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Sample records for plasmids results obtained

  1. Experimental results obtained at GANIL

    International Nuclear Information System (INIS)

    Borrel, V.

    1993-01-01

    A review of experimental results obtained at GANIL on the study of nuclear structure and nuclear reactions with secondary radioactive beams is presented. Mass measurements by means of the GANIL cyclotrons are described. The possibilities of GANIL/LISE3 for the production and separation of radioactive beams are illustrated through a large variety of experiments. (author). 19 refs., 8 figs

  2. Overview of galactic results obtained by MAGIC

    Energy Technology Data Exchange (ETDEWEB)

    Zanin, Roberta

    2013-06-15

    MAGIC is a system of two atmospheric Cherenkov telescopes which explores the very-high-energy sky, from some tens of GeV up to tens of TeV. Located in the Canary island of La Palma, MAGIC has the lowest energy threshold among the instruments of its kind, well suited to study the still poorly explored energy band below 100 GeV. Although the space-borne gamma-ray telescope Fermi/LAT is sensitive up to 300 GeV, gamma-ray rates drop fast with increasing energy, so γ-ray collection areas larger than 10{sup 4}m{sup 2}, as those provided by grounds-based instruments, are crucial above a few GeV. The combination of MAGIC and Fermi/LAT observations have provided the first astrophysical spectra sampled in the inverse Compton peak region, resulting in a complete coverage from MeV up to TeV energies, as well as the discovery of a pulsed emission in the very-high-energy band. This paper focuses on the latest results on Galactic sources obtained by MAGIC which are highlighted by the detection of the pulsed gamma-ray emission from the Crab pulsar up to 400 GeV. In addition, we will present the morphological study on the W51 complex which allowed to pinpoint the location of the majority of the emission around the interaction point between the supernova remnant W51C and the star forming region W51B, but also to find a possible contribution from the associated pulsar wind nebula. Other important scientific achievements involve the Crab Nebula with an unprecedented spectrum covering three decades in energy starting from 50 GeV and a morphological study of the unidentified source HESS J1857+026 which supports the pulsar wind nebula scenario. Finally we will report on the searches of very-high-energy signals from gamma-ray binaries, mainly LS I 303+ and HESS J0632+057.

  3. Results obtained on dileptons and prospects

    International Nuclear Information System (INIS)

    Roche, G.

    1992-01-01

    Experimental results (mostly from the DLS collaboration at LBL) and model calculations of dielectron production in pp/pd/pBe (1-5 GeV) and CaCa (1-2 GeV/A) are presented. Dileptons appear to be a suitable probe to approach in-medium properties of hadrons in nuclear matter. At the present time, the ρ/ω region looks the most promising. Oncoming programs are briefly presented, in particular the HADES project at GSI. 7 figs

  4. pTC Plasmids from Sulfolobus Species in the Geothermal Area of Tengchong, China: Genomic Conservation and Naturally-Occurring Variations as a Result of Transposition by Mobile Genetic Elements.

    Science.gov (United States)

    Xiang, Xiaoyu; Huang, Xiaoxing; Wang, Haina; Huang, Li

    2015-02-12

    Plasmids occur frequently in Archaea. A novel plasmid (denoted pTC1) containing typical conjugation functions has been isolated from Sulfolobus tengchongensis RT8-4, a strain obtained from a hot spring in Tengchong, China, and characterized. The plasmid is a circular double-stranded DNA molecule of 20,417 bp. Among a total of 26 predicted pTC1 ORFs, 23 have homologues in other known Sulfolobus conjugative plasmids (CPs). pTC1 resembles other Sulfolobus CPs in genome architecture, and is most highly conserved in the genomic region encoding conjugation functions. However, attempts to demonstrate experimentally the capacity of the plasmid for conjugational transfer were unsuccessful. A survey revealed that pTC1 and its closely related plasmid variants were widespread in the geothermal area of Tengchong. Variations of the plasmids at the target sites for transposition by an insertion sequence (IS) and a miniature inverted-repeat transposable element (MITE) were readily detected. The IS was efficiently inserted into the pTC1 genome, and the inserted sequence was inactivated and degraded more frequently in an imprecise manner than in a precise manner. These results suggest that the host organism has evolved a strategy to maintain a balance between the insertion and elimination of mobile genetic elements to permit genomic plasticity while inhibiting their fast spreading.

  5. pTC Plasmids from Sulfolobus Species in the Geothermal Area of Tengchong, China: Genomic Conservation and Naturally-Occurring Variations as a Result of Transposition by Mobile Genetic Elements

    Directory of Open Access Journals (Sweden)

    Xiaoyu Xiang

    2015-02-01

    Full Text Available Plasmids occur frequently in Archaea. A novel plasmid (denoted pTC1 containing typical conjugation functions has been isolated from Sulfolobus tengchongensis RT8-4, a strain obtained from a hot spring in Tengchong, China, and characterized. The plasmid is a circular double-stranded DNA molecule of 20,417 bp. Among a total of 26 predicted pTC1 ORFs, 23 have homologues in other known Sulfolobus conjugative plasmids (CPs. pTC1 resembles other Sulfolobus CPs in genome architecture, and is most highly conserved in the genomic region encoding conjugation functions. However, attempts to demonstrate experimentally the capacity of the plasmid for conjugational transfer were unsuccessful. A survey revealed that pTC1 and its closely related plasmid variants were widespread in the geothermal area of Tengchong. Variations of the plasmids at the target sites for transposition by an insertion sequence (IS and a miniature inverted-repeat transposable element (MITE were readily detected. The IS was efficiently inserted into the pTC1 genome, and the inserted sequence was inactivated and degraded more frequently in an imprecise manner than in a precise manner. These results suggest that the host organism has evolved a strategy to maintain a balance between the insertion and elimination of mobile genetic elements to permit genomic plasticity while inhibiting their fast spreading.

  6. Role of integrons, plasmids and SXT elements in multidrug resistance of Vibrio cholerae and Providencia vermicola obtained from a clinical isolate of diarrhea

    Directory of Open Access Journals (Sweden)

    Neha eRajpara

    2015-02-01

    Full Text Available The isolates of Vibrio cholerae and Providencia vermicola obtained from a diarrhoeal patient were investigated for genetic elements governing their drug resistance phenotypes. Out of fourteen antibiotics tested, V. cholerae Vc IDH02365 isolate showed resistance to nine antibiotics, while P. vermicola Pv NBA2365 was found to be resistant to all the antibiotics except polymyxin B. Though SXT integrase was depicted in both the bacteria, class 1 integron was found to be associated only with Pv NBA2365. Integrons in Pv NBA2365 conferred resistance to β-lactams, aminoglycosides and trimethoprim. Pv NBA2365 carried two transformable plasmids imparting distinct antibiotic resistance traits to their Escherichia coli transformants. In rabbit ileal loop assays, Pv NBA2365 did not show any fluid accumulation in contrast with Vc IDH02365 that showed high fluid accumulation. To the best of our knowledge, this is the first report of a highly drug resistant P.vermicola and additionally co-existence of multidrug resistant V. cholerae and P. vermicola. Both the microbes appeared to possess a wide array of mobile genetic elements for a large spectrum of antimicrobial agents, some of which are being used in the treatment of acute diarrhoea.

  7. A toxicological study of inhalable particulates in an industrial region of Lanzhou City, northwestern China: Results from plasmid scission assay

    Science.gov (United States)

    Xiao, Zhenghui; Shao, Longyi; Zhang, Ning; Wang, Jing; Chuang, Hsiao-Chi; Deng, Zhenzhen; Wang, Zhen; BéruBé, Kelly

    2014-09-01

    The city of Lanzhou in northwestern China experiences serious air pollution episodes in the form of PM10 that is characterized by having high levels of heavy metals. The Xigu District represents the industrial core area of Lanzhou City and is denoted by having the largest petrochemical bases in western China. This study investigates heavy metal compositions and oxidative potential of airborne PM10 (particulate matter with aerodynamic diameter of 10 μm or less) collected in Xigu District in the summer and winter of 2010. An in vitro plasmid scission assay (PSA) was employed to study the oxidative potential of airborne PM10 and inductively coupled plasma-mass spectrometry (ICP-MS) was used to examine heavy metal compositions. Transmission electron microscopy coupled with energy-dispersive X-ray spectrometry (TEM/EDX) was used to investigate elemental compositions and mixing states of PM10. The average mass concentrations of PM10 collected in Xigu District were generally higher than the national standard for daily PM10 (150 μg/m3). Cr, Zn, Pb and Mn were the most abundant metals in the intact whole particles of PM10. Zn, Mn and As was the most abundant metal in the water-soluble fraction, while Cr, Pb, and V existed primarily in insoluble forms. TD20 values (i.e. toxic dosage of PM10 causing 20% of plasmid DNA damage) varied considerably in both winter and summer (from 19 μg/mL to >1000 μg/mL) but were typically higher in summer, suggesting that the winter PM10 exhibited greater bioreactivity. In addition, the PM10 collected during a dust storm episode had a highest TD20 value and thus the least oxidative damage to supercoiled plasmid DNA, while the particles collected on a hazy day had a lowest TD20 value and thus the highest oxidative damage to supercoiled plasmid DNA. The particles collected on the first day after snow fall and on a day of cold air intrusion exhibited minor oxidative potential (i.e. caused limited DNA damage). The water-soluble Zn, Mn, As, and

  8. Test Beam Results Obtained with the Q4 Prototype

    Energy Technology Data Exchange (ETDEWEB)

    Aguilar-Benitez, M.; Alberdi, J.; Cerrada, M.; Colino, N.; Daniel, M.; Fouz, M. c.; Marin, J.; Mocholi, J.; Oller, J. C.; Puerta, J.; Romero, L.; Salicio, J. M.

    2000-07-01

    A prototype of the CMS Barrel Muon Detector incorporating all the features of the final chambers was built at CIEMAT using the mass production assembly procedures and tools. The performance of this prototype was studied in a muon test beam at CERN and the results obtained are presented here. (Author)

  9. Test beam results obtained with the Q4 prototype

    International Nuclear Information System (INIS)

    Aguilar-Benitez, M.; Alberdi, J.; Cerrada, M.; Colino, N.; Daniel, M.; Fouz, M.C.; Marin, J.; Mocholi, J.; Oller, J. C.; Puerta, J.; Romero, L.; Salicio, J. M.; Willmott, C.

    2000-10-01

    A prototype of the CMS Barrel Muon Detector incorporating all the features of the final chambers was built at CIEMAT using the mass production assembly procedures and tools. The performance of this prototype was studied in a muon test beam at CERN and the results obtained are presented here. (Author)

  10. DIFFERENCES IN RESULTS OBTAINED BY STUDENTS OF DIFFERENT FACULTIES

    Directory of Open Access Journals (Sweden)

    OTAVOVÁ, Miroslava

    2016-03-01

    Full Text Available The paper presents and discusses the results of statistical analysis of differences among scores obtained by students of different faculties of the University of Economics in Prague. The analysed dataset contains the scores for 2256 students that took basic mathematics course during the academic year 2013/2014. A two way analysis of variance was performed with semester and faculty as main factors. The interaction between these two factors was also considered. Students have to take two tests. At first, the sum of the scores obtained from both tests is analysed and then, the two tests are analysed separately. It turns out that the significance of factors is the same in the three analyses. The assumptions of linear models are verified. Due to problem of heteroscedasticity, weighted least squares are used and the possibility of using Box-Cox transformation is also discussed, as the errors are not normally distributed. Finally, the differences between the faculties are described.

  11. Application of methylation in improving plasmid transformation into Helicobacter pylori.

    Science.gov (United States)

    Zhao, Huilin; Xu, Linlin; Rong, Qianyu; Xu, Zheng; Ding, Yunfei; Zhang, Ying; Wu, Yulong; Li, Boqing; Ji, Xiaofei

    2018-05-23

    Helicobacter pylori is an important gastrointestinal pathogen. Its strains possess different levels of powerful restriction modification systems, which are significant barriers to genetic tools used for studying the role of functional genes in its pathogenesis. Methylating vectors in vitro was reported as an alternative to overcome this barrier in several bacteria. In this study we used two H. pylori-E. coli shuttle plasmids and several single/double-crossover homologous recombination gene-targeting plasmids, to test the role of methylation in H. pylori transformation. According to our results, transformants could be obtained only after shuttle plasmids were methylated before transformation. It is helpful in gene complementation and over-expression although at a low frequency. The frequency of gene-targeting transformation was also increased after methylation, especially for the single-crossover recombination plasmids, the transformants of which could only be obtained after methylation. For the double-crossover recombination targeting plasmids, the initial yield of transformants was 0.3-0.8 × 10 2 CFUs per microgram plasmid DNA. With the help of methylation, the yield was increased to 0.4-1.3 × 10 2 CFUs per microgram plasmid DNA. These results suggest that in vitro methylation can improve H. pylori transformation by different plasmids, which will benefit the pathogenic mechanism research. Copyright © 2018. Published by Elsevier B.V.

  12. Auto-assembly of nanometer thick, water soluble layers of plasmid DNA complexed with diamines and basic amino acids on graphite: Greatest DNA protection is obtained with arginine

    Energy Technology Data Exchange (ETDEWEB)

    Khalil, T.T.; Boulanouar, O. [Université de Bourgogne Franche-Comté, UMR CNRS 6249 Chrono-Environnement, 16, Route de Gray, 25030 Besançon Cedex (France); Heintz, O. [Université de Bourgogne Franche-Comté, UMR CNRS 6303Laboratoire Interdisciplinaire Carnot de Bourgogne, DTAI/Centre de micro/nano caractérisation, 9 Av. A. Savary, BP 47870, F-21078 DIJON Cedex (France); Fromm, M., E-mail: michel.fromm@univ-fcomte.fr [Université de Bourgogne Franche-Comté, UMR CNRS 6249 Chrono-Environnement, 16, Route de Gray, 25030 Besançon Cedex (France)

    2017-02-01

    We have investigated the ability of diamines as well as basic amino acids to condense DNA onto highly ordered pyrolytic graphite with minimum damage after re-dissolution in water. Based on a bibliographic survey we briefly summarize DNA binding properties with diamines as compared to basic amino acids. Thus, solutions of DNA complexed with these linkers were drop-cast in order to deposit ultra-thin layers on the surface of HOPG in the absence or presence of Tris buffer. Atomic Force Microscopy analyses showed that, at a fixed ligand-DNA mixing ratio of 16, the mean thickness of the layers can be statistically predicted to lie in the range 0–50 nm with a maximum standard deviation ± 6 nm, using a simple linear law depending on the DNA concentration. The morphology of the layers appears to be ligand-dependent. While the layers containing diamines present holes, those formed in the presence of basic amino acids, except for lysine, are much more compact and dense. X-ray Photoelectron Spectroscopy measurements provide compositional information indicating that, compared to the maximum number of DNA sites to which the ligands may bind, the basic amino acids Arg and His are present in large excess. Conservation of the supercoiled topology of the DNA plasmids was studied after recovery of the complex layers in water. Remarkably, arginine has the best protection capabilities whether Tris was present or not in the initial solution. - Highlights: • Characterization of nanometer scaled layers composed of pUC21 plasmid DNA • Relation between nature of the ligand and structure of the layers • Capacities of the ligands to protect plasmids from strand break depending on their nature.

  13. Lambing results obtained with imported Ile de France rarn semen

    African Journals Online (AJOL)

    insemination was carried out either 54,5 h (fixed time single ... Table 1 Conception rate obtained using two batches of frozen lle de France ... Pregnant mare serum gonadotrophin (Fostim, 300 ... Effect of initial freezing temperature, addition.

  14. Differences in the stability of the plasmids of Yersinia pestis cultures in vitro: impact on virulence

    Directory of Open Access Journals (Sweden)

    TC Leal-Balbino

    2004-11-01

    Full Text Available Plasmid and chromosomal genes encode determinants of virulence for Yersinia pestis, the causative agent of plague. However, in vitro, Y. pestis genome is very plastic and several changes have been described. To evaluate the alterations in the plasmid content of the cultures in vitro and the impact of the alterations to their pathogenicity, three Y. pestis isolates were submitted to serial subculture, analysis of the plasmid content, and testing for the presence of characteristic genes in each plasmid of colonies selected after subculture. Different results were obtained with each strain. The plasmid content of one of them was shown to be stable; no apparent alteration was produced through 32 subcultures. In the other two strains, several alterations were observed. LD50 in mice of the parental strains and the derived cultures with different plasmid content were compared. No changes in the virulence plasmid content could be specifically correlated with changes in the LD50.

  15. Preliminary results obtained from a dense plasma focus

    International Nuclear Information System (INIS)

    Sinman, S.; Sinman, A.

    1982-01-01

    In this study, for the data processing in our Dense Plasma Focus (DPF) system, a multiparameters numerical hierarchy obtained from the fundamental equations based on the snowplows model and its conclusions have been developed. Evaluating the data along the transients of the total plasma current, the anode current and the focus voltage recorded by an oscilloscope, they have been possible to determine the time domain plasma parameters such as the shock thickness, the sheath temperature, velocity and electron density, the pressure at the focus phase and the plasma temperature. Besides, the dissipated energy through the focus notch or in other words, the plasma temperature has also been calculated by means of the numerical integration. In the text, a performance chart together with some other correlation curves for the optimization of the DPF systems are submitted and discussed. (author)

  16. In Vivo Transmission of an IncA/C Plasmid in Escherichia coli Depends on Tetracycline Concentration, and Acquisition of the Plasmid Results in a Variable Cost of Fitness.

    Science.gov (United States)

    Johnson, Timothy J; Singer, Randall S; Isaacson, Richard E; Danzeisen, Jessica L; Lang, Kevin; Kobluk, Kristi; Rivet, Bernadette; Borewicz, Klaudyna; Frye, Jonathan G; Englen, Mark; Anderson, Janet; Davies, Peter R

    2015-05-15

    IncA/C plasmids are broad-host-range plasmids enabling multidrug resistance that have emerged worldwide among bacterial pathogens of humans and animals. Although antibiotic usage is suspected to be a driving force in the emergence of such strains, few studies have examined the impact of different types of antibiotic administration on the selection of plasmid-containing multidrug resistant isolates. In this study, chlortetracycline treatment at different concentrations in pig feed was examined for its impact on selection and dissemination of an IncA/C plasmid introduced orally via a commensal Escherichia coli host. Continuous low-dose administration of chlortetracycline at 50 g per ton had no observable impact on the proportions of IncA/C plasmid-containing E. coli from pig feces over the course of 35 days. In contrast, high-dose administration of chlortetracycline at 350 g per ton significantly increased IncA/C plasmid-containing E. coli in pig feces (P IncA/C plasmid to other indigenous E. coli hosts. There was no evidence of conjugal transfer of the IncA/C plasmid to bacterial species other than E. coli. In vitro competition assays demonstrated that bacterial host background substantially impacted the cost of IncA/C plasmid carriage in E. coli and Salmonella. In vitro transfer and selection experiments demonstrated that tetracycline at 32 μg/ml was necessary to enhance IncA/C plasmid conjugative transfer, while subinhibitory concentrations of tetracycline in vitro strongly selected for IncA/C plasmid-containing E. coli. Together, these experiments improve our knowledge on the impact of differing concentrations of tetracycline on the selection of IncA/C-type plasmids. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  17. Results Obtained in Training Romanian Sport Horses for Jumping Competitions

    Directory of Open Access Journals (Sweden)

    Flavia Luminiţa Bochiş

    2012-10-01

    Full Text Available A number of 29 equines, Romanian Sport Horse stallions, were taken as green horses, break into the saddle, andteach into basic training while later train as jumpers over three years. The obstacle courses results had been obtainedin our country, in Hungary and in Serbia. There were courses jumped in the national championship, somedemonstrations organized in Romania or in the two foreign countries, or periodically test pattern courses run on theown arena. The best results, respectively more than 50% obstacle courses with no fault points over 3 years, were runby five stallions, and the same percent for 2 seasons, was reached by nine stallions.

  18. Paternity tests in Mexico: Results obtained in 3005 cases.

    Science.gov (United States)

    García-Aceves, M E; Romero Rentería, O; Díaz-Navarro, X X; Rangel-Villalobos, H

    2018-04-01

    National and international reports regarding the paternity testing activity scarcely include information from Mexico and other Latin American countries. Therefore, we report different results from the analysis of 3005 paternity cases analyzed during a period of five years in a Mexican paternity testing laboratory. Motherless tests were the most frequent (77.27%), followed by trio cases (20.70%); the remaining 2.04% included different cases of kinship reconstruction. The paternity exclusion rate was 29.58%, higher but into the range reported by the American Association of Blood Banks (average 24.12%). We detected 65 mutations, most of them involving one-step (93.8% and the remaining were two-step mutations (6.2%) thus, we were able to estimate the paternal mutation rate for 17 different STR loci: 0.0018 (95% CI 0.0005-0.0047). Five triallelic patterns and 12 suspected null alleles were detected during this period; however, re-amplification of these samples with a different Human Identification (HID) kit confirmed the homozygous genotypes, which suggests that most of these exclusions actually are one-step mutations. HID kits with ≥20 STRs detected more exclusions, diminishing the rate of inconclusive results with isolated exclusions (Powerplex 21 kit (20 STRs) and Powerplex Fusion kit (22 STRs) offered similar PI (p = 0.379) and average number of exclusions (PE) (p = 0.339) when a daughter was involved in motherless tests. In brief, besides to report forensic parameters from paternity tests in Mexico, results describe improvements to solve motherless paternity tests using HID kits with ≥20 STRs instead of one including 15 STRs. Copyright © 2018 Elsevier Ltd and Faculty of Forensic and Legal Medicine. All rights reserved.

  19. Results obtained by Trillo Nuclear Power Plant in 2001

    International Nuclear Information System (INIS)

    Solar, V.

    2002-01-01

    Unification of the Almaraz and Trillo plant managements was given a strong boost with unification of the central offices and management computer platforms (economic and technical) and with signature of an agreement with Madrid's trade unions providing for appropriate management of surpluses and the transfer of knowledge between people leaving the company and those taking their place. The operating results have been maintained at a high level although they were lower than last year, which was exceptional. The plant was on line for 7,979 hours, most of them at full load. (Author)

  20. Measurement results obtained from air quality monitoring system

    Energy Technology Data Exchange (ETDEWEB)

    Turzanski, P.K.; Beres, R. [Provincial Inspection of Environmental Protection, Cracow (Poland)

    1995-12-31

    An automatic system of air pollution monitoring operates in Cracow since 1991. The organization, assembling and start-up of the network is a result of joint efforts of the US Environmental Protection Agency and the Cracow environmental protection service. At present the automatic monitoring network is operated by the Provincial Inspection of Environmental Protection. There are in total seven stationary stations situated in Cracow to measure air pollution. These stations are supported continuously by one semi-mobile (transportable) station. It allows to modify periodically the area under investigation and therefore the 3-dimensional picture of creation and distribution of air pollutants within Cracow area could be more intelligible.

  1. The similia principle: results obtained in a cellular model system.

    Science.gov (United States)

    Wiegant, Fred; Van Wijk, Roeland

    2010-01-01

    This paper describes the results of a research program focused on the beneficial effect of low dose stress conditions that were applied according to the similia principle to cells previously disturbed by more severe stress conditions. In first instance, we discuss criteria for research on the similia principle at the cellular level. Then, the homologous ('isopathic') approach is reviewed, in which the initial (high dose) stress used to disturb cellular physiology and the subsequent (low dose) stress are identical. Beneficial effects of low dose stress are described in terms of increased cellular survival capacity and at the molecular level as an increase in the synthesis of heat shock proteins (hsps). Both phenomena reflect a stimulation of the endogenous cellular self-recovery capacity. Low dose stress conditions applied in a homologous approach stimulate the synthesis of hsps and enhance survival in comparison with stressed cells that were incubated in the absence of low dose stress conditions. Thirdly, the specificity of the low dose stress condition is described where the initial (high dose) stress is different in nature from the subsequently applied (low dose) stress; the heterologous or 'heteropathic' approach. The results support the similia principle at the cellular level and add to understanding of how low dose stress conditions influence the regulatory processes underlying self-recovery. In addition, the phenomenon of 'symptom aggravation' which is also observed at the cellular level, is discussed in the context of self-recovery. Finally, the difference in efficiency between the homologous and the heterologous approach is discussed; a perspective is indicated for further research; and the relationship between studies on the similia principle and the recently introduced concept of 'postconditioning hormesis' is emphasized. Copyright 2009 The Faculty of Homeopathy. Published by Elsevier Ltd. All rights reserved.

  2. Results obtained using the pellet charge exchange diagnostic on TFTR

    International Nuclear Information System (INIS)

    McChesney, J.M.; Fisher, R.K.; Parks, P.B.; Duong, H.H.; Mansfield, D.K.; Medley, S.S.; Roquemore, A.L.; Petrov, M.P.

    1994-05-01

    Experiments are underway on TFTR to measure the confined alpha particle distribution functions using small low-Z pellets injected into the plasma. Upon entering the plasma, the pellet ablates, forming a plasma ablation cloud, elongated in the magnetic field direction, that travels alongside the pellet. A small fraction of the fusion produced 3.5 MeV alpha particles incident on the cloud are converted to helium neutrals. By measuring the resultant helium neutrals escaping from the plasma by means of a mass and energy resolving charge exchange analyzer, the energy distribution of the alpha particles incident on the cloud can be inferred. Preliminary experiments to observe neutrals from the 100-1000 keV He tail produced during ICRF minority heating experiments were successful. However, no significant alpha particle signals have been observed during D-T operation on TFTR. The authors attribute this lack of signal to stochastic toroidal field ripple loss in the outer regions of the plasma. They are studying ways to improve the pellet penetration so that the pellet penetrates into the central regions of the plasma where ripple induced losses are small and the alpha population is high

  3. Antimicrobial susceptibility pattern and plasmid-mediated ...

    African Journals Online (AJOL)

    negative Staphylococci (CoNS) were isolated from clinical samples and isolates subjected to antibiotic susceptibility testing, plasmid curing and plasmid DNA isolation. Result: The highest percentages isolates were recovered from urine samples and ...

  4. Comparative genomics of the IncA/C multidrug resistance plasmid family.

    Science.gov (United States)

    Fricke, W Florian; Welch, Timothy J; McDermott, Patrick F; Mammel, Mark K; LeClerc, J Eugene; White, David G; Cebula, Thomas A; Ravel, Jacques

    2009-08-01

    Multidrug resistance (MDR) plasmids belonging to the IncA/C plasmid family are widely distributed among Salmonella and other enterobacterial isolates from agricultural sources and have, at least once, also been identified in a drug-resistant Yersinia pestis isolate (IP275) from Madagascar. Here, we present the complete plasmid sequences of the IncA/C reference plasmid pRA1 (143,963 bp), isolated in 1971 from the fish pathogen Aeromonas hydrophila, and of the cryptic IncA/C plasmid pRAx (49,763 bp), isolated from Escherichia coli transconjugant D7-3, which was obtained through pRA1 transfer in 1980. Using comparative sequence analysis of pRA1 and pRAx with recent members of the IncA/C plasmid family, we show that both plasmids provide novel insights into the evolution of the IncA/C MDR plasmid family and the minimal machinery necessary for stable IncA/C plasmid maintenance. Our results indicate that recent members of the IncA/C plasmid family evolved from a common ancestor, similar in composition to pRA1, through stepwise integration of horizontally acquired resistance gene arrays into a conserved plasmid backbone. Phylogenetic comparisons predict type IV secretion-like conjugative transfer operons encoded on the shared plasmid backbones to be closely related to a group of integrating conjugative elements, which use conjugative transfer for horizontal propagation but stably integrate into the host chromosome during vegetative growth. A hipAB toxin-antitoxin gene cluster found on pRA1, which in Escherichia coli is involved in the formation of persister cell subpopulations, suggests persistence as an early broad-spectrum antimicrobial resistance mechanism in the evolution of IncA/C resistance plasmids.

  5. Photoinduced silver nanoparticles/nanorings on plasmid DNA scaffolds.

    Science.gov (United States)

    Liu, Jianhua; Zhang, Xiaoliang; Yu, Mei; Li, Songmei; Zhang, Jindan

    2012-01-23

    Biological scaffolds are being actively explored for the synthesis of nanomaterials with novel structures and unexpected properties. Toroidal plasmid DNA separated from the Bacillus host is applied as a sacrificial mold for the synthesis of silver nanoparticles and nanorings. The photoirradiation method is applied to reduce Ag(I) on the plasmid. The nanoparticles are obtained by varying the concentration of the Ag(I) ion solution and the exposure time of the plasmid-Ag(I) complex under UV light at 254 nm and room temperature. It is found that the plasmid serves not only as a template but also as a reductant to drive the silver nucleation and deposition. The resulting nanoparticles have a face-centered cubic (fcc) crystal structure and 20-30 nm average diameter. The detailed mechanism is discussed, and other metals or alloys could also be synthesized with this method. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. Degenerate primer MOB typing of multiresistant clinical isolates of E. coli uncovers new plasmid backbones.

    Science.gov (United States)

    Garcillán-Barcia, M Pilar; Ruiz del Castillo, Belén; Alvarado, Andrés; de la Cruz, Fernando; Martínez-Martínez, Luis

    2015-01-01

    Degenerate Primer MOB Typing is a PCR-based protocol for the classification of γ-proteobacterial transmissible plasmids in five phylogenetic relaxase MOB families. It was applied to a multiresistant E. coli collection, previously characterized by PCR-based replicon-typing, in order to compare both methods. Plasmids from 32 clinical isolates of multiresistant E. coli (19 extended spectrum beta-lactamase producers and 13 non producers) and their transconjugants were analyzed. A total of 95 relaxases were detected, at least one per isolate, underscoring the high potential of these strains for antibiotic-resistance transmission. MOBP12 and MOBF12 plasmids were the most abundant. Most MOB subfamilies detected were present in both subsets of the collection, indicating a shared mobilome among multiresistant E. coli. The plasmid profile obtained by both methods was compared, which provided useful data upon which decisions related to the implementation of detection methods in the clinic could be based. The phylogenetic depth at which replicon and MOB-typing classify plasmids is different. While replicon-typing aims at plasmid replication regions with non-degenerate primers, MOB-typing classifies plasmids into relaxase subfamilies using degenerate primers. As a result, MOB-typing provides a deeper phylogenetic depth than replicon-typing and new plasmid groups are uncovered. Significantly, MOB typing identified 17 plasmids and an integrative and conjugative element, which were not detected by replicon-typing. Four of these backbones were different from previously reported elements. Copyright © 2014 Elsevier Inc. All rights reserved.

  7. Behavior of IncQ Plasmids in Agrobacterium tumefaciens

    NARCIS (Netherlands)

    Hille, Jacques; Schilperoort, Rob

    1981-01-01

    Inc-Q plasmids were introduced into Agrobacterium tumefuciens, by mobilization from Escherichia coli with an Inc-P plasmid, or by transformation with purified plasmid DNA. It was found that they were stably maintained. The presence of an Inc-Q plasmid did not influence tumorigenicity. These results

  8. Characterization of epidemic IncI1-Iγ plasmids harboring ambler class A and C genes in Escherichia coli and Salmonella enterica from animals and humans.

    Science.gov (United States)

    Smith, Hilde; Bossers, Alex; Harders, Frank; Wu, Guanghui; Woodford, Neil; Schwarz, Stefan; Guerra, Beatriz; Rodríguez, Irene; van Essen-Zandbergen, Alieda; Brouwer, Michael; Mevius, Dik

    2015-09-01

    The aim of the study was to identify the plasmid-encoded factors contributing to the emergence and spread of epidemic IncI1-Iγ plasmids obtained from Escherichia coli and Salmonella enterica isolates from animal and human reservoirs. For this, 251 IncI1-Iγ plasmids carrying various extended-spectrum β-lactamase (ESBL) or AmpC β-lactamase genes were compared using plasmid multilocus sequence typing (pMLST). Thirty-two of these plasmids belonging to different pMLST types were sequenced using Roche 454 and Illumina platforms. Epidemic IncI1-Iγ plasmids could be assigned to various dominant clades, whereas rarely detected plasmids clustered together as a distinct clade. Similar phylogenetic trees were obtained using only the plasmid backbone sequences, showing that the differences observed between the plasmids belonging to distinct clades resulted mainly from differences between their backbone sequences. Plasmids belonging to the various clades differed particularly in the presence/absence of genes encoding partitioning and addiction systems, which contribute to stable inheritance during cell division and plasmid maintenance. Despite this, plasmids belonging to the various phylogenetic clades also showed marked resistance gene associations, indicating the circulation of successful plasmid-gene combinations. The variation in traY and excA genes found in IncI1-Iγ plasmids is conserved within pMLST sequence types and plays a role in incompatibility, although functional study is needed to elucidate the role of these genes in plasmid epidemiology. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  9. [Replication of Streptomyces plasmids: the DNA nucleotide sequence of plasmid pSB 24.2].

    Science.gov (United States)

    Bolotin, A P; Sorokin, A V; Aleksandrov, N N; Danilenko, V N; Kozlov, Iu I

    1985-11-01

    The nucleotide sequence of DNA in plasmid pSB 24.2, a natural deletion derivative of plasmid pSB 24.1 isolated from S. cyanogenus was studied. The plasmid amounted by its size to 3706 nucleotide pairs. The G-C composition was equal to 73 per cent. The analysis of the DNA structure in plasmid pSB 24.2 revealed the protein-encoding sequence of DNA, the continuity of which was significant for replication of the plasmid containing more than 1300 nucleotide pairs. The analysis also revealed two A-T-rich areas of DNA, the G-C composition of which was less than 55 per cent and a DNA area with a branched pin structure. The results may be of value in investigation of plasmid replication in actinomycetes and experimental cloning of DNA with this plasmid as a vector.

  10. Horizontal gene transfer of a ColV plasmid has resulted in a dominant avian clonal type of Salmonella enterica serovar Kentucky.

