of an Ignatian spiritual and mystical experience?

  3. Nuclear architecture and gene silencing in olfactory sensory neurons. (United States)

    Armelin-Correa, Lucia M; Nagai, Maíra H; Leme Silva, Artur G; Malnic, Bettina


    Odorants are discriminated by hundreds of odorant receptor (OR) genes, which are dispersed throughout the mammalian genome. The OR genes are expressed in a highly specialized type of cell, the olfactory sensory neuron. Each one of these neurons expresses one of the 2 alleles from one single OR gene type. The mechanisms underlying OR gene expression are unclear. Here we describe recent work demonstrating that the olfactory sensory neuron shows a particular nuclear architecture, and that the genomic OR loci are colocalized in silencing heterochromatin compartments within the nucleus. These discoveries highlight the important role played by epigenetic modifications and nuclear genome organization in the regulation of OR gene expression.

  4. Suitability of Varicose Veins for Endovenous Treatments

    International Nuclear Information System (INIS)

    Goode, S. D.; Kuhan, G.; Altaf, N.; Simpson, R.; Beech, A.; Richards, T.; MacSweeney, S. T.; Braithwaite, B. D.


    The aim of the study was to assess the suitability of radiofrequency ablation (RFA), endovenous laser ablation (EVLA), and foam sclerotherapy (FS) for patients with symptomatic varicose veins (VVs). The study comprised 403 consecutive patients with symptomatic VVs. Data on 577 legs from 403 consecutive patients with symptomatic VVs were collected for the year 2006. Median patient age was 55 years (interquartile range 45-66), and 62% patients were women. A set of criteria based on duplex ultrasonography was used to select patients for each procedure. Great saphenous vein (GSV) reflux was present in 77% (446 of 577) of legs. Overall, 328 (73%) of the legs were suitable for at least one of the endovenous options. Of the 114 legs with recurrent GSV reflux disease, 83 (73%) were suitable to receive endovenous therapy. Patients with increasing age were less likely to be suitable for endovenous therapy (P = 0.03). Seventy-three percent of patients with VVs caused by GSV incompetence are suitable for endovenous therapy.

  5. Suitability of NAA for certification of reference materials for multielements

    International Nuclear Information System (INIS)

    Tian Weizhi; Ni Bangfa; Wang Pingsheng; Nie Huiling


    Certifications of trace elements in existing CRMs, especially biological CRMs, are far from satisfactory. Neutron activation analysis (NAA) for its inherent advantages combined with newly established parametric standardization, may contribute to improve this situation. The continuing progress of the hybrid extended k 0 -relative NAA technique developed in our laboratory is discussed. Examples are given to show the reliability of the method in certification of multielements. RNAA is still one of the best methods, or even the method of choice, in analysis at sub-μg/g concentration levels. The suitability of the technique for this purpose has been studied through the determination of rare earth elements at ng/g concentration level in two Chinese biological CRMs using both RNAA and ICPMS. Sampling behaviors of multielements in CRMs have been studied by INAA in an effort to develop CRMs suitable for analysis with small sample sizes. (author)

  6. Additional media studies for site suitability criteria

    International Nuclear Information System (INIS)

    Donich, T.R.; Kaufman, A.M.; Sauter, G.D.; Steinborn, T.L.; Towse, D.F.


    Site suitability studies have been made previously at LLL on bedded salt and shale. In the present study domed salt, basalt, and crystalline rock are compared with bedded salt and shale and with each other as possible repositories. The level of effort required to develop models for these media that are similar in quality to those available for bedded salt and shale is evaluated. The effort necessary to develop data bases on the physical and chemical properties comparable to that available for bedded salt and shale is also estimated. Each medium is evaluated as a suitable repository environment. The funding necessary for model and data base development is estimated

  7. From Suitable Weak Solutions to Entropy Viscosity

    KAUST Repository

    Guermond, Jean-Luc


    This paper focuses on the notion of suitable weak solutions for the three-dimensional incompressible Navier-Stokes equations and discusses the relevance of this notion to Computational Fluid Dynamics. The purpose of the paper is twofold (i) to recall basic mathematical properties of the three-dimensional incompressible Navier-Stokes equations and to show how they might relate to LES (ii) to introduce an entropy viscosity technique based on the notion of suitable weak solution and to illustrate numerically this concept. © 2010 Springer Science+Business Media, LLC.

  8. Uncertainty in Various Habitat Suitability Models and Its Impact on Habitat Suitability Estimates for Fish


    Lin, Yu-Pin; Lin, Wei-Chih; Wu, Wei-Yao


    Species distribution models (SDMs) are extensively used to project habitat suitability of species in stream ecological studies. Owing to complex sources of uncertainty, such models may yield projections with varying degrees of uncertainty. To better understand projected spatial distributions and the variability between habitat suitability projections, this study uses five SDMs that are based on the outputs of a two-dimensional hydraulic model to project the suitability of habitats and to eval...

  9. Live-cell imaging to compare the transfection and gene silencing efficiency of calcium phosphate nanoparticles and a liposomal transfection agent. (United States)

    Chernousova, S; Epple, M


    The processing of DNA (for transfection) and short interfering RNA (siRNA; for gene silencing), introduced into HeLa cells by triple-shell calcium phosphate nanoparticles, was followed by live-cell imaging. For comparison, the commercial liposomal transfection agent Lipofectamine was used. The cells were incubated with these delivery systems, carrying either enhanced green fluorescent protein (eGFP)-encoding DNA or siRNA against eGFP. In the latter case, HeLa cells that stably expressed eGFP were used. The expression of eGFP started after 5 h in the case of nanoparticles and after 4 h in the case of Lipofectamine. The corresponding times for gene silencing were 5 h (nanoparticles) and immediately after incubation (Lipofectamine). The expression of eGFP was notably enhanced 2-3 h after cell division (mitosis). In general, the transfection and gene silencing efficiencies of the nanoparticles were lower than those of Lipofectamime, even at a substantially higher dose (factor 20) of nucleic acids. However, the cytotoxicity of the nanoparticles was lower than that of Lipofectamine, making them suitable vectors for in vivo application.

  10. Sexuality and 'silence' among Khasi youth of Meghalaya, Northeast India. (United States)

    War, Ryntihlin Jennifer; Albert, Sandra


    The importance of sex education has been well documented in the literature, but there exists a lack of research involving indigenous youth in India. This paper describes perceptions, knowledge and attitudes towards sex education, sexuality, pre-marital sex, rape and homosexuality among indigenous students from the matrilineal Khasi tribe attending a university in Meghalaya in northeast India. Qualitative and quantitative data were collected during and after reproductive health, sexuality and life skills courses. Despite the impression of sexual permissiveness of indigenous peoples that exists in India, students reported a societal silence on issues related to sexuality. Lack of appropriate words in the indigenous language potentially contributes to this silence. Although co-habitation is common and culturally acceptable, students disapproved of pre-marital sex. The influence of Christianisation was also perceived in the frequent reference to sin and guilt associated with masturbation, homosexuality, pre-marital sex and abortion. Students reported that the sex education received in school was 'childish' and inadequate for their adult needs. Many had unrealistic images of what constituted 'normal' sex and also blamed women for rape. The majority of indigenous students expressed the need for non-judgmental fora for discussions on sexual health and for sexuality education.

  11. ABCE1 is a highly conserved RNA silencing suppressor.

    Directory of Open Access Journals (Sweden)

    Kairi Kärblane

    Full Text Available ATP-binding cassette sub-family E member 1 (ABCE1 is a highly conserved protein among eukaryotes and archaea. Recent studies have identified ABCE1 as a ribosome-recycling factor important for translation termination in mammalian cells, yeast and also archaea. Here we report another conserved function of ABCE1. We have previously described AtRLI2, the homolog of ABCE1 in the plant Arabidopsis thaliana, as an endogenous suppressor of RNA silencing. In this study we show that this function is conserved: human ABCE1 is able to suppress RNA silencing in Nicotiana benthamiana plants, in mammalian HEK293 cells and in the worm Caenorhabditis elegans. Using co-immunoprecipitation and mass spectrometry, we found a number of potential ABCE1-interacting proteins that might support its function as an endogenous suppressor of RNA interference. The interactor candidates are associated with epigenetic regulation, transcription, RNA processing and mRNA surveillance. In addition, one of the identified proteins is translin, which together with its binding partner TRAX supports RNA interference.

  12. Characterization and subcellular localization of an RNA silencing suppressor encoded by Rice stripe tenuivirus

    International Nuclear Information System (INIS)

    Xiong Ruyi; Wu Jianxiang; Zhou Yijun; Zhou Xueping


    Rice stripe virus (RSV) is a single-stranded (ss) RNA virus belonging to the genus Tenuivirus. RSV is present in many East Asian countries and causes severe diseases in rice fields, especially in China. In this study, we analyzed six proteins encoded by the virus for their abilities to suppress RNA silencing in plant using a green fluorescent protein (GFP)-based transient expression assay. Our results indicate that NS3 encoded by RSV RNA3, but not other five RSV encoded proteins, can strongly suppress local GFP silencing in agroinfiltrated Nicotiana benthamiana leaves. NS3 can reverse the GFP silencing, it can also prevent long distance spread of silencing signals which have been reported to be necessary for inducing systemic silencing in host plants. The NS3 protein can significantly reduce the levels of small interfering RNAs (siRNAs) in silencing cells, and was found to bind 21-nucleotide ss-siRNA, siRNA duplex and long ssRNA but not long double-stranded (ds)-RNA. Both N and C terminal of the NS3 protein are critical for silencing suppression, and mutation of the putative nuclear localization signal decreases its local silencing suppression efficiency and blocks its systemic silencing suppression. The NS3-GFP fusion protein and NS3 were shown to accumulate predominantly in nuclei of onion, tobacco and rice cells through transient expression assay or immunocytochemistry and electron microscopy. In addition, transgenic rice and tobacco plants expressing the NS3 did not show any apparent alteration in plant growth and morphology, although NS3 was proven to be a pathogenicity determinant in the PVX heterogenous system. Taken together, our results demonstrate that RSV NS3 is a suppressor of RNA silencing in planta, possibly through sequestering siRNA molecules generated in cells that are undergoing gene silencing.

  13. A Suitable Artificial Intelligence Model for Inventory Level Optimization

    Directory of Open Access Journals (Sweden)

    Tereza Sustrova


    Full Text Available Purpose of the article: To examine suitable methods of artificial neural networks and their application in business operations, specifically to the supply chain management. The article discusses construction of an artificial neural networks model that can be used to facilitate optimization of inventory level and thus improve the ordering system and inventory management. For the data analysis from the area of wholesale trade with connecting material is used. Methodology/methods: Methods used in the paper consists especially of artificial neural networks and ANN-based modelling. For data analysis and preprocessing, MS Office Excel software is used. As an instrument for neural network forecasting MathWorks MATLAB Neural Network Tool was used. Deductive quantitative methods for research are also used. Scientific aim: The effort is directed at finding whether the method of prediction using artificial neural networks is suitable as a tool for enhancing the ordering system of an enterprise. The research also focuses on finding what architecture of the artificial neural networks model is the most suitable for subsequent prediction. Findings: Artificial neural networks models can be used for inventory management and lot-sizing problem successfully. A network with the TRAINGDX training function and TANSIG transfer function and 6-8-1 architecture can be considered the most suitable for artificial neural network, as it shows the best results for subsequent prediction. Conclusions: It can be concluded that the created model of artificial neural network can be successfully used for predicting order size and therefore for improving the order cycle of an enterprise. Conclusions resulting from the paper are beneficial for further research.

  14. Nanoparticles for siRNA-Based Gene Silencing in Tumor Therapy. (United States)

    Babu, Anish; Muralidharan, Ranganayaki; Amreddy, Narsireddy; Mehta, Meghna; Munshi, Anupama; Ramesh, Rajagopal


    Gene silencing through RNA interference (RNAi) has emerged as a potential strategy in manipulating cancer causing genes by complementary base-pairing mechanism. Small interfering RNA (siRNA) is an important RNAi tool that has found significant application in cancer therapy. However due to lack of stability, poor cellular uptake and high probability of loss-of-function due to degradation, siRNA therapeutic strategies seek safe and efficient delivery vehicles for in vivo applications. The current review discusses various nanoparticle systems currently used for siRNA delivery for cancer therapy, with emphasis on liposome based gene delivery systems. The discussion also includes various methods availed to improve nanoparticle based-siRNA delivery with target specificity and superior efficiency. Further this review describes challenges and perspectives on the development of safe and efficient nanoparticle based-siRNA-delivery systems for cancer therapy.


    African Journals Online (AJOL)

    Ifedotun Aina

    characterize and assess the suitability of the soils for oil palm and plantain production using the linear and square root parametric models. The delineated mapping units classified as Kandiudalfs were deep (>100 cm in depth), well drained with loamy sand to sandy loam surface horizons overlying sandy clay loam to sandy ...

  16. Asymptomatic Malaria Parasitaemia - A Suitable Index for

    African Journals Online (AJOL)

    Orogade AA, Ogala WN, Aikhionbare HA. Asymptomatic Malaria Parasitaemia - A Suitable. Index for Evaluation of Malaria Vector Control Measures. Nigerian journal of Paediatrics 2002;. 29: 23. To evaluate the impact of malaria vector control measures which form an integral part of the ational guidelines for malaria control, ...

  17. Suitability of Ostrich eggshell for radiocarbon dating

    CSIR Research Space (South Africa)

    Vogel, JC


    Full Text Available Ostrich eggshell from archaeological sites remains largely free of more recent carbon contamination and as such is suitable material for radiocarbon dating. The carbonate fraction of the shell does, however, display an initial deficit in C14, which...

  18. Suitability of obstetric ultrasonographic parameters in determining ...

    African Journals Online (AJOL)

    Objective: A prospective study aimed at assessing the suitability of Biparietal Diameter (BPD), Femoral Length (FL), Transverse abdominal Diameter (TAD) and Composite value by ultrasonography, in determining foetal or gestational age derived by last menstrual period (LMP) in our environment was undertaken.

  19. Web page classification on child suitability

    NARCIS (Netherlands)

    C. Eickhoff (Carsten); P. Serdyukov; A.P. de Vries (Arjen)


    htmlabstractChildren spend significant amounts of time on the Internet. Recent studies showed, that during these periods they are often not under adult supervision. This work presents an automatic approach to identifying suitable web pages for children based on topical and non-topical web page

  20. Suitability analysis in a changing climate

    NARCIS (Netherlands)

    Romeijn, Harmen; Faggian, Robert; Pinto Nunes Nogueira Diogo, Vasco; Sposito, Victor


    Land suitability analysis is employed to evaluate the appropriateness of land for a particular purpose whilst integrating both qualitative and quantitative inputs, which can be continuous in nature. However, in agricultural modelling there is often a disregard of this contiguous aspect. Therefore,


    African Journals Online (AJOL)

    SUITABILITY OF CARAMBOLA ( AVERRHOA CARAMBOLA ) FRUIT JUICE AS A SUBSTRATE FOR WINE FERMENTATION. ... The analysis of the sugars showed glucose, fructose and sucrose as the predominant sugars present. Vitamin C content was substantially high (35 mg/100 g), but fat content was low (0.25%).


    Directory of Open Access Journals (Sweden)

    Boris Đurđević


    Full Text Available A total of 17404 soil samples (2003rd-2009th year were analysed in the eastern Croatia. The largest number of soil samples belongs to the Osijek-Baranya county, which together with both Eastern sugar beet Factories (Osijek and Županja, conduct the soil fertility control (~4200 samples/yr.. Computer model suitability assessment for crops, supported by GIS, proved to be fast, efficient enough reliable in terms of the number of analyzed soil samples. It allows the visualization of the agricultural area and prediction of its production properties for the purposes of analysis, planning and rationalization of agricultural production. With more precise data about the soil (soil, climate and reliable Digital Soil Map of Croatia, the model could be an acceptable, not only to evaluate the suitability for growing different crops but also their need for fertilizer, necessary machinery, repairs (liming, and other measures of organic matter input. The abovementioned aims to eliminate or reduce effects of limiting factors in primary agricultural production. Assessment of the relative benefits of soil presented by computer model for the crops production and geostatistical method kriging in the Osijek-Baranya county showed: 1 Average soil suitability being 60.06 percent. 2 Kriging predicted that 51751 ha (17.16% are of limited resources (N1 for growing crops whereas a 86142 ha (28.57% of land is limited suitably (S3, b 132789 ha (44.04% are moderately suitable (S2 and c 30772 ha (10.28% are of excellent fertility (S1. A large number of eastern Croatian land data showed that the computer-geostatistical model for determination of soil benefits for growing crops was automated, fast and simple to use and suitable for the implementation of GIS and automatically downloading the necessary benefit indicators from the input base (land, analytical and climate as well as data from the digital soil maps able to: a visualize the suitability for soil tillage, b predict the

  3. ADAM33 gene silencing by promoter hypermethylation as a molecular marker in breast invasive lobular carcinoma

    Directory of Open Access Journals (Sweden)

    de Souza Emanuel M


    Full Text Available Abstract Background ADAM33 protein is a member of the family of transmembrane glycoproteins composed of multidomains. ADAM family members have different activities, such as proteolysis and adhesion, making them good candidates to mediate the extracellular matrix remodelling and changes in cellular adhesion that characterise certain pathologies and cancer development. It was reported that one family member, ADAM23, is down-regulated by promoter hypermethylation. This seems to correlate with tumour progression and metastasis in breast cancer. In this study, we explored the involvement of ADAM33, another ADAM family member, in breast cancer. Methods First, we analysed ADAM33 expression in breast tumour cell lines by RT-PCR and western blotting. We also used 5-aza-2'-deoxycytidine (5azadCR treatment and DNA bisulphite sequencing to study the promoter methylation of ADAM33 in breast tumour cell lines. We evaluated ADAM33 methylation in primary tumour samples by methylation specific PCR (MSP. Finally, ADAM33 promoter hypermethylation was correlated with clinicopathological data using the chi-square test and Fisher's exact test. Results The expression analysis of ADAM33 in breast tumour cell lines by RT-PCR revealed gene silencing in 65% of tumour cell lines. The corresponding lack of ADAM33 protein was confirmed by western blotting. We also used 5-aza-2'-deoxycytidine (5-aza-dCR demethylation and bisulphite sequencing methodologies to confirm that gene silencing is due to ADAM33 promoter hypermethylation. Using MSP, we detected ADAM33 promoter hypermethylation in 40% of primary breast tumour samples. The correlation between methylation pattern and patient's clinicopathological data was not significantly associated with histological grade; tumour stage (TNM; tumour size; ER, PR or ERBB2 status; lymph node status; metastasis or recurrence. Methylation frequency in invasive lobular carcinoma (ILC was 76.2% compared with 25.5% in invasive ductal carcinoma

  4. Prediction of suitable amount of water addition for wet granulation. (United States)

    Miwa, A; Yajima, T; Itai, S


    The purpose of this study was to predict the amounts of water addition suitable for pharmaceutical formulations in wet granulation, using a high-speed mixer or a fluidized bed granulator, before granulation trials. In order to determine the suitable amount of water addition, each excipient was first subjected to kneading with water in a mortar and a refractive near-infrared moisture sensor (IR sensor) measured the amount of water at the powder surface. Further by analysis the plot (output value of the IR sensor vs. amount of added water) for each excipient, the amount of water addition for granulation was determined for it. As a second step, two model formulations were designed and suitable amounts of water for granulation were predicted by summation of the obtained excipient values. The predicted value was compared with the experimental value for high-speed mixer granulation. The predicted and experimental amounts of water addition corresponded for the two model formulations, suggesting that the above method is useful for estimating suitable amounts of addition of water for formulations before granulation trials.