    Directory of Open Access Journals (Sweden)

    Timothy J Johnson

    Full Text Available Salmonella enterica continues to be a significant cause of foodborne gastrointestinal illness in humans. A wide variety of Salmonella serovars have been isolated from production birds and from retail poultry meat. Recently, though, S. enterica subsp. enterica serovar Kentucky has emerged as one of the prominent Salmonella serovars isolated from broiler chickens. Recent work suggests that its emergence apparently coincides with its acquisition of a ColV virulence plasmid. In the present study, we examined 902 Salmonella isolates belonging to 59 different serovars for the presence of this plasmid. Of the serovars examined, the ColV plasmid was found only among isolates belonging to the serovars Kentucky (72.9%, Typhimurium (15.0% and Heidelberg (1.7%. We demonstrated that a single PFGE clonal type of S. Kentucky harbors this plasmid, and acquisition of this plasmid by S. Kentucky significantly increased its ability to colonize the chicken cecum and cause extraintestinal disease. Comparison of the completed sequences of three ColV plasmids from S. Kentucky isolated from different geographical locales, timepoints and sources revealed a nearly identical genetic structure with few single nucleotide changes or insertions/deletions. Overall, it appears that the ColV plasmid was recently acquired by a single clonal type S. Kentucky and confers to its host enhanced colonization and fitness capabilities. Thus, the potential for horizontal gene transfer of virulence and fitness factors to Salmonella from other enteric bacteria exists in poultry, representing a potential human health hazard.

  11. Plasmid segregation mechanisms

    DEFF Research Database (Denmark)

    Ebersbach, G.; Gerdes, Kenn

    2005-01-01

    Bacterial plasmids encode partitioning (par) loci that ensure ordered plasmid segregation prior to cell division. par loci come in two types: those that encode actin-like ATPases and those that encode deviant Walker-type ATPases. ParM, the actin-like ATPase of plasmid R1, forms dynamic filaments...... that segregate plasmids paired at mid-cell to daughter cells. Like microtubules, ParM filaments exhibit dynamic instability (i.e., catastrophic decay) whose regulation is an important component of the DNA segregation process. The Walker box ParA ATPases are related to MinD and form highly dynamic, oscillating...... filaments that are required for the subcellular movement and positioning of plasmids. The role of the observed ATPase oscillation is not yet understood. However, we propose a simple model that couples plasmid segregation to ParA oscillation. The model is consistent with the observed movement...

  12. Plasmid DNA Analysis of Pasteurella multocida Serotype B isolated from Haemorrhagic Septicaemia outbreaks in Malaysia

    Directory of Open Access Journals (Sweden)

    Jamal, H.

    2005-01-01

    Full Text Available A total of 150 purified isolates of Pasteurella multocida serotype B were used (Salmah, 2004 for plasmid DNA curing experiment to determine hyaluronidase activity, antibiotic resistance pattern (ARP and mice lethality test (LD50 for their role of pathogenicity. A plasmid curing experiment was carried out by using the intercalating agent; ethidium bromide and rifampicin, where it was found all the plasmids had been cured (plasmidless from Pasteurella multocida. All of these plasmidless isolates maintained their phenotypic characteristics. They showed the same antibiotic resistancepattern as before curing, produced hyaluronidase and possessed lethality activity in mice when injected intraperitoneally(i.p. Based on this observation, the antibiotic resistance, hyaluronidase activity and mice virulence could probably be chromosomal-mediated. Plasmids were detected 100% in all P. multocida isolates with identical profile of 2 plasmids size 3.0 and 5.5 kb. No large plasmids could be detected in all isolates. Since all the isolates appeared to have identicalplasmid profiles, they were subjected to restriction enzyme(RE analysis. From RE analysis results obtained, it can be concluded that the plasmid DNA in serotype B isolates are identical. Only 4 of 32 REs were found to cleave these plasmids with identical restriction fingerprints; BglII, HaeIII, RsaI and SspI. From RE analysis results, it can be concluded that the plasmid DNA isolates are identical. This plasmid might not played any role in pathogenicity of Pasteurella multocida serotype B, however this information is important for the construction of shuttle vectors in genetic studies of the pathogenicity of haemorrhagic septicaemia(HS.

  13. Immunization with a dicistronic plasmid expressing a truncated form of bovine herpesvirus-1 glycoprotein D and the amino-terminal subunit of glycoprotein B results in reduced gB-specific immune responses

    International Nuclear Information System (INIS)

    Manoj, Sharmila; Babiuk, Lorne A.; Drunen Littel van-Hurk, Sylvia van den

    2003-01-01

    As an approach to create a divalent DNA vaccine, a truncated secreted version of bovine herpesvirus-1 (BHV-1) glycoprotein D (tgD) and the amino-terminal subunit of glycoprotein B (gBb) were expressed from a dicistronic plasmid, designated pSLIAtgD-IRES-gBb. Intradermal immunization of mice with pSLIAtgD-IRES-gBb or a mixture of plasmids encoding tgD (pSLIAtgD) and gBb (pSLIAgBb) by needle injection or gene gun elicited strong tgD-specific immune responses. However, a significant reduction in gBb-specific immune responses was observed upon immunization of mice with pSLIAtgD-IRES-gBb or a mixture of pSLIAtgD and pSLIAgBb in comparison to immunization with pSLIAgBb alone. This reduction in gBb-specific immune responses induced by pSLIAtgD-IRES-gBb was due to production of low amounts of gBb from pSLIAtgD-IRES-gBb, inefficient processing and transport of gBb, and possibly competition for antigen-presenting cells by tgD and gBb. These results indicate that, although divalent plasmids may be used to express different antigens, the efficacy of vaccination with such plasmids may be influenced by the plasmid design and the characteristics of the expressed antigens

  14. Construction of Biologically Functional Bacterial Plasmids In Vitro

    Science.gov (United States)

    Cohen, Stanley N.; Chang, Annie C. Y.; Boyer, Herbert W.; Helling, Robert B.

    1973-01-01

    The construction of new plasmid DNA species by in vitro joining of restriction endonuclease-generated fragments of separate plasmids is described. Newly constructed plasmids that are inserted into Escherichia coli by transformation are shown to be biologically functional replicons that possess genetic properties and nucleotide base sequences from both of the parent DNA molecules. Functional plasmids can be obtained by reassociation of endonuclease-generated fragments of larger replicons, as well as by joining of plasmid DNA molecules of entirely different origins. Images PMID:4594039

  15. Novel Plasmid Transformation Method Mediated by Chrysotile, Sliding Friction, and Elastic Body Exposure

    Directory of Open Access Journals (Sweden)

    Naoto Yoshida

    2007-01-01

    Full Text Available Escherichia coli as a plasmid recipient cell was dispersed in a chrysotile colloidal solution, containing chrysotile adsorbed to plasmid DNA (chrysotile-plasmid cell mixture. Following this, the chrysotile-plasmid cell mixture was dropped onto the surface of an elastic body, such as agarose, and treated physically by sliding a polystyrene streak bar over the elastic body to create friction. Plasmid DNA was easily incorporated into E. coli, and antibiotic resistance was conferred by transformation. The transformation efficiency of E. coli cultured in solid medium was greater than that of E. coli cultured in broth. To obtain greater transformation efficiency, we attempted to determine optimal transformation conditions. The following conditions resulted in the greatest transformation efficiency: the recipient cell concentration within the chrysotileplasmid cell mixture had an optical density greater than or equal to 2 at 550 nm, the vertical reaction force applied to the streak bar was greater than or equal to 40 g, and the rotation speed of the elastic body was greater than or equal to 34 rpm. Under these conditions, we observed a transformation efficiency of 107 per μg plasmid DNA. The advantage of achieving bacterial transformation using the elastic body exposure method is that competent cell preparation of the recipient cell is not required. In addition to E. coli, other Gram negative bacteria are able to acquire plasmid DNA using the elastic body exposure method.

  16. Comparative symbiotic plasmid analysis indicates that symbiosis gene ancestor type affects plasmid genetic evolution.

    Science.gov (United States)

    Wang, X; Zhao, L; Zhang, L; Wu, Y; Chou, M; Wei, G

    2018-07-01

    Rhizobial symbiotic plasmids play vital roles in mutualistic symbiosis with legume plants by executing the functions of nodulation and nitrogen fixation. To explore the gene composition and genetic constitution of rhizobial symbiotic plasmids, comparison analyses of 24 rhizobial symbiotic plasmids derived from four rhizobial genera was carried out. Results illustrated that rhizobial symbiotic plasmids had higher proportion of functional genes participating in amino acid transport and metabolism, replication; recombination and repair; carbohydrate transport and metabolism; energy production and conversion and transcription. Mesorhizobium amorphae CCNWGS0123 symbiotic plasmid - pM0123d had similar gene composition with pR899b and pSNGR234a. All symbiotic plasmids shared 13 orthologous genes, including five nod and eight nif/fix genes which participate in the rhizobia-legume symbiosis process. These plasmids contained nod genes from four ancestors and fix genes from six ancestors. The ancestral type of pM0123d nod genes was similar with that of Rhizobium etli plasmids, while the ancestral type of pM0123d fix genes was same as that of pM7653Rb. The phylogenetic trees constructed based on nodCIJ and fixABC displayed different topological structures mainly due to nodCIJ and fixABC ancestral type discordance. The study presents valuable insights into mosaic structures and the evolution of rhizobial symbiotic plasmids. This study compared 24 rhizobial symbiotic plasmids that included four genera and 11 species, illuminating the functional gene composition and symbiosis gene ancestor types of symbiotic plasmids from higher taxonomy. It provides valuable insights into mosaic structures and the evolution of symbiotic plasmids. © 2018 The Society for Applied Microbiology.

  17. Self-propelled prospection methodology: first results obtained in the northeast basin (Uruguay)

    International Nuclear Information System (INIS)

    Goso, H; Spoturno, J; Peciozzi, F.

    2008-01-01

    This report refers about uranium prospection methodology in Uruguay and its first results obtained in the northeast basin.The preliminary radiometric researching is carried out four works phases: material preparation, radiometric, statistics analysis, anomalies revision

  18. Conjugal properties of the Sinorhizobium meliloti plasmid mobilome.

    Science.gov (United States)

    Pistorio, Mariano; Giusti, María A; Del Papa, María F; Draghi, Walter O; Lozano, Mauricio J; Tejerizo, Gonzalo Torres; Lagares, Antonio

    2008-09-01

    The biology and biochemistry of plasmid transfer in soil bacteria is currently under active investigation because of its central role in prokaryote adaptation and evolution. In this work, we examined the conjugal properties of the cryptic plasmids present in a collection of the N(2)-fixing legume-symbiont Sinorhizobium meliloti. The study was performed on 65 S. meliloti isolates recovered from 25 humic soils of Argentina, which were grouped into 22 plasmid-profile types [i.e. plasmid operational taxonomic units (OTUs)]. The cumulative Shannon index calculated for the observed plasmid profiles showed a clear saturation plateau, thus indicating an adequate representation of the S. meliloti plasmid-profile types in the isolates studied. The results show that isolates of nearly 14% of the plasmid OTUs hosted transmissible plasmids and that isolates of 29% of the plasmid OTUs were able to retransfer the previously characterized mobilizable-cryptic plasmid pSmeLPU88b to a third recipient strain. It is noteworthy that isolates belonging to 14% of the plasmid OTUs proved to be refractory to the entrance of the model plasmid pSmeLPU88b, suggesting either the presence of surface exclusion phenomena or the occurrence of restriction incompatibility with the incoming replicon. Incompatibility for replication between resident plasmids and plasmid pSmeLPU88b was observed in c. 20% of the OTUs. The results reported here reveal a widespread compatibility among the conjugal functions of the cryptic plasmids in S. meliloti, and this fact, together with the observed high proportion of existing donor genotypes, points to the extrachromosomal compartment of the species as being an extremely active plasmid mobilome.

  19. CIEMAT interlaboratories comparison of the results obtained in the proficiency test run by IAEA

    International Nuclear Information System (INIS)

    Gasco, C.; Anton, M. P.; Alvarez, A.; Navarro, N.; Meral, J.; Gonzalez, A.; Higueras Lafaja, E.

    2000-01-01

    This report contains the results obtained by two different laboratories from CIEMAT after participating in the Proficiency Test organised by IAEA (International Atomic Energy Agency) in 1999. This test involves the analysis of fly ashes containing natural radionuclides and different amounts of added transuranics. The extraction techniques, counting methods and results obtained are detailed. This type of test are used for the labs to achieve their accreditation and check the reliability of the procedures routinely employed. (Author) 4 refs

  20. Analysis of the results obtained by optical measurements in plasma focus equipments

    International Nuclear Information System (INIS)

    Bilbao, L.; Bruzzone, H.

    1982-01-01

    The results obtained by shadow and Schlieren photographies and interferometry in a 1MJ plasma focus device of Frascati (CNEN-EURATOM, Italy) are compared with a great number of similar measurement results in several devices of the world. A critical analysis of the above results is presented. (L.C.) [pt

  1. Chlamydial plasmids and bacteriophages.

    Science.gov (United States)

    Pawlikowska-Warych, Małgorzata; Śliwa-Dominiak, Joanna; Deptuła, Wiesław

    2015-01-01

    Chlamydia are absolute pathogens of humans and animals; despite being rather well recognised, they are still open for discovery. One such discovery is the occurrence of extrachromosomal carriers of genetic information. In prokaryotes, such carriers include plasmids and bacteriophages, which are present only among some Chlamydia species. Plasmids were found exclusively in Chlamydia (C.) trachomatis, C. psittaci, C. pneumoniae, C. suis, C. felis, C. muridarum and C. caviae. In prokaryotic organisms, plasmids usually code for genes that facilitate survival of the bacteria in the environment (although they are not essential). In chlamydia, their role has not been definitely recognised, apart from the fact that they participate in the synthesis of glycogen and encode proteins responsible for their virulence. Furthermore, in C. suis it was evidenced that the plasmid is integrated in a genomic island and contains the tetracycline-resistance gene. Bacteriophages specific for chlamydia (chlamydiaphages) were detected only in six species: C. psittaci, C. abortus, C. felis, C. caviae C. pecorum and C. pneumoniae. These chlamydiaphages cause inhibition of the developmental cycle, and delay transformation of reticulate bodies (RBs) into elementary bodies (EBs), thus reducing the possibility of infecting other cells in time. Plasmids and bacteriophages can be used in the diagnostics of chlamydioses; although especially in the case of plasmids, they are already used for detection of chlamydial infections. In addition, bacteriophages could be used as therapeutic agents to replace antibiotics, potentially addressing the problem of increasing antibiotic-resistance among chlamydia.

  2. Comparison of the results of radiation transport calculation obtained by means of different programs

    International Nuclear Information System (INIS)

    Gorbatkov, D.V.; Kruchkov, V.P.

    1995-01-01

    Verification of calculational results of radiation transport, obtained by the known, programs and constant libraries (MCNP+ENDF/B, ANISN+HILO, FLUKA92) by means of their comparison with the precision results calculations through ROZ-6N+Sadko program constant complex and with experimental data, is carried out. Satisfactory agreement is shown with the MCNP+ENDF/B package data for the energy range of E<14 MeV. Analysis of the results derivations, obtained trough the ANISN-HILO package for E<400 MeV and the FLUKA92 programs of E<200 GeV is carried out. 25 refs., 12 figs., 3 tabs

  3. Brief review of topmost scientific results obtained in 2016 at the Joint Institute for Nuclear Research

    International Nuclear Information System (INIS)

    Kravchenko, E.I.; Sabaeva, E.V.

    2017-01-01

    This brief review presents the topmost scientific results obtained in 2016 at the Joint Institute for Nuclear Research in such fields as theoretical and experimental physics, radiation and radiobiological research, accelerators, information technology and computer physics. It also provides information about the publications by JINR staff members and activities carried out at the JINR University Centre in 2016. [ru

  4. Brief review of topmost scientific results obtained in 2015 at the Joint Institute for Nuclear Research

    International Nuclear Information System (INIS)

    Sabaeva, E.V.; Krupko, E.I.

    2016-01-01

    This brief review presents the topmost scientific results obtained in 2015 at the Joint Institute for Nuclear Research in such fields as theoretical and experimental physics, radiation and radiobiological research, accelerators, information technology and computer physics. It also provides information about the publications by JINR staff members, awards given to JINR scientists, and activities carried out at the JINR University Centre in 2015. [ru

  5. An overview of results obtained in intercomparison exercises for determination of actinides

    International Nuclear Information System (INIS)

    Benedik, L.

    2013-01-01

    In recent years the Jožef Stefan Institute participated in numerous intercomparison exercises for determination of natural and man-made radionuclides. The reported values were mostly in good agreement with the resulting reference values. This paper describes an analysis of the measurement results obtained in intercomparison exercises for determination of actinides in the period from 2009 to 2011, organised by the National Physical Laboratory and the Bundesamt für Strahlenschutz. The review covers neptunium, plutonium and americium radioisotopes over this period. - Highlights: • The goal of this study is an overview of results obtained in intercomparison exercises for determination of actinides. • The disagreement of results for Np-237 was evaluated. • Improvements in separation procedures were done

  6. Near-Infrared Scintillation of Liquid Argon: Recent Results Obtained with the NIR Facility at Fermilab

    Energy Technology Data Exchange (ETDEWEB)

    Escobar, C. O. [Fermilab; Rubinov, P. [Fermilab; Tilly, E. [Sewanee U.

    2018-03-19

    After a short review of previous attempts to observe and measure the near-infrared scintillation in liquid argon, we present new results obtained with NIR, a dedicated cryostat at the Fermilab Proton Assembly Building (PAB). The new results give confidence that the near-infrared light can be used as the much needed light signal in large liquid argon time projection chambers.11 pages,

  7. Drug resistance plasmids in Lactobacillus acidophilus and Lactobacillus reuteri.

    OpenAIRE

    Vescovo, M; Morelli, L; Bottazzi, V

    1982-01-01

    Sixteen strains of Lactobacillus reuteri and 20 strains of Lactobacillus acidophilus were tested for resistance to 22 antibiotics by using commercially available sensitivity disks. Evidence suggesting linkage of these resistances to plasmids was obtained by "curing" experiments with acridine dyes and high growth temperatures. Examination of plasmid patterns of agarose gel electrophoresis provided further evidence of loss in plasmid DNA under curing conditions in some of the strains examined.

  8. Plasmid-mediated mineralization of 4-chlorobiphenyl

    International Nuclear Information System (INIS)

    Shields, M.S.; Hooper, S.W.; Sayler, G.S.

    1985-01-01

    Strains of Alcaligenes and Acinetobacter spp. were isolated from a mixed culture already proven to be proficient at complete mineralization of monohalogenated biphenyls. These strains were shown to harbor a 35 x 10(6)-dalton plasmid mediating a complete pathway for 4-chlorobiphenyl (4CB) oxidation. Subsequent plasmid curing of these bacteria resulted in the abolishment of the 4CB mineralization phenotype and loss of even early 4CB metabolism by Acinetobacter spp. Reestablishment of the Alcaligenes plasmid, denoted pSS50, in the cured Acinetobacter spp. via filter surface mating resulted in the restoration of 4CB mineralization abilities. 4CB mineralization, however, proved to be an unstable characteristic in some subcultured strains. Such loss was not found to coincide with any detectable alteration in plasmid size. Cultures capable of complete mineralization, as well as those limited to partial metabolism of 4CB, produced 4-chlorobenzoate as a metabolite. Demonstration of mineralization of a purified 14 C-labeled chlorobenzoate showed it to be a true intermediate in 4CB mineralization. Unlike the mineralization capability, the ability to produce a metabolite has proven to be stable on subculture. These results indicate the occurrence of a novel plasmid, or evolved catabolic plasmid, that mediates the complete mineralization of 4CB

  9. LEP and results obtained by DELPHI after four years of operation

    International Nuclear Information System (INIS)

    Blocki, J.

    1993-10-01

    We characterize the most important problems of modern elementary particles physics, for the solution of which the LEP (Large Electron Positron) accelerator was built. We present the characteristics of this accelerator. The structure and properties of the DELPHI detector are described with special emphasis on the contribution of Polish groups. The most important results obtained so far in the LEP accelerator are discussed. (author). 12 refs, 17 figs, 1 tab

  10. Brief review of topmost scientific results obtained in 2013 at the Joint Institute for Nuclear Research

    International Nuclear Information System (INIS)

    Sabaeva, E.V.; Kravchenko, E.I.

    2014-01-01

    This brief review presents the topmost scientific results obtained in 2013 at the Joint Institute for Nuclear Research in such areas as theoretical physics, experimental physics, radiation and radiobiological research, accelerators, information technology and computer physics. It also provides information on the number of publications by JINR staff members, awards given to JINR scientists, and activities carried out at the JINR University Centre in 2013.

  11. Brief review of topmost scientific results obtained in 2014 at the Joint Institute for Nuclear Research

    International Nuclear Information System (INIS)

    Bulatova, V.V.; Sabaeva, E.V.

    2015-01-01

    This brief review presents the topmost scientific results obtained in 2014 at the Joint Institute for Nuclear Research in such fields as theoretical and experimental physics, radiation and radiobiological research, accelerators, information technology and computer physics. It also provides information about the publications by JINR staff members, patents for inventions, awards given to JINR scientists, and activities carried out at the JINR University Centre in 2014. [ru

  12. Obtaining patient test results from clinical laboratories: a survey of state law for pharmacists.

    Science.gov (United States)

    Witry, Matthew J; Doucette, William R

    2009-01-01

    To identify states with laws that restrict to whom clinical laboratories may release copies of laboratory test results and to describe how these laws may affect pharmacists' ability to obtain patient laboratory test results. Researchers examined state statutes and administrative codes for all 50 states and the District of Columbia at the University of Iowa Law Library between June and July 2007. Researchers also consulted with lawyers, state Clinical Laboratory Improvement Amendments officers, and law librarians. Laws relating to the study objective were analyzed. 34 jurisdictions do not restrict the release of laboratory test results, while 17 states have laws that restrict to whom clinical laboratories can send copies of test results. In these states, pharmacists will have to use alternative sources, such as physician offices, to obtain test results. Pharmacists must consider state law before requesting copies of laboratory test results from clinical laboratories. This may be an issue that state pharmacy associations can address to increase pharmacist access to important patient information.

  13. STUDY REGARDING EFFICIENCY OF INDUCED GENETIC TRANSFORMATION IN BACILLUS LICHENIFORMIS WITH PLASMID DNA

    Directory of Open Access Journals (Sweden)

    T. VINTILĂ

    2007-05-01

    Full Text Available A strain of Bacillus licheniformis was subject to genetic transformation with plasmid vectors (pLC1 and pNC61, using electroporation technique, protoplast transformation and bivalent cations (CaCl2 mediated transformation. In the case of transformation by electroporation of Bacillus licheniformis B40, the highest number of transformed colonies (3 were obtained only after a 1,79 KV electric shock, for 2,2 milliseconds. Using this transformation technique we have obtained six kanamycin resistant transformants. The frequency of Bacillus licheniformis B40 protoplasts transformation using pLC1 and pNC61 plasmid vectors is approximately 10% (TF = 10%. As a result of pLC1 plasmid integration in Bacillus licheniformis protoplasts, six kanamycin resistant transformants were obtained. The pNC61 plasmid, which confers trimethoprim resistance, does not integrate in receiver cells by protoplast transformation. The direct genetic transformation in the presence of bivalent cations (CaCl2, mediated by pLC1 and pNC61 plasmid vectors, produce a low transformation frequency. Using this technique, we have obtained three trimethoprim resistant colonies and four kanamycin resistant colonies. The chemical way of transformation is the only technique, which realizes the integration of pNC61 in B. licheniformis B40 cells.

  14. Results of Investigative Tests of Gas Turbine Engine Compressor Blades Obtained by Electrochemical Machining

    Science.gov (United States)

    Kozhina, T. D.; Kurochkin, A. V.

    2016-04-01

    The paper highlights results of the investigative tests of GTE compressor Ti-alloy blades obtained by the method of electrochemical machining with oscillating tool-electrodes, carried out in order to define the optimal parameters of the ECM process providing attainment of specified blade quality parameters given in the design documentation, while providing maximal performance. The new technological methods suggested based on the results of the tests; in particular application of vibrating tool-electrodes and employment of locating elements made of high-strength materials, significantly extend the capabilities of this method.

  15. Plasmid fermentation process for DNA immunization applications.

    Science.gov (United States)

    Carnes, Aaron E; Williams, James A

    2014-01-01

    Plasmid DNA for immunization applications must be of the highest purity and quality. The ability of downstream purification to efficiently produce a pure final product is directly influenced by the performance of the upstream fermentation process. While several clinical manufacturing facilities already have validated fermentation processes in place to manufacture plasmid DNA for use in humans, a simple and inexpensive laboratory-scale fermentation process can be valuable for in-house production of plasmid DNA for use in animal efficacy studies. This chapter describes a simple fed-batch fermentation process for producing bacterial cell paste enriched with high-quality plasmid DNA. A constant feeding strategy results in a medium cell density culture with continuously increasing plasmid amplification towards the end of the process. Cell banking and seed culture preparation protocols, which can dramatically influence final product yield and quality, are also described. These protocols are suitable for production of research-grade plasmid DNA at the 100 mg-to-1.5 g scale from a typical 10 L laboratory benchtop fermentor.

  16. The effect of uncertainty of reactor parameters obtained using k0-NAA on result of analysis

    International Nuclear Information System (INIS)

    Sasajima, Fumio

    2006-01-01

    Neutron Activation Analysis using the k 0 method is a useful method allowing convenient and accurate simultaneous analysis of plural elements, eliminating the need for the use of comparative reference samples. As already well known, it is essential for the correct result of an analysis to obtain the α-factor and f-factor for a neutron spectrum in an irradiation field accurately when an attempt is made to use the k 0 method. For this reason, based on data obtained from the experiment conducted in the JRR-3 PN-3 system, how uncertainty of the measured values for α-factor and f-factor affects the result of an analysis was evaluated. The process of evaluation involved intentionally varying the values for reactor parameters followed by making an analysis of environmental reference samples (NIST SRM-1632c) using the k 0 method to examine the effect of these factors on the concentrations of 19 elements. The result of the evaluation revealed that the degree of the effect of uncertainty on the concentrations of 19 elements was at best approx. 1% under the condition of this experiment assuming that the factor α, a reactor parameter, had uncertainty of approx. 200%. (author)

  17. Plasmid Flux in Escherichia coli ST131 Sublineages, Analyzed by Plasmid Constellation Network (PLACNET), a New Method for Plasmid Reconstruction from Whole Genome Sequences

    Science.gov (United States)

    Garcillán-Barcia, M. Pilar; Mora, Azucena; Blanco, Jorge; Coque, Teresa M.; de la Cruz, Fernando

    2014-01-01

    Bacterial whole genome sequence (WGS) methods are rapidly overtaking classical sequence analysis. Many bacterial sequencing projects focus on mobilome changes, since macroevolutionary events, such as the acquisition or loss of mobile genetic elements, mainly plasmids, play essential roles in adaptive evolution. Existing WGS analysis protocols do not assort contigs between plasmids and the main chromosome, thus hampering full analysis of plasmid sequences. We developed a method (called plasmid constellation networks or PLACNET) that identifies, visualizes and analyzes plasmids in WGS projects by creating a network of contig interactions, thus allowing comprehensive plasmid analysis within WGS datasets. The workflow of the method is based on three types of data: assembly information (including scaffold links and coverage), comparison to reference sequences and plasmid-diagnostic sequence features. The resulting network is pruned by expert analysis, to eliminate confounding data, and implemented in a Cytoscape-based graphic representation. To demonstrate PLACNET sensitivity and efficacy, the plasmidome of the Escherichia coli lineage ST131 was analyzed. ST131 is a globally spread clonal group of extraintestinal pathogenic E. coli (ExPEC), comprising different sublineages with ability to acquire and spread antibiotic resistance and virulence genes via plasmids. Results show that plasmids flux in the evolution of this lineage, which is wide open for plasmid exchange. MOBF12/IncF plasmids were pervasive, adding just by themselves more than 350 protein families to the ST131 pangenome. Nearly 50% of the most frequent γ–proteobacterial plasmid groups were found to be present in our limited sample of ten analyzed ST131 genomes, which represent the main ST131 sublineages. PMID:25522143

  18. Plasmid flux in Escherichia coli ST131 sublineages, analyzed by plasmid constellation network (PLACNET), a new method for plasmid reconstruction from whole genome sequences.

    Science.gov (United States)

    Lanza, Val F; de Toro, María; Garcillán-Barcia, M Pilar; Mora, Azucena; Blanco, Jorge; Coque, Teresa M; de la Cruz, Fernando

    2014-12-01

    Bacterial whole genome sequence (WGS) methods are rapidly overtaking classical sequence analysis. Many bacterial sequencing projects focus on mobilome changes, since macroevolutionary events, such as the acquisition or loss of mobile genetic elements, mainly plasmids, play essential roles in adaptive evolution. Existing WGS analysis protocols do not assort contigs between plasmids and the main chromosome, thus hampering full analysis of plasmid sequences. We developed a method (called plasmid constellation networks or PLACNET) that identifies, visualizes and analyzes plasmids in WGS projects by creating a network of contig interactions, thus allowing comprehensive plasmid analysis within WGS datasets. The workflow of the method is based on three types of data: assembly information (including scaffold links and coverage), comparison to reference sequences and plasmid-diagnostic sequence features. The resulting network is pruned by expert analysis, to eliminate confounding data, and implemented in a Cytoscape-based graphic representation. To demonstrate PLACNET sensitivity and efficacy, the plasmidome of the Escherichia coli lineage ST131 was analyzed. ST131 is a globally spread clonal group of extraintestinal pathogenic E. coli (ExPEC), comprising different sublineages with ability to acquire and spread antibiotic resistance and virulence genes via plasmids. Results show that plasmids flux in the evolution of this lineage, which is wide open for plasmid exchange. MOBF12/IncF plasmids were pervasive, adding just by themselves more than 350 protein families to the ST131 pangenome. Nearly 50% of the most frequent γ-proteobacterial plasmid groups were found to be present in our limited sample of ten analyzed ST131 genomes, which represent the main ST131 sublineages.

  19. Plasmid flux in Escherichia coli ST131 sublineages, analyzed by plasmid constellation network (PLACNET, a new method for plasmid reconstruction from whole genome sequences.

    Directory of Open Access Journals (Sweden)

    Val F Lanza

    2014-12-01

    Full Text Available Bacterial whole genome sequence (WGS methods are rapidly overtaking classical sequence analysis. Many bacterial sequencing projects focus on mobilome changes, since macroevolutionary events, such as the acquisition or loss of mobile genetic elements, mainly plasmids, play essential roles in adaptive evolution. Existing WGS analysis protocols do not assort contigs between plasmids and the main chromosome, thus hampering full analysis of plasmid sequences. We developed a method (called plasmid constellation networks or PLACNET that identifies, visualizes and analyzes plasmids in WGS projects by creating a network of contig interactions, thus allowing comprehensive plasmid analysis within WGS datasets. The workflow of the method is based on three types of data: assembly information (including scaffold links and coverage, comparison to reference sequences and plasmid-diagnostic sequence features. The resulting network is pruned by expert analysis, to eliminate confounding data, and implemented in a Cytoscape-based graphic representation. To demonstrate PLACNET sensitivity and efficacy, the plasmidome of the Escherichia coli lineage ST131 was analyzed. ST131 is a globally spread clonal group of extraintestinal pathogenic E. coli (ExPEC, comprising different sublineages with ability to acquire and spread antibiotic resistance and virulence genes via plasmids. Results show that plasmids flux in the evolution of this lineage, which is wide open for plasmid exchange. MOBF12/IncF plasmids were pervasive, adding just by themselves more than 350 protein families to the ST131 pangenome. Nearly 50% of the most frequent γ-proteobacterial plasmid groups were found to be present in our limited sample of ten analyzed ST131 genomes, which represent the main ST131 sublineages.

  20. Deep-sequencing protocols influence the results obtained in small-RNA sequencing.

    Directory of Open Access Journals (Sweden)

    Joern Toedling

    Full Text Available Second-generation sequencing is a powerful method for identifying and quantifying small-RNA components of cells. However, little attention has been paid to the effects of the choice of sequencing platform and library preparation protocol on the results obtained. We present a thorough comparison of small-RNA sequencing libraries generated from the same embryonic stem cell lines, using different sequencing platforms, which represent the three major second-generation sequencing technologies, and protocols. We have analysed and compared the expression of microRNAs, as well as populations of small RNAs derived from repetitive elements. Despite the fact that different libraries display a good correlation between sequencing platforms, qualitative and quantitative variations in the results were found, depending on the protocol used. Thus, when comparing libraries from different biological samples, it is strongly recommended to use the same sequencing platform and protocol in order to ensure the biological relevance of the comparisons.

  1. Results obtained with the passive radiation detectors in the ICCHIBAN-4 experiment

    International Nuclear Information System (INIS)

    Bilski, P.; Horwacik, T.