  5. Transcription-dependent silencing of inducible convergent transgenes in transgenic mice

    Directory of Open Access Journals (Sweden)

    Calero-Nieto Fernando J


    Full Text Available Abstract Background Silencing of transgenes in mice is a common phenomenon typically associated with short multi-copy transgenes. We have investigated the regulation of the highly inducible human granulocyte-macrophage colony-stimulating-factor gene (Csf2 in transgenic mice. Results In the absence of any previous history of transcriptional activation, this transgene was expressed in T lineage cells at the correct inducible level in all lines of mice tested. In contrast, the transgene was silenced in a specific subset of lines in T cells that had encountered a previous episode of activation. Transgene silencing appeared to be both transcription-dependent and mediated by epigenetic mechanisms. Silencing was accompanied by loss of DNase I hypersensitive sites and inability to recruit RNA polymerase II upon stimulation. This pattern of silencing was reflected by increased methylation and decreased acetylation of histone H3 K9 in the transgene. We found that silenced lines were specifically associated with a single pair of tail-to-tail inverted repeated copies of the transgene embedded within a multi-copy array. Conclusions Our study suggests that epigenetic transgene silencing can result from convergent transcription of inverted repeats which can lead to silencing of an entire multi-copy transgene array. This mechanism may account for a significant proportion of the reported cases of transgene inactivation in mice.

  6. Protocol: using virus-induced gene silencing to study the arbuscular mycorrhizal symbiosis in Pisum sativum

    DEFF Research Database (Denmark)

    Grønlund, Mette; Olsen, Anne; Johansen, Elisabeth


    Virus-induced gene silencing (VIGS) is an alternative reverse genetics tool for silencing of genes in some plants, which are difficult to transform. The pea early-browning virus (PEBV) has been developed as a VIGS vector and used in pea for functional analysis of several genes. However, the avail...

  7. Self-Silencing, Emotional Awareness, and Eating Behaviors in College Women (United States)

    Shouse, Sarah H.; Nilsson, Johanna


    Self-silencing (or the suppression of expressing one's thoughts, feelings, and needs) can have a negative impact on the mental health of women, from depression to disordered eating behaviors. The authors examined the relationship between self-silencing and disordered eating as well as intuitive eating. The authors also explored whether emotional…

  8. Identification and characterization of two RNA silencing suppressors encoded by ophioviruses

    NARCIS (Netherlands)

    Robles Luna, Gabriel; Reyes, Carina A.; Peña, Eduardo J.; Ocolotobiche, Eliana; Baeza, Cecilia; Borniego, Maria Belén; Kormelink, Richard; García, María Laura


    Citrus psorosis virus and Mirafiori lettuce big-vein virus are two members of the genus Ophiovirus, family Ophioviridae. So far, how these viruses can interfere in the antiviral RNA silencing pathway is not known. In this study, using a local GFP silencing assay on Nicotiana benthamiana, the

  9. Distinct Effects of p19 RNA Silencing Suppressor on Small RNA Mediated Pathways in Plants.

    Directory of Open Access Journals (Sweden)

    Levente Kontra


    Full Text Available RNA silencing is one of the main defense mechanisms employed by plants to fight viruses. In change, viruses have evolved silencing suppressor proteins to neutralize antiviral silencing. Since the endogenous and antiviral functions of RNA silencing pathway rely on common components, it was suggested that viral suppressors interfere with endogenous silencing pathway contributing to viral symptom development. In this work, we aimed to understand the effects of the tombusviral p19 suppressor on endogenous and antiviral silencing during genuine virus infection. We showed that ectopically expressed p19 sequesters endogenous small RNAs (sRNAs in the absence, but not in the presence of virus infection. Our presented data question the generalized model in which the sequestration of endogenous sRNAs by the viral suppressor contributes to the viral symptom development. We further showed that p19 preferentially binds the perfectly paired ds-viral small interfering RNAs (vsiRNAs but does not select based on their sequence or the type of the 5' nucleotide. Finally, co-immunoprecipitation of sRNAs with AGO1 or AGO2 from virus-infected plants revealed that p19 specifically impairs vsiRNA loading into AGO1 but not AGO2. Our findings, coupled with the fact that p19-expressing wild type Cymbidium ringspot virus (CymRSV overcomes the Nicotiana benthamiana silencing based defense killing the host, suggest that AGO1 is the main effector of antiviral silencing in this host-virus combination.

  10. Horreur et silence : le personnage de Pierre Cange dans l’œuvre narrative de Vercors

    Directory of Open Access Journals (Sweden)

    Elisabetta Sibilio


    Full Text Available Vercors, who achieved fame thanks to his first novel, Le silence de la mer, published undercover in 1942, makes silence one of the main themes in his work. Silence, which is the paradoxical expression of what Vercors calls “la qualité d’homme”, appears in his work according to two major categories: on the one hand the proud and resilient silence of the protagonist of Silence de la mer or of Arnaud, the main character of the short story Le démenti, who loses his life for having maintained an obstinate silence in the hands of his torturers; on the other hand, the guilty and desperate shameful silence of those who had lost their own humanity when faced with the horror of the camps or had hoped to save their lives by col-laborating with the enemy. This paper will examine three of Vercors’ texts, Les armes de la nuit (1946, La puissance du jour (1951 and Le tigre d’Anvers (1986, which have Pierre Cange as protagonist, a member of the Resistance, deported, who through his own experience is faced with the funda-mental theme of silence.

  11. Performing Silence: Gender, Violence, and Resistance in Women's Narratives from Lahaul, India (United States)

    Bhattacharya, Himika


    This article presents two different ways of understanding silence, through a discussion of women's narratives of violence from Lahaul, India. Here I illustrate how feminist ethnography works its way into re-conceptualizing silence as a tool women use to resist existing patriarchal discourses of honor, tribe and nation. (Contains 1 note.)

  12. Giving Voice: Narrating silence, history and memory in André Brink ...

    African Journals Online (AJOL)

    This essay examines André Brink's two most recent novels, The Other Side of Silence (2002) and Before I Forget (2004), in terms of their voicing of silence and the rewriting of history and memory. Each has a theme familiar to Brink's readers – an historical story of colonial violence and violation avenged; and the recounting ...

  13. Analysing the significance of silence in qualitative interviewing: questioning and shifting power relations

    DEFF Research Database (Denmark)

    Bengtsson, Tea Torbenfeldt; Fynbo, Lars


    In this article we analyse the significance of silence in qualitative interviews with 36 individuals interviewed about high-risk, illegal activities. We describe how silence expresses a dynamic power relationship between interviewer and interviewee. In the analysis, we focus on two different type...

  14. Down-Regulation of Gene Expression by RNA-Induced Gene Silencing (United States)

    Travella, Silvia; Keller, Beat

    Down-regulation of endogenous genes via post-transcriptional gene silencing (PTGS) is a key to the characterization of gene function in plants. Many RNA-based silencing mechanisms such as post-transcriptional gene silencing, co-suppression, quelling, and RNA interference (RNAi) have been discovered among species of different kingdoms (plants, fungi, and animals). One of the most interesting discoveries was RNAi, a sequence-specific gene-silencing mechanism initiated by the introduction of double-stranded RNA (dsRNA), homologous in sequence to the silenced gene, which triggers degradation of mRNA. Infection of plants with modified viruses can also induce RNA silencing and is referred to as virus-induced gene silencing (VIGS). In contrast to insertional mutagenesis, these emerging new reverse genetic approaches represent a powerful tool for exploring gene function and for manipulating gene expression experimentally in cereal species such as barley and wheat. We examined how RNAi and VIGS have been used to assess gene function in barley and wheat, including molecular mechanisms involved in the process and available methodological elements, such as vectors, inoculation procedures, and analysis of silenced phenotypes.

  15. Preliminary geological suitability assessment for LILW disposal

    International Nuclear Information System (INIS)

    Tomse, P.; Mele, I.


    Due to the growing need for a final disposal of LILW, the final solution for the short-lived LILW is the key issue of radioactive waste management in Slovenia at the moment. ARAO - the Slovenian Agency for Radwaste Management - is intensely involved in the re-initiated site selection process for a LILW repository. In this new process we are trying to combine as best as possible the technical, geologically-led and the advocacy-site selection processes. By a combination of technical and volunteer approach to the site selection we wish to guarantee high public involvement and sufficient flexibility of the process to adapt to specific conditions or new circumstances while the project is ongoing. In the technical phase, our tendency is to retain a larger number of potential areas/sites. We also keep open the possibility of choosing the type of repository. The decision between the surface and underground option will be made only once the site has been defined. In accordance with the IAEA recommendations the site selection process is divided into four stages: the conceptual and planning stage, area survey stage, site characterisation stage and site confirmation stage. Last year the area survey stage was started. In the preliminary geological suitability assessment the required natural predisposition of Slovene territory was assessed in order to locate geologically suitable formations. The assessment of natural conditions of the system was based on consideration of the main geological, hydro-geological and seismotectonic conditions. It was performed with ARC/INFO technology. The results are compiled in a map, showing potential areas for underground and surface disposal of LILW in Slovenia. It has been established that there is a potential suitability for both surface and underground disposal on about 10 000 km 2 of the Slovenian territory, which represents almost half of the entire Slovenian territory. These preliminary results are now being carefully re-examined. As an

  16. Achievement report for fiscal 1998. Research and development on a new manufacturing method for functional thin films suitable for recycling, and their application to colored glasses (the second year); 1998 nendo seika hokokusho. Recycle ni tekishita kinosei usumaku no shinki seizoho to chakushoku glass eno oyo ni kansuru kenkyu kaihatsu (dai 2 nendo)

    Energy Technology Data Exchange (ETDEWEB)



    A new thin film manufacturing method is established to add a function to glass material surface, as a new material technology which harmonizes with global environment, and is suitable for resource re-utilization and energy conservation. It is intended to develop a leading technology to promote recycling of colored glasses by applying this technical method to colored glasses. Fiscal 1998 has implemented subsequently to fiscal 1997 the following subjects in the three research items composed of a new manufacturing method of functional thin films, application of the functional thin films to colored glasses, and the comprehensive investigative studies: establishment of an industrial manufacturing method for color coating liquid and evaluation of basic characteristics of the colored functional thin films, optimization of element technology for photo-sensitive gel films by means of chemically modifying metallic alkoxide, tests of forming films on glass bottles and plate glasses by using a coating machine installed in fiscal 1997, design and prototype fabrication of a new demonstration coating machine, and analysis on thermal decomposition of the colored thin films. Optimization was performed on the element technology for manufacturing sol-gel functional thin films, and a survey was carried out on recycling systems of colored glasses adopted in Europe. (NEDO)

  17. Improved apple latent spherical virus-induced gene silencing in multiple soybean genotypes through direct inoculation of agro-infiltratedNicotiana benthamianaextract. (United States)

    Gedling, C R; Ali, E M; Gunadi, A; Finer, J J; Xie, K; Liu, Y; Yoshikawa, N; Qu, F; Dorrance, A E


    Virus induced gene silencing (VIGS) is a powerful genomics tool for interrogating the function of plant genes. Unfortunately, VIGS vectors often produce disease symptoms that interfere with the silencing phenotypes of target genes, or are frequently ineffective in certain plant genotypes or tissue types. This is especially true in crop plants like soybean [ Glycine max (L.) Merr]. To address these shortcomings, we modified the inoculation procedure of a VIGS vector based on Apple latent spherical virus (ALSV). The efficacy of this new procedure was assessed in 19 soybean genotypes using a soybean Phytoene desaturase ( GmPDS1 ) gene as the VIGS target. Silencing of GmPDS1 was easily scored as photo-bleached leaves and/or stems. In this report, the ALSV VIGS vector was modified by mobilizing ALSV cDNAs into a binary vector compatible with Agrobacterium tumefaciens -mediated delivery, so that VIGS-triggering ALSV variants could be propagated in agro-infiltrated Nicotiana benthamiana leaves. Homogenate of these N. benthamiana leaves was then applied directly onto the unifoliate of young soybean seedlings to initiate systemic gene silencing. This rapid inoculation method bypassed the need for a particle bombardment apparatus. Among the 19 soybean genotypes evaluated with this new method, photo-bleaching indicative of GmPDS1 silencing was observed in nine, with two exhibiting photo-bleaching in 100% of the inoculated individuals. ALSV RNA was detected in pods, embryos, stems, leaves, and roots in symptomatic plants of four genotypes. This modified protocol allowed for inoculation of soybean plants via simple mechanical rubbing with the homogenate of N. benthamiana leaves agro-infiltrated with ALSV VIGS constructs. More importantly, inoculated plants showed no apparent virus disease symptoms which could otherwise interfere with VIGS phenotypes. This streamlined procedure expanded this functional genomics tool to nine soybean genotypes.

  18. A semiotic approach to suitability for psychotherapy. (United States)

    Meltzer, J D


    At the beginning of my career as a psychotherapist I interviewed a number of patients in an effort to find one who seemed to be a good prospect for long-term psychotherapy. While interviewing with this purpose in mind, I was struck by the fact that a number of the people I spoke with gave me a sense of being extremely suitable for this sort of therapy, but that others seemed quite unsuitable. An informal survey among my peers as well as among more senior therapists revealed that they were not unfamiliar with the phenomenon. The present empirical study of the semiotic aspects of suitability for psychotherapy grew out of this early experience.

  19. Global habitat suitability models of terrestrial mammals. (United States)

    Rondinini, Carlo; Di Marco, Moreno; Chiozza, Federica; Santulli, Giulia; Baisero, Daniele; Visconti, Piero; Hoffmann, Michael; Schipper, Jan; Stuart, Simon N; Tognelli, Marcelo F; Amori, Giovanni; Falcucci, Alessandra; Maiorano, Luigi; Boitani, Luigi


    Detailed large-scale information on mammal distribution has often been lacking, hindering conservation efforts. We used the information from the 2009 IUCN Red List of Threatened Species as a baseline for developing habitat suitability models for 5027 out of 5330 known terrestrial mammal species, based on their habitat relationships. We focused on the following environmental variables: land cover, elevation and hydrological features. Models were developed at 300 m resolution and limited to within species' known geographical ranges. A subset of the models was validated using points of known species occurrence. We conducted a global, fine-scale analysis of patterns of species richness. The richness of mammal species estimated by the overlap of their suitable habitat is on average one-third less than that estimated by the overlap of their geographical ranges. The highest absolute difference is found in tropical and subtropical regions in South America, Africa and Southeast Asia that are not covered by dense forest. The proportion of suitable habitat within mammal geographical ranges correlates with the IUCN Red List category to which they have been assigned, decreasing monotonically from Least Concern to Endangered. These results demonstrate the importance of fine-resolution distribution data for the development of global conservation strategies for mammals.

  20. Postpartum Depression: How Childbirth Educators Can Help Break the Silence (United States)

    Zauderer, Cheryl


    The voices of women suffering from postpartum depression are often silent. Women are reluctant to reveal to others that they are unhappy after the birth of their babies. Much has been written on possible causes, risk factors, and treatments for postpartum depression, but little has been done to investigate why women take so long to seek help. Early detection and treatment are key to a full recovery. Childbirth educators are in the position to offer anticipatory guidance on possible complications of the postpartum period, including postpartum depression. This article explores why women with postpartum depression choose to suffer in silence and suggests how childbirth educators can help new mothers find their voices. PMID:20190853

  1. Sonic memory interventions against politics of urban silencing

    Directory of Open Access Journals (Sweden)

    Hofman Ana


    Full Text Available We discuss the political implications of the noise/silence dialectic in order to reflect on the urban and social materialities of sonic memory activism in the post- Yugoslav space. We see the privatization of public space as one of the defining issues of current socio-political tensions and we strive to offer a more nuanced model for thinking about grassroots practices of musicking and listening in the context of resistance and power and control redistribution. Discussing sonic interventions in Ljubljana and Belgrade enables us both to uncover how important global processes are reflected in these local contexts and to locate diversity of present practices of resistance. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. 177004: Serbian Musical Identities Within Local and Global Frameworks: Traditions, Changes, Challenges

  2. SOCS3 Silencing Attenuates Eosinophil Functions in Asthma Patients (United States)

    Zafra, Mª Paz; Cañas, Jose A.; Mazzeo, Carla; Gámez, Cristina; Sanz, Veronica; Fernández-Nieto, Mar; Quirce, Santiago; Barranco, Pilar; Ruiz-Hornillos, Javier; Sastre, Joaquín; del Pozo, Victoria


    Eosinophils are one of the key inflammatory cells in asthma. Eosinophils can exert a wide variety of actions through expression and secretion of multiple molecules. Previously, we have demonstrated that eosinophils purified from peripheral blood from asthma patients express high levels of suppressor of cytokine signaling 3 (SOCS3). In this article, SOCS3 gene silencing in eosinophils from asthmatics has been carried out to achieve a better understanding of the suppressor function in eosinophils. SOCS3 siRNA treatment drastically reduced SOCS3 expression in eosinophils, leading to an inhibition of the regulatory transcription factors GATA-3 and FoxP3, also interleukin (IL)-10; in turn, an increased STAT3 phosphorilation was observed. Moreover, SOCS3 abrogation in eosinophils produced impaired migration, adhesion and degranulation. Therefore, SOCS3 might be regarded as an important regulator implicated in eosinophil mobilization from the bone marrow to the lungs during the asthmatic process. PMID:25764157

  3. Silencing of voices in a Swedish science classroom (United States)

    Ramos de Robles, S. Lizette


    From a sociocultural perspective, I discuss data from a Swedish science classroom presented in María Gómez's article "Student Explanations of their Science Teachers' Assessments, Grading Practices, and How they learn Science". In this discussion, I focus on the need to change existing conceptions of assessment in the teaching and learning of science. Next, I talk about the importance of taking into consideration the dialectic between agency and passivity as filters in order to understand what student silence may signify in science classes as well as in relation to their perceptions of assessment. I conclude with the importance of the teacher's role in developing formative assessment, along with the challenges in developing assessments which transform science education into a relevant field of knowledge for both students and society at large.

  4. Manipulation of Cell Physiology Enables Gene Silencing in Well-differentiated Airway Epithelia

    Directory of Open Access Journals (Sweden)

    Sateesh Krishnamurthy


    Full Text Available The application of RNA interference-based gene silencing to the airway surface epithelium holds great promise to manipulate host and pathogen gene expression for therapeutic purposes. However, well-differentiated airway epithelia display significant barriers to double-stranded small-interfering RNA (siRNA delivery despite testing varied classes of nonviral reagents. In well-differentiated primary pig airway epithelia (PAE or human airway epithelia (HAE grown at the air–liquid interface (ALI, the delivery of a Dicer-substrate small-interfering RNA (DsiRNA duplex against hypoxanthine–guanine phosphoribosyltransferase (HPRT with several nonviral reagents showed minimal uptake and no knockdown of the target. In contrast, poorly differentiated cells (2–5-day post-seeding exhibited significant oligonucleotide internalization and target knockdown. This finding suggested that during differentiation, the barrier properties of the epithelium are modified to an extent that impedes oligonucleotide uptake. We used two methods to overcome this inefficiency. First, we tested the impact of epidermal growth factor (EGF, a known enhancer of macropinocytosis. Treatment of the cells with EGF improved oligonucleotide uptake resulting in significant but modest levels of target knockdown. Secondly, we used the connectivity map (Cmap database to correlate gene expression changes during small molecule treatments on various cells types with genes that change upon mucociliary differentiation. Several different drug classes were identified from this correlative assessment. Well-differentiated epithelia treated with DsiRNAs and LY294002, a PI3K inhibitor, significantly improved gene silencing and concomitantly reduced target protein levels. These novel findings reveal that well-differentiated airway epithelia, normally resistant to siRNA delivery, can be pretreated with small molecules to improve uptake of synthetic oligonucleotide and RNA interference (RNAi responses.