    2005-05-01

    In frame of the InterComparison of Cosmic rays with Heavy Ions Beams at NIRS (ICCHIBAN) organized at the HIMAC accelerator in Chiba several types of the thermoluminescent detectors (TLD), as well as CR-39 track detectors, were exposed. Four different types of TLDs were used: MTS-7 ( 7 LiF:Mg,Ti), MTS-6 ( 6 LiF:Mg,Ti), MCP-7 ( 7 LiF:Mg,Cu,P) and MTT-7 ( 7 LiF:Mg,Ti with changed activator composition. All TLDs were manufactured at the Institute of Nuclear Physics (INP) in Cracow. The detectors were irradiated with various doses of He, C, Ne and Fe ions. Part of exposures were done in unknown conditions, to test measuring capabilities of the detectors. For analyses of these results, the method of obtaining information on ionisation density of an unknown radiation field, which is based on ratios of responses of different LiF detectors, was successfully used. (author)

  2. Experimental Results Obtained with Air Liquide Cold Compression System CERN LHC and SNS Projects

    CERN Document Server

    Delcayre, F; Hamber, F; Hilbert, B; Monneret, E; Toia, J L

    2006-01-01

    Large scale collider facilities will make intensive use of superconducting magnets, operating below 2.0 K. This dictates high‐capacity refrigeration systems operating below 2.0 K. These systems, making use of cryogenic centrifugal compressors in a series arrangement with room temperature screw compressors will be coupled to a refrigerator, providing a certain power at 4.5 K. A first Air Liquide Cold Compression System (CCS) unit was built and delivered to CERN in 2001. Installed at the beginning of 2002, it was commissioned and tested successfully during year 2002. A series of four sets of identical CCS were then tested in 2004. Another set of four cryogenic centrifugal compressors (CCC) has been delivered to Thomas Jefferson National Accelerator Facility (JLAB) for the Spallation Neutron Source (SNS) in 2002. These compressors were tested and commissioned from December 2004 to July 2005. The experimental results obtained with these systems will be presented and discussed: the characteristics of the CCC wil...

  3. First results obtained from the Cello liquid argon end cap calorimeters

    International Nuclear Information System (INIS)

    Le Diberder, F.

    1981-05-01

    The Cello liquid argon calorimeter is presented in the first part of this thesis. The cryogenic system has to supply three cryostats filled with liquid argon: one cylindrical cryostat of 25 m 3 volume contains 2x8 separate modules; each of the two symmetric end cap cryostats contains two half cylindrical modules. Each module in the end cap part consists of 42 layers of lead strips interleaved with 43 full plates. The strips are alternatively vertical, horizontal and circular. In front of the lead calorimeter are 4 planes of copper foils glued on epoxy for dE/dx measurement. The electronics, signal processing and data acquisition system are described. In the second part, the performance and analysis of data measured by the end cap calorimeters are reported: study of Bhabha scattering e + e - → e + e - ; preliminary results obtained in two photon physics e + e - → e + e - γγ → e + e - X [fr

  4. Optimization of plasmid electrotransformation into Escherichia coli ...

    African Journals Online (AJOL)

    In order to improve electroporation, optical density of bacteria, recovery time and electrical parameter (field strength and capacitance) were optimized using the Taguchi statistical method. ANOVA of obtained data indicated that the optimal conditions of electrotransformation of pET-28a (+) plasmid into Escherichia coli ...

  5. Report of test and research results on atomic energy obtained in national institutes in fiscal 1985

    International Nuclear Information System (INIS)

    1986-01-01

    As for the test and research on the utilization of atomic energy in national institutes, the budget was appropriated for the first time in fiscal year 1956, and since then, the many valuable results of research have been obtained so far in the diversified fields of nuclear fusion, safety research, the irradiation of foods, medicine and others, thus the test and research accomplished the large role for promoting the utilization of atomic energy in Japan. In this report, the gists of the results of the test and research on the utilization of atomic energy carried out by national institutes in fiscal year 1985 are collected. No.1 of this report was published in 1960, and this is No.26. It is desired to increase the understanding about the recent trend and the results of the test and research on atomic energy utilization with this book. The researches on nuclear fusion, engineering safety and environmental radioactivity safety, the irradiation of foods, the countermeasures against cancer, fertilized soil, the quality improvement of brewing and farm products, the protection of farm products and the improvement of breeding, diagnosis and medical treatment, pharmaceuticals, environmental hygiene, the application to physiology and pathology, radiochemistry, radiation measurement, process analysis, nuclear reactor materials, nuclear powered ships, civil engineering, radioactivation analysis and injury prevention are reported. (Kako, I.)

  6. Direct repeat sequences are essential for function of the cis-acting locus of transfer (clt) of Streptomyces phaeochromogenes plasmid pJV1.

    Science.gov (United States)

    Franco, Bernardo; González-Cerón, Gabriela; Servín-González, Luis

    2003-11-01

    The functionality of direct and inverted repeat sequences inside the cis acting locus of transfer (clt) of the Streptomyces plasmid pJV1 was determined by testing the effect of different deletions on plasmid transfer. The results show that the single most important element for pJV1 clt function is a series of evenly spaced 9 bp long direct repeats which match the consensus CCGCACA(C/G)(C/G), since their deletion caused a dramatic reduction in plasmid transfer. The presence of these repeats in the absence of any other clt sequences allowed plasmid transfer to occur at a frequency that was at least two orders of magnitude higher than that obtained in the complete absence of clt. A database search revealed regions with a similar organization, and in the same position, in Streptomyces plasmids pSN22 and pSLS, which have transfer proteins homologous to those of pJV1.

  7. Summary of personal neutron dosemeter results obtained within the EVIDOS project

    International Nuclear Information System (INIS)

    Luszik-Bhadra, M.; Reginatto, M.; Schuhmacher, H.; Bolognese-Milsztajn, T.; Lacoste, V.; Boschung, M.; Fiechtner, A.; Coeck, M.; Vanhavere, F.; Curzio, G.; Errico d', F.; Kylloenen, J.-E.; Lindborg, L.; Bartlett, D.; Tanner, R.

    2005-01-01

    Full text: Within the EC project EVIDOS ('Evaluation of Individual Dosimetry in Mixed Neutron and Photon Radiation Fields'), different types of neutron personal dosemeters were irradiated in workplace fields in nuclear installations in Europe. Mostly electronic personal neutron dosemeters were tested, among them commercial devices (Thermo Electron EPD-N, Aloka PDM-313), devices from first industrial prototype series (Thermo Electron EPD-N2, Saphydose-n) and laboratory prototypes which were already in the stage of lightweight battery-operated instruments (PTB DOS-2002). In addition, dosemeters with (almost) immediate readout (BTI bubble detectors, Rados DIS-N) and passive dosemeters which needed no fielddependent calibration factors (PADC track detectors from PSI and NRPB) were used, as well as those TLD albedo dosemeters which are routinely used in the facilities visited. The results of measurements obtained within the EVIDOS project in workplace fields in nuclear installations in Europe, i.e. at Kruemmel (boiling water reactor, transport cask), at Mol (Venus research reactor, fuel facility Belgonucleaire) and at Ringhals (pressurized water reactor, transport cask) are presented and compared to reference values of personal dose equivalent H p (10) determined by means of Bonner spheres and novel directional spectrometers. In fields with strong back-scatter and moderation of neutrons, several dosemeters showed overreadings by more than a factor of two. In strongly directed fields with neutron dose equivalent peaked at about 1 MeV (MOX fuel), the same dosemeters showed under-readings of about a factor of two. These under- and over-readings in the workplace fields can be explained in terms of the dosemeter response functions for mono-energetic neutrons, which show over-responses in the thermal and intermediate neutron energy region and under-responses at about 1 MeV neutron energy. The dosemeter readings obtained in the workplace fields were checked for consistency by

  8. Experimental results obtained with the simulated cold moderator system. System characteristics and technical issues

    CERN Document Server

    Aso, T; Hino, R; Kaminaga, M; Kinoshita, H; Takahashi, T

    2002-01-01

    The Japan Atomic Energy Research Institute and the High Energy Accelerator Research Organization have been developing a Mega-Watt scale spallation target system. In the system, neutrons generated in a target are sorted out their energy to the proper values in liquid-hydrogen moderators. Then, the liquid-hydrogen is forced to circulate in order to suppress hydrogen temperature increase. In the operation of moderators, it is very important to establish a safety protection system against emergency shutdown of the accelerator or accidents of the cold moderator system. In order to obtain a technical data for design and safety review of the liquid-hydrogen system, we have fabricated an experimental apparatus simulated the cold moderator system using liquid nitrogen (max. 1.5 MPa, mini. 77 K) instead of liquid hydrogen. The experiments on a controllability of the system were carried out to investigate dynamic characteristics of the system. This report presents the experimental results and technical issues for the co...

  9. Experimental Results Obtained with Air Liquide Cold Compression System: CERN LHC and SNS Projects

    Science.gov (United States)

    Delcayre, F.; Courty, J.-C.; Hamber, F.; Hilbert, B.; Monneret, E.; Toia, J.-L.

    2006-04-01

    Large scale collider facilities will make intensive use of superconducting magnets, operating below 2.0 K. This dictates high-capacity refrigeration systems operating below 2.0 K. These systems, making use of cryogenic centrifugal compressors in a series arrangement with room temperature screw compressors will be coupled to a refrigerator, providing a certain power at 4.5 K. A first Air Liquide Cold Compression System (CCS) unit was built and delivered to CERN in 2001. Installed at the beginning of 2002, it was commissioned and tested successfully during year 2002. A series of four sets of identical CCS were then tested in 2004. Another set of four cryogenic centrifugal compressors (CCC) has been delivered to Thomas Jefferson National Accelerator Facility (JLAB) for the Spallation Neutron Source (SNS) in 2002. These compressors were tested and commissioned from December 2004 to July 2005. The experimental results obtained with these systems will be presented and discussed: the characteristics of the CCC will be detailed. The principles of control for the CCC in series will be detailed.

  10. The results obtained by INR-Pitesti to an international in-situ intercomparison exercise

    International Nuclear Information System (INIS)

    Dobrin, Relu; Dulama, Cristian; Toma, Alexandru

    2008-01-01

    Full text: The determination of soil contamination, dose rate measurements and in-situ gamma spectrometry are well established and widely used measurement procedures, especially after large scale nuclear incidents. To support decision makers and first responders with a more comprehensive and accurate overview immediately after a large scale nuclear or radiological emergency, as releases from a nuclear power plant or a terrorist attack with a dirty bomb, a fast and clear presentation of measurement data is indispensable. In 2007, the Austrian Research Centers GmbH - ARC (Seibersdorf) in cooperation with IAEA and the Austrian NBC Defense School organized an international measurement campaign, 'In-Situ Intercomparison Scenario' ISIS 2007, with the focus on In-Situ Gamma Spectrometry and Dose Rate Measurements in Emergency Situations. 56 teams from 25 countries, worldwide, took part to this exercise. The only Romanian team was 'CROWN'. The CROWN Laboratory is part of the Radiation Protection Laboratory of the Institute for Nuclear Research - Pitesti, certified for characterization of radioactive wastes and nuclear materials. The paper presents results obtained during different tasks of the exercise: 'Dose rate mapping'; 'Localization - Drive by'; 'Simulation of a contamination'; 'Identification Shielded; 'Identification/Quantification'; 'Buried sources; 'Environmental measurement'. (authors)

  11. Experimental results obtained with the simulated cold moderator system. System characteristics and technical issues

    Energy Technology Data Exchange (ETDEWEB)

    Aso, Tomokazu; Kaminaga, Masanori; Haga, Katsuhiro; Kinoshita, Hidetaka; Takahashi, Toshio; Hino, Ryutaro [Japan Atomic Energy Research Inst., Tokai, Ibaraki (Japan). Tokai Research Establishment

    2002-12-01

    The Japan Atomic Energy Research Institute and the High Energy Accelerator Research Organization have been developing a Mega-Watt scale spallation target system. In the system, neutrons generated in a target are sorted out their energy to the proper values in liquid-hydrogen moderators. Then, the liquid-hydrogen is forced to circulate in order to suppress hydrogen temperature increase. In the operation of moderators, it is very important to establish a safety protection system against emergency shutdown of the accelerator or accidents of the cold moderator system. In order to obtain a technical data for design and safety review of the liquid-hydrogen system, we have fabricated an experimental apparatus simulated the cold moderator system using liquid nitrogen (max. 1.5 MPa, mini. 77 K) instead of liquid hydrogen. The experiments on a controllability of the system were carried out to investigate dynamic characteristics of the system. This report presents the experimental results and technical issues for the construction of a practical liquid-hydrogen moderator system of the Mega-Watt scale target system. (author)

  12. Effect of degradative plasmid CAM-OCT on responses of Pseudomonas bacteria to UV light

    International Nuclear Information System (INIS)

    McBeth, D.L.

    1989-01-01

    The effect of plasmid CAM-OCT on responses to UV irradiation was compared in Pseudomonas aeruginosa, in Pseudomonas putida, and in Pseudomonas putida mutants carrying mutations in UV response genes. CAM-OCT substantially increased both survival and mutagenesis in the two species. P. aeruginosa strains without CAM-OCT exhibited much higher UV sensitivity than did P. putida strains. UV-induced mutagenesis of plasmid-free P. putida was easily detected in three different assays (two reversion assays and one forward mutation assay), whereas UV mutagenesis of P. aeruginosa without CAM-OCT was seen only in the forward mutation assay. These results suggest major differences in DNA repair between the two species and highlight the presence of error-prone repair functions on CAM-OCT. A number of P. putida mutants carrying chromosomal mutations affecting either survival or mutagenesis after UV irradiation were isolated, and the effect of CAM-OCT on these mutants was determined. All mutations producing a UV-sensitive phenotype in P. putida were fully suppressed by the plasmid, whereas the plasmid had a more variable effect on mutagenesis mutations, suppressing some and producing no suppression of others. On the basis of the results reported here and results obtained by others with plasmids carrying UV response genes, it appears that CAM-OCT may differ either in regulation or in the number and functions of UV response genes encoded

  13. Plasmids in Gram negatives: molecular typing of resistance plasmids.

    Science.gov (United States)

    Carattoli, Alessandra

    2011-12-01

    A plasmid is defined as a double stranded, circular DNA molecule capable of autonomous replication. By definition, plasmids do not carry genes essential for the growth of host cells under non-stressed conditions but they have systems which guarantee their autonomous replication also controlling the copy number and ensuring stable inheritance during cell division. Most of the plasmids confer positively selectable phenotypes by the presence of antimicrobial resistance genes. Plasmids evolve as an integral part of the bacterial genome, providing resistance genes that can be easily exchanged among bacteria of different origin and source by conjugation. A multidisciplinary approach is currently applied to study the acquisition and spread of antimicrobial resistance in clinically relevant bacterial pathogens and the established surveillance can be implemented by replicon typing of plasmids. Particular plasmid families are more frequently detected among Enterobacteriaceae and play a major role in the diffusion of specific resistance genes. For instance, IncFII, IncA/C, IncL/M, IncN and IncI1 plasmids carrying extended-spectrum beta-lactamase genes and acquired AmpC genes are currently considered to be "epidemic resistance plasmids", being worldwide detected in Enterobacteriaceae of different origin and sources. The recognition of successful plasmids is an essential first step to design intervention strategies preventing their spread. Copyright © 2011 Elsevier GmbH. All rights reserved.

  14. Preliminary observations and results obtained with the ultraviolet spectrometer and polarimeter

    International Nuclear Information System (INIS)

    Tandberg-Hanssen, E.; Athay, R.G.; Beckers, J.M.; Brandt, J.C.; Bruner, E.C.; Chapman, R.D.; Cheng, C.C.; Burman, J.G.; Henze, W.; Hyder, C.L.; Michalitsianos, A.G.; Shine, R.A.; Schoolman, S.A.; Woodgate, B.E.

    1981-01-01

    We present new observations with the Ultraviolet Spectrometer and Polarimeter (UVSP) of a number of manifestations of solar activity obtained during the first 3 months of Solar Maximum Mission operations

  15. A rapid method for screening arrayed plasmid cDNA library by PCR

    International Nuclear Information System (INIS)

    Hu Yingchun; Zhang Kaitai; Wu Dechang; Li Gang; Xiang Xiaoqiong

    1999-01-01

    Objective: To develop a PCR-based method for rapid and effective screening of arrayed plasmid cDNA library. Methods: The plasmid cDNA library was arrayed and screened by PCR with a particular set of primers. Results: Four positive clones were obtained through about one week. Conclusion: This method can be applied to screening not only normal cDNA clones, but also cDNA clones-containing small size fragments. This method offers significant advantages over traditional screening method in terms of sensitivity, specificity and efficiency

  16. Yeast transformation mediated by Agrobacterium strains harboring an Ri plasmid: comparative study between GALLS of an Ri plasmid and virE of a Ti plasmid.

    Science.gov (United States)

    Kiyokawa, Kazuya; Yamamoto, Shinji; Sato, Yukari; Momota, Naoto; Tanaka, Katsuyuki; Moriguchi, Kazuki; Suzuki, Katsunori

    2012-07-01

    Agrobacterium strains containing a Ti plasmid can transfer T-DNA not only to plants but also to fungi, including the yeast Saccharomyces cerevisiae. However, no Agrobacterium strain harboring an Ri plasmid has been evaluated in fungal transformation. Some Ri plasmids have GALLS , instead of virE1 and virE2. GALLS protein can functionally substitute in plant transformation for a structurally different protein VirE2. In this study, we compared the yeast transformation ability among Agrobacterium donors: a strain containing a Ti plasmid, strains harboring either an agropine-type or a mikimopine-type Ri plasmid, and a strain having a modified Ri plasmid supplemented with a Ti plasmid type virE operon. Agrobacterium strains possessing GALLS transformed yeast cells far less efficiently than the strain containing virE operon. Production of GALLS in recipient yeast cells improved the yeast transformation mediated by an Agrobacterium strain lacking neither GALLS nor virE operon. A reporter assay to detect mobilization of the proteins fused with Cre recombinase revealed that VirE2 protein is much more abundant in yeast cells than GALLS. Based on these results, we concluded that the low yeast transformability mediated by Agrobacterium strains having the Ri plasmid is because of low amount of mobilized GALLS in yeast cells. © 2012 The Authors Journal compilation © 2012 by the Molecular Biology Society of Japan/Blackwell Publishing Ltd.

  17. Preliminary observations and results obtained with the ultraviolet spectrometer and polarimeter. [for Solar Maximum Mission

    Science.gov (United States)

    Tandberg-Hassen, E.; Cheng, C. C.; Athay, R. G.; Beckers, J. M.; Brandt, J. C.; Chapman, R. D.; Bruner, E. C.; Henze, W.; Hyder, C. L.; Gurman, J. B.

    1981-01-01

    New observation with the Ultraviolet Spectrometer and Polarimeter (UVSP) of a number of manifestations of solar activity obtained during the first three months of Solar Maximum Mission operations are presented. Attention is given to polarimetry in sunspots, oscillations above sunspots, density diagnostics of transition-zone plasmas in active regions, and the eruptive prominence - coronal transient link.

  18. Plasmids in Vibrio parahemolyticus strains isolated in Japan and Bangladesh with special reference to different distributions.

    Science.gov (United States)

    Arai, T; Ando, T; Kusakabe, A; Ullah, M A

    1983-01-01

    We surveyed plasmids in naturally occurring Vibrio parahemolyticus strains isolated in Japan and Bangladesh. Among the strains isolated in Japan, about half of the strains isolated from stools of patients of domestic diarrhea outbreaks as well as of travelers returning from East Asia were found to have plasmids, but no strains from foods had plasmids. In contrast, among the strains isolated in Bangladesh, none of the four strains isolated from patients had plasmids, but two out of eight strains isolated from water had plasmids, suggesting that plasmids are common in strains from the water in Bangladesh. All plasmids so far reported in V. parahemolyticus were detected in strains isolated from stools of patients. Incidences of plasmids in this organism were not so high in either area. In Japan, all plasmids were detected in strains from human intestines at 37 C, but in Bangladesh, where the temperature is around 30-40 C, the plasmids were detected in strains from the natural environment. These results suggested the possibility that these plasmids can come from different bacteria under rather high temperatures and that incidences of plasmids are influenced by the incidences of plasmids in bacteria present in the vicinity of V. parahemolyticus strains. None of these plasmids were found to have any relation to the biological characters tested.

  19. STUDY REGARDING EFFICIENCY OF INDUCED GENETIC TRANSFORMATION IN BACILLUS LICHENIFORMIS WITH PLASMID DNA

    Directory of Open Access Journals (Sweden)

    VINTILĂ T.

    2007-01-01

    Full Text Available A strain of Bacillus licheniformis was subject to genetic transformation with plasmidvectors (pLC1 and pNC61, using electroporation technique, protoplasttransformation and bivalent cations (CaCl2 mediated transformation. In the case oftransformation by electroporation of Bacillus licheniformis B40, the highest numberof transformed colonies (3 were obtained only after a 1,79 KV electric shock, for 2,2milliseconds. Using this transformation technique we have obtained six kanamycinresistant transformants. The frequency of Bacillus licheniformis B40 protoplaststransformation using pLC1 and pNC61 plasmid vectors is approximately 10% (TF =10%. As a result of pLC1 plasmid integration in Bacillus licheniformis protoplasts,six kanamycin resistant transformants were obtained. The pNC61 plasmid, whichconfers trimethoprim resistance, does not integrate in receiver cells by protoplasttransformation. The direct genetic transformation in the presence of bivalent cations(CaCl2, mediated by pLC1 and pNC61 plasmid vectors, produce a lowtransformation frequency. Using this technique, we have obtained three trimethoprimresistant colonies and four kanamycin resistant colonies. The chemical way oftransformation is the only technique, which realizes the integration of pNC61 in B.licheniformis B40 cells.

  20. Persistence Mechanisms of Conjugative Plasmids

    DEFF Research Database (Denmark)

    Bahl, Martin Iain; Hansen, Lars H.; Sørensen, Søren Johannes

    2009-01-01

    Are plasmids selfish parasitic DNA molecules or an integrated part of the bacterial genome? This chapter reviews the current understanding of the persistence mechanisms of conjugative plasmids harbored by bacterial cells and populations. The diversity and intricacy of mechanisms affecting the suc...

  1. Analysis of results obtained from field tracing test under natural rain condition

    International Nuclear Information System (INIS)

    Mukai, M.; Kamiyama, H.; Tanaka, T.; Wang Zhiming; Zhao Yingjie; Li Zhengtang

    1993-01-01

    As one of the tests arranged by the cooperative research between CIRP and JAERI, field tracing tests using 3 H, 60 Co, 85 Sr and 134 Cs were conducted in pits at the CIRP's field test site located on a loess tableland under natural rain condition. Precipitation amount and evaporation rate were measured to study complicated spatial-temporal behavior of soil water movement under that condition. The evaporation rate was obtained through an analysis on the measured data by a combined method of heat balance and eddy correlation. Numerical model, that is based on piston flow assumption of soil water movement, was developed and applied to determine the behavior of the soil water movement in the pits. Using the determined water movement, 3 H migration was evaluated by numerical simulation. Change of 3 H distribution as a function of elapsed time as well explained by careful evaluation of the soil water movement that carried out before the analysis. (5 figs.)

  2. Preliminary Results from Duplex Procedure for Obtain of Fe Based Materials for Automotive Applications

    Science.gov (United States)

    Crăciun, R. C.; Stanciu, S.; Geantă, V.; Voiculescu, I.; Manole, V.; Gârneţ, I. A.; Alexandru, A.; Cimpoesu, N.; Săndulache, F.

    2017-06-01

    Abstract Iron based materials still represent a high percentage from metallic materials used in industry, in general, and in automotive industry, in particular. In this case we used a duplex process in order to obtain the FeMnSiAl experimental alloy for a more efficient use of various units. In the first stage iron, manganese, silicon and aluminum were melted and mixed together using arc melting technology and for the second stage the alloy was re-melt for homogeneity in an induction furnace. Chemical composition, after each melting step, was analyzed using EDS Bruker detector for various areas and microstructural characterization using SEM, VegaTescan LMH II with SE detector, equipment. This alloy is proposed as a metallic approach of mechanical dumpers used in automotive industry for low and medium impact contacts.

  3. Sequence analysis and characterization of rolling-circle replicating plasmid pVCM01 from Salmonella enterica

    Directory of Open Access Journals (Sweden)

    Penido, A. F. B.

    2013-12-01

    Full Text Available Aims: Characterization of cryptic plasmid pVCM01 (accession number JX133088 isolated from Salmonella enterica Enteritidis. Methodology and results: The complete sequence of pVCM01 was obtained. This plasmid possesses 1981 bp, with G+C content of 57% in agreement of the range of Salmonella genomic DNA. pVCM01 has a high degree of similarity to pB and pJ plasmids. It possesses six main open reading frames, only one have a very high degree of amino acid identity with protein involved in the rolling-circle-like replication (RCR. Based on the sequence similarities, pVCM01 plasmid belonged to the pC194/pUB110 rolling-circle replicating plasmid family. The Rep pVCM01 possesses the motifs: FLTLTVRN, HPHFHTL, SGDGYVKHERW, which were present in all Rep proteins. Conclusion, significance and impact of study: The small size of pVCM01 plasmid and its stability in E. coli cells, make it an attractive candidate to develop new vectors, such as cloning and/or expression vector.

  4. Vehicle Animation Software (VAS) to Animate Results Obtained from Vehicle Handling and Rollover Simulations and Tests

    Science.gov (United States)

    1991-04-01

    Results from vehicle computer simulations usually take the form of numeric data or graphs. While these graphs provide the investigator with the insight into vehicle behavior, it may be difficult to use these graphs to assess complex vehicle motion. C...

  5. Explanatory chapter: how plasmid preparation kits work.

    Science.gov (United States)

    Koontz, Laura

    2013-01-01

    To isolate plasmid DNA from bacteria using a commercial plasmid miniprep kit (if interested, compare this protocol with Isolation of plasmid DNA from bacteria). Copyright © 2013 Elsevier Inc. All rights reserved.

  6. Movement and equipositioning of plasmids by ParA filament disassembly

    DEFF Research Database (Denmark)

    Ringgaard, Simon; van Zon, Jeroen; Howard, Martin

    2009-01-01

    , plasmids consistently migrate behind disassembling ParA cytoskeletal structures, suggesting that ParA filaments pull plasmids by depolymerization. The perpetual cycles of ParA assembly and disassembly result in continuous relocation of plasmids, which, on time averaging, results in equidistribution...

  7. Acoustic emission results obtained from testing the ZB-1 intermediate scale pressure vessel

    International Nuclear Information System (INIS)

    Hutton, P.H.; Kurtz, R.J.; Pappas, R.A.; Dawson, J.F.; Dake, L.S.; Skorpik, J.R.

    1985-09-01

    Acoustic emission (AE) monitoring of flaw growth in an intermediate scale vessel during cyclic loading at 65 0 C and 288 0 C is described in this report. The report deals with background, methodology, and results. The work discussed is of major significance in a program supported by NRC to develop and demonstrate application of AE monitoring for continuous surveillance of reactor pressure boundaries to detect and evaluate growing flaws. Several areas of technical concern are addressed. Results support the feasibility of effective continuous monitoring

  8. Occupational radioprotection program at Nuclear Engineering Institute -IEN: results obtained in 1991

    International Nuclear Information System (INIS)

    Fajardo, P.W.; Pastura, V.F.S.; Soares, M.L.; LeRoy, C.L.; Teixeira, M.V.; Santos, I.H.T.; Pujol Filho, S.V.

    1992-01-01

    The results of occupational radioprotection program at Nuclear Engineering Institute-IEN- in 1991 are presented. The personnel monitoring, the routine monitoring of limited areas, the operational monitoring during the operation and the cyclotron CV-28 maintenance, the radioisotope processing and Argonauta Reactor operation, the control of radioprotection equipment and the control of radiation sources are included. (C.G.C.)

  9. Alecto - results obtained with homogeneous critical experiments on plutonium 239, uranium 235 and uranium 233

    International Nuclear Information System (INIS)

    Bruna, J.G.; Brunet, J.P.; Caizegues, R.; Clouet d'Orval, Ch.; Kremser, J.; Tellier, H.; Verriere, Ph.

    1965-01-01

    In this report are given the results of the homogeneous critical experiments ALECTO, made on plutonium 239, uranium 235 and uranium 233. After a brief description of the equipment, the critical masses for cylinders of diameters varying from 25 to 42 cm, are given and compared with other values (foreign results, criticality guide). With respect to the specific conditions of neutron reflection in the ALECTO experiments the minimal values of critical masses are: Pu239 M c = 910 ± 10 g, U235 M c = 1180 ± 12 g and U233 M c = 960 ± 10 g. Experiments relating to cross sections and constants to be used on these materials are presented. Lastly, kinetic experiments allow to compare pulsed neutron methods to fluctuation methods [fr

  10. Macromolecular crystallographic results obtained using a 2048x2048 CCD detector at CHESS

    International Nuclear Information System (INIS)

    Thiel, D.J.; Ealick, S.E.; Tate, M.W.; Gruner, S.M.; Eikenberry, E.F.

    1996-01-01

    We present results of macromolecular crystallographic experiments performed at the Cornell High Energy Synchrotron Source (CHESS) with a new CCD-based detector. This detector, installed in January 1995, complements a 1024x1024 CCD detector that has been in continuous operation at CHESS since December 1993. The new detector is based on a 4-port, 2048x2048 pixel CCD that is directly coupled to a Gd 2 O 2 S:Tb phosphor by a 3:1 tapered fiber optic. The active area of the phosphor is a square 82 mm on an edge. The readout time is 7 seconds. In the standard mode of operation, the pixel size at the active area is 41 μm on the edge leading to the capability of resolving approximately 200 orders of diffraction across the detector face. The detector also operates in a 1024x1024 mode in which the pixel size is electronically increased by a factor of 4 in area resulting in smaller data files and faster detector readout but at the expense of spatial resolution. Most of the data that has been collected by this detector has been collected in this mode. Dozens of data sets have been collected by many experimenters using this detector at CHESS during the four month period from its installation until the start of the six-month down period of the storage ring. The capabilities of the detector will be illustrated with results from various crystallographic measurements including experiments in which the recorded diffraction patterns extend in resolution as far as 1 A. The results demonstrate that this detector is capable of collecting data of quality at least equal to that of imaging plates but, in many circumstances, with much greater beamline efficiency. copyright 1996 American Institute of Physics

  11. Irradiation devices for fusion reactor materials results obtained from irradiated lithium aluminate at the OSIRIS reactor

    International Nuclear Information System (INIS)

    Lefevre, F.; Thevenot, G.; Rasneur, B.; Botter, F.

    1986-06-01

    Studies about controlled fusion reactor of the Tokamak type require the examination of the radiation effects on the behaviour of various potential materials. Thus, in the first part of this paper, are presented the devices adapted to these materials studies and used in the OSIRIS reactor. In a second part, is described an experiment of irradiation ceramics used as candidates for breeding material and are given the first results

  12. Report of test and research results on atomic energy obtained in national institutes in fiscal 1987

    International Nuclear Information System (INIS)

    1989-01-01

    The test and research regarding the utilization of atomic energy carried out in national institutions have produced many valuable results in diverse fields so far, such as nuclear fusion, safety research, food irradiation and medicine, since the budget had been appropriated for the first time in 1956. It has accomplished large role in the promotion of atomic energy utilization in Japan. This report is volume 28, in which the results of the test and research on atomic energy utilization carried out by national institutions in fiscal year 1987 are summarized. It is hoped that the understanding about the recent trend and the results of the test and research on atomic energy utilization is further promoted by this report. The contents of this report are nuclear fusion; the research on engineering safety and environmental radioactivity safety; food irradiation; the countermeasures against cancer; fertilized soil, the improvement of quality, the protection of plants and the improvement of breeding in agriculture and fishery fields; diagnosis and medical treatment, pharmaceuticals, environmental hygiene and the application to physiology and pathology in medical field; radiation measurement and process analysis in mining and industry fields; nuclear reactor materials and nuclear-powered ships; civil engineering; radioactivation analysis; the research on the prevention of injuries; and the basic researches on materials and acessment and reduction of irradiation risk. (J.P.N.)

  13. Report of test and research results on atomic energy obtained in national institutes in fiscal 1986

    International Nuclear Information System (INIS)

    1988-01-01

    The test and research regarding the utilization of atomic energy carried out in national institutions have produced many valuable results in diverse fields so far, such as nuclear fusion, safety research, food irradiation and medicine, since the budget had been appropriated for the first time in 1956. It has accomplished large role in the promotion of atomic energy utilization in Japan. This report is volume 27, in which the results of the test and research on atomic energy utilization carried out by national institutions in fiscal year 1986 are summarized. It is hoped that the understanding about the recent trend and the results of the test and research on atomic energy utilization is further promoted by this report. The contents of this report are nuclear fusion; the research on engineering safety and environmental radioactivity safety; food irradiation; the countermeasures against cancer; fertilized soil, the improvement of quality, the protection of plants and the improvement of breeding in agriculture and fishery fields; diagnosis and medical treatment, pharmaceuticals, environmental hygiene and the application to physiology and pathology in medical field; radiation chemistry and radiation measurement in mining and industry fields; nuclear reactor materials and nuclear-powered ships; civil engineering; radioactivation analysis; and the research on the prevention of injuries. (Kako, I.)

  14. Report of test and research results on atomic energy obtained in national institutes in fiscal 1984

    International Nuclear Information System (INIS)

    1985-01-01

    The test and research regarding the utilization of atomic energy carried out in national institutions have produced many valuable results in diverse fields so far, such as nuclear fusion, safety research, food irradiation and medicine, since the budget had been appropriated for the first time in 1956. It has accomplished large role in the promotion of atomic energy utilization in Japan. This report is Volume 25, in which the results of the test and research on atomic energy utilization carried out by national institutions in fiscal year 1984 are summarized. It is hoped that the understanding about the recent trend and the results of the test and research on atomic energy utilization is further promoted by this report. The contents of this report are nuclear fusion; the research on engineering safety and environmental radioactivity safety; food irradiation; the countermeasures against cancer; fertilized soil, the improvement of quality, the protection of plants and the improvement of breeding in agriculture and fishery fields; diagnosis and medical treatment, pharmaceuticals, environmental hygiene and the application to physiology and pathology in medical field; radiation chemistry and radiation measurement in mining and industry fields; nuclear reactor materials and nuclear-powered ships; civil engineering; radioactivation analysis; and the research on the prevention of injuries. (Kako, I.)

  15. Radon in Estonian buildings. Establishment of a measurement system and obtained results

    International Nuclear Information System (INIS)

    Pahapill, L.; Rulkov, A.; Swedjemark, G.A.