  5. High-throughput sequencing of RNA silencing-associated small RNAs in olive (Olea europaea L..

    Directory of Open Access Journals (Sweden)

    Livia Donaire

    Full Text Available Small RNAs (sRNAs of 20 to 25 nucleotides (nt in length maintain genome integrity and control gene expression in a multitude of developmental and physiological processes. Despite RNA silencing has been primarily studied in model plants, the advent of high-throughput sequencing technologies has enabled profiling of the sRNA component of more than 40 plant species. Here, we used deep sequencing and molecular methods to report the first inventory of sRNAs in olive (Olea europaea L.. sRNA libraries prepared from juvenile and adult shoots revealed that the 24-nt class dominates the sRNA transcriptome and atypically accumulates to levels never seen in other plant species, suggesting an active role of heterochromatin silencing in the maintenance and integrity of its large genome. A total of 18 known miRNA families were identified in the libraries. Also, 5 other sRNAs derived from potential hairpin-like precursors remain as plausible miRNA candidates. RNA blots confirmed miRNA expression and suggested tissue- and/or developmental-specific expression patterns. Target mRNAs of conserved miRNAs were computationally predicted among the olive cDNA collection and experimentally validated through endonucleolytic cleavage assays. Finally, we use expression data to uncover genetic components of the miR156, miR172 and miR390/TAS3-derived trans-acting small interfering RNA (tasiRNA regulatory nodes, suggesting that these interactive networks controlling developmental transitions are fully operational in olive.

  6. High-Throughput Sequencing of RNA Silencing-Associated Small RNAs in Olive (Olea europaea L.) (United States)

    Donaire, Livia; Pedrola, Laia; de la Rosa, Raúl; Llave, César


    Small RNAs (sRNAs) of 20 to 25 nucleotides (nt) in length maintain genome integrity and control gene expression in a multitude of developmental and physiological processes. Despite RNA silencing has been primarily studied in model plants, the advent of high-throughput sequencing technologies has enabled profiling of the sRNA component of more than 40 plant species. Here, we used deep sequencing and molecular methods to report the first inventory of sRNAs in olive (Olea europaea L.). sRNA libraries prepared from juvenile and adult shoots revealed that the 24-nt class dominates the sRNA transcriptome and atypically accumulates to levels never seen in other plant species, suggesting an active role of heterochromatin silencing in the maintenance and integrity of its large genome. A total of 18 known miRNA families were identified in the libraries. Also, 5 other sRNAs derived from potential hairpin-like precursors remain as plausible miRNA candidates. RNA blots confirmed miRNA expression and suggested tissue- and/or developmental-specific expression patterns. Target mRNAs of conserved miRNAs were computationally predicted among the olive cDNA collection and experimentally validated through endonucleolytic cleavage assays. Finally, we use expression data to uncover genetic components of the miR156, miR172 and miR390/TAS3-derived trans-acting small interfering RNA (tasiRNA) regulatory nodes, suggesting that these interactive networks controlling developmental transitions are fully operational in olive. PMID:22140484

  7. Flexible tools for gene expression and silencing in tomato. (United States)

    Fernandez, Ana I; Viron, Nicolas; Alhagdow, Moftah; Karimi, Mansour; Jones, Matthew; Amsellem, Ziva; Sicard, Adrien; Czerednik, Anna; Angenent, Gerco; Grierson, Donald; May, Sean; Seymour, Graham; Eshed, Yuval; Lemaire-Chamley, Martine; Rothan, Christophe; Hilson, Pierre


    As a genetic platform, tomato (Solanum lycopersicum) benefits from rich germplasm collections and ease of cultivation and transformation that enable the analysis of biological processes impossible to investigate in other model species. To facilitate the assembly of an open genetic toolbox designed to study Solanaceae, we initiated a joint collection of publicly available gene manipulation tools. We focused on the characterization of promoters expressed at defined time windows during fruit development, for the regulated expression or silencing of genes of interest. Five promoter sequences were captured as entry clones compatible with the versatile MultiSite Gateway format: PPC2, PG, TPRP, and IMA from tomato and CRC from Arabidopsis (Arabidopsis thaliana). Corresponding transcriptional fusions were made with the GUS gene, a nuclear-localized GUS-GFP reporter, and the chimeric LhG4 transcription factor. The activity of the promoters during fruit development and in fruit tissues was confirmed in transgenic tomato lines. Novel Gateway destination vectors were generated for the transcription of artificial microRNA (amiRNA) precursors and hairpin RNAs under the control of these promoters, with schemes only involving Gateway BP and LR Clonase reactions. Efficient silencing of the endogenous phytoene desaturase gene was demonstrated in transgenic tomato lines producing a matching amiRNA under the cauliflower mosaic virus 35S or PPC2 promoter. Lastly, taking advantage of the pOP/LhG4 two-component system, we found that well-characterized flower-specific Arabidopsis promoters drive the expression of reporters in patterns generally compatible with heterologous expression. Tomato lines and plasmids will be distributed through a new Nottingham Arabidopsis Stock Centre service unit dedicated to Solanaceae resources.

  8. Polyphenoloxidase Silencing Affects Latex Coagulation in Taraxacum Species1[W (United States)

    Wahler, Daniela; Gronover, Christian Schulze; Richter, Carolin; Foucu, Florence; Twyman, Richard M.; Moerschbacher, Bruno M.; Fischer, Rainer; Muth, Jost; Prüfer, Dirk


    Latex is the milky sap that is found in many different plants. It is produced by specialized cells known as laticifers and can comprise a mixture of proteins, carbohydrates, oils, secondary metabolites, and rubber that may help to prevent herbivory and protect wound sites against infection. The wound-induced browning of latex suggests that it contains one or more phenol-oxidizing enzymes. Here, we present a comprehensive analysis of the major latex proteins from two dandelion species, Taraxacum officinale and Taraxacum kok-saghyz, and enzymatic studies showing that polyphenoloxidase (PPO) is responsible for latex browning. Electrophoretic analysis and amino-terminal sequencing of the most abundant proteins in the aqueous latex fraction revealed the presence of three PPO-related proteins generated by the proteolytic cleavage of a single precursor (pre-PPO). The laticifer-specific pre-PPO protein contains a transit peptide that can target reporter proteins into chloroplasts when constitutively expressed in dandelion protoplasts, perhaps indicating the presence of structures similar to plastids in laticifers, which lack genuine chloroplasts. Silencing the PPO gene by constitutive RNA interference in transgenic plants reduced PPO activity compared with wild-type controls, allowing T. kok-saghyz RNA interference lines to expel four to five times more latex than controls. Latex fluidity analysis in silenced plants showed a strong correlation between residual PPO activity and the coagulation rate, indicating that laticifer-specific PPO plays a major role in latex coagulation and wound sealing in dandelions. In contrast, very little PPO activity is found in the latex of the rubber tree Hevea brasiliensis, suggesting functional divergence of latex proteins during plant evolution. PMID:19605551

  9. Dismantling the Silence: LGBTQ Aging Emerging From the Margins. (United States)

    Fredriksen-Goldsen, Karen I


    Historical, environmental, and cultural contexts intersect with aging, sexuality, and gender across communities and generations. My scholarship investigates health and well-being over the life course across marginalized communities, including LGBTQ (lesbian, gay, bisexual, transgender, and queer) midlife and older adults, native communities experiencing cardiovascular risk, and families in China living with HIV, in order to balance the realities of unique lives in contemporary society. By probing the intersection of age, sexuality, and gender, my analysis is informed by both personal and professional experiences. With the death of my partner occurring at a time of profound invisibility and silence before HIV/AIDS, I found my life out of sync, experiencing a loss without a name. My life was thrust into a paradox: My relationship was defined by a world that refused to recognize it. This essay provides an opportunity for me to weave together how such critical turning points in my own life helped shape my approach to gerontology and how gerontology has informed my work and life. Reflecting on this journey, I illustrate the ways in which historical, structural, environmental, psychosocial, and biological factors affect equity, and the health-promoting and adverse pathways to health and well-being across marginalized communities. Although gerontology as a discipline has historically silenced the lives of marginalized older adults, it has much to learn from these communities. The growing and increasingly diverse older adult population provides us with unique opportunities to better understand both cultural variations and shared experiences in aging over the life course. © The Author 2017. Published by Oxford University Press on behalf of The Gerontological Society of America. All rights reserved. For permissions, please e-mail:

  10. Silencing Intersectin 1 Slows Orthotopic Neuroblastoma Growth in Mice. (United States)

    Harris, Jamie; Herrero-Garcia, Erika; Russo, Angela; Kajdacsy-Balla, Andre; O'Bryan, John P; Chiu, Bill


    Neuroblastoma accounts for 15% of all pediatric cancer deaths. Intersectin 1 (ITSN1), a scaffold protein involved in phosphoinositide 3-kinase (PI3K) signaling, regulates neuroblastoma cells independent of MYCN status. We hypothesize that by silencing ITSN1 in neuroblastoma cells, tumor growth will be decreased in an orthotopic mouse tumor model. SK-N-AS neuroblastoma cells transfected with empty vector (pSR), vectors expressing scrambled shRNA (pSCR), or shRNAs targeting ITSN1 (sh#1 and sh#2) were used to create orthotopic neuroblastoma tumors in mice. Volume was monitored weekly with ultrasound. End-point was tumor volume >1000 mm. Tumor cell lysates were analyzed with anti-ITSN1 antibody by Western blot. Orthotopic tumors were created in all cell lines. Twenty-five days post injection, pSR tumor size was 917.6±247.7 mm, pSCR was 1180±159.9 mm, sh#1 was 526.3±212.8 mm, and sh#2 was 589.2±74.91 mm. sh#1-tumors and sh#2-tumors were smaller than pSCR (P=0.02), no difference between sh#1 and sh#2. Survival was superior in sh#2-tumors (P=0.02), trended towards improved survival in sh#1-tumors (P=0.09), compared with pSCR-tumors, no difference in pSR tumors. Western blot showed decreased ITSN1 expression in sh#1 and sh#2 compared with pSR and pSCR. Silencing ITSN1 in neuroblastoma cells led to decreased tumor growth in an orthotopic mouse model. Orthotopic animal models can provide insight into the role of ITSN1 pathways in neuroblastoma tumorigenesis.

  11. AMFR gene silencing inhibits the differentiation of porcine preadipocytes. (United States)

    Chen, C Z; Zhu, Y N; Chai, M L; Dai, L S; Gao, Y; Jiang, H; Zhang, L J; Ding, Y; Liu, S Y; Li, Q Y; Lu, W F; Zhang, J B


    Our study clarifies the role of the autocrine motility factor receptor (AMFR) gene in porcine preadipocyte differentiation. AMFR-siRNA was transfected into porcine preadipocytes and the preadipocytes were induced to differentiation. Subsequently, qRT-PCR was conducted to examine changes in mRNA expression of a series of genes in porcine preadipocytes, including AMFR, sterol-regulatory element-binding protein-1a (SREBP1a), SREBP2, insulin-induced gene 1 (Insig1), and Insig2. Expression changes in the mRNA of genes regulating adipocyte differentiation were also analyzed using qRT-PCR, including peroxisome proliferator-activated receptor gamma (PPARγ), CCAAT/enhancer-binding protein alpha (C/EBPα), and Kruppel-like factor 2 (KLF2). Western blot analysis was conducted to examine the changes in AMFR protein expression in porcine preadipocytes. Additionally, morphological changes in differentiated porcine preadipocytes were examined by oil red O staining, and changes in optical density (OD) values were measured using an ultraviolet spectrophotometer. At 24 h after transfection with AMFR-siRNA, AMFR mRNA expression significantly reduced (P SREBP1a, SREBP2, Insig1, and C/EBPα was significantly reduced (P < 0.01), whereas the expression of KLF2 mRNA was significantly elevated (P < 0.01). After induction of preadipocyte differentiation, the number of lipid droplets decreased in the AMFR-silenced group, and the OD value markedly reduced (P < 0.05). In addition, the expression of C/EBPα mRNA significantly decreased (P < 0.05), whereas the expression of KLF2 mRNA considerably increased (P < 0.05). Taken together, silencing of the AMFR gene inhibits the differentiation of porcine preadipocytes.

  12. Hypoxia Silences Retrotrapezoid Nucleus Respiratory Chemoreceptors via Alkalosis (United States)

    Basting, Tyler M.; Burke, Peter G.R.; Kanbar, Roy; Viar, Kenneth E.; Stornetta, Daniel S.; Stornetta, Ruth L.


    In conscious mammals, hypoxia or hypercapnia stimulates breathing while theoretically exerting opposite effects on central respiratory chemoreceptors (CRCs). We tested this theory by examining how hypoxia and hypercapnia change the activity of the retrotrapezoid nucleus (RTN), a putative CRC and chemoreflex integrator. Archaerhodopsin-(Arch)-transduced RTN neurons were reversibly silenced by light in anesthetized rats. We bilaterally transduced RTN and nearby C1 neurons with Arch (PRSx8-ArchT-EYFP-LVV) and measured the cardiorespiratory consequences of Arch activation (10 s) in conscious rats during normoxia, hypoxia, or hyperoxia. RTN photoinhibition reduced breathing equally during non-REM sleep and quiet wake. Compared with normoxia, the breathing frequency reduction (ΔfR) was larger in hyperoxia (65% FiO2), smaller in 15% FiO2, and absent in 12% FiO2. Tidal volume changes (ΔVT) followed the same trend. The effect of hypoxia on ΔfR was not arousal-dependent but was reversed by reacidifying the blood (acetazolamide; 3% FiCO2). ΔfR was highly correlated with arterial pH up to arterial pH (pHa) 7.5 with no frequency inhibition occurring above pHa 7.53. Blood pressure was minimally reduced suggesting that C1 neurons were very modestly inhibited. In conclusion, RTN neurons regulate eupneic breathing about equally during both sleep and wake. RTN neurons are the first putative CRCs demonstrably silenced by hypocapnic hypoxia in conscious mammals. RTN neurons are silent above pHa 7.5 and increasingly active below this value. During hyperoxia, RTN activation maintains breathing despite the inactivity of the carotid bodies. Finally, during hypocapnic hypoxia, carotid body stimulation increases breathing frequency via pathways that bypass RTN. PMID:25589748

  13. Dismantling the Silence: LGBTQ Aging Emerging From the Margins (United States)

    Fredriksen-Goldsen, Karen I.


    Historical, environmental, and cultural contexts intersect with aging, sexuality, and gender across communities and generations. My scholarship investigates health and well-being over the life course across marginalized communities, including LGBTQ (lesbian, gay, bisexual, transgender, and queer) midlife and older adults, native communities experiencing cardiovascular risk, and families in China living with HIV, in order to balance the realities of unique lives in contemporary society. By probing the intersection of age, sexuality, and gender, my analysis is informed by both personal and professional experiences. With the death of my partner occurring at a time of profound invisibility and silence before HIV/AIDS, I found my life out of sync, experiencing a loss without a name. My life was thrust into a paradox: My relationship was defined by a world that refused to recognize it. This essay provides an opportunity for me to weave together how such critical turning points in my own life helped shape my approach to gerontology and how gerontology has informed my work and life. Reflecting on this journey, I illustrate the ways in which historical, structural, environmental, psychosocial, and biological factors affect equity, and the health-promoting and adverse pathways to health and well-being across marginalized communities. Although gerontology as a discipline has historically silenced the lives of marginalized older adults, it has much to learn from these communities. The growing and increasingly diverse older adult population provides us with unique opportunities to better understand both cultural variations and shared experiences in aging over the life course. PMID:28053011

  14. Niche suitability affects development: skull asymmetry increases in less suitable areas.

    Directory of Open Access Journals (Sweden)

    Renan Maestri

    Full Text Available For conservation purposes, it is important to take into account the suitability of a species to particular habitats; this information may predict the long-term survival of a species. In this sense, morphological measures of developmental stress, such as fluctuating asymmetry, can be proxies for an individual's performance in different regions. In this study, we conducted tests to determine whether areas with different levels of suitability for a species (generated by ecological niche models were congruent with morphological markers that reflect environmental stress and morphological variance. We generated a Maxent niche model and compared the suitability assessments of several areas with the skull morphology data (fluctuating asymmetry and morphological disparity of populations of the Atlantic forest endemic to Brazil rodent Akodon cursor. Our analyses showed a significant negative relationship between suitability levels and fluctuating asymmetry levels, which indicates that in less suitable areas, the individuals experience numerous disturbances during skull ontogeny. We have not found an association between morphological variance and environmental suitability. As expected, these results suggest that in environments with a lower suitability, developmental stress is increased. Such information is helpful in the understanding of the species evolution and in the selection of priority areas for the conservation of species.

  15. Position-Dependent Methylation and Transcriptional Silencing of Transgenes in Inverted T-DNA Repeats: Implications for Posttranscriptional Silencing of Homologous Host Genes in Plants (United States)

    Stam, Maike; Viterbo, Ada; Mol, Joseph N. M.; Kooter, Jan M.


    Posttranscriptional silencing of chalcone synthase (Chs) genes in petunia transformants occurs by introducing T-DNAs that contain a promoter-driven or promoterless Chs transgene. With the constructs we used, silencing occurs only by T-DNA loci which are composed of two or more T-DNA copies that are arranged as inverted repeats (IRs). Since we are interested in the mechanism by which these IR loci induce silencing, we have analyzed different IR loci and nonsilencing single-copy (S) T-DNA loci with respect to the expression and methylation of the transgenes residing in these loci. We show that in an IR locus, the transgenes located proximal to the IR center are much more highly methylated than are the distal genes. A strong silencing locus composed of three inverted T-DNAs bearing promoterless Chs transgenes was methylated across the entire locus. The host Chs genes in untransformed plants were moderately methylated, and no change in methylation was detected when the genes were silenced. Run-on transcription assays showed that promoter-driven transgenes located proximal to the center of a particular IR are transcriptionally more repressed than are the distal genes of the same IR locus. Transcription of the promoterless Chs transgenes could not be detected. In the primary transformant, some of the IR loci were detected together with an unlinked S locus. We observed that the methylation and expression characteristics of the transgenes of these S loci were comparable to those of the partner IR loci, suggesting that there has been cross talk between the two types of loci. Despite the similar features, S loci are unable to induce silencing, indicating that the palindromic arrangement of the Chs transgenes in the IR loci is critical for silencing. Since transcriptionally silenced transgenes in IRs can trigger posttranscriptional silencing of the host genes, our data are most consistent with a model of silencing in which the transgenes physically interact with the

  16. Agroforestry suitability analysis based upon nutrient availability mapping: a GIS based suitability mapping

    Directory of Open Access Journals (Sweden)

    Firoz Ahmad


    Full Text Available Agroforestry has drawn the attention of researchers due to its capacity to reduce the poverty and land degradation, improve food security and mitigate the climate change. However, the progress in promoting agroforestry is held back due to the lack of reliable data sets and appropriate tools to accurately map and to have an adequate decision making system for agroforestry modules. Agroforestry suitability being one special form of land suitability is very pertinent to study in the current times when there is tremendous pressure on the land as it is a limited commodity. The study aims for applying the geo-spatial tools towards visualizing various soil and environmental data to reveal the trends and interrelationships and to achieve a nutrient availability and agroforestry suitability map. Using weight matrix and ranks, individual maps were developed in ArcGIS 10.1 platform to generate nutrient availability map, which was later used to develop agroforestry suitability map. Watersheds were delineated using DEM in some part of the study area and were evaluated for prioritizing it and agroforestry suitability of the watersheds were also done as per the schematic flowchart. Agroforestry suitability regions were delineated based upon the weight and ranks by integrated mapping. The total open area was identified 42.4% out of which 21.6% area was found to have high suitability towards agroforestry. Within the watersheds, 22 village points were generated for creating buffers, which were further evaluated showing its proximity to high suitable agroforestry sites thus generating tremendous opportunity to the villagers to carry out agroforestry projects locally. This research shows the capability of remote sensing in studying agroforestry practices and in estimating the prominent factors for its optimal productivity. The ongoing agroforestry projects can be potentially diverted in the areas of high suitability as an extension. The use of ancillary data in GIS

  17. Simultaneous silencing of multiple genes in the apple scab fungus, Venturia inaequalis, by expression of RNA with chimeric inverted repeats

    NARCIS (Netherlands)

    Fitzgerald, A.; Kan, van J.A.L.; Plummer, K.M.