    1996-12-01

    One purpose of this project was the establishment of a radon monitoring programme inside the state environmental monitoring programme. Another purpose was to investigate regions, expected to have high radon levels indoors. A new method for the long-term measurement of indoor radon was established and the staff for these measurements was trained. The results of the measurement can be used by Estonian decision-makers to work out rules and standards. There is no legislative act in the field of radiation in Estonian at this time. To summarize the results of the measurements we can say that indoor radon concentrations vary by region. The radon investigations must be continued to identify the risk areas and types of housing construction. The results of the state radon monitoring are provided to the municipalities, who advice the owners of planned new houses to select the right construction for the house. A new project will follow with an investigation of radon in randomly selected dwellings, training and equipment for radon measurement in soil, and general advice with regard to radon, as well as assistance in preparing information about radon. 7 refs, 5 figs

  16. Experimental results on advanced inertial fusion schemes obtained within the HiPER project

    International Nuclear Information System (INIS)

    Batani, Dimitri; Santos, Jorge J.; Schurtz, Guy; Hulin, Sebastien; Ribeyre, Xavier; Nicolai, Philippe; Vauzour, Benjamin; Dorchies, Fabien; Gizzi, Leonida A.; Koester, Petra; Labate, Luca; Honrubia, Javier; Antonelli, Luca; Morace, Alessio; Volpe, Luca; Nazarov, Wiger; Pasley, John; Richetta, Maria; Lancaster, Kate; Spindloe, Christopher; Tolley, Martin; Neely, David; Kozlova, Michaela; Nejdl, Jaroslav; Rus, Bedrich; Wolowski, Jerzy; Badziak, Jan

    2012-01-01

    This paper presents the results of experiments conducted within the Work Package 10 (fusion experimental programme) of the HiPER project. The aim of these experiments was to study the physics relevant for advanced ignition schemes for inertial confinement fusion, i.e. the fast ignition and the shock ignition. Such schemes allow to achieve a higher fusion gain compared to the indirect drive approach adopted in the National Ignition Facility in United States, which is important for the future inertial fusion energy reactors and for realising the inertial fusion with smaller facilities. (authors)

  17. In Silico Detection and Typing of Plasmids using PlasmidFinder and Plasmid Multilocus Sequence Typing

    DEFF Research Database (Denmark)

    Carattoli, Alessandra; Zankari, Ea; García-Fernández, Aurora

    2014-01-01

    In the work presented here, we designed and developed two easy-to-use Web tools for in silico detection and characterization of whole-genome sequence (WGS) and whole-plasmid sequence data from members of the family Enterobacteriaceae. These tools will facilitate bacterial typing based on draft...... genomes of multidrug-resistant Enterobacteriaceae species by the rapid detection of known plasmid types. Replicon sequences from 559 fully sequenced plasmids associated with the family Enterobacteriaceae in the NCBI nucleotide database were collected to build a consensus database for integration...... sequences identified in the 559 fully sequenced plasmids. For plasmid multilocus sequence typing (pMLST) analysis, a database that is updated weekly was generated from www.pubmlst.org and integrated into a Web tool called pMLST. Both databases were evaluated using draft genomes from a collection...

  18. Using the Viking biology experimental results to obtain chemical information about Martian regolith

    Science.gov (United States)

    Plumb, Robert C.

    1992-01-01

    Although initially formulated as biology experiments, most of the results produced by the Viking Labeled Release (LR), Gas Exchange (GEX), and Pyrolytic Release (PR) experiments have been reproduced by chemical means. The experiments do not need more study as 'biological' phenomena, but they do deserve much more careful consideration from a chemical viewpoint. They are the only 'wet-chemical' experiments that scientists have performed on another planet, but they have not found very general use as sources of scientific information. There is a large set of potentially useful chemical observations, e.g., the three resolvable and precisely measured kinetic components of the release of C-14-labeled gases, the thermal sensitivity and magnitudes of the oxidation reaction(s) of the LR experiments, the kinetics and magnitude of the O2 and CO2 release of the GEX experiments, the thermal sensitivity of the GEX results, the differences between the thermal sensitivity of the GEX and the thermal sensitivity of the LR responses, and the kinetics and magnitudes of the LR successive injection reabsorption effect. It should be possible to test many chemical aspects of hypothetical martian phenomena in experiments using the biology experimental configurations and derive much valuable information by comparisons with the Viking observations.

  19. Report of test and research results on atomic energy obtained in national institutes in fiscal 1990

    International Nuclear Information System (INIS)

    1991-01-01

    The tests and researches on the development and utilization of atomic energy in national laboratories were begun in 1956, and have accomplished the great role for the advance of the development and utilization of atomic energy in Japan by having produced many valuable results so far. Atomic energy has been utilized in diverse fields, and also in national laboratories, the research for expanding the development and utilization of atomic energy in food irradiation, medicine, agriculture, forestry, fishery and others in addition to the basic research on nuclear fusion and safety have been advanced. Further expecting the pervasive effect to general science and technology, the development of basic technology and integrated research are promoted from the viewpoint of new techical innovation and creative technology. This is 31st report in which the results of the tests and researches carried out by national laboratories in fiscal year 1990 are summarized. Nuclear fusion, safety research, food irradiation, cancer countermeasures, agriculture, forestry, fishery, medicine, mining and manufacture, power utilization, construction, radioactivation analysis and so on were the main subjects. (K.I.)

  20. Report of test and research results on atomic energy obtained in national institutes in fiscal 1982

    International Nuclear Information System (INIS)

    1983-01-01

    As for the test and research on the utilization of atomic energy by national organizations, the budget was appropriated for the first time in fiscal year 1956. Since then, many valuable results of research have been produced in the diverse fields of nuclear fusion, safety research, food irradiation, medicine and others, in this way, the test and research have played large roles in the promotion of the utilization of atomic energy in Japan. This is the 23rd report, in which the results of the test and research on the utilization of atomic energy carried out in fiscal year 1982 by national organizations are summarized. 5 researches on nuclear fusion, 12 researches on engineering safety, 5 researches on environmental radioactivity safety, 3 researches on food irradiation, 5 researches on the countermeasures to cancer, 8 researches on soil fertilization, 4 researches on quality improvement, 7 researches on crop protection, 5 researches on the improvement of breeding, 8 researches on diagnosis and treatment, 8 researches on pharmaceuticals, 10 researches on the application to pathology, 6 researches on mining and industry, 6 researches on power reactors and nuclear ships, 1 research on underground water, 6 researches on activation analysis and 3 researches on injury prevention are reported. (Kako, I.)

  1. Report of test and research results on atomic energy obtained in national institutes in fiscal 1983

    International Nuclear Information System (INIS)

    1984-01-01

    As for the test and research on the utilization of atomic energy by national organizations, the budget was appropriated for the first time in fiscal year 1956. Since then, many valuable results of research have been produced in the diverse fields of nuclear fusion, safety research, food irradiation, medicine and others, in this way, the test and research have played large roles in the promotion of the utilization of atomic energy in Japan. This is the 24th report, in which the results of the test and research on the utilization of atomic energy carried out in fiscal year 1983 by national organizations are summarized. 5 researches on nuclear fusion, 19 researches on engineering safety and environmental radioactivity safety, 3 researches on food irradiation, 6 researches on the countermeasures to cancer, 19 researches on agriculture, forestry and fishery, 30 researches on medicine, pharmaceuticals and environmental hygiene, 6 researches on mining and industry, 6 researches on power reactors and nuclear ships, 1 research on agricultural water, 7 researches on activation analysis and 4 researches on injury prevention are reported. (Kako, I.)

  2. Aircrew Exposure To Cosmic Radiation Evaluated By Means Of Several Methods; Results Obtained In 2006

    International Nuclear Information System (INIS)

    Ploc, Ondrej; Spurny, Frantisek; Jadrnickova, Iva; Turek, Karel

    2008-01-01

    Routine evaluation of aircraft crew exposure to cosmic radiation in the Czech Republic is performed by means of calculation method. Measurements onboard aircraft work as a control tool of the routine method, as well as a possibility of comparison of results measured by means of several methods. The following methods were used in 2006: (1) mobile dosimetry unit (MDU) type Liulin--a spectrometer of energy deposited in Si-detector; (2) two types of LET spectrometers based on the chemically etched track detectors (TED); (3) two types of thermoluminescent detectors; and (4) two calculation methods. MDU represents currently one of the most reliable equipments for evaluation of the aircraft crew exposure to cosmic radiation. It is an active device which measures total energy depositions (E dep ) in the semiconductor unit, and, after appropriate calibration, is able to give a separate estimation for non-neutron and neutron-like components of H*(10). This contribution consists mostly of results acquired by means of this equipment; measurements with passive detectors and calculations are mentioned because of comparison. Reasonably good agreement of all data sets could be stated

  3. Report of test and research results on atomic energy obtained in national institutes in fiscal 1992

    International Nuclear Information System (INIS)

    1994-01-01

    The tests and researches on the development and utilization of atomic energy in national laboratories were begun in 1956, and have accomplished the great role for the advance of the development and utilization of atomic energy in Japan by having produced many valuable results so far. Atomic energy has been utilized not only in the field of nuclear power but also in diverse fields, and in national laboratories, the research for expanding the development and utilization of atomic energy in medicine, agriculture, forestry, fishery, radioactivation analysis and others in addition the basic research on nuclear fusion have been advanced. Further expecting the pervasive effect to general science and technology, the development of integrated research are promoted from the viewpoint of new technical innovation and creative technology. The safety research of nuclear facilities have been carried out to keep them high level on the basis of the yearly program enacted by Nuclear Safety Commission. This is the report No. 33, in which the results of the test and research in the fields of nuclear fusion safety research, food irradiation, cancer countermeasures, agriculture, forestry, fishery, medicine, mining and manufacture, power utilization, construction, radioactivation analysis carried on in fiscal 1992 are summarized. (J.P.N.)

  4. Operating results obtained in a nuclear power plant with a sensor surveillance prototype

    International Nuclear Information System (INIS)

    Jacquot, J.P.; Poujol, A.; Beaubatie, J.; Ciaramitaro, W.

    1983-03-01

    Surveillance methods have been validated and specific equipment have been built to measure the response time of sensors from a nuclear power plant protection channel. The reason of the choice of this parameter is twofold: the sensor response time is representative of the sensor physical status and is also part of the overall channel response time. Two surveillance methods are used: noise analysis (by AR or PSD modeling), and loop current step response (for resistance thermometer detectors only). The methods were validated on test facilities and on nuclear power plants. Two test equipments were built and tested on plants. Results are represented and conclusions are drawn on the feasibility of such methods for sensor surveillance [fr

  5. 2D and 3D core-collapse supernovae simulation results obtained with the CHIMERA code

    Energy Technology Data Exchange (ETDEWEB)

    Bruenn, S W; Marronetti, P; Dirk, C J [Physics Department, Florida Atlantic University, 777 W. Glades Road, Boca Raton, FL 33431-0991 (United States); Mezzacappa, A; Hix, W R [Physics Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831-6354 (United States); Blondin, J M [Department of Physics, North Carolina State University, Raleigh, NC 27695-8202 (United States); Messer, O E B [Center for Computational Sciences, Oak Ridge National Laboratory, Oak Ridge, TN 37831-6354 (United States); Yoshida, S, E-mail: bruenn@fau.ed [Max-Planck-Institut fur Gravitationsphysik, Albert Einstein Institut, Golm (Germany)

    2009-07-01

    Much progress in realistic modeling of core-collapse supernovae has occurred recently through the availability of multi-teraflop machines and the increasing sophistication of supernova codes. These improvements are enabling simulations with enough realism that the explosion mechanism, long a mystery, may soon be delineated. We briefly describe the CHIMERA code, a supernova code we have developed to simulate core-collapse supernovae in 1, 2, and 3 spatial dimensions. We then describe the results of an ongoing suite of 2D simulations initiated from a 12, 15, 20, and 25 M{sub o-dot} progenitor. These have all exhibited explosions and are currently in the expanding phase with the shock at between 5,000 and 20,000 km. We also briefly describe an ongoing simulation in 3 spatial dimensions initiated from the 15 M{sub o-dot} progenitor.

  6. THE ANALYSIS OF SEVERAL RESULTS OBTAINED BY ROMANIA IN THE FIELD OF SUSTAINABLE TRANSPORT

    Directory of Open Access Journals (Sweden)

    Perțicaș Diana Claudia

    2015-07-01

    Full Text Available We know that between the transport sector and all other branches of economy there is a strong interdependence link but also between it and the surrounding environment, being one of the most polluting sectors of activity. Transport is considered a primary field in any national economy development context, especially if we take into account its interdependence with other branches of national economy. Developing of transports also includes improving road, rail, river and sea services, as well as air transports. The objectives of the EU aim especially to modernize the transport infrastructures, be them by road, ship or by air, which would result in increasing the speed of freight transport, fluidizing traffic, attracting new foreign investors in various areas, accelerating the renewal of the auto park and decommissioning morally and physically worn vehicles which are extremely pollutant, the revival of maritime transports through Romanian ports, progressive completion of imposed performances through standards and regulations on the transport market, etc. All these objectives have as a main purpose the reducing of energy consumption, reducing transport costs as well as increasing competition in the national transport system.The development of transport has the role of stimulating public transport services and to guarantee a minimum general accessibility to public services for all citizens. Children, the elderly, disabled people or other vulnerable categories of people are not and will not be forgotten, for which certain standards set by the European Union must be respected.The present paper wishes to analyze a part of the results, either positive or negative, in the field of transports, made by our country.

  7. Comparison of different scenarios for the deployment of fast reactors in France. Results obtained with COSI

    International Nuclear Information System (INIS)

    Coquelet-Pascal, Christine; Meyer, Maryan; Girieud, Richard

    2011-01-01

    In the frame of the French Act for waste management, scenarios studies are carried out with the simulation software COSI to compare different options of evolution of the French reactor fleet, possibilities of plutonium recycling, and options of separation and transmutation of minor actinides. The goal of these studies is to evaluate the sustainability of Sodium cooled Fast Reactors (SFR) deployment from plutonium availability viewpoint, as well as the interest of minor actinides transmutation options in SFR and associated impacts (decay heat and toxicity) on cycle facilities and geological storage. Each option has been evaluated separately in dynamic scenarios taking into account the transition between the current nuclear reactor fleet and a generation IV fleet, with the deployment of SFR in replacement of PWR. In this paper, the results of three types of scenarios, in the continuity of the paper of GLOBAL 2009, are discussed: 1 - plutonium recycling in SFR; 2 - plutonium recycling and minor actinides transmutation in SFR; 3 - plutonium recycling and americium transmutation in SFR. MA transmutation in heterogeneous mode (named 'het.') and in homogeneous mode (named 'hom.') are distinguished. By comparison with the previous paper, new scenarios and extended results are added and a global comparison between transmutation performances is done. Scenarios have been optimized to minimize the impacts on fuel cycle: stabilization of plants capacities over 40 years, limitation of minor actinides storage in the case of heterogeneous transmutation, limitation of MA content in the case of homogeneous transmutation. The impact of MA transmutation on decay heat of fresh and spent fuel are also assessed. The selected scenarios bring also elements on the impact of a SFR deployment delayed from 2040 to 2080, with or without MA transmutation, and the impact of an increase in the total power capacity installed by maximizing the SFR share in a symbiotic fleet PWR-SFR. Whatever the

  8. Gravity Changes and Internal Processes: Some Results Obtained from Observations at Three Volcanoes

    Science.gov (United States)

    Jentzsch, Gerhard; Weise, Adelheid; Rey, Carlos; Gerstenecker, Carl

    Temporal gravity changes provide information about mass and/or density variations within and below the volcano edifice. Three active volcanoes have been under investigation; each of them related to a plate boundary: Mayon/Luzon/Philippines, Merapi/Java/Indonesia, and Galeras/Colombia. The observed gravity changes are smaller than previously expected but significant. For the three volcanoes under investigation, and within the observation period, mainly the increase of gravity is observed, ranging from 1,000 nm-2 to 1,600 nms-2. Unexpectedly, the gravity increase is confined to a rather small area with radii of 5 to 8 km around the summit. At Mayon and Merapi the parallel GPS measurements yield no significant elevation changes. This is crucial for the interpretation, as the internal pressure variations do not lead to significant deformation at the surface. Thus the classical Mogi-model for a shallow extending magma reservoir cannot apply. To confine the possible models, the attraction due to changes of groundwater level or soil moisture is estimated along the slope of Merapi exemplarily by 2-D modelling. Mass redistribution or density changes were evaluated within the vent as well as deeper fluid processes to explain the gravity variations; the results are compared to the model incorporating the additional effect of elastic deformation.

  9. Development, calibration and experimental results obtained with an innovative calorimeter (CALMOS) for nuclear heating measurements

    International Nuclear Information System (INIS)

    Carcreff, H.; Cloute-Cazalaa, V.; Salmon, L.

    2011-01-01

    Nuclear heating inside an MTR reactor has to be known in order to be able to control samples temperature during irradiation experiments. An R and D program has been carried out at CEA to design a new type of in-core calorimetric system. This new development, started in 2002, has for main objective to manufacture a calorimeter suitable to monitoring nuclear heating inside the 70 MWth OSIRIS material testing reactor operated by CEA's Nuclear Energy Div. at the Saclay research center. An innovative calorimetric probe, associated to a specific handling system, has been designed to provide access to measurements both along the fissile height and on the upper part of the core, where nuclear heating still remains high. Two mock-ups of the probe were manufactured and tested in 2005 and 2009 in ex-core area of OSIRIS reactor for process validation, while a displacement system has been especially studied to move the probe along a given axial measurement range. This paper deals with the development, tests on preliminary mock-ups and the finalization of the probe. Main modeling and experimental results are presented. Moreover, alternative methods to calibration for nuclear heating rate measurements which are now possible with this new calorimeter are presented and discussed. (authors)

  10. Development, calibration, and experimental results obtained with an innovative calorimeter (CALMOS) for nuclear heating measurements

    International Nuclear Information System (INIS)

    Carcreff, Hubert; Cloute-Cazalaa, Veronique; Salmon, Laurent

    2012-01-01

    Nuclear heating inside an MTR reactor has to be known in order to be able to control samples temperature during irradiation experiments. An R and D program has been carried out at CEA to design a new type of in-core calorimetric system. This new development, started in 2002, has for main objective to manufacture a calorimeter suitable to monitoring nuclear heating inside the 70 MWth OSIRIS material testing reactor operated by CEA's Nuclear Energy Division at the Saclay research center. An innovative calorimetric probe, associated to a specific handling system, has been designed to provide access to measurements both along the fissile height and on the upper part of the core, where nuclear heating still remains high. Two mock-ups of the probe were manufactured and tested in 2005 and 2009 in ex-core area of OSIRIS reactor for process validation, while a displacement system has been especially studied to move the probe along a given axial measurement range. This paper deals with the development, tests on preliminary mock-ups and the finalization of the probe. Main modeling and experimental results are presented. Moreover, alternative methods to calibration for nuclear heating rate measurements which are now possible with this new calorimeter are presented and discussed. (authors)

  11. Preliminary physico-chemical results obtained on water using new data acquisition systems for deep wells

    International Nuclear Information System (INIS)

    Vinson, J.M.; Peyrus, J.C.

    1984-02-01

    Data acquisition systems recently developed in the context of research on deep storage facilities have provided with an initial set of interesting observations for the physico-chemical study of boreholes. It is possible to make correlations between the chemical compositions of water, pH and the nature of the substrate. The sampling done at Auriat with a Gerhardt-Owen probe shows the variability in the composition of water as a function of depth. The variation in calcium content, following that of pH, is particularly notable. Examination of pH measurements is of particular interest. A general gradient correlates exactly with the nature of the substrate. Whereas steel piping has a very alkaline pH, distinct pH values correspond to the two types of granite substrate. In this general gradient, series of disturbances can be seen which correspond perfectly to fracturation zones or large fractures. These most promising preliminary results lead to believe that in situ physico-chemical measurements should be continued and developed with a view to improved evaluation of the safety of deep storage facilities

  12. Latest results of SEE measurements obtained by the STRURED demonstrator ASIC

    Energy Technology Data Exchange (ETDEWEB)

    Candelori, A. [INFN, Section of Padova, Via Marzolo 8, c.a.p. 35131, Padova (Italy); De Robertis, G. [INFN Section of Bari, Via Orabona 4, c.a.p. 70126, Bari (Italy); Gabrielli, A. [Physics Department, University of Bologna, Viale Berti Pichat 6/2, c.a.p. 40127, Bologna (Italy); Mattiazzo, S.; Pantano, D. [INFN, Section of Padova, Via Marzolo 8, c.a.p. 35131, Padova (Italy); Ranieri, A., E-mail: antonio.ranieri@ba.infn.i [INFN Section of Bari, Via Orabona 4, c.a.p. 70126, Bari (Italy); Tessaro, M. [INFN, Section of Padova, Via Marzolo 8, c.a.p. 35131, Padova (Italy)

    2011-01-21

    With the perspective to develop a radiation-tolerant circuit for High Energy Physics (HEP) applications, a test digital ASIC VLSI chip, called STRURED, has been designed and fabricated using a standard-cell library of commercial 130 nm CMOS technology by implementing three different radiation-tolerant architectures (Hamming, Triple Modular Redundancy and Triple Time Redundancy) in order to correct circuit malfunctions induced by the occurrence of Soft Errors (SEs). SEs are one of the main reasons of failures affecting electronic digital circuits operating in harsh radiation environments, such as in experiments performed at HEP colliders or in apparatus to be operated in space. In this paper we present and discuss the latest results of SE cross-section measurements performed using the STRURED digital device, exposed to high energy heavy ions at the SIRAD irradiation facility of the INFN National Laboratories of Legnaro (Padova, Italy). In particular the different behaviors of the input part and the core of the three radiation-tolerant architectures are analyzed in detail.

  13. Restriction Fragment Length Polymorphisms of Virulence Plasmids in Rhodococcus equi

    Science.gov (United States)

    Takai, Shinji; Shoda, Masato; Sasaki, Yukako; Tsubaki, Shiro; Fortier, Guillaume; Pronost, Stephane; Rahal, Karim; Becu, Teotimo; Begg, Angela; Browning, Glenn; Nicholson, Vivian M.; Prescott, John F.

    1999-01-01

    Virulent Rhodococcus equi, which is a well-known cause of pyogranulomatous pneumonia in foals, possesses a large plasmid encoding virulence-associated 15- to 17-kDa antigens. Foal and soil isolates from five countries—Argentina, Australia, Canada, France, and Japan—were investigated for the presence of 15- to 17-kDa antigens by colony blotting, using the monoclonal antibody 10G5, and the gene coding for 15- to 17-kDa antigens by PCR. Plasmid DNAs extracted from positive isolates were digested with restriction endonucleases BamHI, EcoRI, EcoT22I, and HindIII, and the digestion patterns that resulted divided the plasmids of virulent isolates into five closely related types. Three of the five types had already been reported in Canadian and Japanese isolates, and the two new types had been found in French and Japanese isolates. Therefore, we tentatively designated these five types 85-kb type I (pREAT701), 85-kb type II (a new type), 87-kb type I (EcoRI and BamHI type 2 [V. M. Nicholson and J. F. Prescott, J. Clin. Microbiol. 35:738–740, 1997]), 87-kb type II (a new type), and 90-kb (pREL1) plasmids. The 85-kb type I plasmid was found in isolates from Argentina, Australia, Canada, and France. Plasmid 87-kb type I was isolated in specimens from Argentina, Canada, and France. The 85-kb type II plasmid appeared in isolates from France. On the other hand, plasmids 87-kb type II and 90-kb were found only in isolates from Japan. These results revealed geographic differences in the distribution of the virulence plasmids found in the five countries and suggested that the restriction fragment length polymorphism of virulence plasmids might be useful to elucidate the molecular epidemiology of virulent R. equi in the world. PMID:10488224

  14. Multi-Grid detector for neutron spectroscopy: results obtained on time-of-flight spectrometer CNCS

    Science.gov (United States)

    Anastasopoulos, M.; Bebb, R.; Berry, K.; Birch, J.; Bryś, T.; Buffet, J.-C.; Clergeau, J.-F.; Deen, P. P.; Ehlers, G.; van Esch, P.; Everett, S. M.; Guerard, B.; Hall-Wilton, R.; Herwig, K.; Hultman, L.; Höglund, C.; Iruretagoiena, I.; Issa, F.; Jensen, J.; Khaplanov, A.; Kirstein, O.; Lopez Higuera, I.; Piscitelli, F.; Robinson, L.; Schmidt, S.; Stefanescu, I.

    2017-04-01

    The Multi-Grid detector technology has evolved from the proof-of-principle and characterisation stages. Here we report on the performance of the Multi-Grid detector, the MG.CNCS prototype, which has been installed and tested at the Cold Neutron Chopper Spectrometer, CNCS at SNS. This has allowed a side-by-side comparison to the performance of 3He detectors on an operational instrument. The demonstrator has an active area of 0.2 m2. It is specifically tailored to the specifications of CNCS. The detector was installed in June 2016 and has operated since then, collecting neutron scattering data in parallel to the He-3 detectors of CNCS. In this paper, we present a comprehensive analysis of this data, in particular on instrument energy resolution, rate capability, background and relative efficiency. Stability, gamma-ray and fast neutron sensitivity have also been investigated. The effect of scattering in the detector components has been measured and provides input to comparison for Monte Carlo simulations. All data is presented in comparison to that measured by the 3He detectors simultaneously, showing that all features recorded by one detector are also recorded by the other. The energy resolution matches closely. We find that the Multi-Grid is able to match the data collected by 3He, and see an indication of a considerable advantage in the count rate capability. Based on these results, we are confident that the Multi-Grid detector will be capable of producing high quality scientific data on chopper spectrometers utilising the unprecedented neutron flux of the ESS.

  15. Simbol-X Hard X-ray Focusing Mirrors: Results Obtained During the Phase A Study

    International Nuclear Information System (INIS)

    Tagliaferri, G.; Basso, S.; Civitani, M.; Conconi, P.; Cotroneo, V.; Pareschi, G.; Spiga, D.; Borghi, G.; Garoli, D.; Mattarello, V.; Orlandi, A.; Valsecchi, G.; Vernani, D.; Burkert, W.; Freyberg, M.; Hartner, G.; Citterio, O.; Gorenstein, P.; Romaine, S.

    2009-01-01

    Simbol-X will push grazing incidence imaging up to 80 keV, providing a strong improvement both in sensitivity and angular resolution compared to all instruments that have operated so far above 10 keV. The superb hard X-ray imaging capability will be guaranteed by a mirror module of 100 electroformed Nickel shells with a multilayer reflecting coating. Here we will describe the technogical development and solutions adopted for the fabrication of the mirror module, that must guarantee an Half Energy Width (HEW) better than 20 arcsec from 0.5 up to 30 keV and a goal of 40 arcsec at 60 keV. During the phase A, terminated at the end of 2008, we have developed three engineering models with two, two and three shells, respectively. The most critical aspects in the development of the Simbol-X mirrors are i) the production of the 100 mandrels with very good surface quality within the timeline of the mission, ii) the replication of shells that must be very thin (a factor of 2 thinner than those of XMM-Newton) and still have very good image quality up to 80 keV, iii) the development of an integration process that allows us to integrate these very thin mirrors maintaining their intrinsic good image quality. The Phase A study has shown that we can fabricate the mandrels with the needed quality and that we have developed a valid integration process. The shells that we have produced so far have a quite good image quality, e.g. HEW < or approx. 30 arcsec at 30 keV, and effective area. However, we still need to make some improvements to reach the requirements. We will briefly present these results and discuss the possible improvements that we will investigate during phase B.

  16. Simbol-X Hard X-ray Focusing Mirrors: Results Obtained During the Phase A Study

    Science.gov (United States)

    Tagliaferri, G.; Basso, S.; Borghi, G.; Burkert, W.; Citterio, O.; Civitani, M.; Conconi, P.; Cotroneo, V.; Freyberg, M.; Garoli, D.; Gorenstein, P.; Hartner, G.; Mattarello, V.; Orlandi, A.; Pareschi, G.; Romaine, S.; Spiga, D.; Valsecchi, G.; Vernani, D.

    2009-05-01

    Simbol-X will push grazing incidence imaging up to 80 keV, providing a strong improvement both in sensitivity and angular resolution compared to all instruments that have operated so far above 10 keV. The superb hard X-ray imaging capability will be guaranteed by a mirror module of 100 electroformed Nickel shells with a multilayer reflecting coating. Here we will describe the technogical development and solutions adopted for the fabrication of the mirror module, that must guarantee an Half Energy Width (HEW) better than 20 arcsec from 0.5 up to 30 keV and a goal of 40 arcsec at 60 keV. During the phase A, terminated at the end of 2008, we have developed three engineering models with two, two and three shells, respectively. The most critical aspects in the development of the Simbol-X mirrors are i) the production of the 100 mandrels with very good surface quality within the timeline of the mission, ii) the replication of shells that must be very thin (a factor of 2 thinner than those of XMM-Newton) and still have very good image quality up to 80 keV, iii) the development of an integration process that allows us to integrate these very thin mirrors maintaining their intrinsic good image quality. The Phase A study has shown that we can fabricate the mandrels with the needed quality and that we have developed a valid integration process. The shells that we have produced so far have a quite good image quality, e.g. HEW <~30 arcsec at 30 keV, and effective area. However, we still need to make some improvements to reach the requirements. We will briefly present these results and discuss the possible improvements that we will investigate during phase B.

  17. Comparison of NDT techniques to evaluate CFRP. Results obtained in a MAIzfp round robin test

    Energy Technology Data Exchange (ETDEWEB)

    Grosse, Christian U. [Technische Univ. Muenchen (Germany). Chair of Non-destructive Testing; Goldammer, Matthias; Grager, Jan-Carl [Siemens AG Corporate Technology, Muenchen (Germany); and others

    2016-10-01

    Fiber reinforced polymeric materials are used for lightweight constructions and are an integral part of cars, airplanes or rotor blades of wind turbines. Nondestructive testing (NDT) methods play an increasing role concerning the manufacturing process and the inspection during lifetime. The selection of the best NDT technique for a certain application depends - of course - on many factors including the type, position and size of the defect to be detected but also on secondary issues like accessibility, automation, testing costs, reliability and resolution to mention only some. For the more technical-scientific part of these issues, the determination of the probability of detection (PoD) plays a significant role. Early in the design process questions should be raised concerning the probability with which certain attribute of interest (a defect that has an effect on the structural behavior) can be detected (and localized) in a certain construction. Several defect types have been identified to be critical like impact damages, undulations and porosity. Test samples out of differently processed Carbon Fiber-Reinforced Polymers (CFRP) as used in the automotive or aeronautical industry have been produced including defects of different type and size. In order to determine the PoD and to check whether a technique is applicable the different partners applied a broad variety of selected NDT techniques including Micro CT, Ultrasound (including phased-array and air-coupled UT), Active Thermography, Eddy Current, Vibration and Visual Analysis and Local Acoustic Resonance Spectroscopy (LARS). The presentation will summarize some of the results of the experiments and ongoing data analysis.

  18. Comparison of NDT techniques to evaluate CFRP. Results obtained in a MAIzfp round robin test

    International Nuclear Information System (INIS)

    Grosse, Christian U.

    2016-01-01

    Fiber reinforced polymeric materials are used for lightweight constructions and are an integral part of cars, airplanes or rotor blades of wind turbines. Nondestructive testing (NDT) methods play an increasing role concerning the manufacturing process and the inspection during lifetime. The selection of the best NDT technique for a certain application depends - of course - on many factors including the type, position and size of the defect to be detected but also on secondary issues like accessibility, automation, testing costs, reliability and resolution to mention only some. For the more technical-scientific part of these issues, the determination of the probability of detection (PoD) plays a significant role. Early in the design process questions should be raised concerning the probability with which certain attribute of interest (a defect that has an effect on the structural behavior) can be detected (and localized) in a certain construction. Several defect types have been identified to be critical like impact damages, undulations and porosity. Test samples out of differently processed Carbon Fiber-Reinforced Polymers (CFRP) as used in the automotive or aeronautical industry have been produced including defects of different type and size. In order to determine the PoD and to check whether a technique is applicable the different partners applied a broad variety of selected NDT techniques including Micro CT, Ultrasound (including phased-array and air-coupled UT), Active Thermography, Eddy Current, Vibration and Visual Analysis and Local Acoustic Resonance Spectroscopy (LARS). The presentation will summarize some of the results of the experiments and ongoing data analysis.

  19. The main tasks and obtained results within soil protection working group of the Danube countries

    International Nuclear Information System (INIS)

    Dzatko, M.