    RNA-mediated gene silencing has been demonstrated in plants, animals, and more recently in filamentous fungi. Here, we report high frequency, RNA-mediated gene silencing in the apple scab fungus, Venturia inaequalis. The green fluorescent protein (GFP) transgene was silenced in a GFP-expressing

  18. Ways to Promote the Classroom Participation of International Students by Understanding the Silence of Japanese University Students (United States)

    Kim, Soonhyang; Ates, Burcu; Grigsby, Yurimi; Kraker, Stefani; Micek, Timothy A.


    The authors explored the role of silence and deciphered its meaning and usefulness as a teaching and learning strategy for Japanese students through a survey of Japanese university students in their home country. This study has revealed that participant responses were evenly divided among comfortable with silence, uncomfortable with silence, and…

  19. Delineation Lithuanian agricultural land for agro-ecological suitability for farming using soil and terrain criteria

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    Jarasiunas Gintaras


    Full Text Available The aim of investigation is to analyse and classify the state of agricultural land affected by naturalbiophysical, that is, soil and terrain-slope, handicaps on its overall agro-ecological suitability for agricultural use. For the classification of land for suitability with respect to sustainable use and efficient protection, the following actual criteria were selected: soil texture, soil drainage and terrainslope. For identifying the relatively homogeneous areas, the Ward hierarchical cluster method was used. According to our estimates, Lithuanian agricultural land with unfavourable soil texture, poorly soil drainage and steep slopes covers an area of 33.59, 4.76 and 1.03% of total agricultural area, respectively. On the basis of functional classification of state of agro-ecological conditions of Lithuania, two orders of suitability (S-suitable, N-not suitable and five land suitability classes were identified and delineated: S1 (highly suitable included 10 district municipalities, S2 (moderately suitable included 12 district municipalities, S3 (marginally suitable included 15 district municipalities, N1 (currently not suitable included 10 district municipalities and N2 (permanently not suitable included 4 district municipalities. S3 occupies the largest (29.80% share of the Lithuanian territory and N2 the least (5.66%. The land suitable for agriculture means with suitability classes S1, S2 and S3 was found on an area of 2,960,562 ha, which is 81.6% of the total land. In addition, soil texture was the most important factor causing differences in the classes of suitability. On the basis of natural limiting factors from an agronomic and eco-environmental perspective, the optimal sustainable development in agrosphere and a balanced-practical concept of land management and proper land use policy is presented. It is a reasonable base for agroecological regionalisation of Lithuania.

  20. Optimization of a Virus-Induced Gene Silencing System with Soybean yellow common mosaic virus for Gene Function Studies in Soybeans

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    Kil Hyun Kim


    Full Text Available Virus-induced gene silencing (VIGS is an effective tool for the study of soybean gene function. Successful VIGS depends on the interaction between virus spread and plant growth, which can be influenced by environmental conditions. Recently, we developed a new VIGS system derived from the Soybean yellow common mosaic virus (SYCMV. Here, we investigated several environmental and developmental factors to improve the efficiency of a SYCMV-based VIGS system to optimize the functional analysis of the soybean. Following SYCMV: Glycine max-phytoene desaturase (GmPDS infiltration, we investigated the effect of photoperiod, inoculation time, concentration of Agrobacterium inoculm, and growth temperature on VIGS efficiency. In addition, the relative expression of GmPDS between non-silenced and silenced plants was measured by qRT-PCR. We found that gene silencing efficiency was highest at a photoperiod of 16/8 h (light/dark at a growth temperature of approximately 27°C following syringe infiltration to unrolled unifoliolate leaves in cotyledon stage with a final SYCMV:GmPDS optimal density (OD₆₀₀ of 2.0. Using this optimized protocol, we achieved high efficiency of GmPDS-silencing in various soybean germplasms including cultivated and wild soybeans. We also confirmed that VIGS occurred in the entire plant, including the root, stem, leaves, and flowers, and could transmit GmPDS to other soybean germplasms via mechanical inoculation. This optimized protocol using a SYCMV-based VIGS system in the soybean should provide a fast and effective method to elucidate gene functions and for use in large-scale screening experiments.

  1. An unbiased method for the quantitation of disease phenotypes using a custom-built macro plugin for the program ImageJ. (United States)

    Abd-El-Haliem, Ahmed


    Accurate evaluation of disease phenotypes is considered a key step to study plant-microbe interactions, as the rate of host colonization by the pathogenic microbe directly reflects whether the defense response of the plant is compromised. Although several techniques were developed to quantitate the amount of infection, only a few of them are inherently suitable for large disease screens. Here, I describe an unbiased method to quantitate disease phenotypes which manifest themselves by visible symptoms contrasting with the remaining unaffected parts of the host tissue. The method utilizes a macro plugin written for the image processing program "ImageJ" to calculate two values which determine the disease index for a specific treatment. In case the disease symptoms are not clear, a transgenic pathogenic fungus expressing the GUS gene is suitable for high-throughput disease screens, since staining for GUS activity facilitates an easy detection of the blue-stained pathogen. I illustrate the versatility of this method by analyzing a data set from a functional silencing screening experiment in resistant tomato that was inoculated with a GUS-expressing strain of the fungus Cladosporium fulvum. The method calculates a disease index for each silenced plant and thereby provides a basis for the unbiased identification of candidate host genes required for full resistance to this fungus.

  2. Adaptation of the Four Forms of Employee Silence Scale in a Polish sample

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    Krystyna Adamska


    Full Text Available Background Silence is understood as a decision not to speak up in situations of observed irregularities both in productivity and ethics. The study examined the validity of the Four Forms of Employee Silence Scale (FFESS in the Polish population. The scale is a four-factor measure designed to capture differently motivated tendencies to be silent in organizations. The scale distinguishes acquiescent, quiescent, prosocial and opportunistic silence. Employee silence has been linked to many important individual outcomes: failure to react to ethical transgressions, stress and depression, and lower creativity and productivity. Participants and procedure A total of 1044 employees of various organizations working for at least six months at a given position provided the responses for the validation study. Results The results confirmed the superiority of the four-factor model shown by adequate fit indexes: The FFESS has adequate internal consistency at both the scale and item levels. The criterion-related validity of the scale was established by correlating four forms of silence with measures of emotional attitude toward organization, procedural justice, relational contract and turnover intention. Conclusions The four forms of employee silence are empirically distinct concepts in the Polish sample. The scale may be used as the measurement of individual differences. It can also serve as a tool for diagnosing a climate of silence in an organization.

  3. RNA Silencing in Plants: Mechanisms, Technologies and Applications in Horticultural Crops. (United States)

    Guo, Qigao; Liu, Qing; Smith, Neil A; Liang, Guolu; Wang, Ming-Bo


    Understanding the fundamental nature of a molecular process or a biological pathway is often a catalyst for the development of new technologies in biology. Indeed, studies from late 1990s to early 2000s have uncovered multiple overlapping but functionally distinct RNA silencing pathways in plants, including the posttranscriptional microRNA and small interfering RNA pathways and the transcriptional RNA-directed DNA methylation pathway. These findings have in turn been exploited for developing artificial RNA silencing technologies such as hairpin RNA, artificial microRNA, intrinsic direct repeat, 3' UTR inverted repeat, artificial trans-acting siRNA, and virus-induced gene silencing technologies. Some of these RNA silencing technologies, such as the hairpin RNA technology, have already been widely used for genetic improvement of crop plants in agriculture. For horticultural plants, RNA silencing technologies have been used to increase disease and pest resistance, alter plant architecture and flowering time, improve commercial traits of fruits and flowers, enhance nutritional values, remove toxic compounds and allergens, and develop high-value industrial products. In this article we aim to provide an overview of the RNA silencing pathways in plants, summarize the existing RNA silencing technologies, and review the current progress in applying these technologies for the improvement of agricultural crops particularly horticultural crops.

  4. A translational silencing function of MCPIP1/Regnase-1 specified by the target site context. (United States)

    Behrens, Gesine; Winzen, Reinhard; Rehage, Nina; Dörrie, Anneke; Barsch, Monika; Hoffmann, Anne; Hackermüller, Jörg; Tiedje, Christopher; Heissmeyer, Vigo; Holtmann, Helmut


    The expression of proteins during inflammatory and immune reactions is coordinated by post-transcriptional mechanisms. A particularly strong suppression of protein expression is exerted by a conserved translational silencing element (TSE) identified in the 3' UTR of NFKBIZ mRNA, which is among the targets of the RNA-binding proteins Roquin-1/2 and MCPIP1/Regnase-1. We present evidence that in the context of the TSE MCPIP1, so far known for its endonuclease activity toward mRNAs specified by distinct stem-loop (SL) structures, also suppresses translation. Overexpression of MCPIP1 silenced translation in a TSE-dependent manner and reduced ribosome occupancy of the mRNA. Correspondingly, MCPIP1 depletion alleviated silencing and increased polysomal association of the mRNA. Translationally silenced NFKBIZ or reporter mRNAs were mostly capped, polyadenylated and ribosome associated. Furthermore, MCPIP1 silenced also cap-independent, CrPV-IRES-dependent translation. This suggests that MCPIP1 suppresses a post-initiation step. The TSE is predicted to form five SL structures. SL4 and 5 resemble target structures reported for MCPIP1 and together were sufficient for MCPIP1 binding and mRNA destabilization. Translational silencing, however, required SL1-3 in addition. Thus the NFKBIZ TSE functions as an RNA element in which sequences adjacent to the site of interaction with MCPIP1 and dispensable for accelerated mRNA degradation extend the functional repertoire of MCPIP1 to translational silencing.

  5. Generosity or genocide? Identity implications of silence in American Thanksgiving commemorations. (United States)

    Kurtiş, Tuğçe; Adams, Glenn; Bird, Michael Yellow


    This paper investigates the identity implications of silence about genocide in commemorations of American Thanksgiving. In Study 1 we assessed the co-occurrence of national glorification themes with different forms of silence in commemoration products by conducting a content analysis of presidential Thanksgiving proclamations. In Study 2 we examined the extent to which different commemoration products are infused with particular beliefs and desires by measuring participants' reactions to different Thanksgiving commemorations-a literal-silence condition that did not mention Indigenous Peoples, an interpretive-silence condition that mentioned Indigenous Peoples but did not explicitly mention genocidal conquest, and an anti-silence condition that did mention genocidal conquest-as a function of national glorification. In Study 3 we manipulated exposure to different Thanksgiving commemorations (with associated forms of silence) and assessed the impact on national glorification and identity-relevant action. Results provide evidence for the hypothesised, bi-directional relationship between national glorification and silence about genocide in commemorations of American Thanksgiving.

  6. Language matters: towards an understanding of silence and humour in medical education. (United States)

    Lingard, Lorelei


    This paper considers the state of the science regarding language matters in medical education, with particular attention to two informal language practices: silence and humour. Silence and humour pervade clinical training settings, although we rarely attend explicitly to them. This paper considers the treatment of these topics in our field to date and introduces a selection of the scholarship on silence and humour from other fields, including philosophy, sociology, anthropology, linguistics and rhetoric. Particular attention is paid to distilling the theoretical and methodological possibilities for an elaborated research agenda around silence and humour in medical education. These two language practices assume a variety of forms and serve a range of social functions. Episodes of silence and humour are intimately tied to their relational and institutional contexts. Power often figures centrally, although not predictably. A rich theoretical and methodological basis exists on which to elaborate a research agenda around silence and humour in medical education. Such research promises to reveal more fully the contributions of silence and humour to socialisation in clinical training settings. © Blackwell Publishing Ltd 2013.

  7. RNA-Interference Components Are Dispensable for Transcriptional Silencing of the Drosophila Bithorax-Complex

    KAUST Repository

    Cernilogar, Filippo M.


    Background:Beyond their role in post-transcriptional gene silencing, Dicer and Argonaute, two components of the RNA interference (RNAi) machinery, were shown to be involved in epigenetic regulation of centromeric heterochromatin and transcriptional gene silencing. In particular, RNAi mechanisms appear to play a role in repeat induced silencing and some aspects of Polycomb-mediated gene silencing. However, the functional interplay of RNAi mechanisms and Polycomb group (PcG) pathways at endogenous loci remains to be elucidated.Principal Findings:Here we show that the endogenous Dicer-2/Argonaute-2 RNAi pathway is dispensable for the PcG mediated silencing of the homeotic Bithorax Complex (BX-C). Although Dicer-2 depletion triggers mild transcriptional activation at Polycomb Response Elements (PREs), this does not induce transcriptional changes at PcG-repressed genes. Moreover, Dicer-2 is not needed to maintain global levels of methylation of lysine 27 of histone H3 and does not affect PRE-mediated higher order chromatin structures within the BX-C. Finally bioinformatic analysis, comparing published data sets of PcG targets with Argonaute-2-bound small RNAs reveals no enrichment of these small RNAs at promoter regions associated with PcG proteins.Conclusions:We conclude that the Dicer-2/Argonaute-2 RNAi pathway, despite its role in pairing sensitive gene silencing of transgenes, does not have a role in PcG dependent silencing of major homeotic gene cluster loci in Drosophila. © 2013 Cernilogar et al.

  8. The Luteovirus P4 Movement Protein Is a Suppressor of Systemic RNA Silencing. (United States)

    Fusaro, Adriana F; Barton, Deborah A; Nakasugi, Kenlee; Jackson, Craig; Kalischuk, Melanie L; Kawchuk, Lawrence M; Vaslin, Maite F S; Correa, Regis L; Waterhouse, Peter M


    The plant viral family Luteoviridae is divided into three genera: Luteovirus , Polerovirus and Enamovirus . Without assistance from another virus, members of the family are confined to the cells of the host plant's vascular system. The first open reading frame (ORF) of poleroviruses and enamoviruses encodes P0 proteins which act as silencing suppressor proteins (VSRs) against the plant's viral defense-mediating RNA silencing machinery. Luteoviruses, such as barley yellow dwarf virus-PAV (BYDV-PAV), however, have no P0 to carry out the VSR role, so we investigated whether other proteins or RNAs encoded by BYDV-PAV confer protection against the plant's silencing machinery. Deep-sequencing of small RNAs from plants infected with BYDV-PAV revealed that the virus is subjected to RNA silencing in the phloem tissues and there was no evidence of protection afforded by a possible decoy effect of the highly abundant subgenomic RNA3. However, analysis of VSR activity among the BYDV-PAV ORFs revealed systemic silencing suppression by the P4 movement protein, and a similar, but weaker, activity by P6. The closely related BYDV-PAS P4, but not the polerovirus potato leafroll virus P4, also displayed systemic VSR activity. Both luteovirus and the polerovirus P4 proteins also showed transient, weak local silencing suppression. This suggests that systemic silencing suppression is the principal mechanism by which the luteoviruses BYDV-PAV and BYDV-PAS minimize the effects of the plant's anti-viral defense.

  9. Apparent ploidy effects on silencing are post-transcriptional at HML and telomeres in Saccharomyces cerevisiae.

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    Jenny M McLaughlan

    Full Text Available The repression of genes in regions of heterochromatin is known as transcriptional silencing. It occurs in a wide range of organisms and can have importance in adaptation to the environment, developmental changes and disease. The model organism Saccharomyces cerevisiae has been used for many years to study transcriptional silencing, but until recently no study has been made in relation to ploidy. The aim of this work was to compare transcriptional silencing in haploids and diploids at both telomeres and the hidden mating-type (HM loci. Transcriptional silencing was assayed, by growth on 5-fluoroorotic acid (5-FOA media or by flow cytometry, on strains where a telomere or HM locus was marked. RNA levels were measured by quantitative RT-PCR to confirm that effects were transcriptional. 5-FOA assays and flow cytometry were consistent with transcriptional silencing at telomeres and at HML being reduced as ploidy increases which agreed with conclusions in previous publications. However, QRT-PCR revealed that transcriptional silencing was unaffected by ploidy and thus protein levels were increasing independently of RNA levels. At telomere XI left (XI-L, changes in protein level were strongly influenced by mating-type, whereas at HML mating-type had much less influence. The post-transcriptional effects seen in this study, illustrate the often ignored need to measure RNA levels when assaying transcriptional silencing in Saccharomyces cerevisiae.

  10. Reconstructing Climate Change: Not All Glaciers Suitable (United States)

    Yde, Jacob C.; Paasche, Øyvind


    Glaciers are among the most trusted indicators of climate change, not just because they retreat due to the current rise in global temperatures but also because of their central role in reconstructing past climates. Glaciers come in many forms, and their sensitivity to climate change depends partly on the physics governing the individual glacier, implying that a response can be fast or slow, straightforward or complex, which in sum suggests that not all glaciers are equally suitable for reconstructing past and present climate conditions. In particular, surging and debris-covered glaciers may especially yield misleading results (Figure 1).

  11. Reference Genes in the Pathosystem Phakopsora pachyrhizi/ Soybean Suitable for Normalization in Transcript Profiling

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    Daniela Hirschburger


    Full Text Available Phakopsora pachyrhizi is a devastating pathogen on soybean, endangering soybean production worldwide. Use of Host Induced Gene Silencing (HIGS and the study of effector proteins could provide novel strategies for pathogen control. For both approaches quantification of transcript abundance by RT-qPCR is essential. Suitable stable reference genes for normalization are indispensable to obtain accurate RT-qPCR results. According to the Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE guidelines and using algorithms geNorm and NormFinder we tested candidate reference genes from P. pachyrhizi and Glycine max for their suitability in normalization of transcript levels throughout the infection process. For P. pachyrhizi we recommend a combination of CytB and PDK or GAPDH for in planta experiments. Gene expression during in vitro stages and over the whole infection process was found to be highly unstable. Here, RPS14 and UbcE2 are ranked best by geNorm and NormFinder. Alternatively CytB that has the smallest Cq range (Cq: quantification cycle could be used. We recommend specification of gene expression relative to the germ tube stage rather than to the resting urediospore stage. For studies omitting the resting spore and the appressorium stages a combination of Elf3 and RPS9, or PKD and GAPDH should be used. For normalization of soybean genes during rust infection Ukn2 and cons7 are recommended.

  12. Reference Genes in the Pathosystem Phakopsora pachyrhizi/ Soybean Suitable for Normalization in Transcript Profiling. (United States)

    Hirschburger, Daniela; Müller, Manuel; Voegele, Ralf T; Link, Tobias


    Phakopsora pachyrhizi is a devastating pathogen on soybean, endangering soybean production worldwide. Use of Host Induced Gene Silencing (HIGS) and the study of effector proteins could provide novel strategies for pathogen control. For both approaches quantification of transcript abundance by RT-qPCR is essential. Suitable stable reference genes for normalization are indispensable to obtain accurate RT-qPCR results. According to the Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines and using algorithms geNorm and NormFinder we tested candidate reference genes from P. pachyrhizi and Glycine max for their suitability in normalization of transcript levels throughout the infection process. For P. pachyrhizi we recommend a combination of CytB and PDK or GAPDH for in planta experiments. Gene expression during in vitro stages and over the whole infection process was found to be highly unstable. Here, RPS14 and UbcE2 are ranked best by geNorm and NormFinder. Alternatively CytB that has the smallest Cq range (Cq: quantification cycle) could be used. We recommend specification of gene expression relative to the germ tube stage rather than to the resting urediospore stage. For studies omitting the resting spore and the appressorium stages a combination of Elf3 and RPS9, or PKD and GAPDH should be used. For normalization of soybean genes during rust infection Ukn2 and cons7 are recommended.