    1997-01-01

    In the frame of the Danube Countries Working Community activities was in 1993 constituted independent Soil Protection Working Group (SPWG). Its primary task is to elaborate principles and common soil protection concept in given countries accepted on the level of governments and related authorities, and also to the solution of the solution of the problems on regional levels. Final objective is to implement such concept of soil protection policy, which is able to maintain its quality and productivity potential for next generations also. Based on four years activities coordination could be significance and the SPWG relevance expressed in following topics: (1) Soil pollution and soil degradation, particularly in most pos-communistic countries has been attaining high degree, and in many locations also the threshold of ecological be arability. As reclamation and revitalization of the degraded and polluted soils require long time and considerable financial means, it is not only moral;, even also economically more effective to protect preventively than subsequent reclamation. (2) Main objective of the correct soil protection policy should be its high quality conservation also for the next generations. To this is joined also the protection not only productional, but also non-productional functions, particularly filtrational, transformational and buffering capacibility, including its role and significance, as land and environment. From, in this way considered relationships reality is resulting that the soil protection objectives are not only laws and prohibition approvement, but also active relationships harmonization between the man requirements and soil productivity potential, in order of the sustainable land resources use for the next generations. (3) Based on mentioned realities and relationships we consider the SPWG as an active gremium for elaboration of the the uniform soil protection concepts for governments and responsible organisations that in financial consequence

  20. Implantation of peritoneal catheters by laparotomy: nephrologists obtained similar results to general surgeons

    Directory of Open Access Journals (Sweden)

    Restrepo CA

    2014-10-01

    Full Text Available Cesar A Restrepo, Carlos Alberto Buitrago, Cielo Holguin Division of Nephrology, Department of Health Sciences, Caldas University, Caldas, ColombiaPurpose: To analyze the complications and costs of minilaparotomies performed by a nephrologist (group A compared with conventional laparotomies performed by a surgeon (group B for peritoneal catheter implantation.Setting: Two university hospitals (Santa Sofia and Caldas in Manizales, Caldas, Colombia.Methods: The study included stage 5 chronic kidney disease patients, with indication of renal replacement therapy, who were candidates for peritoneal dialysis and gave informed consent for a peritoneal catheter implant. Minilaparotomies were performed by a nephrologist in a minor surgery room under local anesthesia. Conventional laparotomies were performed by a surgeon in an operating room under general anesthesia.Results: Two nephrologists inserted 157 peritoneal catheters, and seven general surgeons inserted 185 peritoneal catheters. The groups had similar characteristics: the mean age was 55 years, 49.5% were men, and the primary diagnoses were diabetic nephropathy, hypertensive nephropathy, and unknown etiology. The implant was successful for 98.09% of group A and 99.46% of group B. There was no procedure-related mortality. The most frequent complications in the first 30 days postsurgery in group A versus group B, respectively, were: peritonitis (6.37% versus 3.78%, exit-site infection (3.82% versus 2.16%, tunnel infection (0% versus 0.54%, catheter entrapment by omentum (1.27% versus 3.24%, peritoneal effluent spillover (1.91% versus 2.16%, draining failure (4.46% versus 6.49%, hematoma (0% versus 1.08%, catheter migration with kinking (3.18% versus 2.70%, hemoperitoneum (1.27% versus 0%, and hollow viscera accidental puncture (1.91% versus 0.54%. There were no statistically significant differences in the number of complications between groups. In 2013, the cost of a surgeon-implanted peritoneal

  1. Study liquid length penetration results obtained with a direct acting piezo electric injector

    International Nuclear Information System (INIS)

    Payri, Raul; Gimeno, Jaime; Bardi, Michele; Plazas, Alejandro H.

    2013-01-01

    Highlights: ► A direct acting injector capable of controlling needle lift has been used to determine liquid phase penetration. ► The influence of injection pressure, chamber density and chamber temperature have been measured. ► When needle lift is reduced the stabilized liquid length is shortened. ► The relationship between needle lift and liquid length makes needle lift as a new way to control the injection event. - Abstract: A state of the art prototype common rail injector featuring direct control of the needle by means of a piezo stack (direct acting) has been tested. Liquid phase penetration of the sprays in diesel engine-like conditions has been studied via imaging technique in a novel continuous flow test chamber that allows an accurate control on a wide range of thermodynamic conditions (up to 1000 K and 15 MPa). This state of the art injector fitted with a 7-hole nozzle, allows a fully flexible control on the nozzle needle movement, enabling various fuel injection rate typologies. The temporal evolution of the seven sprays has been studied recording movies of the injection event in evaporative conditions via Mie scattering imaging technique and using a high speed camera. The results showed a strong influence of needle position on the stabilized liquid length while the effect of the injection pressure is negligible: the decrease of the needle lift causes a pressure drop in the needle seat and thus a reduction in the effective pressure upstream of the orifices (in the nozzle sac). According to known literature the stabilized liquid-length depends mainly on effective diameter, spray cone-angle and fuel/air properties and does not depend on fuel velocity at the orifice outlet. Therefore, due to small change in the spray cone-angle, higher injection pressures give slightly lower liquid length. However, partial needle lifts has an opposite effect: when needle is partially lifted a dramatic increase of the spray cone-angle and a consequent reduction of

  2. Results from CrIS-ATMS Obtained Using the AIRS Science Team Retrieval Methodology

    Science.gov (United States)

    Susskind, Joel; Kouvaris, Louis C.; Iredell, Lena

    2013-01-01

    which significantly improved results of AIRS Version-6. Version-5.70 CrIS/ATMS temperature profile and surface skin temperature retrievals are of very good quality, and are better than AIRS Version-5 retrievals, but are still significantly poorer than those of AIRS Version-6. CrIS/ATMS retrievals should improve when a Neural-Net start-up system is ready for use. We also examined CrIS/ATMS retrievals generated by NOAA using their NUCAPS retrieval algorithm, which is based on earlier versions of the AIRS Science Team retrieval algorithms. We show that the NUCAPS algorithm as currently configured is not well suited for climate monitoring purposes.

  3. Plasmid and chromosome partitioning: surprises from phylogeny

    DEFF Research Database (Denmark)

    Gerdes, Kenn; Møller-Jensen, Jakob; Bugge Jensen, Rasmus

    2000-01-01

    Plasmids encode partitioning genes (par) that are required for faithful plasmid segregation at cell division. Initially, par loci were identified on plasmids, but more recently they were also found on bacterial chromosomes. We present here a phylogenetic analysis of par loci from plasmids and chr...

  4. A plasmid containing the human metallothionein II gene can function as an antibody-assisted electrophoretic biosensor for heavy metals.

    Science.gov (United States)

    Wooten, Dennis C; Starr, Clarise R; Lyon, Wanda J

    2016-01-01

    Different forms of heavy metals affect biochemical systems in characteristic ways that cannot be detected with typical metal analysis methods like atomic absorption spectrometry. Further, using living systems to analyze interaction of heavy metals with biochemical systems can be laborious and unreliable. To generate a reliable easy-to-use biologically-based biosensor system, the entire human metallothionein-II (MT-II) gene was incorporated into a plasmid (pUC57-MT) easily replicated in Escherichia coli. In this system, a commercial polyclonal antibody raised against human metal-responsive transcription factor-1 protein (MTF-1 protein) could modify the electrophoretic migration patterns (i.e. cause specific decreases in agarose gel electrophoretic mobility) of the plasmid in the presence or absence of heavy metals other than zinc (Zn). In the study here, heavy metals, MTF-1 protein, and polyclonal anti-MTF-1 antibody were used to assess pUC57-MT plasmid antibody-assisted electrophoretic mobility. Anti-MTF-1 antibody bound both MTF-1 protein and pUC57-MT plasmid in a non-competitive fashion such that it could be used to differentiate specific heavy metal binding. The results showed that antibody-inhibited plasmid migration was heavy metal level-dependent. Zinc caused a unique mobility shift pattern opposite to that of other metals tested, i.e. Zn blocked the antibody ability to inhibit plasmid migration, despite a greatly increased affinity for DNA by the antibody when Zn was present. The Zn effect was reversed/modified by adding MTF-1 protein. Additionally, antibody inhibition of plasmid mobility was resistant to heat pre-treatment and trypsinization, indicating absence of residual DNA extraction-resistant bacterial DNA binding proteins. DNA binding by anti-DNA antibodies may be commonly enhanced by xenobiotic heavy metals and elevated levels of Zn, thus making them potentially effective tools for assessment of heavy metal bioavailability in aqueous solutions and

  5. AAVS1-Targeted Plasmid Integration in AAV Producer Cell Lines.

    Science.gov (United States)

    Luo, Yuxia; Frederick, Amy; Martin, John M; Scaria, Abraham; Cheng, Seng H; Armentano, Donna; Wadsworth, Samuel C; Vincent, Karen A

    2017-06-01

    Adeno-associated virus (AAV) producer cell lines are created via transfection of HeLaS3 cells with a single plasmid containing three components (the vector sequence, the AAV rep and cap genes, and a selectable marker gene). As this plasmid contains both the cis (Rep binding sites) and trans (Rep protein encoded by the rep gene) elements required for site-specific integration, it was predicted that plasmid integration might occur within the AAVS1 locus on human chromosome 19 (chr19). The objective of this study was to investigate whether integration in AAVS1 might be correlated with vector yield. Plasmid integration sites within several independent cell lines were assessed via Southern, fluorescence in situ hybridization (FISH) and PCR analyses. In the Southern analyses, the presence of fragments detected by both rep- and AAVS1-specific probes suggested that for several mid- and high-producing lines, plasmid DNA had integrated into the AAVS1 locus. Analysis with puroR and AAVS1-specific probes suggested that integration in AAVS1 was a more widespread phenomenon. High-producing AAV2-secreted alkaline phosphatase (SEAP) lines (masterwell 82 [MW82] and MW278) were evaluated via FISH using probes specific for the plasmid, AAVS1, and a chr19 marker. FISH analysis detected two plasmid integration sites in MW278 (neither in AAVS1), while a total of three sites were identified in MW82 (two in AAVS1). An inverse PCR assay confirmed integration within AAVS1 for several mid- and high-producing lines. In summary, the FISH, Southern, and PCR data provide evidence of site-specific integration of the plasmid within AAVS1 in several AAV producer cell lines. The data also suggest that integration in AAVS1 is a general phenomenon that is not necessarily restricted to high producers. The results also suggest that plasmid integration within the AAVS1 locus is not an absolute requirement for a high vector yield.

  6. Plasmid pVAX1-NH36 purification by membrane and bead perfusion chromatography.

    Science.gov (United States)

    Franco-Medrano, Diana Ivonne; Guerrero-Germán, Patricia; Montesinos-Cisneros, Rosa María; Ortega-López, Jaime; Tejeda-Mansir, Armando

    2017-03-01

    The demand for plasmid DNA (pDNA) has increased in response to the rapid advances in vaccines applications to prevent and treat infectious diseases caused by virus, bacteria or parasites, such as Leishmania species. The immunization protocols require large amounts of supercoiled plasmid DNA (sc-pDNA) challenging the development of efficient and profitable processes for capturing and purified pDNA molecules from large volumes of lysates. A typical bioprocess involves four steps: fermentation, primary recovery, intermediate recovery and final purification. Ion-exchange chromatography is one of the key operations in the purification schemes of pDNA owing the chemical structure of these macromolecules. The goal of this research was to compare the performance of the final purification step of pDNA using ion-exchange chromatography on columns packed with Mustang Q membranes or perfusive beads POROS 50 HQ. The experimental results showed that both matrixes could separate the plasmid pVAX1-NH36 (3936 bp) from impurities in clarified Escherichia coli lysates with an adequate resolution. In addition, a 24- and 21-fold global purification factor was obtained. An 88 and 63% plasmid recuperation was achieved with ion-exchange membranes and perfusion beads, respectively. A better understanding of perfusion-based matrices for the purification of pDNA was developed in this research.

  7. Complete nucleotide sequence of CTX-M-15-plasmids from clinical Escherichia coli isolates: insertional events of transposons and insertion sequences.

    Directory of Open Access Journals (Sweden)

    Annemieke Smet

    Full Text Available BACKGROUND: CTX-M-producing Escherichia coli strains are regarded as major global pathogens. METHODOLOGY/PRINCIPAL FINDINGS: The nucleotide sequence of three plasmids (pEC_B24: 73801-bp; pEC_L8: 118525-bp and pEC_L46: 144871-bp from Escherichia coli isolates obtained from patients with urinary tract infections and one plasmid (pEC_Bactec: 92970-bp from an Escherichia coli strain isolated from the joint of a horse with arthritis were determined. Plasmid pEC_Bactec belongs to the IncI1 group and carries two resistance genes: bla(TEM-1 and bla(CTX-M-15. It shares more than 90% homology with a previously published bla(CTX-M-plasmid from E. coli of human origin. Plasmid pEC_B24 belongs to the IncFII group whereas plasmids pEC_L8 and pEC_L46 represent a fusion of two replicons of type FII and FIA. On the pEC_B24 backbone, two resistance genes, bla(TEM-1 and bla(CTX-M-15, were found. Six resistance genes, bla(TEM-1, bla(CTX-M-15, bla(OXA-1, aac6'-lb-cr, tetA and catB4, were detected on the pEC_L8 backbone. The same antimicrobial drug resistance genes, with the exception of tetA, were also identified on the pEC_L46 backbone. Genome analysis of all 4 plasmids studied provides evidence of a seemingly frequent transposition event of the bla(CTX-M-15-ISEcp1 element. This element seems to have a preferred insertion site at the tnpA gene of a bla(TEM-carrying Tn3-like transposon, the latter itself being inserted by a transposition event. The IS26-composite transposon, which contains the bla(OXA-1, aac6'-lb-cr and catB4 genes, was inserted into plasmids pEC_L8 and pEC_L46 by homologous recombination rather than a transposition event. Results obtained for pEC_L46 indicated that IS26 also plays an important role in structural rearrangements of the plasmid backbone and seems to facilitate the mobilisation of fragments from other plasmids. CONCLUSIONS: Collectively, these data suggests that IS26 together with ISEcp1 could play a critical role in the evolution of

  8. Pipeline inspection - failures selection and results obtained; Inspeccion interna de ductos - seleccion de fallas y resultados obtenidos

    Energy Technology Data Exchange (ETDEWEB)

    Motylicki, Juan Enrique [Oleoductos del Valle S.A. (OLDEVAL S.A.), (Argentina)

    2003-07-01

    The main objective of the pipelines maintenance is to assure the integrity of them in order to have a reliable operation and with an acceptable level of risk that minimizes the impact to people, to the environment and the business. The objective of this work is to expose the experiences obtained before by Oldelval and after the passages of the intelligent pig, carried out in the years 1999/2000. In the same one they will be described: How the flaws were selected to verify / to repair. Results obtained in digs. Laboratory studies to determine like they affect to the integrity of the pipeline, the different types of opposing plane defects. Conclusions. (author)

  9. The characteristics of micrococcus (deinococcus) radiodurans sark plasmids

    International Nuclear Information System (INIS)

    Sjarief, Sri Hariani; Kikuchi, Masahiro; Watanabe, Hiroshi.

    1994-01-01

    The characterization of micrococcus (deinococcus) radiodurans sark plasmids. This bacterium has been classified as a new genus deinococcus radiodurans which is resistant to gamma-rays. It can repair itself completely almost all of DNA damages including double strand breaks induced by gamma-rays up to about 5 KGy. To reveal the repair mechanism, several investigations had been done to develop a cloning vector available for the genetic analysis. For this purpose D. radiodurans Sark are to be prepared as a vector by studying the characteristics of its plasmid. Plasmids were isolated by electrophoresis using 0.6% low-melting-temperature agarose in TAE and run for 5.5 hours, followed by the identification. An antibiotic marker was also carried out in this experiment to identify its location in the genetic materials of the cell, beside making a restriction map of the plasmid. Results have shown that D. radiodurans Sark has 4 plasmids (P1, P2, P3, and P4) and the refampicin resistant genes were not found in the plasmid. (authors). 14 refs; 4 figs

  10. New high-definition thickness data obtained at tropical glaciers: preliminary results from Antisana volcano (Ecuador) using GPR prospection

    Science.gov (United States)

    Zapata, Camilo; Andrade, Daniel; Córdova, Jorge; Maisincho, Luis; Carvajal, Juan; Calispa, Marlon; Villacís, Marcos

    2014-05-01

    The study of tropical glaciers has been a significant contribution to the understanding of glacier dynamics and climate change. Much of the data and results have been obtained by analyzing plan-view images obtained by air- and space-borne sensors, as well as depth data obtained by diverse methodologies at selected points on the glacier surface. However, the measurement of glacier thicknesses has remained an elusive task in tropical glaciers, often located in rough terrains where the application of geophysical surveys (i.e. seismic surveys) requires logistics sometimes hardly justified by the amount of obtained data. In the case of Ecuador, however, where most glaciers have developed on active volcanoes and represent sources/reservoirs of fresh water, the precise knowledge of such information is fundamental for scientific research but also in order to better assess key aspects for the society. The relatively recent but fast development of the GPR technology has helped to obtain new highdefinition thickness data at Antisana volcano that will be used to: 1) better understand the dynamics and fate of tropical glaciers; 2) better estimate the amount of fresh water stored in the glaciers; 3) better assess the hazards associated with the sudden widespread melting of glaciers during volcanic eruptions. The measurements have been obtained at glaciers 12 and 15 of Antisana volcano, with the help of a commercial GPR equipped with a 25 MHz antenna. A total of 30 transects have been obtained, covering a distance of more than 3 km, from the glacier ablation zone, located at ~ 4600 masl, up to the level of 5200 masl. The preliminary results show a positive correlation between altitude and glacier thickness, with maximum and minimum calculated values reaching up to 80 m, and down to 15 m, respectively. The experience gained at Antisana volcano will be used to prepare a more widespread GPR survey in the glaciers of Cotopaxi volcano, whose implications in terms of volcanic hazards

  11. Plasmid profilling and similarities in identities of probable microbes isolated from crude oil contaminated agricultural soil

    Directory of Open Access Journals (Sweden)

    Toochukwu Ekwutosi OGBULIE

    2013-05-01

    Full Text Available Plasmid analysis of bacteria isolated from agricultural soil experimentally contaminated with crude oil was carried out and the resultant bands’ depicting the different molecular sizes of the plasmid DNA molecules per isolate was obtained. There was no visible band observed for Klebsiella indicating that the organism lack plasmid DNA that confers degradative ability to it, possibly the gene could be borne on the chromosomal DNA which enabled its persistence in the polluted soil. Molecular characterization was undertaken to confirm the identities of the possible microorganisms that may be present in crude oil-contaminated soil. The result of the DNA extracted and amplified in a PCR using EcoRI and EcoRV restriction enzymes for cutting the DNA of the bacterial cells indicated no visible band for cuts made with EcoRV restriction enzyme showing that the enzyme is not specific for bacterial DNA of isolates in the samples, hence there was no amplification. By contrast though, visible bands of amplicons were observed using EcoRI restriction enzymes. The resultant visible bands of microbial profile obtained using the universal RAPD primer with nucleotide sequence of 5’—CTC AAA GCA TCT AGG TCC A---3’ showed that only Pseudomonas fluorescens and Bacillus mycoides had visible bands at identical position on the gel indicating that both species possibly had identical sequence or genes of negligible differences coding for degradation of hydrocarbons as shown by similar values in molecular weight and positions in the gel electrophoresis field.

  12. Pregnancy does not affect HIV incidence test results obtained using the BED capture enzyme immunoassay or an antibody avidity assay.

    Directory of Open Access Journals (Sweden)

    Oliver Laeyendecker

    2010-10-01

    Full Text Available Accurate incidence estimates are needed for surveillance of the HIV epidemic. HIV surveillance occurs at maternal-child health clinics, but it is not known if pregnancy affects HIV incidence testing.We used the BED capture immunoassay (BED and an antibody avidity assay to test longitudinal samples from 51 HIV-infected Ugandan women infected with subtype A, C, D and intersubtype recombinant HIV who were enrolled in the HIVNET 012 trial (37 baseline samples collected near the time of delivery and 135 follow-up samples collected 3, 4 or 5 years later. Nineteen of 51 women were also pregnant at the time of one or more of the follow-up visits. The BED assay was performed according to the manufacturer's instructions. The avidity assay was performed using a Genetic Systems HIV-1/HIV-2 + O EIA using 0.1M diethylamine as the chaotropic agent.During the HIVNET 012 follow-up study, there was no difference in normalized optical density values (OD-n obtained with the BED assay or in the avidity test results (% when women were pregnant (n = 20 results compared to those obtained when women were not pregnant (n = 115; for BED: p = 0.9, generalized estimating equations model; for avidity: p = 0.7, Wilcoxon rank sum. In addition, BED and avidity results were almost exactly the same in longitudinal samples from the 18 women who were pregnant at only one study visit during the follow-up study (p = 0.6, paired t-test.These results from 51 Ugandan women suggest that any changes in the antibody response to HIV infection that occur during pregnancy are not sufficient to alter results obtained with the BED and avidity assays. Confirmation with larger studies and with other HIV subtypes is needed.

  13. Method of forecasting pollutant transfer in an aquifer initial results obtained in a sandy medium (Barp site, Gironde)

    International Nuclear Information System (INIS)

    Madoz-Escande, C.; Peyrus, J.-C.

    1979-01-01

    Hydrogeological studies are undertaken in the context of the radiological safety of nuclear plants to forecast consequences of accidental releases of radioactive pollutants into an aquifer (transfer time, concentration at points of emergence). This quantitative forecast is obtained with the aid of a mathematical model with sequential emission. This requires a knowledge of the physical parameters of the aquifer and of the behavior of the pollutant in relation to the water-bearing medium. The physical parameters of a saturated porous medium are presented with the aid of radioactive tracer tests on a model and also in the field. The initial results obtained in a sandy medium are presented. In view of the difficulty of extrapolating to field conditions the conclusions of tests on models, it was necessary to set up a mobile laboratory with which in situ studies could be undertaken. The behavior of the pollutant in relation to the water-bearing medium is the subject of preliminary laboratory research on the laws of adsorption under different pH and temperature conditions. The numerical results obtained call for confirmation in the field. A description is given of a method which should enable the distribution coefficients to be evaluated in situ

  14. Assessment of the French environment radiological status in 2009. Synthesis of the results obtained by the IRSN survey networks

    International Nuclear Information System (INIS)

    2009-01-01

    After a brief recall of the IRSN's missions, this report describes why and how radioactivity is controlled in France. It presents the national network for environment radioactivity measurements, and indicates the objectives of this radiological survey. It comments the status and levels of the natural and artificial radioactivity in France, discusses the results of the territory general survey (atmosphere, inland, coast, regions), the results of fuel cycle related sites (nuclear sites, old mining sites, front end industries, electricity production plants, back end industries, waste storage sites), the results concerning the nuclear medicine radioactive releases, the results obtained during selective expertises, and discusses the assessment of the exposure of the French population to ionizing radiations. It finally indicates how those data are published

  15. New dynamics information from experimental results obtained in d-Au collisions at RHIC-BNL energies

    International Nuclear Information System (INIS)

    Jipa, Alexandru

    2004-01-01

    During the last runs d-Au collisions at √s NN = 200 GeV have been investigated at RHIC-BNL using the BRAHMS Experiment. Some interesting experimental results on charged particle multiplicities, rapidity distributions, transverse momentum spectra, antiparticle to particle ratios, participant spectator evolution have been obtained. In this work the most interesting results are presented for different rapidity and collision centrality ranges. Taking into account the importance of the collision geometry and collision symmetry in the collision dynamics, comparisons with the similar experimental results obtained in Au-Au collisions have been done. New interesting results can be reported. The most significant are related to the evolution of the nuclear modification factor with rapidity and collision centrality. The high transverse momentum suppression and the behaviours in different rapidity and centrality ranges suggest strong initial state effects. These effects could be related to the gluonic structure of the colliding nuclei. Some insights on the Color Glass Condensate formation are possible. (author)

  16. Origin and Evolution of Rickettsial Plasmids.

    Directory of Open Access Journals (Sweden)

    Khalid El Karkouri

    Full Text Available Rickettsia species are strictly intracellular bacteria that have undergone a reductive genomic evolution. Despite their allopatric lifestyle, almost half of the 26 currently validated Rickettsia species have plasmids. In order to study the origin, evolutionary history and putative roles of rickettsial plasmids, we investigated the evolutionary processes that have shaped 20 plasmids belonging to 11 species, using comparative genomics and phylogenetic analysis between rickettsial, microbial and non-microbial genomes.Plasmids were differentially present among Rickettsia species. The 11 species had 1 to 4 plasmid (s with a size ranging from 12 kb to 83 kb. We reconstructed pRICO, the last common ancestor of the current rickettsial plasmids. pRICO was vertically inherited mainly from Rickettsia/Orientia chromosomes and diverged vertically into a single or multiple plasmid(s in each species. These plasmids also underwent a reductive evolution by progressive gene loss, similar to that observed in rickettsial chromosomes, possibly leading to cryptic plasmids or complete plasmid loss. Moreover, rickettsial plasmids exhibited ORFans, recent gene duplications and evidence of horizontal gene transfer events with rickettsial and non-rickettsial genomes mainly from the α/γ-proteobacteria lineages. Genes related to maintenance and plasticity of plasmids, and to adaptation and resistance to stress mostly evolved under vertical and/or horizontal processes. Those involved in nucleotide/carbohydrate transport and metabolism were under the influence of vertical evolution only, whereas genes involved in cell wall/membrane/envelope biogenesis, cycle control, amino acid/lipid/coenzyme and secondary metabolites biosynthesis, transport and metabolism underwent mainly horizontal transfer events.Rickettsial plasmids had a complex evolution, starting with a vertical inheritance followed by a reductive evolution associated with increased complexity via horizontal gene

  17. Matching the results of a theoretical model with failure rates obtained from a population of non-nuclear pressure vessels

    International Nuclear Information System (INIS)

    Harrop, L.P.

    1982-02-01

    Failure rates for non-nuclear pressure vessel populations are often regarded as showing a decrease with time. Empirical evidence can be cited which supports this view. On the other hand theoretical predictions of PWR type reactor pressure vessel failure rates have shown an increasing failure rate with time. It is shown that these two situations are not necessarily incompatible. If adjustments are made to the input data of the theoretical model to treat a non-nuclear pressure vessel population, the model can produce a failure rate which decreases with time. These adjustments are explained and the results obtained are shown. (author)

  18. Aero thermal test results obtained on the n. C 5 EL 4 Cluster in the atmospheric pressure cell

    International Nuclear Information System (INIS)

    Gasc, B.

    1964-01-01

    In the framework of thermal studies on the EL-4 cluster, the full-scale tests at atmospheric pressure are designed to permit measurement of local values of the wall temperature, of the velocity and of the temperature in the fluid. The experimental results, obtained with the help of an original measuring apparatus, make it possible to follow the changes in these values along the cluster and to predict in much detail the in-pile thermal behaviour. In particular it is shown that changes in the wall temperature along the cluster are greatly influenced by disruption of the flow caused by grids and supports. (author) [fr

  19. Comparison between dosimeter films and electronic dosimeters results obtained in F-18 production practices at IEN/CNEN

    International Nuclear Information System (INIS)

    Paulo, Osvaldir; Carlos, Luiz R.J.; Mendes, Milton

    2008-01-01

    Full text: The aim of the present work is the comparison of the dose rates values obtained from dosimeter films with the values obtained from electronic dosimeters used by radiological protection technician involved in practices with fluorine-18 production in CV-28 cyclotron at IEN/CNEN. The motivation for this work was the increase on the demand of this radiopharmaceutical. Nowadays there is an increase demand on the number of clinics and hospitals which had started to use the technique of radiodiagnostic using positron-emission tomography (PET). Therefore, the produced radionuclide activities (physics quantity) and also the complexity of the practices of radiological protection in the areas of that facility had increased a lot. The conventional statistical methods will be used to evaluate the results obtained in the analysis of the data obtained in the reading of the dosimeters. This comparative method is however a first step to taking decisions regarding radiological protection service. Therefore that the radiometric surveys of routine are taken in consideration, as well as the information from the technician of radiological protection that is working in those areas. All this data will give support to improve and implement methods and practices on the accelerators' facilities. In addition, the use of these electronic dosimeters will makes possible the technician read these dose rates received immediately during the practices and use these information you make decisions. In case of dose rates values to present above the expected the practice will be ploughed. Another point to be considered is that this work will propitiate the future updates of the procedures designated to those practices and to conduct at the radiological protection optimization. (author)

  20. Conjugative plasmids: Vessels of the communal gene pool

    DEFF Research Database (Denmark)

    Norman, Anders; Hansen, Lars H.; Sørensen, Søren Johannes

    2009-01-01

    to the hosts and, potentially, other resident prokaryotes within specific environmental niches. Insight into the evolution of plasmid modules therefore contributes to our knowledge of gene dissemination and evolution within prokaryotic communities. This communal pool provides the prokaryotes with an important...... mechanistic framework for obtaining adaptability and functional diversity that alleviates the need for large genomes of specialized ‘private genes'....

  1. Plasmid marker rescue transformation proceeds by breakage-reunion in Bacillus subtilis

    International Nuclear Information System (INIS)

    Weinrauch, Y.; Dubnau, D.

    1987-01-01

    Bacillus subtilis carrying a plasmid which replicates with a copy number of about 1 was transformed with linearized homologous plasmid DNA labeled with the heavy isotopes 2 H and 15 N, in the presence of 32 Pi and 6-(p-hydroxyphenylazo)-uracil to inhibit DNA replication. Plasmid DNA was isolated from the transformed culture and fractionated in cesium chloride density gradients. The distribution of total and donor plasmid DNA was examined, using specific hybridization probes. The synthesis of new DNA, associated with the integration of donor moiety, was also monitored. Donor-specific sequences were present at a density intermediate between that of light and hybrid DNA. This recombinant DNA represented 1.4% of total plasmid DNA. The latter value corresponded well with the transforming activity (1.7%) obtained for the donor marker. Newly synthesized material associated with plasmid DNA at the recombinant density amounted to a minor portion of the recombinant plasmid DNA. These data suggest that, like chromosomal transformation, plasmid marker rescue transformation does not require replication for the integration of donor markers and, also like chromosomal transformation, proceeds by a breakage-reunion mechanism. The extent of donor DNA replacement of recipient DNA per plasmid molecule of 54 kilobases (27 kilobase pairs) was estimated as 16 kilobases

  2. Plasmid DNA Delivery: Nanotopography Matters.

    Science.gov (United States)

    Song, Hao; Yu, Meihua; Lu, Yao; Gu, Zhengying; Yang, Yannan; Zhang, Min; Fu, Jianye; Yu, Chengzhong

    2017-12-20

    Plasmid DNA molecules with unique loop structures have widespread bioapplications, in many cases relying heavily on delivery vehicles to introduce them into cells and achieve their functions. Herein, we demonstrate that control over delicate nanotopography of silica nanoparticles as plasmid DNA vectors has significant impact on the transfection efficacy. For silica nanoparticles with rambutan-, raspberry-, and flower-like morphologies composed of spike-, hemisphere-, and bowl-type subunit nanotopographies, respectively, the rambutan-like nanoparticles with spiky surfaces demonstrate the highest plasmid DNA binding capability and transfection efficacy of 88%, higher than those reported for silica-based nanovectors. Moreover, it is shown that the surface spikes of rambutan nanoparticles provide a continuous open space to bind DNA chains via multivalent interactions and protect the gene molecules sheltered in the spiky layer against nuclease degradation, exhibiting no significant transfection decay. This unique protection feature is in great contrast to a commercial transfection agent with similar transfection performance but poor protection capability against enzymatic cleavage. Our study provides new understandings in the rational design of nonviral vectors for efficient gene delivery.

  3. Potential theory for stationary Schrödinger operators: a survey of results obtained with non-probabilistic methods

    Directory of Open Access Journals (Sweden)

    Marco Bramanti

    1992-05-01

    Full Text Available In this paper we deal with a uniformly elliptic operator of the kind: Lu  Au + Vu, where the principal part A is in divergence form, and V is a function assumed in a “Kato class”. This operator has been studied in different contexts, especially using probabilistic techniques. The aim of the present work is to give a unified and simplified presentation of the results obtained with non probabilistic methods for the operator L on a bounded Lipschitz domain. These results regard: continuity of the solutions of Lu=0; Harnack inequality; estimates on the Green's function and L-harmonic measure; boundary behavior of positive solutions of Lu=0, in particular a “Fatou's theorem”.

  4. Analysis of results obtained with different cutting techniques and associated filtration systems for the dismantling of radioactive metallic components

    International Nuclear Information System (INIS)

    Bach, F.W.; Steiner, H.; Schreck, G.

    1993-01-01

    The present joint study performed by the Commissariat a l'energie atomique and the Universitaet Hannover and coordinated by the Commission of the European Communities was intended to analyse the results generated in a number of research contracts concerned with cutting tests in air and underwater, with consideration of the prevailing working conditions. The analysis has led to a large database, giving broadly-assessed information for the dismantling of radioactive components. The range of study was enlarged, where possible, to include recently obtained results outside the present research programme, consideration also being given to supplementary cutting tools and filtration systems not covered by the present programme. Data was concentrated in structured information packages on practical experience available for a series of cutting tools and filters. These were introduced into a computerized user-friendly databank, to be considered as a first-stage development, which should be continuously updated and possibly oriented in the future to an expert system

  5. Determination of averaged axisymmetric flow surfaces according to results obtained by numerical simulation of flow in turbomachinery

    Directory of Open Access Journals (Sweden)

    Bogdanović-Jovanović Jasmina B.