  13. Lack of pairing during meiosis triggers multigenerational transgene silencing in Caenorhabditis elegans (United States)

    Leopold, Luciana E.; Heestand, Bree N.; Seong, Soobin; Shtessel, Ludmila; Ahmed, Shawn


    Single-copy transgenes in Caenorhabditis elegans can be subjected to a potent, irreversible silencing process termed small RNA-induced epigenetic silencing (RNAe). RNAe is promoted by the Piwi Argonaute protein PRG-1 and associated Piwi-interacting RNAs (piRNAs), as well as by proteins that promote and respond to secondary small interfering RNA (siRNA) production. Here we define a related siRNA-mediated silencing process, termed “multigenerational RNAe,” which can occur for transgenes that are maintained in a hemizygous state for several generations. We found that transgenes that contain either GFP or mCherry epitope tags can be silenced via multigenerational RNAe, whereas a transgene that possesses GFP and a perfect piRNA target site can be rapidly and permanently silenced via RNAe. Although previous studies have shown that PRG-1 is typically dispensable for maintenance of RNAe, we found that both initiation and maintenance of multigenerational RNAe requires PRG-1 and the secondary siRNA biogenesis protein RDE-2. Although silencing via RNAe is irreversible, we found that transgene expression can be restored when hemizygous transgenes that were silenced via multigenerational RNAe become homozygous. Furthermore, multigenerational RNAe was accelerated when meiotic pairing of the chromosome possessing the transgene was abolished. We propose that persistent lack of pairing during meiosis elicits a reversible multigenerational silencing response, which can lead to permanent transgene silencing. Multigenerational RNAe may be broadly relevant to single-copy transgenes used in experimental biology and to shaping the epigenomic landscape of diverse species, where genomic polymorphisms between homologous chromosomes commonly result in unpaired DNA during meiosis. PMID:25941370

  14. Driving in unheard silence: Disability and the politics of shutting up. (United States)

    Lourens, Heidi


    The politics of silence is central to disability experience and the field of disability studies. In this analytical autoethnography, I write about my silences as a visually impaired woman. I explore and make sense of personal life stories through a theoretical perspective. The analysis of these personal experiences lead me to argue that disability-related silences are mostly created through the confluence of inaccessible physical and social environments and the psychological internalisation of these worlds. I also discuss the ways in which I am currently regaining my voice. Further research on resistance by disabled persons is recommended.

  15. Empathy and silence in pastoral care for traumatic grief and loss. (United States)

    Capretto, Peter


    This paper evaluates silence as a therapeutic practice in pastoral care for traumatic grief and loss. Informed by the history of attachment and mourning theory, its research considers the basic effect that empathy has upon the therapeutic relationship around psychic difference. The study appraises the potential resources and detriments that empathic language may have for the grief process. Offering clinical examples in hospice chaplaincy, it refutes the idea that silence is formulaic tool to be used. It instead offers silence as the acceptance of the limits of empathic language and the affirmation of psychological difference and theological wholeness.

  16. Shifting suitability for malaria vectors across Africa with warming climates

    Directory of Open Access Journals (Sweden)

    Peterson A Townsend


    Full Text Available Abstract Background Climates are changing rapidly, producing warm climate conditions globally not previously observed in modern history. Malaria is of great concern as a cause of human mortality and morbidity, particularly across Africa, thanks in large part to the presence there of a particularly competent suite of mosquito vector species. Methods I derive spatially explicit estimates of human populations living in regions newly suitable climatically for populations of two key Anopheles gambiae vector complex species in Africa over the coming 50 years, based on ecological niche model projections over two global climate models, two scenarios of climate change, and detailed spatial summaries of human population distributions. Results For both species, under all scenarios, given the changing spatial distribution of appropriate conditions and the current population distribution, the models predict a reduction of 11.3–30.2% in the percentage of the overall population living in areas climatically suitable for these vector species in coming decades, but reductions and increases are focused in different regions: malaria vector suitability is likely to decrease in West Africa, but increase in eastern and southern Africa. Conclusion Climate change effects on African malaria vectors shift their distributional potential from west to east and south, which has implications for overall numbers of people exposed to these vector species. Although the total is reduced, malaria is likely to pose novel public health problems in areas where it has not previously been common.

  17. Increase in quantity and quality of suitable areas for invasive species as climate changes. (United States)

    Bertelsmeier, Cleo; Luque, Gloria M; Courchamp, Franck


    As climatically suitable range projections become increasingly used to assess distributions of species, we recommend systematic assessments of the quality of habitat in addition to the classical binary classification of habitat. We devised a method to assess occurrence probability, captured by a climatic suitability index, through which we could determine variations in the quality of potential habitat. This relative risk assessment circumvents the use of an arbitrary suitability threshold. We illustrated our method with 2 case studies on invasive ant species. We estimated invasion potential of the destroyer ant (Monomorium destructor) and the European fire ant (Myrmica rubra) on a global scale currently and by 2080 with climate change. We found that 21.1% of the world's landmass currently has a suitable climate for the destroyer ant and 16% has a suitable climate for European fire ant. Our climatic suitability index showed that both ant species would benefit from climate change, but in different ways. The size of the potential distribution increased by 35.8% for the destroyer ant. Meanwhile, the total area of potential distribution remained the same for the European fire ant (>0.05%), but the level of climatic suitability within this range increased greatly and led to an improvement in habitat quality (i.e., of invasive species' establishment likelihood). Either through quantity or quality of suitable areas, both invasive ant species are likely to increase the extent of their invasion in the future, following global climate change. Our results show that species may increase their range if either more areas become suitable or if the available areas present improved suitability. Studies in which an arbitrary suitability threshold was used may overlook changes in area quality within climatically suitable areas and as a result reach incorrect predictions. Incremento de la Cantidad y Calidad de Áreas Idóneas para Especies Invasoras a Medida que Cambia el Clima.

  18. Silence as an element of care: A meta-ethnographic review of professional caregivers' experience in clinical and pastoral settings. (United States)

    Bassett, Lynn; Bingley, Amanda F; Brearley, Sarah G


    In interactions between professional caregivers, patients and family members at the end of life, silence often becomes more prevalent. Silence is acknowledged as integral to interpersonal communication and compassionate care but is also noted as a complex and ambiguous phenomenon. This review seeks interdisciplinary experience to deepen understanding of qualities of silence as an element of care. To search for published papers which describe professional caregivers' experience of silence as an element of care, in palliative and other clinical, spiritual and pastoral care settings and to synthesise their findings. Meta-ethnography: employing a systematic search strategy and line-of-argument synthesis. PsycINFO and seven other cross-disciplinary databases, supplemented by hand-search, review of reference lists and citation tracking. No date range was imposed. Inclusion criteria focused on reported experience of silence in professional caregiving. Selected papers ( n = 18) were appraised; none were rejected on grounds of quality. International, interdisciplinary research and opinion endorses the value of silence in clinical care. As a multi-functional element of interpersonal relationships, silence operates in partnership with speech to support therapeutic communication. As a caregiving practice, silence is perceived as particularly relevant in spiritual and existential dimensions of care when words may fail. Experience of silence as an element of care was found in palliative and spiritual care, psychotherapy and counselling supporting existing recognition of the value of silence as a skill and practice. Because silence can present challenges for caregivers, greater understanding may offer benefits for clinical practice.

  19. Controlling low frequency noise using a passive silencer

    Energy Technology Data Exchange (ETDEWEB)

    DeGagne, D.C.; Faszer, A.C. [Noise Solutions Inc., Calgary, AB (Canada)


    Nearly 85 per cent of all the oil and natural gas recovered in Canada comes from Alberta where noise emissions from energy facilities are regulated by the Energy Resources Conservation Board (ERCB) and the Alberta Utilities Commission (AUC) through Noise Control Directive D-038. Operators of energy facilities in Alberta must meet the Permissible Sound Level (PSL) established for industrial facilities that are established for receptor locations such as nearby residences. Despite efforts to manage noise, complaints from nearby residents continue to occur even if the facility is in compliance. This is because high and mid frequency components can decay or be absorbed by air and ground conditions leaving mostly low frequency noise (LFN) at the residence. This paper explored the significance of LFN and how some facility operators are dealing with this problem. The impacts of LFN can range from creating a sensation of pressure in the ear, disturbing normal conversation, to creating secondary vibrating effects within homes. If severe enough, it can also result in potential behavioural dysfunction such as task performance deterioration, sleep disturbance and headaches. In 2006, Noise Solutions Inc. was approached by Hunt Oil Company regarding the installation of a new compressor at their Caroline site. The proposal posed a serious noise problem for the community which requested that the new compressor unit be developed without any additional noise impact on the area. Noise Solutions Inc. used a multi-phased approach to develop the most effective LFN silencer using best practical technology and materials. In an effort to fully attenuate both the high frequency noise (dBA) and the low frequency noise (dBC) of the new compressor unit, Noise Solutions and Hunt Oil agreed that a significant level of sound-suppression would be necessary. The entire noise-suppression unit was specially designed to compensate for the sheer size of the compressor building. The first step to

  20. Le silence, ce cri qui résonne dans l’écriture de Viviane Forrester

    Directory of Open Access Journals (Sweden)

    Amelia Peral Crespo


    Full Text Available The Holocaust Literature is tied to Silence from its beginning as the writing to silence. Write about the Holocaust and relate the experiences lived in first person or transmit them to the coming generations means leaving the silence which, for years, was the ideal refuge. This research focuses on the literary production of Viviane Forrester. The different expressions of silence that characterize her writings are analyzed. Silence is the cry that underlies in the most profound of the human being that return from the concentration camps, as Lazarus from the dead. It is a silence that grows to a cry because Forrester’s writing, almost in parallel to the fragmentary writing of Blanchot, an-nounces livre à venir, expressing the cry in silence without the ending of the word

  1. Optimized Audio Classification and Segmentation Algorithm by Using Ensemble Methods

    Directory of Open Access Journals (Sweden)

    Saadia Zahid


    Full Text Available Audio segmentation is a basis for multimedia content analysis which is the most important and widely used application nowadays. An optimized audio classification and segmentation algorithm is presented in this paper that segments a superimposed audio stream on the basis of its content into four main audio types: pure-speech, music, environment sound, and silence. An algorithm is proposed that preserves important audio content and reduces the misclassification rate without using large amount of training data, which handles noise and is suitable for use for real-time applications. Noise in an audio stream is segmented out as environment sound. A hybrid classification approach is used, bagged support vector machines (SVMs with artificial neural networks (ANNs. Audio stream is classified, firstly, into speech and nonspeech segment by using bagged support vector machines; nonspeech segment is further classified into music and environment sound by using artificial neural networks and lastly, speech segment is classified into silence and pure-speech segments on the basis of rule-based classifier. Minimum data is used for training classifier; ensemble methods are used for minimizing misclassification rate and approximately 98% accurate segments are obtained. A fast and efficient algorithm is designed that can be used with real-time multimedia applications.

  2. Novel siRNA delivery system using a ternary polymer complex with strong silencing effect and no cytotoxicity. (United States)

    Kodama, Yukinobu; Shiokawa, Yumi; Nakamura, Tadahiro; Kurosaki, Tomoaki; Aki, Keisei; Nakagawa, Hiroo; Muro, Takahiro; Kitahara, Takashi; Higuchi, Norihide; Sasaki, Hitoshi


    We developed a novel small interfering RNA (siRNA) delivery system using a ternary complex with polyethyleneimine (PEI) and γ-polyglutamic acid (γ-PGA), which showed silencing effect and no cytotoxicity. The binary complexes of siRNA with PEI were approximately 73-102 nm in particle size and 45-52 mV in ζ-potential. The silencing effect of siRNA/PEI complexes increased with an increase of PEI, and siRNA/PEI complexes with a charge ratio greater than 16 showed significant luciferase knockdown in a mouse colon carcinoma cell line regularly expressing luciferase (Colon26/Luc cells). However, strong cytotoxicity and blood agglutination were observed in the siRNA/Lipofectamine complex and siRNA/PEI16 complex. Recharging cationic complexes with an anionic compound was reported to be a promising method for overcoming these toxicities. We therefore prepared ternary complexes of siRNA with PEI (charge ratio 16) by the addition of γ-PGA to reduce cytotoxicity and deliver siRNA. As expected, the cytotoxicity of the ternary complexes decreased with an increase of γ-PGA content, which decreased the ζ-potential of the complexes. A strong silencing effect comparable to siRNA/Lipofectamine complex was discovered in ternary complexes including γ-PGA with an anionic surface charge. The high incorporation of ternary complexes into Colon26/Luc cells was confirmed with fluorescence microcopy. Having achieved knockdown of an exogenously transfected gene, the ability of the complex to mediate knockdown of an endogenous housekeeping gene, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), was assessed in B16-F10 cells. The ternary complex (siRNA/PEI16/γ-PGA12 complex) exhibited a significant GAPDH knockdown effect. Thus, we developed a useful siRNA delivery system.

  3. RNAi-mediated endogene silencing in strawberry fruit: detection of primary and secondary siRNAs by deep sequencing. (United States)

    Härtl, Katja; Kalinowski, Gregor; Hoffmann, Thomas; Preuss, Anja; Schwab, Wilfried


    RNA interference (RNAi) has been exploited as a reverse genetic tool for functional genomics in the nonmodel species strawberry (Fragaria × ananassa) since 2006. Here, we analysed for the first time different but overlapping nucleotide sections (>200 nt) of two endogenous genes, FaCHS (chalcone synthase) and FaOMT (O-methyltransferase), as inducer sequences and a transitive vector system to compare their gene silencing efficiencies. In total, ten vectors were assembled each containing the nucleotide sequence of one fragment in sense and corresponding antisense orientation separated by an intron (inverted hairpin construct, ihp). All sequence fragments along the full lengths of both target genes resulted in a significant down-regulation of the respective gene expression and related metabolite levels. Quantitative PCR data and successful application of a transitive vector system coinciding with a phenotypic change suggested propagation of the silencing signal. The spreading of the signal in strawberry fruit in the 3' direction was shown for the first time by the detection of secondary small interfering RNAs (siRNAs) outside of the primary targets by deep sequencing. Down-regulation of endogenes by the transitive method was less effective than silencing by ihp constructs probably because the numbers of primary siRNAs exceeded the quantity of secondary siRNAs by three orders of magnitude. Besides, we observed consistent hotspots of primary and secondary siRNA formation along the target sequence which fall within a distance of less than 200 nt. Thus, ihp vectors seem to be superior over the transitive vector system for functional genomics in strawberry fruit. © 2016 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

  4. TGIF1 Gene Silencing in Tendon-Derived Stem Cells Improves the Tendon-to-Bone Insertion Site Regeneration

    Directory of Open Access Journals (Sweden)

    Liyang Chen


    Full Text Available Background/Aims: The slow healing process of tendon-to-bone junctions can be accelerated via implanted tendon-derived stem cells (TDSCs with silenced transforming growth interacting factor 1 (TGIF1 gene. Tendon-to-bone insertion site is the special form of connective tissues derivatives of common connective progenitors, where TGF-β plays bidirectional effects (chondrogenic or fibrogenic through different signaling pathways at different stages. A recent study revealed that TGF-β directly induces the chondrogenic gene Sox9. However, TGIF1 represses the expression of the cartilage master Sox9 gene and changes its expression rate against the fibrogenesis gene Scleraxis (Scx. Methods: TGIF1 siRNA was transduced or TGIF1 was over-expressed in tendon-derived stem cells. Following suprapinatus tendon repair, rats were either treated with transduced TDSCs or nontransduced TDSCs. Histologic examination and Western blot were performed in both groups. Results: In this study, the silencing of TGIF1 significantly upregulated the chondrogenic genes and markers. Similarly, TGIF1 inhibited TDSC differentiation into cartilage via interactions with TGF-β-activated Smad2 and suppressed the phosphorylation of Smad2. The area of fibrocartilage at the tendon-bone interface was significantly increased in the TGIF1 (- group compared with the control and TGIF1-overexpressing groups in the early stages of the animal model. The interface between the tendon and bone showed a increase of new bone and fibrocartilage in the TGIF1 (- group at 4 weeks. Fibrovascular scar tissue was observed in the TGIF1-overexpressing group and the fibrin glue only group. Low levels of fibrocartilage and fibrovascular scar tissue were found in the TDSCs group. Conclusion: Collectively, this study shows that the tendon-derived stem cell modified with TGIF1 gene silencing has promising effects on tendon-to-bone healing which can be further explored as a therapeutic tool in regenerative medicine.

  5. Silencing two main isoforms of crustacean hyperglycemic hormone (CHH) induces compensatory expression of two CHH-like transcripts in the red swamp crayfish Procambarus clarkii


    Manfrin, C; Peruzza, L; Bonzi, Lc; Pallavicini, A; Giulianini, Pg


    RNA interference has frequently been applied to modulate gene function in organisms. With the aim of creating new autocidal methods based on neuro-endocrine disruptors for invasive populations of Procambarus clarkii, we silenced the Crustacean Hyperglycemic Hormone (CHH) by injecting the corresponding dsRNA. CHH is a pleiotropic hormone that primarily regulates the mobilization of energy reserves and plays a pivotal role in stress responses. Here, we describe two experiments aimed at testing ...

  6. Evaluation of land suitability for main irrigated crops in the North-Western Region of Libya

    Directory of Open Access Journals (Sweden)

    Salah Hassanien Abd El-Aziz


    Full Text Available Land suitability analysis can help to achieve sustainable crop production with a proper use of the natural resources. The current study was carried out on the soils of north-western area of Libya to assess their morphological, physical and chemical properties and their suitability for growing irrigated crops. The studied area lies between latitudes 32° 30′ 00.9" and 32° 57′ 34.2'' N and between longitudes 11° 35′ 08.4" and 11° 45′ 09.2" E. Two suitability methods (Sys &Verhey and Storie methods were used to assess the land suitability of this area. According to Sys and Verhye method, the soils of the studied area varied in the suitability for irrigation between highly suitable (S1 to marginally suitable (S3. However, according to modified Storie index method the soils productivity, ranged from excellent (grade 1 for agriculture to non-agricultural (grade 6. The modified Storie index method was more effective in assessing the land suitability of this area. The drip irrigation system was also more suitable than surface irrigation method for most of the soils of the studied area. The indeces of soil suitability rating and percentage for growing alfalfa, sorghum, barley, maize, millets, wheat and safflower were higher compared to those for growing soybean, sunflower and sesame. Onion and green pepper crops were moderately suitable to be grown in 42% of the soils of the studied area while the other vegetables were not suitable to be grown in most of the soils of the studied area. The evaluated fruit trees could be arranged according to the soil suitability rating and percentage in the order of date palm > olives > guava > citrus > banana. The results also revealed that the studied area has a good potential to produce the selected crops under irrigation provided that the water requirements for these crops are met. The main limiting factors for land suitability for growing crops are soil texture, soil depth, calcium carbonate, alkaline p

  7. Glass compositions suitable for PFR wastes

    International Nuclear Information System (INIS)

    Boult, K.A.; Dalton, J.T.; Eccles, E.W.; Hough, A.; Marples, J.A.C.; Paige, E.L.; Sutcliffe, P.W.


    In previous work, a glass composition (PFR116) had been identified as suitable for vitrifying high level wastes from the Prototype Fast Reactor (PFR) fuel reprocessing plant. Further work has now shown that: (a) it is tolerant of quite large variations in the proportion of waste in the glass: (b) it could also be used to vitrify waste from the European Demonstration Reprocessing Plant. Current wastes from the PFR reprocessing plant contain Cu and Zn which arise from the use of a sacrificial brass basket to transfer chopped fuel to the dissolver. It was originally thought that the presence of the resulting CuO and ZnO in the glass would lead to increased corrosion rates of the Inconel alloys that are used in vitrification plants. Further compatibility tests have shown that this is not so although some Cu contamination of Inconel 690 does seem to occur. (author)

  8. The suitability of internal versus external successors

    DEFF Research Database (Denmark)

    Boyd, Britta; Royer, Susanne


    that in industries where tacit knowledge forms the basis for competitive advantage, the use of internal successors can help family firms excel after a transition of power has occurred. Theoretical and practical implications are discussed to enhance the long-term perspective for family businesses.......Understanding knowledge transfer in family firm succession is important for the survival of family firms. Previous research has begun to explore the suitability of internal versus external successor in family firms with regard to relevant knowledge types. This paper builds on the contingency model...... of family business succession in order to understand when family successors are preferred because of their family-specific experiential knowledge. A case study analysis from the German-Danish border region explores how a family firm has used internal successors for the last 12 successions. We argue...

  9. Land Suitability Assessment And Precision Farming Prospects For ...

    African Journals Online (AJOL)

    The soils on the farm were sampled and profiles classified using South African Binomial system of soil classification. The farm was divided into suitability classes of highly suitable (S1), suitable (S2), and unsuitable (N1), and permanently unsuitable (N2) classes. Maize-soybean suitability maps were produced based on soil ...