    2012-01-01

    Full Text Available In the increasing need for energy saving worldwide, the designing process of turbomachinery, as an essential part of thermal and hydroenergy systems, goes in the direction of enlarging efficiency. Therefore, the optimization of turbomachinery designing strongly affects the energy efficiency of the entire system. In the designing process of turbomachinery blade profiling, the model of axisymmetric fluid flows is commonly used in technical practice, even though this model suits only the profile cascades with infinite number of infinitely thin blades. The actual flow in turbomachinery profile cascades is not axisymmetric, and it can be fictively derived into the axisymmetric flow by averaging flow parameters in the blade passages according to the circular coordinate. Using numerical simulations of flow in turbomachinery runners, its operating parameters can be preliminarily determined. Furthermore, using the numerically obtained flow parameters in the blade passages, averaged axisymmetric flow surfaces in blade profile cascades can also be determined. The method of determination of averaged flow parameters and averaged meridian streamlines is presented in this paper, using the integral continuity equation for averaged flow parameters. With thus obtained results, every designer can be able to compare the obtained averaged flow surfaces with axisymmetric flow surfaces, as well as the specific work of elementary stages, which are used in the procedure of blade designing. Numerical simulations of flow in an exemplary axial flow pump, used as a part of the thermal power plant cooling system, were performed using Ansys CFX. [Projekat Ministarstva nauke Republike Srbije, br. TR33040: Revitalization of existing and designing new micro and mini hydropower plants (from 100 kW to 1000 kW in the territory of South and Southeast Serbia

  6. Plasmids and rickettsial evolution: insight from Rickettsia felis.

    Directory of Open Access Journals (Sweden)

    Joseph J Gillespie

    2007-03-01

    Full Text Available The genome sequence of Rickettsia felis revealed a number of rickettsial genetic anomalies that likely contribute not only to a large genome size relative to other rickettsiae, but also to phenotypic oddities that have confounded the categorization of R. felis as either typhus group (TG or spotted fever group (SFG rickettsiae. Most intriguing was the first report from rickettsiae of a conjugative plasmid (pRF that contains 68 putative open reading frames, several of which are predicted to encode proteins with high similarity to conjugative machinery in other plasmid-containing bacteria.Using phylogeny estimation, we determined the mode of inheritance of pRF genes relative to conserved rickettsial chromosomal genes. Phylogenies of chromosomal genes were in agreement with other published rickettsial trees. However, phylogenies including pRF genes yielded different topologies and suggest a close relationship between pRF and ancestral group (AG rickettsiae, including the recently completed genome of R. bellii str. RML369-C. This relatedness is further supported by the distribution of pRF genes across other rickettsiae, as 10 pRF genes (or inactive derivatives also occur in AG (but not SFG rickettsiae, with five of these genes characteristic of typical plasmids. Detailed characterization of pRF genes resulted in two novel findings: the identification of oriV and replication termination regions, and the likelihood that a second proposed plasmid, pRFdelta, is an artifact of the original genome assembly.Altogether, we propose a new rickettsial classification scheme with the addition of a fourth lineage, transitional group (TRG rickettsiae, that is unique from TG and SFG rickettsiae and harbors genes from possible exchanges with AG rickettsiae via conjugation. We offer insight into the evolution of a plastic plasmid system in rickettsiae, including the role plasmids may have played in the acquirement of virulence traits in pathogenic strains, and the

  7. Edaphic macrofauna as biological indicator of the conservation/disturbance status of soil. Results obtained in Cuba

    International Nuclear Information System (INIS)

    Cabrera, Grisel

    2012-01-01

    In order to predict the degradation status of a soil, a group of variables comprising its physical, chemical and/or biological properties is used. Macrofauna, which includes soil invertebrates higher than 2 mm of diameter, is a biological component that can be used for such purpose. Its taxonomic richness as well as its density, biomass and functional composition change depending on the effect of diverse land uses and managements. This review reaffirms that the macrofauna characteristics and the results obtained, mainly in Cuba, about its variation in ecosystems with different anthropization levels, support the potential use of this fauna as biological indicator of the soil's conservation status. Future studies should consider a lower taxonomic level in the identification of macrofauna, and relate its taxonomic and functional composition to the climate and pedological factors. (author)

  8. Multiple drug resistant carbapenemases producing Acinetobacter baumannii isolates harbours multiple R-plasmids

    Directory of Open Access Journals (Sweden)

    Rajagopalan Saranathan

    2014-01-01

    Full Text Available Background & objectives: The nosocomial human pathogen Acinetobacter baumannii has high propensity to develop resistance to antimicrobials and to become multidrug resistant (MDR, consequently complicating the treatment. This study was carried out to investigate the presence of resistant plasmids (R-plasmids among the clinical isolates of A. baumannii. In addition, the study was performed to check the presence of common β-lactamases encoding genes on these plasmids. Methods: A total of 55 clinical isolates of A. baumannii were included in the study and all were subjected to plasmid DNA isolation, followed by PCR to check the presence of resistance gene determinants such as blaOXA-23 , blaOXA-51, blaOXA-58 and blaIMP-1 on these plasmids that encode for oxacillinase (OXA and metallo-β-lactamase (MBL type of carbapenemases. Plasmid curing experiments were carried out on selected isolates using ethidium bromide and acridine orange as curing agents and the antibiotic resistance profiles were evaluated before and after curing. Results: All the isolates were identified as A. baumannii by 16SrDNA amplification and sequencing. Plasmid DNA isolated from these isolates showed the occurrence of multiple plasmids with size ranging from 500bp to ≥ 25 kb. The percentage of blaOXA-51 and blaOXA-23 on plasmids were found to be 78 and 42 per cent, respectively and 20 isolates (36% carried blaIMP-1 gene on plasmids. Significant difference was observed in the antibiograms of plasmid cured isolates when compared to their parental ones. The clinical isolates became susceptible to more than two antibiotic classes after curing of plasmids indicating plasmid borne resistance. Interpretation & conclusions: Our study determined the plasmid mediated resistance mechanisms and occurrence of different resistance genes on various plasmids isolated from MDR A. baumannii. The present findings showed the evidence for antibiotic resistance mediated through multiple plasmids in

  9. First In-Core Measurement Results Obtained with the Innovative Mobile Calorimeter CALMOS inside the OSIRIS Material Testing Reactor

    International Nuclear Information System (INIS)

    Carcreff, Hubert; Salmon, Laurent; Courtaux, Cedric

    2013-06-01

    Nuclear heating rate inside an MTR has to be known in order to design and to run irradiation experiments which have to fulfill target temperature constraints. This measurement is usually carried out by calorimetry [1, 2]. An innovative calorimetric system, CALMOS, has been studied and built in 2011 for the 70 MWth OSIRIS reactor operated by CEA. Thanks to a new calorimetric probe, associated to a specific displacement system, it provides measurements along the fissile height and above the core. The development of the calorimetric probe required the manufacturing and the irradiation of mock-ups in the ex-core area, where nuclear heating rate does not exceed 2 W.g -1 . The calorimeter working mode, the different measurement procedures allowed with such a new probe and main modeling and experimental results have been already presented [3, 4]. In this paper, we present the first results obtained during several measurement campaigns carried out in 2012 and 2013 inside the OSIRIS core with the final device. For the first time, this new experimental measurement system was operated in nominal in-core thermo hydraulic conditions with nominal neutron and gamma fluxes (up to 6 W.g -1 ) in several experimental locations. After a brief presentation of the displacement system specificities, first nuclear heating distributions are presented and discussed. Experimental data were also used to upgrade the Finite Element model of the calorimeter in order to match measured temperatures with calculated ones. This model allowed to estimate a Kc correction factor which takes into account small nonlinearities when the heating rate is deduced from the calibration method. A comparison is made between nuclear heating rates determined from the probe calibration and from the zero method. In addition, an evaluation of the global uncertainty associated to the measurements is detailed. Finally, a global comparison is made with available measurements obtained from previous calorimeters. (authors)

  10. First in-core measurement results obtained with the innovative mobile calorimeter CALMOS inside the OSIRIS material testing reactor

    International Nuclear Information System (INIS)

    Carcreff, Hubert; Salmon, Laurent; Courtaux, Cedric

    2014-01-01

    Nuclear heating rate inside an MTR has to be known in order to design and to run irradiation experiments which have to fulfill target temperature constraints. This measurement is usually carried out by calorimetry. An innovative calorimetric system, CALMOS, has been studied and built in 2011 for the 70 MWth OSIRIS reactor operated by CEA. Thanks to a new calorimetric probe, associated to a specific displacement system, it provides measurements along the fissile height and above the core. Development of the calorimetric probe required manufacturing and irradiation of mock-ups in the ex-core area, where nuclear heating rate does not exceed 2 W.g -1 . The calorimeter working mode, the different measurement procedures, main modeling and ex-core experimental results have been already presented in previous papers. In this paper, we present in-core results obtained from 2011 to 2013 with the final device. For the first time, this new experimental measurement system was operated in several experimental locations, with nominal in-core thermal hydraulic conditions, nominal neutron flux and nuclear heating rate up to 6 W.g -1 (in graphite). After a brief presentation of the displacement system specificities, first nuclear heating distributions are presented and discussed. The Finite Element model of the calorimeter was upgraded in order to match calculated temperatures with measured ones. This 'validated' model allowed to estimate a Kc factor which tends to correct small nonlinearities when heating rate is calculated from the 'calibration method'. A comparison is made between nuclear heating rates determined from 'calibration' and 'zero methods'. In addition, an evaluation of the global uncertainty associated to the measurements is detailed. Finally, a comparison is made with available measurements obtained from previous calorimeters. (authors)

  11. Standardization of a fluconazole bioassay and correlation of results with those obtained by high-pressure liquid chromatography.

    Science.gov (United States)

    Rex, J H; Hanson, L H; Amantea, M A; Stevens, D A; Bennett, J E

    1991-01-01

    An improved bioassay for fluconazole was developed. This assay is sensitive in the clinically relevant range (2 to 40 micrograms/ml) and analyzes plasma, serum, and cerebrospinal fluid specimens; bioassay results correlate with results obtained by high-pressure liquid chromatography (HPLC). Bioassay and HPLC analyses of spiked plasma, serum, and cerebrospinal fluid samples (run as unknowns) gave good agreement with expected values. Analysis of specimens from patients gave equivalent results by both HPLC and bioassay. HPLC had a lower within-run coefficient of variation (less than 2.5% for HPLC versus less than 11% for bioassay) and a lower between-run coefficient of variation (less than 5% versus less than 12% for bioassay) and was more sensitive (lower limit of detection, 0.1 micrograms/ml [versus 2 micrograms/ml for bioassay]). The bioassay is, however, sufficiently accurate and sensitive for clinical specimens, and its relative simplicity, low sample volume requirement, and low equipment cost should make it the technique of choice for analysis of routine clinical specimens. PMID:1854166

  12. Prenatal cytogenetic diagnosis in Spain: analysis and evaluation of the results obtained from amniotic fluid samples during the last decade.

    Science.gov (United States)

    Mademont-Soler, Irene; Morales, Carme; Clusellas, Núria; Soler, Anna; Sánchez, Aurora

    2011-08-01

    Chromosome abnormalities are one of the main causes of congenital defects, and establishing their frequency according to the different clinical indications for invasive procedure during pregnancy is especially important for genetic counselling. We analyzed the results of 29,883 amniotic fluid samples referred to our laboratory for cytogenetic studies from 1998 to 2009, which constitutes the largest series of cytogenetic analysis performed on amniotic fluid samples in Spain. The number of samples received tended to increase from 1998 to 2005, but after 2005 it decreased substantially. Cytogenetic results were obtained in 99.5% of the samples, and the detected incidence of chromosome abnormalities was 2.9%. Of these, 48.1% consisted of classical autosomal aneuploidies, trisomy 21 being the most frequent one. The main clinical indications for amniocentesis were positive prenatal screening and advanced maternal age, but referral reasons with highest positive predictive values were, excluding parental chromosome rearrangement, increased nuchal translucency (9.2%) and ultrasound abnormalities (6.6%). In conclusion, performing the karyotype on amniotic fluid samples is a good method for the detection of chromosome abnormalities during pregnancy. The number of cytogenetic studies on amniotic fluid has now decreased, however, due to the implementation of first trimester prenatal screening for the detection of Down syndrome, which allows karyotyping on chorionic villus samples. Our results also show that both ultrasound abnormalities and increased nuchal translucency are excellent clinical indicators for fetal chromosome abnormality. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

  13. Novel plasmids and resistance phenotypes in Yersinia pestis: unique plasmid inventory of strain Java 9 mediates high levels of arsenic resistance.

    Science.gov (United States)

    Eppinger, Mark; Radnedge, Lyndsay; Andersen, Gary; Vietri, Nicholas; Severson, Grant; Mou, Sherry; Ravel, Jacques; Worsham, Patricia L

    2012-01-01

    Growing evidence suggests that the plasmid repertoire of Yersinia pestis is not restricted to the three classical virulence plasmids. The Java 9 strain of Y. pestis is a biovar Orientalis isolate obtained from a rat in Indonesia. Although it lacks the Y. pestis-specific plasmid pMT, which encodes the F1 capsule, it retains virulence in mouse and non-human primate animal models. While comparing diverse Y. pestis strains using subtractive hybridization, we identified sequences in Java 9 that were homologous to a Y. enterocolitica strain carrying the transposon Tn2502, which is known to encode arsenic resistance. Here we demonstrate that Java 9 exhibits high levels of arsenic and arsenite resistance mediated by a novel promiscuous class II transposon, named Tn2503. Arsenic resistance was self-transmissible from Java 9 to other Y. pestis strains via conjugation. Genomic analysis of the atypical plasmid inventory of Java 9 identified pCD and pPCP plasmids of atypical size and two previously uncharacterized cryptic plasmids. Unlike the Tn2502-mediated arsenic resistance encoded on the Y. enterocolitica virulence plasmid; the resistance loci in Java 9 are found on all four indigenous plasmids, including the two novel cryptic plasmids. This unique mobilome introduces more than 105 genes into the species gene pool. The majority of these are encoded by the two entirely novel self-transmissible plasmids, which show partial homology and synteny to other enterics. In contrast to the reductive evolution in Y. pestis, this study underlines the major impact of a dynamic mobilome and lateral acquisition in the genome evolution of the plague bacterium.

  14. Novel plasmids and resistance phenotypes in Yersinia pestis: unique plasmid inventory of strain Java 9 mediates high levels of arsenic resistance.

    Directory of Open Access Journals (Sweden)

    Mark Eppinger

    Full Text Available Growing evidence suggests that the plasmid repertoire of Yersinia pestis is not restricted to the three classical virulence plasmids. The Java 9 strain of Y. pestis is a biovar Orientalis isolate obtained from a rat in Indonesia. Although it lacks the Y. pestis-specific plasmid pMT, which encodes the F1 capsule, it retains virulence in mouse and non-human primate animal models. While comparing diverse Y. pestis strains using subtractive hybridization, we identified sequences in Java 9 that were homologous to a Y. enterocolitica strain carrying the transposon Tn2502, which is known to encode arsenic resistance. Here we demonstrate that Java 9 exhibits high levels of arsenic and arsenite resistance mediated by a novel promiscuous class II transposon, named Tn2503. Arsenic resistance was self-transmissible from Java 9 to other Y. pestis strains via conjugation. Genomic analysis of the atypical plasmid inventory of Java 9 identified pCD and pPCP plasmids of atypical size and two previously uncharacterized cryptic plasmids. Unlike the Tn2502-mediated arsenic resistance encoded on the Y. enterocolitica virulence plasmid; the resistance loci in Java 9 are found on all four indigenous plasmids, including the two novel cryptic plasmids. This unique mobilome introduces more than 105 genes into the species gene pool. The majority of these are encoded by the two entirely novel self-transmissible plasmids, which show partial homology and synteny to other enterics. In contrast to the reductive evolution in Y. pestis, this study underlines the major impact of a dynamic mobilome and lateral acquisition in the genome evolution of the plague bacterium.

  15. Cefotaxime resistant Escherichia coli collected from a healthy volunteer; characterisation and the effect of plasmid loss.

    Directory of Open Access Journals (Sweden)

    Miranda Kirchner

    Full Text Available In this study 6 CTX-M positive E. coli isolates collected during a clinical study examining the effect of antibiotic use in a human trial were analysed. The aim of the study was to analyse these isolates and assess the effect of full or partial loss of plasmid genes on bacterial fitness and pathogenicity. A DNA array was utilised to assess resistance and virulence gene carriage. Plasmids were characterised by PCR-based replicon typing and addiction system multiplex PCR. A phenotypic array and insect virulence model were utilised to assess the effect of plasmid-loss in E. coli of a large multi-resistance plasmid. All six E. coli carrying bla CTX-M-14 were detected from a single participant and were identical by pulse field gel electrophoresis and MLST. Plasmid profiling and arrays indicated absence of a large multi-drug resistance (MDR F-replicon plasmid carrying blaTEM, aadA4, strA, strB, dfrA17/19, sul1, and tetB from one isolate. Although this isolate partially retained the plasmid it showed altered fitness characteristics e.g. inability to respire in presence of antiseptics, similar to a plasmid-cured strain. However, unlike the plasmid-cured or plasmid harbouring strains, the survival rate for Galleria mellonella infected by the former strain was approximately 5-times lower, indicating other possible changes accompanying partial plasmid loss. In conclusion, our results demonstrated that an apparently healthy individual can harbour bla CTX-M-14 E. coli strains. In one such strain, isolated from the same individual, partial absence of a large MDR plasmid resulted in altered fitness and virulence characteristics, which may have implications in the ability of this strain to infect and any subsequent treatment.

  16. An In Vitro Chicken Gut Model Demonstrates Transfer of a Multidrug Resistance Plasmid from Salmonella to Commensal Escherichia coli.

    Science.gov (United States)

    Card, Roderick M; Cawthraw, Shaun A; Nunez-Garcia, Javier; Ellis, Richard J; Kay, Gemma; Pallen, Mark J; Woodward, Martin J; Anjum, Muna F

    2017-07-18

    The chicken gastrointestinal tract is richly populated by commensal bacteria that fulfill various beneficial roles for the host, including helping to resist colonization by pathogens. It can also facilitate the conjugative transfer of multidrug resistance (MDR) plasmids between commensal and pathogenic bacteria which is a significant public and animal health concern as it may affect our ability to treat bacterial infections. We used an in vitro chemostat system to approximate the chicken cecal microbiota, simulate colonization by an MDR Salmonella pathogen, and examine the dynamics of transfer of its MDR plasmid harboring several genes, including the extended-spectrum beta-lactamase bla CTX-M1 We also evaluated the impact of cefotaxime administration on plasmid transfer and microbial diversity. Bacterial community profiles obtained by culture-independent methods showed that Salmonella inoculation resulted in no significant changes to bacterial community alpha diversity and beta diversity, whereas administration of cefotaxime caused significant alterations to both measures of diversity, which largely recovered. MDR plasmid transfer from Salmonella to commensal Escherichia coli was demonstrated by PCR and whole-genome sequencing of isolates purified from agar plates containing cefotaxime. Transfer occurred to seven E. coli sequence types at high rates, even in the absence of cefotaxime, with resistant strains isolated within 3 days. Our chemostat system provides a good representation of bacterial interactions, including antibiotic resistance transfer in vivo It can be used as an ethical and relatively inexpensive approach to model dissemination of antibiotic resistance within the gut of any animal or human and refine interventions that mitigate its spread before employing in vivo studies. IMPORTANCE The spread of antimicrobial resistance presents a grave threat to public health and animal health and is affecting our ability to respond to bacterial infections

  17. Construction of Stable Fluorescent Reporter Plasmids for Use in Staphylococcus aureus

    Directory of Open Access Journals (Sweden)

    Michelle D. Rodriguez

    2017-12-01

    Full Text Available Here, the genes encoding three different fluorescent proteins were cloned into the stably maintained Staphylococcus aureus shuttle vector pKK30. The resulting plasmids were transformed into two S. aureus strains; SH1000 and RN4220. Stability assays illustrated that the three recombinant plasmids retained near 100% maintenance in vitro for 160 generations. S. aureus strain SH1000 expressing green fluorescent protein was then inoculated in an ovine model and in vivo stability for 6 days was demonstrated. In essence, these reporter plasmids represent a useful set of tools for dynamic imaging studies in S. aureus. These three reporter plasmids are available through BEI Resources.

  18. Construction of Stable Fluorescent Reporter Plasmids for Use in Staphylococcus aureus.

    Science.gov (United States)

    Rodriguez, Michelle D; Paul, Zubin; Wood, Charles E; Rice, Kelly C; Triplett, Eric W

    2017-01-01

    Here, the genes encoding three different fluorescent proteins were cloned into the stably maintained Staphylococcus aureus shuttle vector pKK30. The resulting plasmids were transformed into two S. aureus strains; SH1000 and RN4220. Stability assays illustrated that the three recombinant plasmids retained near 100% maintenance in vitro for 160 generations. S. aureus strain SH1000 expressing green fluorescent protein was then inoculated in an ovine model and in vivo stability for 6 days was demonstrated. In essence, these reporter plasmids represent a useful set of tools for dynamic imaging studies in S. aureus . These three reporter plasmids are available through BEI Resources.

  19. Genetic characterization of blaNDM-harboring plasmids in carbapenem-resistant Escherichia coli from Myanmar.

    Directory of Open Access Journals (Sweden)

    Yo Sugawara

    Full Text Available The bacterial enzyme New Delhi metallo-β-lactamase hydrolyzes almost all β-lactam antibiotics, including carbapenems, which are drugs of last resort for severe bacterial infections. The spread of carbapenem-resistant Enterobacteriaceae that carry the New Delhi metallo-β-lactamase gene, blaNDM, poses a serious threat to public health. In this study, we genetically characterized eight carbapenem-resistant Escherichia coli isolates from a tertiary care hospital in Yangon, Myanmar. The eight isolates belonged to five multilocus-sequence types and harbored multiple antimicrobial-resistance genes, resulting in resistance against nearly all of the antimicrobial agents tested, except colistin and fosfomycin. Nine plasmids harboring blaNDM genes were identified from these isolates. Multiple blaNDM genes were found in the distinct Inc-replicon types of the following plasmids: an IncA/C2 plasmid harboring blaNDM-1 (n = 1, IncX3 plasmids harboring blaNDM-4 (n = 2 or blaNDM-7 (n = 1, IncFII plasmids harboring blaNDM-4 (n = 1 or blaNDM-5 (n = 3, and a multireplicon F plasmid harboring blaNDM-5 (n = 1. Comparative analysis highlighted the diversity of the blaNDM-harboring plasmids and their distinct characteristics, which depended on plasmid replicon types. The results indicate circulation of phylogenetically distinct strains of carbapenem-resistant E. coli with various plasmids harboring blaNDM genes in the hospital.

  20. Computational design and characterization of a temperature-sensitive plasmid replicon for gram positive thermophiles

    Directory of Open Access Journals (Sweden)

    Olson Daniel G

    2012-05-01

    Full Text Available Abstract Background Temperature-sensitive (Ts plasmids are useful tools for genetic engineering, but there are currently none compatible with the gram positive, thermophilic, obligate anaerobe, Clostridium thermocellum. Traditional mutagenesis techniques yield Ts mutants at a low frequency, and therefore requires the development of high-throughput screening protocols, which are also not available for this organism. Recently there has been progress in the development of computer algorithms which can predict Ts mutations. Most plasmids currently used for genetic modification of C. thermocellum are based on the replicon of plasmid pNW33N, which replicates using the RepB replication protein. To address this problem, we set out to create a Ts plasmid by mutating the gene coding for the RepB replication protein using an algorithm designed by Varadarajan et al. (1996 for predicting Ts mutants based on the amino-acid sequence of the protein. Results A library of 34 mutant plasmids was designed, synthesized and screened, resulting in 6 mutants which exhibited a Ts phenotype. Of these 6, the one with the most temperature-sensitive phenotype (M166A was compared with the original plasmid. It exhibited lower stability at 48°C and was completely unable to replicate at 55°C. Conclusions The plasmid described in this work could be useful in future efforts to genetically engineer C. thermocellum, and the method used to generate this plasmid may be useful for others trying to make Ts plasmids.

  1. Evaluation of the CDTN's cementation facility using the results obtained in two different sizes: laboratory and facility

    International Nuclear Information System (INIS)

    Haucz, Maria Judite A.; Calabria, Jaqueline A. Almeida; Tello, Cledola Cassia O.; Candido, Francisco Donizete; Seles, Sandro Rogerio Novaes

    2011-01-01

    At CDTN R and D activities generate low-level radioactive wastes, among them the aqueous liquid ones, which need special attention due to their volumes. The usual treatment for these wastes is the volume reduction process using chemicals in order to change the radionuclides of the waste to an insoluble form. The resultant sludge is incorporated into cement in 200-liter drums using an in-line batch mixing system (CDTN Cementation Facility). The process parameters and solidified product quality are evaluated at the Cementation Laboratory (LABCIM), where tests are carried out to determine the viscosity, set time and density of the paste. In addition of the visual examination, compressive strength test and density are also established in the solidified product. In the cementation facility some pastes are also prepared for the immobilization of non-compactable waste, such as dump, scrap metal, wood, and other ones. This paper aims to present the study comparing the results of tests done in the pastes and products prepared in LABCIM and the ones obtained in CDTN's cementation facility, in order to check the efficiency of the cementation system. (author)

  2. Evaluation of the CDTN cementation facility using the results obtained in two different sizes: laboratory and facility

    International Nuclear Information System (INIS)

    Haucz, Maria Judite A.; Calabria, Jaqueline A. Almeida; Tello, Cledola Cassia O.; Candido, Francisco Donizete; Seles, Sandro Rogerio Novaes

    2011-01-01

    At CDTN R and D activities generate low-level radioactive wastes, among them the aqueous liquid ones, which need special attention due to their volumes. The usual treatment for these wastes is the volume reduction process using chemicals in order to change the radionuclides of the waste to an insoluble form. The resultant sludge is incorporated into cement in 200-liter drums using an in-line batch mixing system (CDTN Cementation Facility). The process parameters and solidified product quality are evaluated at the Cementation Laboratory (LABCIM), where tests are carried out to determine the viscosity, set time and density of the paste. In addition of the visual examination, compressive strength test and density are also established in the solidified product. In the cementation facility some pastes are also prepared for the immobilization of non-compactable waste, such as dump, scrap metal, wood, and other ones. This paper aims to present the study comparing the results of tests done in the pastes and products prepared in LABCIM and the ones obtained in CDTN's cementation facility, in order to check the efficiency of the cementation system. (author)

  3. Decontamination in preparation for dismantlement - AREVA's chemical decontamination technologies, projects performed and results obtained in the period 2011-2016

    International Nuclear Information System (INIS)

    Topf, C.; Sempere Belda, L.

    2017-01-01

    As a consequence of the nuclear phase-out decreed by the German government, several nuclear power plants in the country have already ceased operation. The remaining ones will cease operation by 2022. This has turned Germany into one of the most active regions worldwide in the field of nuclear decommissioning, with new and emerging technologies being deployed on the field, and already preexisting technologies being put to the test, optimized and developed into full maturity. The chemistry services division of AREVA GmbH has already performed 5 Full System Decontaminations (FSD) in preparation for decommissioning in this period - three in PWRs and two in BWRs - along with other international projects of relevance for decommissioning operations. During a FSD, the complete primary circuit of a nuclear power plant including auxiliary systems is subject to a chemical treatment; designed to remove radioactive matter accumulated onto system surfaces during operation. Through the effective removal of this radioactive accumulations contact dose rates on the different components of the primary circuit can be consistently reduced by factors larger than 50. This results in much lower ambient dose rates and, hence, in very significant dose savings for subsequent decommissioning activities. Additionally, dismantlement operations of large components are considerably simplified and can be performed under conditions that wouldn't have been possible before. The project specific objectives and challenges, the technologies employed, and the results obtained are presented and commented here. (authors)

  4. Biomonitoring of heavy metal deposition in the south Ural region: some preliminary results obtained by nuclear and related techniques

    International Nuclear Information System (INIS)

    Frontas'eva, M.V.; Steinnes, E.; Lyapunov, S.M.; Cherchintsev, V.D.; Smirnov, L.I.

    1999-01-01

    The first results are reported from the analysis of feather mosses used to study heavy metal atmospheric deposition in the vicinity of Magnitogorsk, the center of the steel industry in Russia. Moss samples collected at sites 30 km to the north-west of the industry were analyzed by instrumental neutron activation analysis using epithermal neutrons (ENAA), Results for a total of 38 elements are reported, including Pb, Cd, and Cu determined by atomic absorption spectroscopy (AAS). The element concentrations in moss samples from this work are compared with relevant literature data for strongly polluted areas in Central and Northern Europe and background values from Norway obtained by the same biomonitoring technique. The concentrations of Sb in the examined area are the highest ever reported for mosses, and also levels of Fe, Cr, and V are found to be particularly high. A scanning electron microscope connected to an XRF analyzer (SEM-XRF) was used to examine the surface of the moss samples. Photographs of identified iron spherules along with other aerosol particles made at magnification of 3500 to 5000 times and corresponding XRF analyses verifying the nature of typical particles are presented

  5. Characterization of epidemic IncI1-Iγ plasmids harboring ambler class A and C genes in Escherichia coli and Salmonella enterica from animals and humans

    NARCIS (Netherlands)

    Smith, Hilde; Bossers, Alex; Harders, Frank; Wu, Guanghui; Woodford, Neil; Schwarz, Stefan; Guerra, Beatriz; Rodríguez, Irene; van Essen-Zandbergen, Alieda; Brouwer, Michael; Mevius, Dik

    The aim of the study was to identify the plasmid-encoded factors contributing to the emergence and spread of epidemic IncI1-Iγ plasmids obtained from Escherichia coli and Salmonella enterica isolates from animal and human reservoirs. For this, 251 IncI1-Iγ plasmids carrying various extended-spectrum

  6. Plasmid-based generation of induced neural stem cells from adult human fibroblasts

    Directory of Open Access Journals (Sweden)

    Philipp Capetian

    2016-10-01

    Full Text Available Direct reprogramming from somatic to neural cell types has become an alternative to induced pluripotent stem cells. Most protocols employ viral expression systems, posing the risk of random genomic integration. Recent developments led to plasmid-based protocols, lowering this risk. However, these protocols either relied on continuous presence of a variety of small molecules or were only able to reprogram murine cells. We therefore established a reprogramming protocol based on vectors containing the Epstein-Barr virus (EBV-derived oriP/EBNA1 as well as the defined expression factors Oct3/4, Sox2, Klf4, L-myc, Lin28, and a small hairpin directed against p53. We employed a defined neural medium in combination with the neurotrophins bFGF, EGF and FGF4 for cultivation without the addition of small molecules. After reprogramming, cells demonstrated a temporary increase in the expression of endogenous Oct3/4. We obtained induced neural stem cells (iNSC 30 days after transfection. In contrast to previous results, plasmid vectors as well as a residual expression of reprogramming factors remained detectable in all cell lines. Cells showed a robust differentiation into neuronal (72% and glial cells (9% astrocytes, 6% oligodendrocytes. Despite the temporary increase of pluripotency-associated Oct3/4 expression during reprogramming, we did not detect pluripotent stem cells or non-neural cells in culture (except occasional residual fibroblasts. Neurons showed electrical activity and functional glutamatergic synapses. Our results demonstrate that reprogramming adult human fibroblasts to iNSC by plasmid vectors and basic neural medium without small molecules is possible and feasible. However, a full set of pluripotency-associated transcription factors may indeed result in the acquisition of a transient (at least partial pluripotent intermediate during reprogramming. In contrast to previous reports, the EBV-based plasmid system remained present and active inside

  7. Role of plasmids in Lactobacillus brevis BSO 464 hop tolerance and beer spoilage.

    Science.gov (United States)

    Bergsveinson, Jordyn; Baecker, Nina; Pittet, Vanessa; Ziola, Barry

    2015-02-01

    Specific isolates of lactic acid bacteria (LAB) can grow in the harsh beer environment, thus posing a threat to brew quality and the economic success of breweries worldwide. Plasmid-localized genes, such as horA, horC, and hitA, have been suggested to confer hop tolerance, a trait required for LAB survival in beer. The presence and expression of these genes among LAB, however, do not universally correlate with the ability to grow in beer. Genome sequencing of the virulent beer spoilage organism Lactobacillus brevis BSO 464 revealed the presence of eight plasmids, with plasmids 1, 2, and 3 containing horA, horC, and hitA, respectively. To investigate the roles that these and the other five plasmids play in L. brevis BSO 464 growth in beer, plasmid curing with novobiocin was used to derive 10 plasmid variants. Multiplex PCRs were utilized to determine the presence or absence of each plasmid, and how plasmid loss affected hop tolerance and growth in degassed (noncarbonated) beer was assessed. Loss of three of the eight plasmids was found to affect hop tolerance and growth in beer. Loss of plasmid 2 (horC and 28 other genes) had the most dramatic effect, with loss of plasmid 4 (120 genes) and plasmid 8 (47 genes) having significant, but smaller, impacts. These results support the contention that genes on mobile genetic elements are essential for bacterial growth in beer and that beer spoilage ability is not dependent solely on the three previously described hop tolerance genes or on the chromosome of a beer spoilage LAB isolate.

  8. Conjugative IncFI plasmids carrying CTX-M-15 among Escherichia coli ESBL producing isolates at a University hospital in Germany

    Directory of Open Access Journals (Sweden)

    Hain Torsten

    2009-06-01

    Full Text Available Abstract Background Multi-drug-resistant, extended-spectrum β-lactamase (ESBL-producing Enterobacteriaceae, constitute an emerging public-health concern. Little data on the molecular epidemiology of ESBL producing Escherichia coli is available in Germany. Here we describe the prevalence and molecular epidemiology of ESBL producing-Escherichia coli isolates at a German University hospital. Methods We analysed 63 non-duplicate clinical ESBL isolates obtained over an 8-month period using PCR and sequence-based ESBL allele typing, plasmid replicon typing, phylogenetic group typing. Pulsed-field gel electrophoresis (PFGE based genotyping and plasmid profiling was performed, as well as confirmatory DNA-based hybridization assays. Results Examination of the 63 Escherichia coli isolates revealed an almost equal distribution among the E. coli phylogenetic groups A, B1, B2 and D. High prevalence (36/63 of the CTX-M-15 gene was observed and an analysis of PFGE-based patterns revealed the presence of this CTX-M allele in multiple clones. Resistance to cefotaxime was a transferable trait and a commonly occurring 145.5 kb conjugative IncFI plasmid was detected in 65% of E. coli carrying the CTX-M-15 allele. The rate of transferable antibiotic resistances for GM, SXT, TET, GM-SXT-TET, SXT-TET and GM-TET was 33%, 61%, 61%, 27%, 44% and 11%, respectively. The remaining strains did not have a common IncFI plasmid but harboured transferable IncFI plasmids with sizes that ranged from 97 to 242.5 kb. Conclusion Our data demonstrate the presence of IncFI plasmids within the prevailing E. coli population in a hospital setting and suggest that the dissemination of CTX-M-15 allele is associated to lateral transfer of these well-adapted, conjugative IncFI plasmids among various E. coli genotypes.

  9. How many replicate tests do I need?$-$ Variability of cookstove performance and emissions has implications for obtaining useful results

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Yungang [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Environmental Energy Technologies Division; Sohn, Michael D. [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Environmental Energy Technologies Division; Gadgil, Ashok J. [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Environmental Energy Technologies Division; Wang, Yilun [ISO Innovative Analytics San Francisco, CA (United States); Lask, Kathleen M. [Univ. of California, Berkeley, CA (United States). College of Engineering Applied Science and Technology Program; Kirchstetter, Thomas W. [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Environmental Energy Technologies Division

    2013-02-01

    Almost half of the world’s population still cooks on biomass cookstoves of poor efficiency and primitive design, such as three stone fires (TSF). Emissions from biomass cookstoves contribute to adverse health effects and climate change. A number of “improved cookstoves” with higher energy efficiency and lower emissions have been designed and promoted across the world. During the design development, and for selection of a stove for dissemination, the stove performance and emissions are commonly evaluated, communicated and compared using the arithmetic average of replicate tests made using a standardized laboratory-based test, commonly the water boiling test (WBT). However, published literature shows different WBT results reported from different laboratories for the same stove technology. Also, there is no agreement in the literature on how many replicate tests should be performed to ensure “significance” in the reported average performance. This matter has not received attention in the rapidly growing literature on stoves, and yet is crucial for estimating and communicating the performance of a stove, and for comparing the performance between stoves. We present results of statistical analyses using data from a number of replicate tests of performance and emission of the Berkeley-Darfur Stove (BDS) and the TSF under well-controlled laboratory conditions. We observed moderate variability in the test results for the TSF and BDS when measuring several characteristics. Here we focus on two as illustrative: time-to-boil and PM2.5 (particulate matter less than or equal to 2.5 micrometers in diameter) emissions. We demonstrate that interpretation of the results comparing these stoves could be misleading if only a small number of replicates had been conducted. We then describe a practical approach, useful to both stove testers and designers, to assess the number of replicates needed to obtain useful data. Caution should be exercised in attaching high credibility to

  10. Seasonal differences in the subjective assessment of outdoor thermal conditions and the impact of analysis techniques on the obtained results

    Science.gov (United States)

    Kántor, Noémi; Kovács, Attila; Takács, Ágnes

    2016-11-01

    Wide research attention has been paid in the last two decades to the thermal comfort conditions of different outdoor and semi-outdoor urban spaces. Field studies were conducted in a wide range of geographical regions in order to investigate the relationship between the thermal sensation of people and thermal comfort indices. Researchers found that the original threshold values of these indices did not describe precisely the actual thermal sensation patterns of subjects, and they reported neutral temperatures that vary among nations and with time of the year. For that reason, thresholds of some objective indices were rescaled and new thermal comfort categories were defined. This research investigates the outdoor thermal perception patterns of Hungarians regarding the Physiologically Equivalent Temperature ( PET) index, based on more than 5800 questionnaires. The surveys were conducted in the city of Szeged on 78 days in spring, summer, and autumn. Various, frequently applied analysis approaches (simple descriptive technique, regression analysis, and probit models) were adopted to reveal seasonal differences in the thermal assessment of people. Thermal sensitivity and neutral temperatures were found to be significantly different, especially between summer and the two transient seasons. Challenges of international comparison are also emphasized, since the results prove that neutral temperatures obtained through different analysis techniques may be considerably different. The outcomes of this study underline the importance of the development of standard measurement and analysis methodologies in order to make future studies comprehensible, hereby facilitating the broadening of the common scientific knowledge about outdoor thermal comfort.

  11. Design, operational experiences and beam results obtained with the SNS H- ion source and LEBT at Berkeley Lab

    International Nuclear Information System (INIS)

    Keller, R.; Thomae, R.; Stockli, M.; Welton, R.

    2002-01-01

    The ion source and Low-Energy Transport (LEBT) system that will provide H - ion beams to the Spallation Neutron Source (SNS)** Front End and the accelerator chain have been developed into a mature unit that fully satisfies the operational requirements through the commissioning and early operating phases of SNS. Compared to the early R and D version, many features of the ion source have been improved, and reliable operation at 6% duty factor has been achieved producing beam currents in the 35-mA range and above. LEBT operation proved that the purely electrostatic focusing principle is well suited to inject the ion beam into the RFQ accelerator, including the steering and pre-chopping functions. This paper will discuss the latest design features of the ion source and LEBT, give performance data for the integrated system, and report on commissioning results obtained with the SNS RFQ and Medium-Energy Beam Transport (MEBT) system. Prospects for further improvements will be outlined in concluding remarks

  12. Functional relationships between plasmids and their significance for metabolism and symbiotic performance of Rhizobium leguminosarum bv. trifolii.

    Science.gov (United States)

    Stasiak, Grażyna; Mazur, Andrzej; Wielbo, Jerzy; Marczak, Małgorzata; Zebracki, Kamil; Koper, Piotr; Skorupska, Anna

    2014-11-01

    Rhizobium leguminosarum bv. trifolii TA1 (RtTA1) is a soil bacterium establishing a highly specific symbiotic relationship with clover, which is based on the exchange of molecular signals between the host plant and the microsymbiont. The RtTA1 genome is large and multipartite, composed of a chromosome and four plasmids, which comprise approximately 65 % and 35 % of the total genome, respectively. Extrachromosomal replicons were previously shown to confer significant metabolic versatility to bacteria, which is important for their adaptation in the soil and nodulation competitiveness. To investigate the contribution of individual RtTA1 plasmids to the overall cell phenotype, metabolic properties and symbiotic performance, a transposon-based elimination strategy was employed. RtTA1 derivatives cured of pRleTA1b or pRleTA1d and deleted in pRleTA1a were obtained. In contrast to the in silico predictions of pRleTA1b and pRleTA1d, which were described as chromid-like replicons, both appeared to be completely curable. On the other hand, for pRleTA1a (symbiotic plasmid) and pRleTA1c, which were proposed to be unessential for RtTA1 viability, it was not possible to eliminate them at all (pRleTA1c) or entirely (pRleTA1a). Analyses of the phenotypic traits of the RtTA1 derivatives obtained revealed the functional significance of individual plasmids and their indispensability for growth, certain metabolic pathways, production of surface polysaccharides, autoaggregation, biofilm formation, motility and symbiotic performance. Moreover, the results allow us to suggest broad functional cooperation among the plasmids in shaping the phenotypic properties and symbiotic capabilities of rhizobia.

  13. Solar heating by radiant floor: Experimental results and emission reduction obtained with a micro photovoltaic–heat pump system

    International Nuclear Information System (INIS)

    Izquierdo, M.; Agustín-Camacho, P. de

    2015-01-01

    Highlights: • This work presents a PVT multicrystalline solar heating system for buildings. • The PV DC electricity generated was converted to AC to drive an air–water heat pump. • Experimental results obtained from December 1, 2012 to April 30, 2013 are detailed. • An environmental study is also presented. - Abstract: An experimental research with a solar photovoltaic thermal (PVT) micro grid feeding a reversible air–water, 6 kW heating capacity heat pump, has been carried out from December 2012 to April 2013. Its purpose is to heat a laboratory that is used as a house prototype for the study of heating/cooling systems. It was built in accordance with the 2013 Spanish CTE, and has an area of 35 m 2 divided into two internal rooms: one of them housing the storage system, the solar controller, the inverter and the control system; the other one is occupied by three people. Its main thermal characteristics are: UA = 125 W/°C and a maximum thermal load about 6.0 kW at the initial time. The PVT field consists of 12 modules, with a total area of 15.7 m 2 and useful area of 14 m 2 . Each module is composed of 48 polycrystalline silicon cells of 243.4 cm 2 , which with a nominal efficiency 14% can generate a power of 180 W, being the total nominal power installed 2.16 kW. The PV system stores electricity in 250 Ah batteries from where is converted from DC to AC through a 3.0 kW inverter that feeds the heat pump. This works supplying 840 l/h of hot water at 35–45 °C to the radiant floor. The data storing system is recording variables such as solar radiation; temperatures; input power to batteries; heat produced; heat transferred by the radiant floor; heat pump’s COP; isolated ratio; and solar fraction. The objective of this work is to present and discuss the experimental results and the emission reduction of CO 2 obtained during the period from 01/12/2012 to 30/04/2013, including the detailed results of two representative days of Madrid’s climate: 28

  14. Infectious alphavirus production from a simple plasmid transfection+

    Directory of Open Access Journals (Sweden)

    Olson Ken E

    2011-07-01

    Full Text Available Abstract We have developed a new method for producing infectious double subgenomic alphaviruses from plasmids transfected into mammalian cells. A double subgenomic Sindbis virus (TE3'2J was transcribed from a cytomegalovirus PolII promoter, which results in the production of infectious virus. Transfection of as little as 125 ng of plasmid is able to produce 1 × 108 plaque forming units/ml (PFU/ml of infectious virus 48 hours post-transfection. This system represents a more efficient method for producing recombinant Sindbis viruses.

  15. Comparative Sequence Analysis of Multidrug-Resistant IncA/C Plasmids from Salmonella enterica.

    Science.gov (United States)

    Hoffmann, Maria; Pettengill, James B; Gonzalez-Escalona, Narjol; Miller, John; Ayers, Sherry L; Zhao, Shaohua; Allard, Marc W; McDermott, Patrick F; Brown, Eric W; Monday, Steven R

    2017-01-01

    Determinants of multidrug resistance (MDR) are often encoded on mobile elements, such as plasmids, transposons, and integrons, which have the potential to transfer among foodborne pathogens, as well as to other virulent pathogens, increasing the threats these traits pose to human and veterinary health. Our understanding of MDR among Salmonella has been limited by the lack of closed plasmid genomes for comparisons across resistance phenotypes, due to difficulties in effectively separating the DNA of these high-molecular weight, low-copy-number plasmids from chromosomal DNA. To resolve this problem, we demonstrate an efficient protocol for isolating, sequencing and closing IncA/C plasmids from Salmonella sp. using single molecule real-time sequencing on a Pacific Biosciences (Pacbio) RS II Sequencer. We obtained six Salmonella enterica isolates from poultry, representing six different serovars, each exhibiting the MDR-Ampc resistance profile. Salmonella plasmids were obtained using a modified mini preparation and transformed with Escherichia coli DH10Br. A Qiagen Large-Construct kit™ was used to recover highly concentrated and purified plasmid DNA that was sequenced using PacBio technology. These six closed IncA/C plasmids ranged in size from 104 to 191 kb and shared a stable, conserved backbone containing 98 core genes, with only six differences among those core genes. The plasmids encoded a number of antimicrobial resistance genes, including those for quaternary ammonium compounds and mercury. We then compared our six IncA/C plasmid sequences: first with 14 IncA/C plasmids derived from S. enterica available at the National Center for Biotechnology Information (NCBI), and then with an additional 38 IncA/C plasmids derived from different taxa. These comparisons allowed us to build an evolutionary picture of how antimicrobial resistance may be mediated by this common plasmid backbone. Our project provides detailed genetic information about resistance genes in

  16. Comparative Sequence Analysis of Multidrug-Resistant IncA/C Plasmids from Salmonella enterica

    Directory of Open Access Journals (Sweden)

    Maria Hoffmann

    2017-08-01

    Full Text Available Determinants of multidrug resistance (MDR are often encoded on mobile elements, such as plasmids, transposons, and integrons, which have the potential to transfer among foodborne pathogens, as well as to other virulent pathogens, increasing the threats these traits pose to human and veterinary health. Our understanding of MDR among Salmonella has been limited by the lack of closed plasmid genomes for comparisons across resistance phenotypes, due to difficulties in effectively separating the DNA of these high-molecular weight, low-copy-number plasmids from chromosomal DNA. To resolve this problem, we demonstrate an efficient protocol for isolating, sequencing and closing IncA/C plasmids from Salmonella sp. using single molecule real-time sequencing on a Pacific Biosciences (Pacbio RS II Sequencer. We obtained six Salmonella enterica isolates from poultry, representing six different serovars, each exhibiting the MDR-Ampc resistance profile. Salmonella plasmids were obtained using a modified mini preparation and transformed with Escherichia coli DH10Br. A Qiagen Large-Construct kit™ was used to recover highly concentrated and purified plasmid DNA that was sequenced using PacBio technology. These six closed IncA/C plasmids ranged in size from 104 to 191 kb and shared a stable, conserved backbone containing 98 core genes, with only six differences among those core genes. The plasmids encoded a number of antimicrobial resistance genes, including those for quaternary ammonium compounds and mercury. We then compared our six IncA/C plasmid sequences: first with 14 IncA/C plasmids derived from S. enterica available at the National Center for Biotechnology Information (NCBI, and then with an additional 38 IncA/C plasmids derived from different taxa. These comparisons allowed us to build an evolutionary picture of how antimicrobial resistance may be mediated by this common plasmid backbone. Our project provides detailed genetic information about

  17. The effects of metal ion PCR inhibitors on results obtained with the Quantifiler(®) Human DNA Quantification Kit.

    Science.gov (United States)

    Combs, Laura Gaydosh; Warren, Joseph E; Huynh, Vivian; Castaneda, Joanna; Golden, Teresa D; Roby, Rhonda K

    2015-11-01

    Forensic DNA samples may include the presence of PCR inhibitors, even after extraction and purification. Studies have demonstrated that metal ions, co-purified at specific concentrations, inhibit DNA amplifications. Metal ions are endogenous to sample types, such as bone, and can be introduced from environmental sources. In order to examine the effect of metal ions as PCR inhibitors during quantitative real-time PCR, 2800 M DNA was treated with 0.0025-18.750 mM concentrations of aluminum, calcium, copper, iron, nickel, and lead. DNA samples, both untreated and metal-treated, were quantified using the Quantifiler(®) Human DNA Quantification Kit. Quantification cycle (Cq) values for the Quantifiler(®) Human DNA and internal PCR control (IPC) assays were measured and the estimated concentrations of human DNA were obtained. Comparisons were conducted between metal-treated and control DNA samples to determine the accuracy of the quantification estimates and to test the efficacy of the IPC inhibition detection. This kit is most resistant to the presence of calcium as compared to all metals tested; the maximum concentration tested does not affect the amplification of the IPC or quantification of the sample. This kit is most sensitive to the presence of aluminum; concentrations greater than 0.0750 mM negatively affected the quantification, although the IPC assay accurately assessed the presence of PCR inhibition. The Quantifiler(®) Human DNA Quantification Kit accurately quantifies human DNA in the presence of 0.5000 mM copper, iron, nickel, and lead; however, the IPC does not indicate the presence of PCR inhibition at this concentration of these metals. Unexpectedly, estimates of DNA quantity in samples treated with 18.750 mM copper yielded values in excess of the actual concentration of DNA in the samples; fluorescence spectroscopy experiments indicated this increase was not a direct interaction between the copper metal and 6-FAM dye used to label the probe that

  18. Uncertainty and Sensitivity Analysis Results Obtained in the 1996 Performance Assessment for the Waste Isolation Pilot Plant

    Energy Technology Data Exchange (ETDEWEB)

    Bean, J.E.; Berglund, J.W.; Davis, F.J.; Economy, K.; Garner, J.W.; Helton, J.C.; Johnson, J.D.; MacKinnon, R.J.; Miller, J.; O' Brien, D.G.; Ramsey, J.L.; Schreiber, J.D.; Shinta, A.; Smith, L.N.; Stockman, C.; Stoelzel, D.M.; Vaughn, P.

    1998-09-01

    The Waste Isolation Pilot Plant (WPP) is located in southeastern New Mexico and is being developed by the U.S. Department of Energy (DOE) for the geologic (deep underground) disposal of transuranic (TRU) waste. A detailed performance assessment (PA) for the WIPP was carried out in 1996 and supports an application by the DOE to the U.S. Environmental Protection Agency (EPA) for the certification of the WIPP for the disposal of TRU waste. The 1996 WIPP PA uses a computational structure that maintains a separation between stochastic (i.e., aleatory) and subjective (i.e., epistemic) uncertainty, with stochastic uncertainty arising from the many possible disruptions that could occur over the 10,000 yr regulatory period that applies to the WIPP and subjective uncertainty arising from the imprecision with which many of the quantities required in the PA are known. Important parts of this structure are (1) the use of Latin hypercube sampling to incorporate the effects of subjective uncertainty, (2) the use of Monte Carlo (i.e., random) sampling to incorporate the effects of stochastic uncertainty, and (3) the efficient use of the necessarily limited number of mechanistic calculations that can be performed to support the analysis. The use of Latin hypercube sampling generates a mapping from imprecisely known analysis inputs to analysis outcomes of interest that provides both a display of the uncertainty in analysis outcomes (i.e., uncertainty analysis) and a basis for investigating the effects of individual inputs on these outcomes (i.e., sensitivity analysis). The sensitivity analysis procedures used in the PA include examination of scatterplots, stepwise regression analysis, and partial correlation analysis. Uncertainty and sensitivity analysis results obtained as part of the 1996 WIPP PA are presented and discussed. Specific topics considered include two phase flow in the vicinity of the repository, radionuclide release from the repository, fluid flow and radionuclide

  19. Uncertainty and Sensitivity Analysis Results Obtained in the 1996 Performance Assessment for the Waste Isolation Pilot Plant

    International Nuclear Information System (INIS)

    Bean, J.E.; Berglund, J.W.; Davis, F.J.; Economy, K.; Garner, J.W.; Helton, J.C.; Johnson, J.D.; MacKinnon, R.J.; Miller, J.; O'Brien, D.G.; Ramsey, J.L.; Schreiber, J.D.; Shinta, A.; Smith, L.N.; Stockman, C.; Stoelzel, D.M.; Vaughn, P.

    1998-01-01

    The Waste Isolation Pilot Plant (WPP) is located in southeastern New Mexico and is being developed by the U.S. Department of Energy (DOE) for the geologic (deep underground) disposal of transuranic (TRU) waste. A detailed performance assessment (PA) for the WIPP was carried out in 1996 and supports an application by the DOE to the U.S. Environmental Protection Agency (EPA) for the certification of the WIPP for the disposal of TRU waste. The 1996 WIPP PA uses a computational structure that maintains a separation between stochastic (i.e., aleatory) and subjective (i.e., epistemic) uncertainty, with stochastic uncertainty arising from the many possible disruptions that could occur over the 10,000 yr regulatory period that applies to the WIPP and subjective uncertainty arising from the imprecision with which many of the quantities required in the PA are known. Important parts of this structure are (1) the use of Latin hypercube sampling to incorporate the effects of subjective uncertainty, (2) the use of Monte Carlo (i.e., random) sampling to incorporate the effects of stochastic uncertainty, and (3) the efficient use of the necessarily limited number of mechanistic calculations that can be performed to support the analysis. The use of Latin hypercube sampling generates a mapping from imprecisely known analysis inputs to analysis outcomes of interest that provides both a display of the uncertainty in analysis outcomes (i.e., uncertainty analysis) and a basis for investigating the effects of individual inputs on these outcomes (i.e., sensitivity analysis). The sensitivity analysis procedures used in the PA include examination of scatterplots, stepwise regression analysis, and partial correlation analysis. Uncertainty and sensitivity analysis results obtained as part of the 1996 WIPP PA are presented and discussed. Specific topics considered include two phase flow in the vicinity of the repository, radionuclide release from the repository, fluid flow and radionuclide

  20. Characterization of a collection of plasmid-containing bacteria isolated from an on-farm biopurification system used for pesticide removal.

    Science.gov (United States)

    Martini, María Carla; Albicoro, Francisco Javier; Nour, Eman; Schlüter, Andreas; van Elsas, Jan Dirk; Springael, Dirk; Smalla, Kornelia; Pistorio, Mariano; Lagares, Antonio; Del Papa, María Florencia

    2015-07-01

    Biopurification systems (BPS) are complex soil-related and artificially-generated environments usually designed for the removal of toxic compounds from contaminated wastewaters. The present study has been conducted to isolate and characterize a collection of cultivable plasmid-carrying bacterial isolates recovered from a BPS established for the decontamination of wastewater generated in a farmyard. Out of 1400 isolates, a collection of 75 plasmid-containing bacteria was obtained, of which 35 representative isolates comprising in total at least 50 plasmids were chosen for further characterization. Bacterial hosts were taxonomically assigned by 16S ribosomal RNA gene sequencing and phenotypically characterized according to their ability to grow in presence of different antibiotics and heavy metals. The study demonstrated that a high proportion of the isolates was tolerant to antibiotics and/or heavy metals, highlighting the on-farm BPS enrichment in such genetic traits. Several plasmids conferring such resistances in the bacterial collection were detected to be either mobilizable or selftransmissible. Occurrence of broad host range plasmids of the incompatibility groups IncP, IncQ, IncN and IncW was examined with positive results only for the first group. Presence of the IS1071 insertion sequence, frequently associated with xenobiotics degradation genes, was detected in DNA obtained from 24 of these isolates, strongly suggesting the presence of yet-hidden catabolic activities in the collection of isolates. The results showed a remarkable diversity in the plasmid mobilome of cultivable bacteria in the BPS with the presence of abundant resistance markers of different types, thus providing a suitable environment to investigate the genetic structure of the mobile genetic pool in a model on-farm biofilter for wastewater decontamination in intensive agricultural production. Copyright © 2015 Elsevier Inc. All rights reserved.

  1. Mutation in ESBL Plasmid from Escherichia coli O104:H4 Leads Autoagglutination and Enhanced Plasmid Dissemination

    Directory of Open Access Journals (Sweden)

    Mickaël Poidevin

    2018-02-01

    Full Text Available Conjugative plasmids are one of the main driving force of wide-spreading of multidrug resistance (MDR bacteria. They are self-transmittable via conjugation as carrying the required set of genes and cis-acting DNA locus for direct cell-to-cell transfer. IncI incompatibility plasmids are nowadays often associated with extended-spectrum beta-lactamases producing Enterobacteria in clinic and environment. pESBL-EA11 was isolated from Escherichia coli O104:H4 outbreak strain in Germany in 2011. During the previous study identifying transfer genes of pESBL-EA11, it was shown that transposon insertion at certain DNA region of the plasmid, referred to as Hft, resulted in great enhancement of transfer ability. This suggested that genetic modifications can enhance dissemination of MDR plasmids. Such ‘superspreader’ mutations have attracted little attention so far despite their high potential to worsen MDR spreading. Present study aimed to gain our understanding on regulatory elements that involved pESBL transfer. While previous studies of IncI plasmids indicated that immediate downstream gene of Hft, traA, is not essential for conjugative transfer, here we showed that overexpression of TraA in host cell elevated transfer rate of pESBL-EA11. Transposon insertion or certain nucleotide substitutions in Hft led strong TraA overexpression which resulted in activation of essential regulator TraB and likely overexpression of conjugative pili. Atmospheric Scanning Electron Microscopy observation suggested that IncI pili are distinct from other types of conjugative pili (such as long filamentous F-type pili and rather expressed throughout the cell surface. High transfer efficiency in the mutant pESBL-EA11 was involved with hyperpiliation which facilitates cell-to-cell adhesion, including autoagglutination. The capability of plasmids to evolve to highly transmissible mutant is alarming, particularly it might also have adverse effect on host pathogenicity.

  2. Results of environmental radiation monitoring and meteorology measurements (material prepared for obtaining the licence for RA reactor experimental operation)

    International Nuclear Information System (INIS)

    1980-10-01

    According to the demands for obtaining the licence for restarting the Ra reactor and the experimental operation this document includes the radiation monitoring measured data in the working space and environment of the RA reactor, i.e. Boris Kidric Institute. The meteorology measured data are included as well. All the measurements are performed according to the radiation protection program applied actually from the first reactor start-up at the end of 1959 [sr

  3. Obtaining a citric tristeza virus p65 protein antibody and preliminary results of p65 in vivo expression

    Directory of Open Access Journals (Sweden)

    Yanneth Torres

    2003-07-01

    Full Text Available The citric tristeza virus (CTV belongs to the Closteroviridae family which indudes the only vegetal viruses possessing genes homologous to HSP70 thermal cellular shock proteins in their genome. Such is the case of the gene encoding for the CTV p65 protein which presents high homology with the HSP70 protein family. It has been shown recently that HSP70h viral proteins (such as CTV p65 are involved both in viral assembly, as a microtubule binding protein, and in cell-cell movement. Since CTV is the most deleterious citrus pathogen, understanding this protein's role in the pathogenesis process is important. Rabbits were immunised with four synthetic peptides (corresponding to CTV p65 thermal shock protein's carboxyl-terminal region to obtain polyclonal antibodies. All the peptides used were immunogenic, even though two of them showed greater response. Whilst none of the antibodies obtained reacted to non-infected plant extract, the p65 proteins was detected in extracts taken from citric plants infected with CTV Based on the antibody's reaction to two Colombian isolates having different serological characteristics, the p65 antibody's immunological behaviour appeared to be independent of the symptomatic severity of the CTV isolates. It was shown that the ORF encoded for the HSP70 homologue in CTV was expressed in vivo, even though the p65 antibody was only detected in concentrated protein extracts taken from infected plants, supporting reports from other studies that the concentration of this protein in plants infected with CTV is low. This is the first time that a polyclonal CTV antibody has been obtained in Colombia against p65 (a protein intervening in viral assembly and movement. Adapting a technique for obtaining p65 antibodies by using synthetic peptides as immunogens could be useful in the future for detecting or diagnosing p65 proteins present in different Colombian CTV isolates, especially in developing studies contributing towards greater

  4. Visualization of atomic distances at the 1MV electron microscope: first results obtained on the Grenoble 1MV microscope

    International Nuclear Information System (INIS)

    Bourret, A.

    1975-01-01

    Practical and theoretical conditions for obtaining high resolution lattice images are presented. The use of a high voltage electron microscope is particularly powerful to visualize distances smaller than 3A. A 2A resolution test on (200) gold planes has been carried out on the Grenoble 1MV microscope. It would be possible at this level to study the crystalline defects such as dislocations or grain boundaries [fr

  5. Ordering the mob: Insights into replicon and MOB typing schemes from analysis of a curated dataset of publicly available plasmids.

    Science.gov (United States)

    Orlek, Alex; Phan, Hang; Sheppard, Anna E; Doumith, Michel; Ellington, Matthew; Peto, Tim; Crook, Derrick; Walker, A Sarah; Woodford, Neil; Anjum, Muna F; Stoesser, Nicole

    2017-05-01

    Plasmid typing can provide insights into the epidemiology and transmission of plasmid-mediated antibiotic resistance. The principal plasmid typing schemes are replicon typing and MOB typing, which utilize variation in replication loci and relaxase proteins respectively. Previous studies investigating the proportion of plasmids assigned a type by these schemes ('typeability') have yielded conflicting results; moreover, thousands of plasmid sequences have been added to NCBI in recent years, without consistent annotation to indicate which sequences represent complete plasmids. Here, a curated dataset of complete Enterobacteriaceae plasmids from NCBI was compiled, and used to assess the typeability and concordance of in silico replicon and MOB typing schemes. Concordance was assessed at hierarchical replicon type resolutions, from replicon family-level to plasmid multilocus sequence type (pMLST)-level, where available. We found that 85% and 65% of the curated plasmids could be replicon and MOB typed, respectively. Overall, plasmid size and the number of resistance genes were significant independent predictors of replicon and MOB typing success. We found some degree of non-concordance between replicon families and MOB types, which was only partly resolved when partitioning plasmids into finer-resolution groups (replicon and pMLST types). In some cases, non-concordance was attributed to ambiguous boundaries between MOBP and MOBQ types; in other cases, backbone mosaicism was considered a more plausible explanation. β-lactamase resistance genes tended not to show fidelity to a particular plasmid type, though some previously reported associations were supported. Overall, replicon and MOB typing schemes are likely to continue playing an important role in plasmid analysis, but their performance is constrained by the diverse and dynamic nature of plasmid genomes. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

  6. Large-scale preparation of plasmid DNA.

    Science.gov (United States)

    Heilig, J S; Elbing, K L; Brent, R

    2001-05-01

    Although the need for large quantities of plasmid DNA has diminished as techniques for manipulating small quantities of DNA have improved, occasionally large amounts of high-quality plasmid DNA are desired. This unit describes the preparation of milligram quantities of highly purified plasmid DNA. The first part of the unit describes three methods for preparing crude lysates enriched in plasmid DNA from bacterial cells grown in liquid culture: alkaline lysis, boiling, and Triton lysis. The second part describes four methods for purifying plasmid DNA in such lysates away from contaminating RNA and protein: CsCl/ethidium bromide density gradient centrifugation, polyethylene glycol (PEG) precipitation, anion-exchange chromatography, and size-exclusion chromatography.

  7. A degenerate primer MOB typing (DPMT method to classify gamma-proteobacterial plasmids in clinical and environmental settings.

    Directory of Open Access Journals (Sweden)

    Andrés Alvarado

    Full Text Available Transmissible plasmids are responsible for the spread of genetic determinants, such as antibiotic resistance or virulence traits, causing a large ecological and epidemiological impact. Transmissible plasmids, either conjugative or mobilizable, have in common the presence of a relaxase gene. Relaxases were previously classified in six protein families according to their phylogeny. Degenerate primers hybridizing to coding sequences of conserved amino acid motifs were designed to amplify related relaxase genes from γ-Proteobacterial plasmids. Specificity and sensitivity of a selected set of 19 primer pairs were first tested using a collection of 33 reference relaxases, representing the diversity of γ-Proteobacterial plasmids. The validated set was then applied to the analysis of two plasmid collections obtained from clinical isolates. The relaxase screening method, which we call "Degenerate Primer MOB Typing" or DPMT, detected not only most known Inc/Rep groups, but also a plethora of plasmids not previously assigned to any Inc group or Rep-type.

  8. Shaking table testing of a HTGR reactor core, comparison with the results obtained using a nonlinear mathematical model

    International Nuclear Information System (INIS)

    Berriaud, C.; Cebe, E.; Livolant, M.; Buland, P.

    1975-01-01

    Two series of horizontal tests have been performed at Saclay on the shaking table VESUVE: sinusoidal test and time history response. Sinusoidal tests have shown the strongly nonlinear dynamic behavior of the core. The resonant frequency of the core is dependent on the level of the excitation. These phenomena have been explained by a computer code, which is a lumped mass nonlinear model. El Centro time history displacement at the level of PCRV was reproduced on the shaking table. The analytical model was applied to this excitation and good comparison was obtained for forces and velocities [fr

  9. Deciphering conjugative plasmid permissiveness in wastewater microbiomes

    DEFF Research Database (Denmark)

    Jacquiod, Samuel Jehan Auguste; Brejnrod, Asker Daniel; Milani, Stefan Morberg

    2017-01-01

    Wastewater treatment plants (WWTPs) are designed to robustly treat polluted water. They are characterized by ceaseless flows of organic, chemical and microbial matter, followed by treatment steps before environmental release. WWTPs are hotspots of horizontal gene transfer between bacteria via...... still remains largely uncharted. Furthermore, current in vitro methods used to assess conjugation in complex microbiomes do not include in situ behaviours of recipient cells, resulting in partial understanding of transfers. We investigated the in vitro conjugation capacities of WWTP microbiomes from...... inlet sewage and outlet treated water using the broad-host range IncP-1 conjugative plasmid, pKJK5. A thorough molecular approach coupling metagenomes to 16S rRNA DNA/cDNA amplicon sequencing was established to characterize microbiomes using the ecological concept of functional response groups. A broad...

  10. Quantification bias caused by plasmid DNA conformation in quantitative real-time PCR assay.

    Science.gov (United States)

    Lin, Chih-Hui; Chen, Yu-Chieh; Pan, Tzu-Ming

    2011-01-01

    Quantitative real-time PCR (qPCR) is the gold standard for the quantification of specific nucleic acid sequences. However, a serious concern has been revealed in a recent report: supercoiled plasmid standards cause significant over-estimation in qPCR quantification. In this study, we investigated the effect of plasmid DNA conformation on the quantification of DNA and the efficiency of qPCR. Our results suggest that plasmid DNA conformation has significant impact on the accuracy of absolute quantification by qPCR. DNA standard curves shifted significantly among plasmid standards with different DNA conformations. Moreover, the choice of DNA measurement method and plasmid DNA conformation may also contribute to the measurement error of DNA standard curves. Due to the multiple effects of plasmid DNA conformation on the accuracy of qPCR, efforts should be made to assure the highest consistency of plasmid standards for qPCR. Thus, we suggest that the conformation, preparation, quantification, purification, handling, and storage of standard plasmid DNA should be described and defined in the Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) to assure the reproducibility and accuracy of qPCR absolute quantification.

  11. Effect of the atmospheric pressure nonequilibrium plasmas on the conformational changes of plasmid DNA

    International Nuclear Information System (INIS)

    Yan Xu; He Guangyuan; Shi Mengjun; Gao Xuan; Li Yin; Ma Fengyun; Yu Men; Wang Changdong; Wang Yuesheng; Yang Guangxiao; Zou Fei; Lu Xinpei; Xiong Qing; Xiong Zilan

    2009-01-01

    The cold atmospheric pressure plasma, which has been widely used for biomedical applications, may potentially affect the conformation of DNA. In this letter, an atmospheric pressure plasma plume is used to investigate its effects on the conformational changes of DNA of plasmid pAHC25. It is found that the plasma plume could cause plasmid DNA topology alteration, resulting in the percentage of the supercoiled plasmid DNA form decreased while that of the open circular and linearized form of plasmid DNA increased as detected by agrose gel electrophoresis. On the other hand, further investigation by using polymerase chain reaction method shows that the atmospheric pressure plasma jet treatments under proper conditions does not affect the genes of the plasmid DNA, which may have potential application in increasing the transformation frequency by genetic engineering.

  12. An easy way to obtain strong duality results in linear, linear semidefinite and linear semi-infinite programming

    NARCIS (Netherlands)

    Pop, P.C.; Still, Georg J.