  10. Sensorless Suitability Analysis of Hybrid PM Machines for Electric Vehicles

    DEFF Research Database (Denmark)

    Matzen, Torben Nørregaard; Rasmussen, Peter Omand


    Electrical machines for traction in electric vehicles are an essential component which attract attention with respect to machine design and control as a part of the emerging renewable industry. For the hybrid electric machine to replace the familiar behaviour of the combustion engine torque......, control seems necessary to implement. For hybrid permanent magnet (PM) machines torque control in an indirect fashion using dq-current control is frequently done. This approach requires knowledge about the machine shaft position which may be obtained sensorless. In this article a method based on accurate...... knowledge of machine flux linkages is proposed for analysing the suitability for sensorless control at zero and low speed....

  11. Preclinical Evaluation of a Lentiviral Vector for Huntingtin Silencing

    Directory of Open Access Journals (Sweden)

    Karine Cambon


    Full Text Available Huntington’s disease (HD is an autosomal dominant neurodegenerative disorder resulting from a polyglutamine expansion in the huntingtin (HTT protein. There is currently no cure for this disease, but recent studies suggest that RNAi to downregulate the expression of both normal and mutant HTT is a promising therapeutic approach. We previously developed a small hairpin RNA (shRNA, vectorized in an HIV-1-derived lentiviral vector (LV, that reduced pathology in an HD rodent model. Here, we modified this vector for preclinical development by using a tat-independent third-generation LV (pCCL backbone and removing the original reporter genes. We demonstrate that this novel vector efficiently downregulated HTT expression in vitro in striatal neurons derived from induced pluripotent stem cells (iPSCs of HD patients. It reduced two major pathological HD hallmarks while triggering a minimal inflammatory response, up to 6 weeks after injection, when administered by stereotaxic surgery in the striatum of an in vivo rodent HD model. Further assessment of this shRNA vector in vitro showed proper processing by the endogenous silencing machinery, and we analyzed gene expression changes to identify potential off-targets. These preclinical data suggest that this new shRNA vector fulfills primary biosafety and efficiency requirements for further development in the clinic as a cure for HD.

  12. Breaking the silence, dismantling taboos: Latino novels on AIDS. (United States)

    Sandoval Sánchez, A


    As AIDS has ravaged the world in the last two decades, literature has played an important role in giving testimony of the epidemic. According to different communities and particular experiences, AIDS literature can offer ways of coping and surviving. In the last few years the U.S. Latino/a population has been highly affected by AIDS. The first genres to register AIDS were poetry and theatre. In this essay I examine how AIDS is represented in the first four Latino novels to deal with the subject matter: Ana Castillo's So Far from God, Daniel Torres's Morirás si da una primavera, Elías Miguel Muñoz's The Greatest Performance, and Jaime Manrique's latin Moon in Manhattan. My critical reading centers on how AIDS constitutes a new articulation of identity, particularly gay and ethnic identity. Most importantly, I propose that these novels contribute to dismantling homophobia and compulsory heterosexuality, deconstructing sexual and cultural taboos, and breaking the silence on AIDS/SIDA.

  13. Sox6 directly silences epsilon globin expression in definitive erythropoiesis.

    Directory of Open Access Journals (Sweden)


    Full Text Available Sox6 is a member of the Sox transcription factor family that is defined by the conserved high mobility group (HMG DNA binding domain, first described in the testis determining gene, Sry. Previous studies have suggested that Sox6 plays a role in the development of the central nervous system, cartilage, and muscle. In the Sox6-deficient mouse, p100H, epsilony globin is persistently expressed, and increased numbers of nucleated red cells are present in the fetal circulation. Transfection assays in GM979 (erythroleukemic cells define a 36-base pair region of the epsilony proximal promoter that is critical for Sox6 mediated repression. Electrophoretic mobility shift assay (EMSA and chromatin immunoprecipitation (ChIP assays demonstrate that Sox6 acts as a repressor by directly binding to the epsilony promoter. The normal expression of Sox6 in wild-type fetal liver and the ectopic expression of epsilony in p100H homozygous fetal liver demonstrate that Sox6 functions in definitive erythropoiesis. The present study shows that Sox6 is required for silencing of epsilony globin in definitive erythropoiesis and suggests a role for Sox6 in erythroid cell maturation. Thus, Sox6 regulation of epsilony globin might provide a novel therapeutical target in the treatment of hemoglobinopathies such as sickle cell anemia and thalassemia.

  14. Grandparents' experiences of childhood cancer, part 1: doubled and silenced. (United States)

    Moules, Nancy J; Laing, Catherine M; McCaffrey, Graham; Tapp, Dianne M; Strother, Douglas


    In this study, the authors examined the experiences of grandparents who have had, or have, a grandchild with childhood cancer. Sixteen grandparents were interviewed using unstructured interviews, and the data were analyzed according to hermeneutic-phenomenological tradition, as guided by the philosophical hermeneutics of Hans-Georg Gadamer. Interpretive findings indicate that grandparents suffer and worry in many complex ways that include a doubled worry for their own children as well as their grandchildren. According to the grandparents in this study, this worry was, at times, silenced in efforts to protect the parents of the grandchild from the burden of concern for the grandparent. Other interpretations include the nature of having one's universe shaken, of having lives put on hold, and a sense of helplessness. The grandparents in this study offer advice to other grandparents as well as to the health care system regarding what kinds of things might have been more helpful to them as one level of the family system, who, like other subsystems of the family, are also profoundly affected by the event of childhood cancer.

  15. Nature to the natural rescue: Silencing microbial chats. (United States)

    Paul, Diby; Gopal, Judy; Kumar, Manish; Manikandan, Muthu


    Communication is the sole means by which effective networking and co-existence is accomplished amongst living beings. Microbes have their own chit-chats. Science has overheard these microbial gossips and have concluded that these aren't just informal communications, but carefully coordinated signals that plan their effective strategies. Tracking one such signal molecule, N-acyl homoserine lactone (AHL), led to a fundamental understanding to microbial quorum sensing (QS). Furtherance of research sought for ways to cut off communication between these virulent forms, so as to hinder their combinatorial attacks through quorum sensing inhibitors (QSIs). A clear understanding of the inhibitors of these microbial communication systems is vital to destroy their networking and co-working. The current review, consolidates the solutions for QSIs offered from natural sources against these micro components, that are capable of slaughtering even nature's most fit entity-man. The applications of effective out sourcing of this QSI technologies and the need for development are discussed. The importance of silencing this microbial chatter to various aspects of human life and their implications are discussed and elaborated. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Silence in liturgy as the space for the Holy Ghost’s activity

    Directory of Open Access Journals (Sweden)

    Adelajda Sielepin


    Full Text Available The aim of this article is to clarify the essential and often neglected function of liturgical silence. In practice silence is usually intended to prepare the participants for listening or reflecting upon the word, especially when proclaimed by the minister. Whereas it is vital to know that silence constitutes a creative element of the liturgy, because the involvement of the Holy Spirit. In His action and cooperation with the participants He enables them to get into communion with Christ the Word and the Person and eventually to join effectively His mission. In that sense silence helps the Holy Ghost to fulfill His function in continuing the dialogue between God and man as well as in enhancing one’s spiritual skills for the fruitful participation in the Mystery of Christ and the Church.

  17. Reticence in Silence. An Examination of Heidegger's Phenomenology of Language in Contributions to Philosophy

    Czech Academy of Sciences Publication Activity Database

    Nitsche, Martin


    Roč. 765, č. 1 (2011), s. 74-79 ISSN 1407-2157 Institutional research plan: CEZ:AV0Z90090514 Keywords : Heidegger * contributions to philosophy * phenomenology * language * reticence in silence Subject RIV: AA - Philosophy ; Religion

  18. Viral suppressors of RNA silencing hinder exogenous and endogenous small RNA pathways in Drosophila.

    Directory of Open Access Journals (Sweden)

    Bassam Berry


    Full Text Available In plants and insects, RNA interference (RNAi is the main responder against viruses and shapes the basis of antiviral immunity. Viruses counter this defense by expressing viral suppressors of RNAi (VSRs. While VSRs in Drosophila melanogaster were shown to inhibit RNAi through different modes of action, whether they act on other silencing pathways remained unexplored.Here we show that expression of various plant and insect VSRs in transgenic flies does not perturb the Drosophila microRNA (miRNA pathway; but in contrast, inhibits antiviral RNAi and the RNA silencing response triggered by inverted repeat transcripts, and injection of dsRNA or siRNA. Strikingly, these VSRs also suppressed transposon silencing by endogenous siRNAs (endo-siRNAs.Our findings identify VSRs as tools to unravel small RNA pathways in insects and suggest a cosuppression of antiviral RNAi and endo-siRNA silencing by viruses during fly infections.

  19. A viral suppressor protein inhibits host RNA silencing by hooking up with Argonautes

    KAUST Repository

    Jin, Hailing


    RNA viruses are particularly vulnerable to RNAi-based defenses in the host, and thus have evolved specific proteins, known as viral suppressors of RNA silencing (VSRs), as a counterdefense. In this issue of Genes & Development, Azevedo and colleagues (pp. 904-915) discovered that P38, the VSR of Turnip crinkle virus, uses its glycine/tryptophane (GW) motifs as an ARGONAUTE (AGO) hook to attract and disarm the host\\'s essential effector of RNA silencing. Several GW motif-containing cellular proteins are known to be important partners of AGOs in RNA silencing effector complexes in yeast, plants, and animals. The GW motif appears to be a versatile and effective tool for regulating the activities of RNA silencing pathways, and the use of GW mimicry to compete for and inhibit host AGOs may be a strategy used by many pathogens to counteract host RNAi-based defenses. © 2010 by Cold Spring Harbor Laboratory Press.

  20. Identifying Silence Climate in Organizations in the Framework of Contemporary Management Approaches

    Directory of Open Access Journals (Sweden)

    Mustafa Emre Civelek


    Full Text Available Dynamic competition conditions in present day, bring about the consequence for businesses to face varied problems with each passing day. At this point, current management approaches include studies that would shed light on the new problems of businesses. Organizational Silence, a concept that has recently been being voiced in business world, has come up in such context. Organizational silence could be expressed as the employee behavior of keeping silent about certain negativities due to various reasons in an organization. Since knowledge sharing in modern organizations is of capital importance in terms of responding hastily to the changes in a competitive environment, spread of this behavior of employees to organization culture and climate presents a threat of indifference. In this study, the concept of Organizational Silence is defined and the effects of conceived silence climate on management of organizations are discussed.

  1. Identifying Silence Climate in Organizations in the Framework of Contemporary Management Approaches

    Directory of Open Access Journals (Sweden)

    Mustafa Emre Civelek


    Full Text Available Dynamic competition conditions in present day bring about the consequence for businesses to face varied problems with each passing day. At this point, current management approaches include studies that would shed light on the new problems of businesses. Organizational Silence, a concept that has recently been being voiced in business world, has come up in such context. Organizational silence could be expressed as the employee behavior of keeping silent about certain negativities due to various reasons in an organization. Since knowledge sharing in modern organizations is of capital importance in terms of responding hastily to the changes in a competitive environment, spread of this behavior of employees to organization culture and climate presents a threat of indifference. In this study, the concept of Organizational Silence is defined and the effects of conceived silence climate on management of organizations are discussed.

  2. Dissection of tomato lycopene biosynthesis through virus-induced gene silencing. (United States)

    Fantini, Elio; Falcone, Giulia; Frusciante, Sarah; Giliberto, Leonardo; Giuliano, Giovanni


    Lycopene biosynthesis in tomato (Solanum lycopersicum) fruits has been proposed to proceed through a poly-cis pathway catalyzed by phytoene synthase (PSY), two desaturases (phytoene desaturase [PDS] and ζ-carotene desaturase [ZDS]), and two cis-trans isomerases (ζ-carotene isomerase [ZISO] and prolycopene isomerase [CrtISO]). The mechanism of action of these enzymes has been studied in Escherichia coli, but a systematic study of their in vivo function is lacking. We studied the function of nine candidate genes (PSY1, PSY2, PSY3, PDS, ZDS, ZISO, CrtISO, CrtISO-Like1, and CrtISO-Like2) using virus-induced gene silencing (VIGS) coupled to high-resolution liquid chromatography coupled with diode array detector and mass spectrometry, which allowed the identification and quantitation of 45 different carotenoid isomers, including linear xanthophylls. The data confirm the confinement of the VIGS signal to the silenced fruits and the similarity of the phenotypes of PSY1- and CrtISO-silenced fruits with those of the yellow flesh and tangerine mutants. Light was able to restore lycopene biosynthesis in ZISO-silenced fruits. Isomeric composition of fruits silenced at different metabolic steps suggested the existence of three functional units, comprising PSY1, PDS/ZISO, and ZDS/CrtISO, and responsible for the synthesis of 15-cis-phytoene, 9,9'-di-cis-ζ-carotene, and all-trans-lycopene, respectively. Silencing of a desaturase (PDS or ZDS) resulted in the induction of the isomerase in the same functional unit (ZISO or CrtISO, respectively). All-trans-ζ-carotene was detectable in nonsilenced fruits, greatly increased in ZDS-silenced ones, and disappeared in CrtISO-Like1-/CrtISO-Like2-silenced ones, suggesting the existence of a metabolic side branch, comprising this compound and initiated by the latter enzymes.

  3. 'Checkmating HIV&AIDS': Using chess to break the silence in the ...

    African Journals Online (AJOL)

    Omar Esau


    Dec 13, 2012 ... Keywords: HIV&AIDS, action research, chess, checkmating, culture of silence, prevention, awareness. Résumé. Cet article est un compte rendu de mon projet de recherche-action « mettre le VIH/SIDA échec et mat », qui avait pour but de briser dans ma classe la « culture du silence » au sujet du VIH/SIDA ...

  4. Silencing of the rotavirus NSP4 protein decreases the incidence of biliary atresia in murine model.

    Directory of Open Access Journals (Sweden)

    Jiexiong Feng

    Full Text Available Biliary atresia is a common disease in neonates which causes obstructive jaundice and progressive hepatic fibrosis. Our previous studies indicate that rotavirus infection is an initiator in the pathogenesis of experimental biliary atresia (BA through the induction of increased nuclear factor-kappaB and abnormal activation of the osteopontin inflammation pathway. In the setting of rotavirus infection, rotavirus nonstructural protein 4 (NSP4 serves as an important immunogen, viral protein 7 (VP7 is necessary in rotavirus maturity and viral protein 4 (VP4 is a virulence determiner. The purpose of the current study is to clarify the roles of NSP4, VP7 and VP4 in the pathogenesis of experimental BA. Primary cultured extrahepatic biliary epithelia were infected with Rotavirus (mmu18006. Small interfering RNA targeting NSP4, VP7 or VP4 was transfected before rotavirus infection both in vitro and in vivo. We analyzed the incidence of BA, morphological change, morphogenesis of viral particles and viral mRNA and protein expression. The in vitro experiments showed NSP4 silencing decreased the levels of VP7 and VP4, reduced viral particles and decreased cytopathic effect. NSP4-positive cells had strongly positive expression of integrin subunit α2. Silencing of VP7 or VP4 partially decreased epithelial injury. Animal experiments indicated after NSP4 silencing, mouse pups had lower incidence of BA than after VP7 or VP4 silencing. However, 33.3% of VP4-silenced pups (N = 6 suffered BA and 50% of pups (N = 6 suffered biliary injury after VP7 silencing. Hepatic injury was decreased after NSP4 or VP4 silencing. Neither VP4 nor VP7 were detected in the biliary ducts after NSP4. All together, NSP4 silencing down-regulates VP7 and VP4, resulting in decreased incidence of BA.

  5. Supervised learning classification models for prediction of plant virus encoded RNA silencing suppressors.

    Directory of Open Access Journals (Sweden)

    Zeenia Jagga

    Full Text Available Viral encoded RNA silencing suppressor proteins interfere with the host RNA silencing machinery, facilitating viral infection by evading host immunity. In plant hosts, the viral proteins have several basic science implications and biotechnology applications. However in silico identification of these proteins is limited by their high sequence diversity. In this study we developed supervised learning based classification models for plant viral RNA silencing suppressor proteins in plant viruses. We developed four classifiers based on supervised learning algorithms: J48, Random Forest, LibSVM and Naïve Bayes algorithms, with enriched model learning by correlation based feature selection. Structural and physicochemical features calculated for experimentally verified primary protein sequences were used to train the classifiers. The training features include amino acid composition; auto correlation coefficients; composition, transition, and distribution of various physicochemical properties; and pseudo amino acid composition. Performance analysis of predictive models based on 10 fold cross-validation and independent data testing revealed that the Random Forest based model was the best and achieved 86.11% overall accuracy and 86.22% balanced accuracy with a remarkably high area under the Receivers Operating Characteristic curve of 0.95 to predict viral RNA silencing suppressor proteins. The prediction models for plant viral RNA silencing suppressors can potentially aid identification of novel viral RNA silencing suppressors, which will provide valuable insights into the mechanism of RNA silencing and could be further explored as potential targets for designing novel antiviral therapeutics. Also, the key subset of identified optimal features may help in determining compositional patterns in the viral proteins which are important determinants for RNA silencing suppressor activities. The best prediction model developed in the study is available as a

  6. Synthetic RNA Silencing in Bacteria – Antimicrobial Discovery and Resistance Breaking


    Good, Liam; Stach, James E. M.


    The increasing incidence and prevalence of antibiotic resistance in bacteria threatens the “antibiotic miracle.” Conventional antimicrobial drug development has failed to replace the armamentarium needed to combat this problem, and novel solutions are urgently required. Here we review both natural and synthetic RNA silencing and its potential to provide new antibacterials through improved target selection, evaluation, and screening. Furthermore, we focus on synthetic RNA silencers as a novel ...

  7. Virus-Induced Gene Silencing as a Tool to Study Tomato Fruit Biochemistry. (United States)

    Fantini, Elio; Giuliano, Giovanni


    Virus-Induced Gene Silencing (VIGS) is an excellent reverse genetic tool for the study of gene function in plants, based on virus infection. In this chapter, we describe a high-throughput approach based on VIGS for the study of tomato fruit biochemistry. It comprises the selection of the sequence for silencing using bioinformatics tools, the cloning of the fragment in the Tobacco Rattle Virus (TRV), and the agroinfiltration of tomato fruits mediated by Agrobacterium tumefaciens.

  8. Development of a PCR assay suitable for Campylobacter spp. mass screening programs in broiler production

    DEFF Research Database (Denmark)

    Bang, Dang Duong; Pedersen, Karl; Madsen, Mogens


    culture techniques since 1998. However, using conventional culture methods is time consuming and laborious, and therefore a Polymerase Chain Reaction (PCR) Campylobacter detection assay suitable for mass screening of cloacal swab samples from broilers was developed. By comparing the PCR detection...... with conventional culture methods, significantly more samples were found positive for Campylobacter with the PCR method. The PCR method is rapid, sensitive and suitable for mass screening for Campylobacter in poultry. Using this PCR method Campylobacter can be detected within 15 h. Notably, the method can...... be applied to detect Campylobacter directly from chicken feces at the species level....

  9. The molecular basis for stability of heterochromatin-mediated silencing in mammals

    Directory of Open Access Journals (Sweden)

    Hiragami-Hamada Kyoko


    Full Text Available Abstract The archetypal epigenetic phenomenon of position effect variegation (PEV in Drosophila occurs when a gene is brought abnormally close to heterochromatin, resulting in stochastic silencing of the affected gene in a proportion of cells that would normally express it. PEV has been instrumental in unraveling epigenetic mechanisms. Using an in vivo mammalian model for PEV we have extensively investigated the molecular basis for heterochromatin-mediated gene silencing. Here we distinguish 'epigenetic effects' from other cellular differences by studying ex vivo cells that are identical, apart from the expression of the variegating gene which is silenced in a proportion of the cells. By separating cells according to transgene expression we show here that silencing appears to be associated with histone H3 lysine 9 trimethylation (H3K9me3, DNA methylation and the localization of the silenced gene to a specific nuclear compartment enriched in these modifications. In contrast, histone H3 acetylation (H3Ac and lysine 4 di or tri methylation (H3K4me2/3 are the predominant modifications associated with expression where we see the gene in a euchromatic compartment. Interestingly, DNA methylation and inaccessibility, rather than H3K9me3, correlated most strongly with resistance to de-repression by cellular activation. These results have important implications for understanding the contribution of specific factors involved in the establishment and maintenance of gene silencing and activation in vivo.

  10. The Nuclear Cap-Binding Complex Mediates Meiotic Silencing by Unpaired DNA

    Directory of Open Access Journals (Sweden)

    Logan M. Decker


    Full Text Available In the filamentous fungus Neurospora crassa, cross walls between individual cells are normally incomplete, making the entire fungal network vulnerable to attack by viruses and selfish DNAs. Accordingly, several genome surveillance mechanisms are maintained to help the fungus combat these repetitive elements. One of these defense mechanisms is called meiotic silencing by unpaired DNA (MSUD, which identifies and silences unpaired genes during meiosis. Utilizing common RNA interference (RNAi proteins, such as Dicer and Argonaute, MSUD targets mRNAs homologous to the unpaired sequence to achieve silencing. In this study, we have identified an additional silencing component, namely the cap-binding complex (CBC. Made up of cap-binding proteins CBP20 and CBP80, CBC associates with the 5′ cap of mRNA transcripts in eukaryotes. The loss of CBC leads to a deficiency in MSUD activity, suggesting its role in mediating silencing. As confirmed in this study, CBC is predominantly nuclear, although it is known to travel in and out of the nucleus to facilitate RNA transport. As seen in animals but not in plants, CBP20’s robust nuclear import depends on CBP80 in Neurospora. CBC interacts with a component (Argonaute of the perinuclear meiotic silencing complex (MSC, directly linking the two cellular factors.

  11. Strategy of gene silencing in cassava for validation of resistance genes

    International Nuclear Information System (INIS)

    Cortes, Simon; Lopez, Camilo


    Cassava (Manihot esculenta) is a major source of food for more than 1000 million people in the world and constitutes an important staple crop. Cassava bacterial blight, caused by the gram negative bacterium Xanthomonas axonopodis pv. manihotis, is one of the most important constraints for this crop. A candidate resistance gene against cassava bacterial blight, named RXam1, has been identified previously. In this work, we employed the gene silencing approach using the African cassava mosaic virus (ACMV) to validate the function of the RXam1 gene. We used as positive control the su gen, which produce photo blanching in leaves when is silenced. Plants from the SG10735 variety were bombardment with the ACMV-A-SU+ACMV-B y ACMV-A-RXam1+ACMV-B constructions. The silencing efficiency employing the su gene was low, only one of seven plants showed photo blanching. In the putative silenced plants for the RXam1 gene, no presence of siRNAs corresponding to RXam1 was observed; although a low diminution of the RXam1 gene expression was obtained. The growth curves for the Xam strain CIO136 in cassava plants inoculated showing a little but no significance difference in the susceptibility in the silenced plants compared to not silenced

  12. Silencing of the FRO1 gene and its effects on iron partition in Nicotiana benthamiana. (United States)

    Gama, Florinda; Saavedra, Teresa; Dandlen, Susana; de Varennes, Amarilis; Correia, Pedro J; Pestana, Maribela; Nolasco, Gustavo


    To evaluate the dynamic role of the ferric-chelate reductase enzyme (FCR) and to identify possible pathways of regulation of its activity in different plant organs an investigation was conducted by virus-induced gene silencing (VIGS) using tobacco rattle virus (TRV) to silence the ferric reductase oxidase gene (FRO1) that encodes the FCR enzyme. Half of Nicotiana benthamiana plants received the VIGS vector and the rest remained as control. Four treatments were imposed: two levels of Fe in the nutrient solution (0 or 2.5 μM of Fe), each one with silenced or non-silenced (VIGS-0; VIGS-2.5) plants. Plants grown without iron (0; VIGS-0) developed typical symptoms of iron deficiency in the youngest leaves. To prove that FRO1 silencing had occurred, resupply of Fe (R) was done by adding 2.5 μM of Fe to the nutrient solution in a subset of chlorotic plants (0-R; VIGS-R). Twelve days after resupply, 0-R plants had recovered from Fe deficiency while plants containing the VIGS vector (VIGS-R) remained chlorotic and both FRO1 gene expression and FCR activity were considerably reduced, consequently preventing Fe uptake. With the VIGS technique we were able to silence the FRO1 gene in N. benthamiana and point out its importance in chlorophyll synthesis and Fe partition. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  13. On the role of RNA silencing in the pathogenicity and evolution of viroids and viral satellites (United States)

    Wang, Ming-Bo; Bian, Xue-Yu; Wu, Li-Min; Liu, Li-Xia; Smith, Neil A.; Isenegger, Daniel; Wu, Rong-Mei; Masuta, Chikara; Vance, Vicki B.; Watson, John M.; Rezaian, Ali; Dennis, Elizabeth S.; Waterhouse, Peter M.


    Viroids and most viral satellites have small, noncoding, and highly structured RNA genomes. How they cause disease symptoms without encoding proteins and why they have characteristic secondary structures are two longstanding questions. Recent studies have shown that both viroids and satellites are capable of inducing RNA silencing, suggesting a possible role of this mechanism in the pathology and evolution of these subviral RNAs. Here we show that preventing RNA silencing in tobacco, using a silencing suppressor, greatly reduces the symptoms caused by the Y satellite of cucumber mosaic virus. Furthermore, tomato plants expressing hairpin RNA, derived from potato spindle tuber viroid, developed symptoms similar to those of potato spindle tuber viroid infection. These results provide evidence suggesting that viroids and satellites cause disease symptoms by directing RNA silencing against physiologically important host genes. We also show that viroid and satellite RNAs are significantly resistant to RNA silencing-mediated degradation, suggesting that RNA silencing is an important selection pressure shaping the evolution of the secondary structures of these pathogens. PMID:14978267

  14. Geophysical remote sensing of water reservoirs suitable for desalinization.

    Energy Technology Data Exchange (ETDEWEB)

    Aldridge, David Franklin; Bartel, Lewis Clark; Bonal, Nedra; Engler, Bruce Phillip


    In many parts of the United States, as well as other regions of the world, competing demands for fresh water or water suitable for desalination are outstripping sustainable supplies. In these areas, new water supplies are necessary to sustain economic development and agricultural uses, as well as support expanding populations, particularly in the Southwestern United States. Increasing the supply of water will more than likely come through desalinization of water reservoirs that are not suitable for present use. Surface-deployed seismic and electromagnetic (EM) methods have the potential for addressing these critical issues within large volumes of an aquifer at a lower cost than drilling and sampling. However, for detailed analysis of the water quality, some sampling utilizing boreholes would be required with geophysical methods being employed to extrapolate these sampled results to non-sampled regions of the aquifer. The research in this report addresses using seismic and EM methods in two complimentary ways to aid in the identification of water reservoirs that are suitable for desalinization. The first method uses the seismic data to constrain the earth structure so that detailed EM modeling can estimate the pore water conductivity, and hence the salinity. The second method utilizes the coupling of seismic and EM waves through the seismo-electric (conversion of seismic energy to electrical energy) and the electro-seismic (conversion of electrical energy to seismic energy) to estimate the salinity of the target aquifer. Analytic 1D solutions to coupled pressure and electric wave propagation demonstrate the types of waves one expects when using a seismic or electric source. A 2D seismo-electric/electro-seismic is developed to demonstrate the coupled seismic and EM system. For finite-difference modeling, the seismic and EM wave propagation algorithms are on different spatial and temporal scales. We present a method to solve multiple, finite-difference physics

  15. Selecting suitable enclosures for digitally printed materials

    International Nuclear Information System (INIS)

    Burge, D; Rima, L


    It cannot be assumed that storage enclosures considered safe for traditionally printed images and documents are suitable for modern, digitally printed materials. In this project, a large variety of digital print types were tested using a modified version of the ISO 18916 Imaging materials-Processed imaging materials-Photographic activity test for enclosure materials standard to assess the risk to digital prints by paper enclosures known to be inert or reactive with traditional photographic prints. The types of enclosures tested included buffered and non-buffered cotton papers, and groundwood paper. In addition, qualitative filter paper that had been wetted and dried with either an acidic or basic solution was also tested to determine the effects of enclosure pH on digitally printed materials. It was determined that, in general, digital prints tended to be less reactive with various enclosure types than traditional prints. Digital prints were most sensitive to paper that contained groundwood. The enclosure reactivity test results were then integrated with previous published work on the tendencies of various enclosure types to abrade, ferrotype, or block to digital prints in order to create a comprehensive set of recommendations for digital print storage enclosures.

  16. Epigenetic silencing of miRNA-9 is associated with HES1 oncogenic activity and poor prognosis of medulloblastoma (United States)

    Fiaschetti, G; Abela, L; Nonoguchi, N; Dubuc, A M; Remke, M; Boro, A; Grunder, E; Siler, U; Ohgaki, H; Taylor, M D; Baumgartner, M; Shalaby, T; Grotzer, M A


    Background: microRNA-9 is a key regulator of neuronal development aberrantly expressed in brain malignancies, including medulloblastoma. The mechanisms by which microRNA-9 contributes to medulloblastoma pathogenesis remain unclear, and factors that regulate this process have not been delineated. Methods: Expression and methylation status of microRNA-9 in medulloblastoma cell lines and primary samples were analysed. The association of microRNA-9 expression with medulloblastoma patients' clinical outcome was assessed, and the impact of microRNA-9 restoration was functionally validated in medulloblastoma cells. Results: microRNA-9 expression is repressed in a large subset of MB samples compared with normal fetal cerebellum. Low microRNA-9 expression correlates significantly with the diagnosis of unfavourable histopathological variants and with poor clinical outcome. microRNA-9 silencing occurs via cancer-specific CpG island hypermethylation. HES1 was identified as a direct target of microRNA-9 in medulloblastoma, and restoration of microRNA-9 was shown to trigger cell cycle arrest, to inhibit clonal growth and to promote medulloblastoma cell differentiation. Conclusions: microRNA-9 is a methylation-silenced tumour suppressor that could be a potential candidate predictive marker for poor prognosis of medulloblastoma. Loss of microRNA-9 may confer a proliferative advantage to tumour cells, and it could possibly contribute to disease pathogenesis. Thus, re-expression of microRNA-9 may constitute a novel epigenetic regulation strategy against medulloblastoma. PMID:24346283

  17. Improved production of heterologous lipase in Trichoderma reesei by RNAi mediated gene silencing of an endogenic highly expressed gene. (United States)

    Qin, Li-Na; Cai, Fu-Rong; Dong, Xin-Rui; Huang, Zhen-Bang; Tao, Yong; Huang, Jian-Zhong; Dong, Zhi-Yang


    A lipase gene (Lip) of the Aspergillus niger was de novo synthesized and expressed in the Trichoderma reesei under the promoter of the cellobiohydrolase I gene (cbh1). RNAi-mediated gene silencing was successfully used to further improve the recombinant lipase production via down-regulation of CBHI which comprised more than 60% of the total extracellular proteins in T. reesei. The gene and protein expression of CBHI and recombinant lipase were analyzed by real-time PCR, SDS-PAGE and activity assay. The results demonstrated that RNAi-mediated gene silencing could effectively suppress cbh1 gene expression and the reduction of CBHI could result in obvious improvement of heterologous lipase production. The reconstructed strains with decreased CBHI production exhibited 1.8- to 3.2-fold increase in lipase activity than that of parental strain. The study herein provided a feasible and advantageous method of increasing heterologous target gene expression in T. reesei through preventing the high expression of a specific endogenenous gene by RNA interference. Copyright © 2012 Elsevier Ltd. All rights reserved.

  18. Silencing by H-NS potentiated the evolution of Salmonella.

    Directory of Open Access Journals (Sweden)

    Sabrina S Ali


    Full Text Available The bacterial H-NS protein silences expression from sequences with higher AT-content than the host genome and is believed to buffer the fitness consequences associated with foreign gene acquisition. Loss of H-NS results in severe growth defects in Salmonella, but the underlying reasons were unclear. An experimental evolution approach was employed to determine which secondary mutations could compensate for the loss of H-NS in Salmonella. Six independently derived S. Typhimurium hns mutant strains were serially passaged for 300 generations prior to whole genome sequencing. Growth rates of all lineages dramatically improved during the course of the experiment. Each of the hns mutant lineages acquired missense mutations in the gene encoding the H-NS paralog StpA encoding a poorly understood H-NS paralog, while 5 of the mutant lineages acquired deletions in the genes encoding the Salmonella Pathogenicity Island-1 (SPI-1 Type 3 secretion system critical to invoke inflammation. We further demonstrate that SPI-1 misregulation is a primary contributor to the decreased fitness in Salmonella hns mutants. Three of the lineages acquired additional loss of function mutations in the PhoPQ virulence regulatory system. Similarly passaged wild type Salmonella lineages did not acquire these mutations. The stpA missense mutations arose in the oligomerization domain and generated proteins that could compensate for the loss of H-NS to varying degrees. StpA variants most able to functionally substitute for H-NS displayed altered DNA binding and oligomerization properties that resembled those of H-NS. These findings indicate that H-NS was central to the evolution of the Salmonellae by buffering the negative fitness consequences caused by the secretion system that is the defining characteristic of the species.


    Directory of Open Access Journals (Sweden)

    Vladimir Vukadinović


    Full Text Available A total of 17405 soil samples (2003rd-2009th years were analyzed in the eastern part of Croatia. The aim of this paper is to assess land suitability for crops i.e. to describe quantitatively land quality and indicate disadvantages of land using system in investigated area. The described mathematical model uses score functions for estimating indicators of soil suitability. Soil suitability assessment computer model for crops, supported by GIS, proved to be fast, efficient and enough reliable. Using GIS tool it is possible to visualize land suitability and present it in different cartographic bases such as maps whereas using geostatistical method – kriging enables to possible to provide regionalization of production area based on quantitative assessment of land suitability for crops.

  20. Identification of suitable sites for mountain ginseng cultivation using GIS and geo-temperature. (United States)

    Kang, Hag Mo; Choi, Soo Im; Kim, Hyun


    This study was conducted to explore an accurate site identification technique using a geographic information system (GIS) and geo-temperature (gT) for locating suitable sites for growing cultivated mountain ginseng (CMG; Panax ginseng), which is highly sensitive to the environmental conditions in which it grows. The study site was Jinan-gun, South Korea. The spatial resolution for geographic data was set at 10 m × 10 m, and the temperatures for various climatic factors influencing CMG growth were calculated by averaging the 3-year temperatures obtained from the automatic weather stations of the Korea Meteorological Administration. Identification of suitable sites for CMG cultivation was undertaken using both a conventional method and a new method, in which the gT was added as one of the most important factors for crop cultivation. The results yielded by the 2 methods were then compared. When the gT was added as an additional factor (new method), the proportion of suitable sites identified decreased by 0.4 % compared with the conventional method. However, the proportion matching real CMG cultivation sites increased by 3.5 %. Moreover, only 68.2 % corresponded with suitable sites identified using the conventional factors; i.e., 31.8 % were newly detected suitable sites. The accuracy of GIS-based identification of suitable CMG cultivation sites improved by applying the temperature factor (i.e., gT) in addition to the conventionally used factors.

  1. Development of a gene silencing DNA vector derived from a broad host range geminivirus

    Directory of Open Access Journals (Sweden)

    Hancock Leandria C


    Full Text Available Abstract Background Gene silencing is proving to be a powerful tool for genetic, developmental, and physiological analyses. The use of viral induced gene silencing (VIGS offers advantages to transgenic approaches as it can be potentially applied to non-model systems for which transgenic techniques are not readily available. However, many VIGS vectors are derived from Gemini viruses that have limited host ranges. We present a new, unipartite vector that is derived from a curtovirus that has a broad host range and will be amenable to use in many non-model systems. Results The construction of a gene silencing vector derived from the geminivirus Beet curly top virus (BCTV, named pWSRi, is reported. Two versions of the vector have been developed to allow application by biolistic techniques or by agro-infiltration. We demonstrate its ability to silence nuclear genes including ribulose bisphosphate carboxylase small subunit (rbcS, transketolase, the sulfur allele of magnesium chelatase (ChlI, and two homeotic transcription factors in spinach or tomato by generating gene-specific knock-down phenotypes. Onset of phenotypes occurred 3 to 12 weeks post-inoculation, depending on the target gene, in organs that developed after the application. The vector lacks movement genes and we found no evidence for significant spread from the site of inoculation. However, viral amplification in inoculated tissue was detected and is necessary for systemic silencing, suggesting that signals generated from active viral replicons are efficiently transported within the plant. Conclusion The unique properties of the pWSRi vector, the ability to silence genes in meristem tissue, the separation of virus and silencing phenotypes, and the broad natural host range of BCTV, suggest that it will have wide utility.

  2. Specific tandem repeats are sufficient for paramutation-induced trans-generational silencing.

    Directory of Open Access Journals (Sweden)

    Christiane L Belele

    Full Text Available Paramutation is a well-studied epigenetic phenomenon in which trans communication between two different alleles leads to meiotically heritable transcriptional silencing of one of the alleles. Paramutation at the b1 locus involves RNA-mediated transcriptional silencing and requires specific tandem repeats that generate siRNAs. This study addressed three important questions: 1 are the tandem repeats sufficient for paramutation, 2 do they need to be in an allelic position to mediate paramutation, and 3 is there an association between the ability to mediate paramutation and repeat DNA methylation levels? Paramutation was achieved using multiple transgenes containing the b1 tandem repeats, including events with tandem repeats of only one half of the repeat unit (413 bp, demonstrating that these sequences are sufficient for paramutation and an allelic position is not required for the repeats to communicate. Furthermore, the transgenic tandem repeats increased the expression of a reporter gene in maize, demonstrating the repeats contain transcriptional regulatory sequences. Transgene-mediated paramutation required the mediator of paramutation1 gene, which is necessary for endogenous paramutation, suggesting endogenous and transgene-mediated paramutation both require an RNA-mediated transcriptional silencing pathway. While all tested repeat transgenes produced small interfering RNAs (siRNAs, not all transgenes induced paramutation suggesting that, as with endogenous alleles, siRNA production is not sufficient for paramutation. The repeat transgene-induced silencing was less efficiently transmitted than silencing induced by the repeats of endogenous b1 alleles, which is always 100% efficient. The variability in the strength of the repeat transgene-induced silencing enabled testing whether the extent of DNA methylation within the repeats correlated with differences in efficiency of paramutation. Transgene-induced paramutation does not require extensive

  3. The Luteovirus P4 Movement Protein Is a Suppressor of Systemic RNA Silencing

    Directory of Open Access Journals (Sweden)

    Adriana F. Fusaro


    Full Text Available The plant viral family Luteoviridae is divided into three genera: Luteovirus, Polerovirus and Enamovirus. Without assistance from another virus, members of the family are confined to the cells of the host plant’s vascular system. The first open reading frame (ORF of poleroviruses and enamoviruses encodes P0 proteins which act as silencing suppressor proteins (VSRs against the plant’s viral defense-mediating RNA silencing machinery. Luteoviruses, such as barley yellow dwarf virus-PAV (BYDV-PAV, however, have no P0 to carry out the VSR role, so we investigated whether other proteins or RNAs encoded by BYDV-PAV confer protection against the plant’s silencing machinery. Deep-sequencing of small RNAs from plants infected with BYDV-PAV revealed that the virus is subjected to RNA silencing in the phloem tissues and there was no evidence of protection afforded by a possible decoy effect of the highly abundant subgenomic RNA3. However, analysis of VSR activity among the BYDV-PAV ORFs revealed systemic silencing suppression by the P4 movement protein, and a similar, but weaker, activity by P6. The closely related BYDV-PAS P4, but not the polerovirus potato leafroll virus P4, also displayed systemic VSR activity. Both luteovirus and the polerovirus P4 proteins also showed transient, weak local silencing suppression. This suggests that systemic silencing suppression is the principal mechanism by which the luteoviruses BYDV-PAV and BYDV-PAS minimize the effects of the plant’s anti-viral defense.

  4. Balanced phono-amps an extension to the 'the sound of silence' editions

    CERN Document Server

    Vogel, Burkhard


    In 12 chapters (Part I) this extension to the two 'The Sound of Silence' editions covers the development, calculation, construction and measurement of the fully differential (= balanced) phono-amp solution 'RIAA Phono-Amp Engine II'. Additionally, the balanced measurement amplifiers & measurement tools, the discussion on BJT gain stages, the 1/f noise calculation methods for BJTs, the calculation of fully-differential amplifiers, the numerous Mathcad worksheets, and the presentation of test and calibration records fill a further 10 chapters of Part II with essential knowhow that will equip the reader to develop his/her own phono-amp solution in the most balanced way - and of course, as low-noise as possible. Engine II offers eight different amplifying paths from the Engine's input to its output - solid-state as well as valve driven. To expand the input possibilities via an additional external input, any kind of other linear input amplifiers can be connected. Further, a selection of six highly diverse exte...

  5. Analytical tools for identifying bicycle route suitability, coverage, and continuity. (United States)


    This report presents new tools created to assess bicycle suitability using geographic information systems (GIS). Bicycle suitability is a rating of how appropriate a roadway is for bicycle travel based on attributes of the roadway, such as vehi...

  6. Application of neutron activation and high resolution X-ray spectrometry to the development of analytical methods suitable for the assay of trace elements in biological materials. Part of a coordinated programme on comparative methods for the study of trace elements in human nutrition

    International Nuclear Information System (INIS)

    Mantel, J.


    A new method for determining trace elements in biological materials by instrumental neutron activation analysis (INAA) has been investigated. This method is based on the measurement of low-energy X-rays emitted by the sample after appropriate activation with thermal neutrons. The X-rays were detected by a Si(Li) diode connected to a multichannel analyser. The background in the energy region of interest was reduced by means of an electromagnet which deflects high-energy beta-particles away from the detector. This method of measurement has been evaluated by application to a number of different biological reference materials. The use of the electromagnet for background reduction was shown to be extremely effective for several radionuclides that are common sources of disturbance in INAA. For 32 P the background was only 0.8% of its original value. For real samples the background reduction depends somewhat on the matrix and therefore varies from one material to another. Practical detection limits for typical biological materials were derived for 18 different trace elements. The use of a perspex absorber was also considered as an alternative to magnetic deflection of the beta rays. In practice, magnetic deflection is preferable for the lower-energy X-ray emitters such as the activation products of Co, Cr, Cu and Zn

  7. miRNA-mediated post-transcriptional silencing of transgenes leads to increased adeno-associated viral vector yield and targeting specificity. (United States)

    Reid, C A; Boye, S L; Hauswirth, W W; Lipinski, D M


    The production of high-titer recombinant adeno-associated virus (rAAV) vector is essential for treatment of genetic diseases affecting the retina and choroid, where anatomical constraints may limit injectable volumes. Problematically, cytotoxicity arising from overexpression of the transgene during vector production frequently leads to a reduction in vector yield. Herein, we evaluate the use of microRNA (miRNA)-mediated silencing to limit overexpression of cytotoxic transgenes during packaging as a method of increasing vector yield. We examined if post-transcriptional regulation of transgenes during packaging via miRNA technology would lead to increased rAAV yields. Our results demonstrate that silencing of cytotoxic transgenes during production resulted in up to a 22-fold increase in vector yield. The inclusion of organ-specific miRNA sequences improved biosafety by limiting off-target expression following systemic rAAV administration. The small size (22-23 bp) of the target site allows for the inclusion of multiple copies into the vector with minimal impact on coding capacity. Taken together, our results suggest that inclusion of miRNA target sites into the 3'-untranslated region of the AAV cassette allow for silencing of cytotoxic transgenes during vector production leading to improved vector yield, in addition to increasing targeting specificity without reliance on cell-specific promoters.

  8. Combination of siRNA-directed Kras oncogene silencing and arsenic-induced apoptosis using a nanomedicine strategy for the effective treatment of pancreatic cancer. (United States)

    Zeng, Linjuan; Li, Jingguo; Wang, Yong; Qian, Chenchen; Chen, Yinting; Zhang, Qiubo; Wu, Wei; Lin, Zhong; Liang, Jianzhong; Shuai, Xintao; Huang, Kaihong


    The synergetic inhibitory effects on human pancreatic cancer by nanoparticle-mediated siRNA and arsenic therapy were investigated both in vitro and in vivo. Poly(ethylene glycol)-block-poly(L-lysine) were prepared to form siRNA-complexed polyplex and poly(ethylene glycol)-block-poly(DL-lactide) were prepared to form arsenic-encapsulated vesicle, respectively. Down-regulation of the mutant Kras gene by siRNA caused defective abilities of proliferation, clonal formation, migration, and invasion of pancreatic cancer cells, as well as cell cycle arrest at the G0/G1 phase, which substantially enhanced the apoptosis-inducing effect of arsenic administration. Consequently, co-administration of the two nanomedicines encapsulating siRNA or arsenic showed ideal tumor growth inhibition both in vitro and in vivo as a result of synergistic effect of the siRNA-directed Kras oncogene silencing and arsenic-induced cell apoptosis. These results suggest that the combination of mutant Kras gene silencing and arsenic therapy using nanoparticle-mediated delivery strategy is promising for pancreatic cancer treatment. Treatment of pancreatic cancer remains a major challenge. These authors demonstrate a method that combines a siRNA-based Kras silencing with arsenic delivery to pancreatic cancer cells using nanoparticles, resulting in enhanced apoptosis induction in the treated cells. © 2013.

  9. Epigenetic silencing of the 3p22 tumor suppressor DLEC1 by promoter CpG methylation in non-Hodgkin and Hodgkin lymphomas

    Directory of Open Access Journals (Sweden)

    Wang Zhaohui


    Full Text Available Abstract Background Inactivaion of tumor suppressor genes (TSGs by promoter CpG methylation frequently occurs in tumorigenesis, even in the early stages, contributing to the initiation and progression of human cancers. Deleted in lung and esophageal cancer 1 (DLEC1, located at the 3p22-21.3 TSG cluster, has been identified frequently silenced by promoter CpG methylation in multiple carcinomas, however, no study has been performed for lymphomas yet. Methods We examined the expression of DLEC1 by semi-quantitative reverse transcription (RT-PCR, and evaluated the promoter methylation of DLEC1 by methylation-specific PCR (MSP and bisulfite genomic sequencing (BGS in common lymphoma cell lines and tumors. Results Here we report that DLEC1 is readily expressed in normal lymphoid tissues including lymph nodes and PBMCs, but reduced or silenced in 70% (16/23 of non-Hodgkin and Hodgkin lymphoma cell lines, including 2/6 diffuse large B-cell (DLBCL, 1/2 peripheral T cell lymphomas, 5/5 Burkitt, 6/7 Hodgkin and 2/3 nasal killer (NK/T-cell lymphoma cell lines. Promoter CpG methylation was frequently detected in 80% (20/25 of lymphoma cell lines and correlated with DLEC1 downregulation/silencing. Pharmacologic demethylation reversed DLEC1 expression in lymphoma cell lines along with concomitant promoter demethylation. DLEC1 methylation was also frequently detected in 32 out of 58 (55% different types of lymphoma tissues, but not in normal lymph nodes. Furthermore, DLEC1 was specifically methylated in the sera of 3/13 (23% Hodgkin lymphoma patients. Conclusions Thus, methylation-mediated silencing of DLEC1 plays an important role in multiple lymphomagenesis, and may serve as a non-invasive tumor marker for lymphoma diagnosis.

  10. Active chromatin marks are retained on X chromosomes lacking gene or repeat silencing despite XIST/Xist expression in somatic cell hybrids.

    Directory of Open Access Journals (Sweden)

    Nancy P Thorogood


    Full Text Available X-chromosome inactivation occurs early in mammalian development and results in the inactive X chromosome acquiring numerous hallmarks of heterochromatin. While XIST is a key player in the inactivation process, the method of action of this ncRNA is yet to be determined.To assess which features of heterochromatin may be directly recruited by the expression and localization of the XIST RNA we have analyzed a mouse/human somatic cell hybrid in which expression of human and mouse XIST/Xist has been induced from the active X by demethylation. Such hybrids had previously been demonstrated to disconnect XIST/Xist expression from gene silencing and we confirm maintenance of X-linked gene expression, even close to the Xist locus, despite the localized expression of mouse Xist.Loss of the active chromatin marks H3 acetylation and H3 lysine 4 methylation was not observed upon XIST/Xist expression, nor was there a gain of DNA methylation; thus these marks of facultative heterochromatin are not solely dependent upon Xist expression. Cot-1 holes, regions of depleted RNA hybridization with a Cot-1 probe, were observed upon Xist expression; however, these were at reduced frequency and intensity in these somatic cells. Domains of human Cot-1 transcription were observed corresponding to the human chromosomes in the somatic cell hybrids. The Cot-1 domain of the X was not reduced with the expression of XIST, which fails to localize to the human X chromosome in a mouse somatic cell background. The human inactive X in a mouse/human hybrid cell also shows delocalized XIST expression and an ongoing Cot-1 domain, despite X-linked gene silencing. These results are consistent with recent reports separating Cot-1 silencing from genic silencing, but also demonstrate repetitive element expression from an otherwise silent X chromosome in these hybrid cells.

  11. Characteristics and suitability of waste dump sites in Owerri, Nigeria ...

    African Journals Online (AJOL)

    A five point criteria screening of the three major waste dumpsites in Owerri Municipality was identified, and a ranking procedure adopted to determine the suitability or otherwise of the dumpsites. The sites were screened and ranked hydro-geologically and geo-technically in order of suitable, moderately suitable, and not ...

  12. Tool for Land Suitability Assessment for Rice Production in

    African Journals Online (AJOL)


    results of the thematic suitability assessment for the three cultivars of rice show that the percent of land area very suitable for rice production doubles from medium matured variety to early matured variety, and from early matured variety to very early matured variety. For suitable areas, and for the same sequence as just.

  13. The Edge Detectors Suitable for Retinal OCT Image Segmentation

    Directory of Open Access Journals (Sweden)

    Su Luo


    Full Text Available Retinal layer thickness measurement offers important information for reliable diagnosis of retinal diseases and for the evaluation of disease development and medical treatment responses. This task critically depends on the accurate edge detection of the retinal layers in OCT images. Here, we intended to search for the most suitable edge detectors for the retinal OCT image segmentation task. The three most promising edge detection algorithms were identified in the related literature: Canny edge detector, the two-pass method, and the EdgeFlow technique. The quantitative evaluation results show that the two-pass method outperforms consistently the Canny detector and the EdgeFlow technique in delineating the retinal layer boundaries in the OCT images. In addition, the mean localization deviation metrics show that the two-pass method caused the smallest edge shifting problem. These findings suggest that the two-pass method is the best among the three algorithms for detecting retinal layer boundaries. The overall better performance of Canny and two-pass methods over EdgeFlow technique implies that the OCT images contain more intensity gradient information than texture changes along the retinal layer boundaries. The results will guide our future efforts in the quantitative analysis of retinal OCT images for the effective use of OCT technologies in the field of ophthalmology.

  14. Multi-criteria Decision Analysis to Model Ixodes ricinus Habitat Suitability. (United States)

    Rousseau, Raphaël; McGrath, Guy; McMahon, Barry J; Vanwambeke, Sophie O


    Tick-borne diseases present a major threat to both human and livestock health throughout Europe. The risk of infection is directly related to the presence of its vector. Thereby it is important to know their distribution, which is strongly associated with environmental factors: the presence and availability of a suitable habitat, of a suitable climate and of hosts. The present study models the habitat suitability for Ixodes ricinus in Ireland, where data on tick distribution are scarce. Tick habitat suitability was estimated at a coarse scale (10 km) with a multi-criteria decision analysis (MCDA) method according to four different scenarios (depending on the variables used and on the weights granted to each of them). The western part of Ireland and the Wicklow mountains in the East were estimated to be the most suitable areas for I. ricinus in the island. There was a good level of agreement between results from the MCDA and recorded tick presence. The different scenarios did not affect the spatial outputs substantially. The current study suggests that tick habitat suitability can be mapped accurately at a coarse scale in a data-scarce context using knowledge-based methods. It can serve as a guideline for future countrywide sampling that would help to determine local risk of tick presence and refining knowledge on tick habitat suitability in Ireland.

  15. C. elegans RNA-dependent RNA polymerases rrf-1 and ego-1 silence Drosophila transgenes by differing mechanisms. (United States)

    Duan, Guowen; Saint, Robert B; Helliwell, Chris A; Behm, Carolyn A; Wang, Ming-Bo; Waterhouse, Peter M; Gordon, Karl H J


    Drosophila possesses the core gene silencing machinery but, like all insects, lacks the canonical RNA-dependent RNA polymerases (RdRps) that in C. elegans either trigger or enhance two major small RNA-dependent gene silencing pathways. Introduction of two different nematode RdRps into Drosophila showed them to be functional, resulting in differing silencing activities. While RRF-1 enhanced transitive dsRNA-dependent silencing, EGO-1 triggered dsRNA-independent silencing, specifically of transgenes. The strain w; da-Gal4; UAST-ego-1, constitutively expressing ego-1, is capable of silencing transgene including dsRNA hairpin upon a single cross, which created a powerful tool for research in Drosophila. In C. elegans, EGO-1 is involved in transcriptional gene silencing (TGS) of chromosome regions that are unpaired during meiosis. There was no opportunity for meiotic interactions involving EGO-1 in Drosophila that would explain the observed transgene silencing. Transgene DNA is, however, unpaired during the pairing of chromosomes in embryonic mitosis that is an unusual characteristic of Diptera, suggesting that in Drosophila, EGO-1 triggers transcriptional silencing of unpaired DNA during embryonic mitosis.

  16. The VP3 factor from viruses of Birnaviridae family suppresses RNA silencing by binding both long and small RNA duplexes.

    Directory of Open Access Journals (Sweden)

    Adrian Valli

    Full Text Available RNA silencing is directly involved in antiviral defense in a wide variety of eukaryotic organisms, including plants, fungi, invertebrates, and presumably vertebrate animals. The study of RNA silencing-mediated antiviral defences in vertebrates is hampered by the overlap with other antiviral mechanisms; thus, heterologous systems are often used to study the interplay between RNA silencing and vertebrate-infecting viruses. In this report we show that the VP3 protein of the avian birnavirus Infectious bursal disease virus (IBDV displays, in addition to its capacity to bind long double-stranded RNA, the ability to interact with double-stranded small RNA molecules. We also demonstrate that IBDV VP3 prevents the silencing mediated degradation of a reporter mRNA, and that this silencing suppression activity depends on its RNA binding ability. Furthermore, we find that the anti-silencing activity of IBDV VP3 is shared with the homologous proteins expressed by both insect- and fish-infecting birnaviruses. Finally, we show that IBDV VP3 can functionally replace the well-characterized HCPro silencing suppressor of Plum pox virus, a potyvirus that is unable to infect plants in the absence of an active silencing suppressor. Altogether, our results support the idea that VP3 protects the viral genome from host sentinels, including those of the RNA silencing machinery.

  17. The VP3 factor from viruses of Birnaviridae family suppresses RNA silencing by binding both long and small RNA duplexes. (United States)

    Valli, Adrian; Busnadiego, Idoia; Maliogka, Varvara; Ferrero, Diego; Castón, José R; Rodríguez, José Francisco; García, Juan Antonio


    RNA silencing is directly involved in antiviral defense in a wide variety of eukaryotic organisms, including plants, fungi, invertebrates, and presumably vertebrate animals. The study of RNA silencing-mediated antiviral defences in vertebrates is hampered by the overlap with other antiviral mechanisms; thus, heterologous systems are often used to study the interplay between RNA silencing and vertebrate-infecting viruses. In this report we show that the VP3 protein of the avian birnavirus Infectious bursal disease virus (IBDV) displays, in addition to its capacity to bind long double-stranded RNA, the ability to interact with double-stranded small RNA molecules. We also demonstrate that IBDV VP3 prevents the silencing mediated degradation of a reporter mRNA, and that this silencing suppression activity depends on its RNA binding ability. Furthermore, we find that the anti-silencing activity of IBDV VP3 is shared with the homologous proteins expressed by both insect- and fish-infecting birnaviruses. Finally, we show that IBDV VP3 can functionally replace the well-characterized HCPro silencing suppressor of Plum pox virus, a potyvirus that is unable to infect plants in the absence of an active silencing suppressor. Altogether, our results support the idea that VP3 protects the viral genome from host sentinels, including those of the RNA silencing machinery.

  18. Reverse electrodialysis : evaluation of suitable electrode systems

    NARCIS (Netherlands)

    Veerman, J.; Saakes, M.; Metz, S. J.; Harmsen, G. J.

    Reverse electrodialysis (RED) is a method for directly extracting electrical energy from salinity gradients, especially from sea and river water. For the commercial implementation of RED, the electrode system is a key component. In this paper, novel electrode systems for RED were compared with

  19. Nicotinamide clearance by Pnc1 directly regulates Sir2-mediated silencing and longevity. (United States)

    Gallo, Christopher M; Smith, Daniel L; Smith, Jeffrey S


    The Saccharomyces cerevisiae Sir2 protein is an NAD(+)-dependent histone deacetylase (HDAC) that functions in transcriptional silencing and longevity. The NAD(+) salvage pathway protein, Npt1, regulates Sir2-mediated processes by maintaining a sufficiently high intracellular NAD(+) concentration. However, another NAD(+) salvage pathway component, Pnc1, modulates silencing independently of the NAD(+) concentration. Nicotinamide (NAM) is a by-product of the Sir2 deacetylase reaction and is a natural Sir2 inhibitor. Pnc1 is a nicotinamidase that converts NAM to nicotinic acid. Here we show that recombinant Pnc1 stimulates Sir2 HDAC activity in vitro by preventing the accumulation of NAM produced by Sir2. In vivo, telomeric, rDNA, and HM silencing are differentially sensitive to inhibition by NAM. Furthermore, PNC1 overexpression suppresses the inhibitory effect of exogenously added NAM on silencing, life span, and Hst1-mediated transcriptional repression. Finally, we show that stress suppresses the inhibitory effect of NAM through the induction of PNC1 expression. Pnc1, therefore, positively regulates Sir2-mediated silencing and longevity by preventing the accumulation of intracellular NAM during times of stress.

  20. [Silence pact from the perspective of caretakers of palliative care patients]. (United States)

    Cejudo López, Ángela; López López, Begoña; Duarte Rodríguez, Miguel; Crespo Serván, María Pilar; Coronado Illescas, Concepción; de la Fuente Rodríguez, Carlota


    To determine why terminally-ill patients, family caregivers and health care providers make a pact of silence about the terminal status of the patient, and to identify the attitudes, experiences and opinions of family caregivers concerning the conspiracy of silence in palliative care. A qualitative phenomenological study based on an interpretive analysis, conducted in Primary Health Care, Seville, Spain. Study dimensions: knowledge of the diagnosis and prognosis of the condition; disclosure of information; reaction to information received, feelings and approach to death; information disclosed to the patient and reasons behind partial disclosure; communication between patients, families, and health care providers. Family caregivers of patients on palliative care suspicious about a pact of silence. Family caregivers hamper professional-patient communication; use of deceit to conceal the truth; suspicion that the patient knows the truth; the clinician conceals the truth; paternalist attitudes; feelings of sadness, grief, resignation, acceptance of the disease. The pact of silence has negative effects on coping with death, quality of life in the last days of life, and mourning. Communication between patients, health care providers, and families should be improved to prevent the pact of silence, and help patients cope with death. Copyright © 2015 Elsevier España, S.L.U. All rights reserved.