    1999-01-01

    In linear programming it is known that an appropriate non-homogeneous Farkas Lemma leads to a short proof of the strong duality results for a pair of primal and dual programs. By using a corresponding generalized Farkas lemma we give a similar proof of the strong duality results for semidefinite

  13. Recent results from Pb - Au collisions at 158 GeV/c per nucleon obtained with the CERES spectrometer

    CERN Document Server

    Agakichiev, G; Baur, R; Braun-Munzinger, P; Ceretto, F; Cherlin, A; Drees, A; Esumi, S C; Faschingbauer, U; Fraenkel, Zeev; Glässel, P; Hering, G; Holl, P; Jung, C; Lenkeit, B C; Marin, A; Messer, M; Miskowiec, D; Panebratsev, Yu A; Pfeiffer, A; Rak, J; Ravinovich, I; Razin, S V; Rehak, P; Richter, M; Saveljic, N; Schmitz, W; Schükraft, Jürgen; Seipp, W; Shimansky, S S; Slivova, J; Socol, E; Specht, H J; Stachel, J J; Tserruya, Itzhak; Ullrich, T S; Voigt, C A; Voloshin, S A; Weber, C; Wessels, J P; Wienold, T; Wurm, J P; Yurevich, V I

    1999-01-01

    During the 1996 lead run time, CERES has accumulated 42 million events, corresponding to a factor of 5 more statistics than in 1995 and 2.5 million events of a special photon-run. We report on the results of the low-mass e/sup +/e/sup -/-pair analysis. Since the most critical item is the poor signal-to-background ratio we also discuss the understanding of this background, in absolute terms, with the help of a detailed Monte Carlo simulation. We show preliminary results of the photon analysis and summarize the results of the hadron analysis preliminarily reported on already at QM'97. (19 refs) .

  14. Selfish restriction modification genes: resistance of a resident R/M plasmid to displacement by an incompatible plasmid mediated by host killing.

    Science.gov (United States)

    Naito, Y; Naito, T; Kobayashi, I

    1998-01-01

    Previous work from this laboratory demonstrated that plasmids carrying a type II restriction-modification gene complex are not easily lost from their bacterial host because plasmid-free segregant cells are killed through chromosome cleavage. Here, we have followed the course of events that takes place when an Escherichia coli rec BC sbcA strain carrying a plasmid coding for the PaeR7I restriction-modification (R/M) gene complex is transformed by a plasmid with an identical origin of replication. The number of transformants that appeared was far fewer than with the restriction-minus (r-) control. Most of the transformants were very small. After prolonged incubation, the number and the size of the colonies increased, but this increase never attained the level of the r- control. Most of the transformed colonies retained the drug-resistance of the resident, r+ m+ plasmid. These results indicate that post-segregational host killing occurs when a plasmid bearing an R/M gene complex is displaced by an incompatible plasmid. Such cell killing eliminates the competitor plasmid along with the host and, thus, would allow persistence of the R/M plasmid in the neighboring, clonal host cells in nature. This phenomenon is reminiscent of mammalian apoptosis and other forms of altruistic cell death strategy against infection. This type of resistance to displacement was also studied in a wild type Escherichia coli strain that was normal for homologous recombination (rec+). A number of differences between the recBC sbcA strain and the rec+ strain were observed and these will be discussed.

  15. Study of the nuclear multifragmentation: recent results obtained with the INDRA detector in the intermediate energy domain

    International Nuclear Information System (INIS)

    Saint-Laurent, F.

    1994-01-01

    The new 4π multidetector INDRA, designed for the study of hot nuclear systems decaying by multifragmentation, is available for experiments since the beginning of 1993. First results emphasize its very high detection capabilities. Preliminary results on multiplicity distributions and elemental charge distributions in the most violent collisions for the 36 Ar + 58 Ni and 129 Xe + nat Sn systems are presented. (author) 5 figs., 22 refs

  16. Characterization of new plasmids from methylotrophic bacteria.

    Science.gov (United States)

    Brenner, V; Holubová, I; Benada, O; Hubácek, J

    1991-07-01

    Several tens of methanol-utilizing bacterial strains isolated from soil were screened for the presence of plasmids. From the obligate methylotroph Methylomonas sp. strain R103a plasmid pIH36 (36 kb) was isolated and its restriction map was constructed. In pink-pigmented facultative methylotrophs (PPFM), belonging to the genus Methylobacterium four plasmids were detected: plasmids pIB200 (200 kb) and pIB14 (14 kb) in the strain R15d and plasmids pWU14 (14 kb) and pWU7 (7.8 kb) in the strain M17. Because of the small size and the presence of several unique REN sites (HindIII, EcoRI, NcoI), plasmid pWU7 was chosen for the construction of a vector for cloning in methylotrophs. Cointegrates pKWU7A and pKWU7B were formed between pWU7 and the E. coli plasmid pK19 Kmr, which were checked for conjugative transfer from E. coli into the methylotrophic host.

  17. Main results obtained in France in the development of the gaseous diffusion process for uranium isotope separation

    International Nuclear Information System (INIS)

    Frejacques, C.; Bilous, O.; Dixmier, J.; Massignon, D.; Plurien, P.

    1958-01-01

    The main problems which occur in the study of uranium isotope separation by the gaseous diffusion process, concern the development of the porous barrier, the corrosive nature of uranium hexafluoride and also the chemical engineering problems related to process design and the choice of best plant and stage characteristics. Porous barriers may be obtained by chemical attack of non porous media or by agglomeration of very fine powders. Examples of these two types of barriers are given. A whole set of measurement techniques were developed for barrier structure studies, to provide control and guidance of barrier production methods. Uranium hexafluoride reactivity and corrosive properties are the source of many difficult technological problems. A high degree of plant leak tightness must be achieved. This necessity creates a special problem in compressor bearing design. Barrier lifetime is affected by the corrosive properties of the gas, which may lead to a change of barrier structure with time. Barrier hexafluoride permeability measurements have helped to make a systematic study of this point. Finally an example of a plant flowsheet, showing stage types and arrangements and based on a minimisation of enriched product costs is also given as an illustration of some of the chemical engineering problems present. (author) [fr

  18. Friction and wear in liquid-metal systems: comparability problems of test results obtained from different test facilities

    International Nuclear Information System (INIS)

    Wild, E.; Mack, K.J.

    1976-01-01

    Operational induced relative movements take place between contacting components in the core region of sodium cooled reactors. To ensure reliable long term functioning of such friction loaded components, materials are needed with good sliding properties and high wear resistance. Therefore, tribological properties of material combinations in liquid metal have been investigated experimentally for many years at various research establishments. However, despite identical boundary conditions, the comparison of results published does not yield a satisfactory agreement. The cause must be seen in the individual design and concept of the test sections used. This discrepancy was investigated. The results show that the elasticity, mass movement, and relative motion characteristic to the system prove to be the most important criteria influencing the test results

  19. Frequency and diversity of small cryptic plasmids in the genus Rahnella

    Directory of Open Access Journals (Sweden)

    Summers David K

    2010-02-01

    Full Text Available Abstract Background Rahnella is a widely distributed genus belonging to the Enterobacteriaceae and frequently present on vegetables. Although Rahnella has interesting agro-economical and industrial properties and several strains possess antibiotic resistances and toxin genes which might spread within microbial communities, little is known about plasmids of this genus. Thus, we isolated a number of Rahnella strains and investigated their complements of small plasmids. Results In total 53 strains were investigated and 11 plasmids observed. Seven belonged to the ColE1 family; one was ColE2-like and three shared homology to rolling circle plasmids. One of them belonged to the pC194/pUB110 family and two showed similarity to poorly characterised plasmid groups. The G+C content of two rolling circle plasmids deviated considerably from that of Rahnella, indicating that their usual hosts might belong to other genera. Most ColE1-like plasmids formed a subgroup within the ColE1 family that seems to be fairly specific for Rahnella. Intriguingly, the multimer resolution sites of all ColE1-like plasmids had the same orientation with respect to the origin of replication. This arrangement might be necessary to prevent inappropriate synthesis of a small regulatory RNA that regulates cell division. Although the ColE1-like plasmids did not possess any mobilisation system, they shared large parts with high sequence identity in coding and non-coding regions. In addition, highly homologous regions of plasmids isolated from Rahnella and the chromosomes of Erwinia tasmaniensis and Photorhabdus luminescens could be identified. Conclusions For the genus Rahnella we observed plasmid-containing isolates at a frequency of 19%, which is in the average range for Enterobacteriaceae. These plasmids belonged to diffent groups with members of the ColE1-family most frequently found. Regions of striking sequence homology of plasmids and bacterial chromosomes highlight the

  20. A new warfarin dosing algorithm including VKORC1 3730 G > A polymorphism: comparison with results obtained by other published algorithms.

    Science.gov (United States)

    Cini, Michela; Legnani, Cristina; Cosmi, Benilde; Guazzaloca, Giuliana; Valdrè, Lelia; Frascaro, Mirella; Palareti, Gualtiero

    2012-08-01

    Warfarin dosing is affected by clinical and genetic variants, but the contribution of the genotype associated with warfarin resistance in pharmacogenetic algorithms has not been well assessed yet. We developed a new dosing algorithm including polymorphisms associated both with warfarin sensitivity and resistance in the Italian population, and its performance was compared with those of eight previously published algorithms. Clinical and genetic data (CYP2C9*2, CYP2C9*3, VKORC1 -1639 G > A, and VKORC1 3730 G > A) were used to elaborate the new algorithm. Derivation and validation groups comprised 55 (58.2% men, mean age 69 years) and 40 (57.5% men, mean age 70 years) patients, respectively, who were on stable anticoagulation therapy for at least 3 months with different oral anticoagulation therapy (OAT) indications. Performance of the new algorithm, evaluated with mean absolute error (MAE) defined as the absolute value of the difference between observed daily maintenance dose and predicted daily dose, correlation with the observed dose and R(2) value, was comparable with or slightly lower than that obtained using the other algorithms. The new algorithm could correctly assign 53.3%, 50.0%, and 57.1% of patients to the low (≤25 mg/week), intermediate (26-44 mg/week) and high (≥ 45 mg/week) dosing range, respectively. Our data showed a significant increase in predictive accuracy among patients requiring high warfarin dose compared with the other algorithms (ranging from 0% to 28.6%). The algorithm including VKORC1 3730 G > A, associated with warfarin resistance, allowed a more accurate identification of resistant patients who require higher warfarin dosage.

  1. Cytogenetical effects of low doses: results obtained by N.N.Luchik and present-day problems

    International Nuclear Information System (INIS)

    Geras'kin, S.A.; Sevan'kaev, A.V.

    1996-01-01

    The analysis of present status of the problem of quantitative assessment of cytogenetic effects low is presented. The importance of works of N.V. Luchik is demonstrated for the development of this field of radiobiology. The results of the author's own experimental and theoretical research on the regularities of induction of cytogenetical damage by low doses of ionising radiation are presented

  2. Detection of occlusal caries lesions using fluorescence: correlation between histology and obtained results for Diagnodent and spectroscopy

    International Nuclear Information System (INIS)

    Rocha-Cabral, Renata Maciel

    2006-01-01

    The aims of this study were to develop and test a method to detect caries lesions in vivo and in vitro, using a portable spectrometer (PS); to analyze the performance of PS as well as the commercial device Diagnodent (Dd); correlate them with the gold standard, their transversal section areas and lesions depth and between themselves. 66 occlusal pre-molars sites were examined in vivo with Dd. Sequentially, fluorescence (λexc ∼ 657 nm) was collected by an optical fiber, conducted to PS and then analyzed as spectra, which were normalized and had calculated the Ratios of their Areas Under the Curves (RAUC) of carious and sound tissues. Experiments were conducted in vitro in the same sites. Gold Standard was obtained by polarized light microscopy. Pearson correlation was used to compare the devices with transversal section area, lesions depth and between themselves. The area under ROC curve, sensitivity, specificity as well as accuracy were calculated and verified with McNemar test. Dd and RAUC showed statistically significant correlation with gold standard (p < 0.01 for Dd and p < 0.05 for RAUC) and between themselves (r = 0,83 in vivo and r = 0,87 in vitro). Although it was significant, the devices showed low correlation with depth of lesions in vivo and in vitro (r = ∼ 0.43). The transversal section area of the lesion had no influence on readings in both devices. Dd showed higher sensitivity (0.76) than PS (0.60) in vivo (p < 0.05), though this fact was not able to improve its performance. In turn, PS showed higher sensitivity (0.88) than Dd (0.79) in vitro, but this difference was not significantly. The other parameters did not show statistically significant differences (p < 0.05) between methods. PS showed positive correlation with Dd, equal correlation with lesions depth and higher ability of detecting the disease in vitro, what suggests that if accompanied with a conic and an angulated probe and a dedicated software, the PS method could be useful in clinics

  3. A classification system for plasmids from Enterococci and other Gram-positive bacteria

    DEFF Research Database (Denmark)

    Jensen, Lars Bogø; Garcia-Migura, Lourdes; Valenzuela, Antonio Jesus Sanchez

    2010-01-01

    A classification system for plasmids isolated from enterococci and other Gram-positive bacteria was developed based on 111 published plasmid sequences from enterococci and other Gram-positive bacteria; mostly staphylococci. Based on PCR amplification of conserved areas of the replication initiating....... Furthermore, conjugation experiments were performed obtaining 30 transconjugants when selecting for antimicrobial resistance. Among them 19 gave no positive amplicons indicating presence of rep-families not tested for in this experimental setup....

  4. Quality control of baby food products on the basis of results obtained using the instrumental neutron-activation analysis technique

    International Nuclear Information System (INIS)

    Mukhammedov, S.; Khaydarov, A.; Pardaev, O.

    2013-01-01

    The purpose of this study was to use the instrumental neutron-activation analysis (INAA) to determine the elemental composition of some kind of imported baby food products (BFP) and to compare the results with the permissible contents. The nuclear reactor WWR-SM of INP has been used to develop INAA to study the mineral composition of some children's food products. The concentration of 26 trace elements, including Mg, Ca, Fe, Zn, etc. was found. The comparison of the results with regulation contents and the daily data on food needs have shown that the investigated group of BFP does not meet the requirements for all trace and macro elements composition. (authors)

  5. Pressure suppression system (PSS) for nuclear ships. Experimental results obtained at the GKSS PSS-test-facillity

    International Nuclear Information System (INIS)

    Aust, E.; Niemann, H.R.; Schwan, H.; Vollbrandt, J.

    1978-01-01

    The PSS-test facility is shortly presented which was designed to show experimentally the operation of the pressure suppression containment for the NCS 80 concept. The results of the experimental LOCA-simulation tests in the PSS-test facility are illustrated by diagrams. The observed phenomena as chugging and pessure oscillations immediately after vent clearing are reported as well as the thermohydraulic loadings of the total system. Finally a short view is given on the future test program

  6. Study of Finite Element Number Influence over the Obtained Results in Finite Element Analyses of a Mechanical Structure

    Directory of Open Access Journals (Sweden)

    Ana-Maria Budai

    2013-05-01

    Full Text Available This paper present the results of a study that was made to establish the influence of finite element number used to determined the real load of a structure. Actually, the study represent a linear static analyze for a link gear control mechanism of a Kaplan turbine. The all analyze was made for the normal condition of functioning having like final scope to determine de life time duration of mentioned mechanism.

  7. Constructing kinetics fatigue diagrams using testing results obtained on a machine with rigid loading for specimens of various thickness

    International Nuclear Information System (INIS)

    Simin'kovich, V.N.; Gladkij, Ya.N.; Deev, N.A.

    1981-01-01

    Bending tests of 40KhS steel specimens, tempered at 200 and 500 deg C, are conducted to investigate the possible effects of specimen thickness on fatigue crack growth. Kinetic fatigue diagrams are constructed using the investigation results. An increase in crack growth with thickness is observed only in high-tempered specimens. Changes in specimen thickness do not affect crack growth in 40KhS low-tempered steel [ru

  8. Characterization of High-Power Turbomachinery Tilting Pad Journal Bearings: First Results Obtained on a Novel Test Bench

    Directory of Open Access Journals (Sweden)

    Enrico Ciulli

    2018-01-01

    Full Text Available Tilting pad journal bearings are usually employed in turbomachines for their stable behavior at high rotational speeds. Devoted test rigs have been realized to validate the predictions of theoretical models. However, the design of new high-performance and large-size bearings needs to be supported by experimental investigations on high-performance large test rigs. The main characteristics of a recently built facility for testing large tilting pad journal bearings with diameters from 150 to 300 mm are described in this work. The test rig is versatile and can be used to test bearings of different size, configurations and to investigate the influence of many parameters, even the effect of misalignment. Sample results of the static characterization of a four-pad high-performance tilting pad journal bearing are reported evidencing some transient effects. A few sample dynamic results are also reported. The presented experimental results demonstrated the capabilities of the rig for investigating the static and the dynamic characteristics of the bearings accurately measuring slow and fast variables.

  9. Porous titanium obtained by a new powder metallurgy technique: Preliminary results of human osteoblast adhesion on surface polished substrates.

    Science.gov (United States)

    Biasotto, M; Ricceri, R; Scuor, N; Schmid, C; Sandrucci, M A; Di Lenarda, R; Matteazzi, P

    2003-01-01

    This study concerns a novel powder metallurgy method for producing porous titanium (pTi) exhibiting high mechanical properties. The preparation procedure consisted of the following stages: first, the preparation of Ti and titanium hydride (TiH2) powder mixtures and their consolidation with a cold isostatic press, followed by a sintering of the green bodies performed with hot isostatic press (HIP) equipment. Thermal decomposition in controlled environment of the TiH2 phase results in the foam structure. The resulting porosity percolates with a volume fraction of approximately 20%. The final material exhibits interesting mechanical properties, comparable to those of full density titanium (between grade 2 and grade 3), with the advantage of a minor density. The samples produced were tested to verify their biological response by studying the effectiveness of osteoblast adhesion and growth. In this preliminary study, osteoblastic cell morphology was investigated and compared to that observed on fully dense commercially pure titanium (Ti-cp) (ASTM, grade 3). The preliminary results were promising regarding cellular adhesion and spreading. (Journal of Applied Biomaterials & Biomechanics 2003; 1: 172-7).

  10. The large universal Pantoea plasmid LPP-1 plays a major role in biological and ecological diversification

    Directory of Open Access Journals (Sweden)

    De Maayer Pieter

    2012-11-01

    Full Text Available Abstract Background Pantoea spp. are frequently isolated from a wide range of ecological niches and have various biological roles, as plant epi- or endophytes, biocontrol agents, plant-growth promoters or as pathogens of both plant and animal hosts. This suggests that members of this genus have undergone extensive genotypic diversification. One means by which this occurs among bacteria is through the acquisition and maintenance of plasmids. Here, we have analyzed and compared the sequences of a large plasmid common to all sequenced Pantoea spp. Results and discussion The Large PantoeaPlasmids (LPP-1 of twenty strains encompassing seven different Pantoea species, including pathogens and endo-/epiphytes of a wide range of plant hosts as well as insect-associated strains, were compared. The LPP-1 plasmid sequences range in size from ~281 to 794 kb and carry between 238 and 750 protein coding sequences (CDS. A core set of 46 proteins, encompassing 2.2% of the total pan-plasmid (2,095 CDS, conserved among all LPP-1 plasmid sequences, includes those required for thiamine and pigment biosynthesis. Phylogenetic analysis reveals that these plasmids have arisen from an ancestral plasmid, which has undergone extensive diversification. Analysis of the proteins encoded on LPP-1 also showed that these plasmids contribute to a wide range of Pantoea phenotypes, including the transport and catabolism of various substrates, inorganic ion assimilation, resistance to antibiotics and heavy metals, colonization and persistence in the host and environment, pathogenesis and antibiosis. Conclusions LPP-1 is universal to all Pantoea spp. whose genomes have been sequenced to date and is derived from an ancestral plasmid. LPP-1 encodes a large array of proteins that have played a major role in the adaptation of the different Pantoea spp. to their various ecological niches and their specialization as pathogens, biocontrol agents or benign saprophytes found in many diverse

  11. High resolution spectrometry: how the analyzer and spectrometer performances and the beam emittance contribute to the results obtained

    International Nuclear Information System (INIS)

    Roussel, P.

    1984-01-01

    Using first order calculations derived for an achromatic system A) (deltaxsub(F)/deltaEsub(i)=0) or an optimised system O) (xsub(F) minima). It is shown that the final resolution measured in the local plane of the spectrometer depends only on the emittance of the accelerator and of the efficient area of the analyser exclusive of the properties of the spectrometer. The use of this result is only limited by higher order terms in the calculation or considerations out of this scope like target effects etc.. [fr

  12. Thermal monitoring of hydrothermal activity by permanent infrared automatic stations: Results obtained at Solfatara di Pozzuoli, Campi Flegrei (Italy)

    Science.gov (United States)

    Chiodini, G.; Vilardo, G.; Augusti, V.; Granieri, D.; Caliro, S.; Minopoli, C.; Terranova, C.

    2007-12-01

    A permanent automatic infrared (IR) station was installed at Solfatara crater, the most active zone of Campi Flegrei caldera. After a positive in situ calibration of the IR camera, we analyze 2175 thermal IR images of the same scene from 2004 to 2007. The scene includes a portion of the steam heated hot soils of Solfatara. The experiment was initiated to detect and quantify temperature changes of the shallow thermal structure of a quiescent volcano such as Solfatara over long periods. Ambient temperature is the main parameter affecting IR temperatures, while air humidity and rain control image quality. A geometric correction of the images was necessary to remove the effects of slow movement of the camera. After a suitable correction the images give a reliable and detailed picture of the temperature changes, over the period October 2004 to January 2007, which suggests that origin of the changes were linked to anthropogenic activity, vegetation growth, and the increase of the flux of hydrothermal fluids in the area of the hottest fumaroles. Two positive temperature anomalies were registered after the occurrence of two seismic swarms which affected the hydrothermal system of Solfatara in October 2005 and October 2006. It is worth noting that these signs were detected in a system characterized by a low level of activity with respect to systems affected by real volcanic crisis where more spectacular results will be expected. Results of the experiment show that this kind of monitoring system can be a suitable tool for volcanic surveillance.

  13. [The medical dissertation--no definitive model. Results of a survey about obtaining a doctorate contradict frequently stated opinions].

    Science.gov (United States)

    Weihrauch, M; Strate, J; Pabst, R

    2003-12-05

    The relevance of medical dissertations is controversial in Germany. The "Wissenschaftsrat" (Science Council) of Germany even made the radical proposal of abolishing medical dissertations, in their present form, in the recently published "Guidelines on writing a thesis". Using a questionnaire with 22 items all students submitting their dissertations within one year at the Medical School in Hannover were asked to participate in a survey about the dissertation, supervision, time spent on it and publishing the results. The answers were evaluated statistically by explorative data analysis. A total of 232 questionnaires were evaluated, which is equivalent to a response rate of 87% (47% were women). A majority of 69% had prepared their thesis in a clinical institution. The supervision during various phases of the dissertation was graded as good. Altogether 90% thought that it had been personally worth-while. In 57% of cases the data of the dissertation had already been published and in 39 % of these cases the author of the dissertation was listed as first author of the publication. In agreement with former studies in German universities an overwhelming majority of 90% of respondents graded dissertations as a significant part of their medical studies, although requiring much additional time. They would recommend such a research phase to younger students as highly valuable. These results are in contrast to many repeatedly made statements.

  14. Overview of experimental results obtained under the Prestressed Concrete Nuclear Pressure Vessel Development Program at the Oak Ridge National Laboratory

    International Nuclear Information System (INIS)

    Naus, D.J.

    1978-01-01

    Under the Prestressed Concrete Nuclear Pressure Vessel Development Program at the Oak Ridge National Laboratory, various aspects of Prestressed Concrete Pressure Vessels (PCPVs) are investigated and evaluated with respect to reliability, structural performance, constructability, and economy. Based upon identified needs, analytical and experimental investigations are conducted. Areas of interest include finite-element analysis development, materials and structural behavior tests, instrumentation evaluation and development, and structural model tests. Studies have been recently completed in the following areas: concrete embedment instrumentation systems for PCPVs, grouted-nongrouted prestressing systems, acoustic emission as a technique for structural integrity monitoring, and model tests of steam-generator cavity closure plugs for a Gas-Cooled Fast Reactor (GCFR). An overview of results is presented

  15. Wear of control rod cluster assemblies and of instrumentation thimbles: first results obtained with the vibrateau wear simulator

    International Nuclear Information System (INIS)

    Zbinden, M.; Hersant, D.

    1993-07-01

    Several REP components are affected by a particular sort of damage called impact/sliding wear. This kind of wear, originating from flow induced vibrations, affects loosely supported tubular structures. The main involved components are: - the RCCAs claddings and the guides tubes, - the instrumentation thimbles, - the fuel rods claddings, - the SG tubes. The R and D Division is concerned with studies aiming to understand and to master the phenomena leading to this wear. The MTC Branch is charged of the study of the wear itself. Tests are carried out on wear rigs to understand and to model wear mechanisms. The following work is related to the two first wear tests campaigns on the VIBRATEAU wear simulator: - a reproducibility test series in order to assess the spreading of the experimental results, - a comparative test series on surface treatments used to improve the components war resistance. (authors). 7 figs., 2 tabs., 4 refs

  16. Short Communication: Testosterone Measured with an Automatic Immunoassay Compares Reasonbly Well to Results Obtained by LC-MS/MS

    DEFF Research Database (Denmark)

    Knudsen, Cindy Søndersø; Højskov, Carsten Schriver; Møller, Holger Jon

    2016-01-01

    Background: Previous studies have reported problems measuring testosterone with immunological assays. Here we explore an automatic second generation immunoassay compared to a LC-MS/MS method. Methods: We collected blood samples from 76 women and measured testosterone, progesterone, gender...... hormonebinding globulin (SHBG), and albumin employing Cobas e601/c501. Testosterone, androstenedione (andro), dehydroepiandrosterone sulphate (DHEAS), and 17-hydroxyprogesterone (17-OHP) concentrations were measured employing LC-MS/MS. We evaluated the difference between testosterone measured by the two methods...... and examined the potential interference from the selected steroids and bindings proteins. Results: Testosterone concentrations measured by the two methods yielded: Cobas e601 = 1.240 x (LC-MS/MS) - 0.197, r = 0.84, for testosterone concentrations between 0.22 - 4.9 nmol/L. A positive correlation was observed...

  17. First results of registering ionospheric disturbances obtained with SibNet network of GNSS receivers in active space experiments

    Science.gov (United States)

    Ishin, Artem; Perevalova, Natalia; Voeykov, Sergey; Khakhinov, Vitaliy

    2017-12-01

    Global and regional networks of GNSS receivers have been successfully used for geophysical research for many years; the number of continuous GNSS stations in the world is steadily growing. The article presents the first results of the use of a new regional network of GNSS stations (SibNet) in active space experiments. The Institute of Solar-Terrestrial Physics of Siberian Branch of Russian Academy of Sciences (ISTP SB RAS) has established this network in the South Baikal region. We describe in detail SibNet, characteristics of receivers in use, parameters of antennas and methods of their installation. We also present the general structure of observation site and the plot of coverage of the receiver operating zone at 50-55° latitudes by radio paths. It is shown that the selected location of receivers allows us to detect ionospheric irregularities of various scales. The purpose of the active space experiments was to reveal and record parameters of the ionospheric irregu larities caused by effects from jet streams of Progress cargo spacecraft. The mapping technique enabled us to identify weak, vertically localized ionospheric irregularities and associate them with the Progress spacecraft engine impact. Thus, it has been shown that SibNet deployed in the Southern Baikal region is an effective instrument for monitoring ionospheric conditions.

  18. Plutonium burning and minor actinides transmutation in fast reactors: first results obtained within the frame of the CAPRA programme

    International Nuclear Information System (INIS)

    Garnier, J.C.; Rouault, J.; Kiefhaber, E.; Sunderland, R.

    1994-01-01

    The CAPRA program gas been established by the CEA in early 1993 with the primary goal of investigating the feasibility of a fast reactor core optimised to burn plutonium. CAPRA is now being jointly pursued by the European Research and Development (R and D) organisations (CEA in France, AEA in the UK and KFK in Germany) and the design companies grouped under the European Fast Reactor Associates umbrella. The first phase of the CAPRA programme is planned to last until the end of 1994. Its goal is to deliver an overall assessment on the feasibility of fast reactor plutonium burner cores. This assessment will also include the minor actinides transmutation capability of such cores. The objective of this paper is to present the progress made so far. After an introduction to the basic physics boundary conditions of burner cores, a description of the studies performed and the main results are given. Then the efforts made towards the definition of an accompanying experimental Research and Development (R and D) program are summarised, followed by the conclusions and an outlook to the future work. (authors). 4 refs., 3 figs., 2 tabs

  19. Isolation/separation of plasmid DNA using hemoglobin modified magnetic nanocomposites as solid-phase adsorbent.

    Science.gov (United States)

    Chen, Xu-Wei; Mao, Quan-Xing; Liu, Jia-Wei; Wang, Jian-Hua

    2012-10-15

    Hemoglobin (Hb) modified magnetic nanocomposites are prepared by immobilization of Hb onto the surface of amino-functionalized Fe(3)O(4)@SiO(2) magnetic nanoparticles via covalent bonding with glutaraldehyde as cross-linker. The obtained nanocomposites are characterized with FT-IR, SEM, XRD and surface charge analysis. A direct solid-phase extraction procedure for the isolation/separation of plasmid DNA using this nanocomposite as a novel adsorbent is thus developed. Some important experimental parameters governing the sorption efficiency, i.e., the pH of sample solution and the ionic strength, are investigated. The Hb modified magnetic nanocomposites provide a sorption capacity of 27.86 mg g(-1) for DNA. By using 2.0mg of the nanocomposites as sorption medium and a suitable acidity of pH 6.1, a sorption efficiency of 93% is achieved for 25 μg mL(-1) of DNA in 1.0 mL of sample solution. Afterwards, the absorbed DNA could be readily recovered by using 1.0 mL of Tris-HCl buffer (pH 8.9, 0.01 mol L(-1)), giving rise to a recovery of ca. 68.3%. The present solid-phased extraction protocol is applied for the isolation of plasmid DNA from Escherichia coli culture, resulting in comparable yield and purity of plasmid DNA with respect to those obtained by using commercial kits. Copyright © 2012 Elsevier B.V. All rights reserved.

  20. Natural plasmid transformation in a high-frequency-of transformation marine Vibrio strain

    International Nuclear Information System (INIS)

    Frischer, M.E.; Thurmond, J.M.; Paul, J.H.

    1990-01-01

    The estuarine bacterium Vibrio strain DI-9 has been shown to be naturally transformable with both broad host range plasmid multimers and homologous chromosomal DNA at average frequencies of 3.5 x 10 -9 and 3.4 x 10 -7 transformants per recipient, respectively. Growth of plasmid transformants in nonselective medium resulted in cured strains that transformed 6 to 42,857 times more frequently than the parental strain, depending on the type of transforming DNA. These high-frequency-of-transformation (HfT) strains were transformed at frequencies ranging from 1.1 x 10 -8 to 1.3 x 10 -4 transformants per recipient with plasmid DNA and at an average frequency of 8.3 x 10 -5 transformants per recipient with homologous chromosomal DNA. The highest transformation frequencies were observed by using multimers of an R1162 derivative carrying the transposon Tn5 (pQSR50). Probing of total DNA preparations from one of the cured strains demonstrated that no plasmid DNA remained in the cured strains which may have provided homology to the transforming DNA. All transformants and cured strains could be differentiated from the parental strains by colony morphology. DNA binding studies indicated that late-log-phase HfT strains bound [ 3 H]bacteriophage lambda DNA 2.1 times more rapidly than the parental strain. These results suggest that the original plasmid transformation event of strain DI-9 was the result of uptake and expression of plasmid DNA by a competent mutant (HfT strain). Additionally, it was found that a strain of Vibrio parahaemolyticus, USFS 3420, could be naturally transformed with plasmid DNA. Natural plasmid transformation by high-transforming mutants may be a means of plasmid acquisition by natural aquatic bacterial populations

  1. Molecular processes as basis for plasmid-mediated bacterial UV-light resistance and mutagenesis

    International Nuclear Information System (INIS)

    Aleshkin, G.I.; Brukhanskij, G.V.; Skavronskaya, A.G.

    1985-01-01

    The increase of UV-resistance and UV-induced mutagenesis by lambda 1 pint intmid as well as molecular-genetic mechanisms of plasmid participation in reparation and DNA replication and its degradation after UV-irradiation in plasmid cells on pKM101 plasmid model have been investigated. Data testifying to the necessity of intmid integration in chromosome as obligatory stage of intmid participation in increasing UV-resistance of bacterial cells are obtained. It has been found that intmid raises UV-resistance of cells and increases respectively the UV-induced reverants efficiency. On the basis of the experiment data the conclusion is drawn that the intmid capacity to raise UV-resistance and, possibly, mutagenesis is bound not only with its integration into chromosome but also with pol A + chromosome replication by dependendent imtmid replication complex. It is shown that pKM101 plasmid ensures functioning in E coli cells of inducible, chloroamphenicol-resistant DNA replication, highly resistant to UV-light harmful effect and that the volume of excision reparation in E. coli cells carrying pKM101 plasmid is increased as compared with the volume of reparation in plasmid legs cells. The combination of the data obtained gives grounds to the authors to assume that inducible replication, inducible reparation of DNA and inducible decrease of DNA degradation determined by pKM101 plasmid may serve as recA + lexA + basis dependent increase of UV-resistance and mutagenesis and that these processes provide the possibility of functioning of integrative replication mechanism of plasmid participation in ensuring UV-resistance and mutagenesis of plants

  2. Reconstructing the complex evolutionary history of mobile plasmids in red algal genomes

    Science.gov (United States)

    Lee, JunMo; Kim, Kyeong Mi; Yang, Eun Chan; Miller, Kathy Ann; Boo, Sung Min; Bhattacharya, Debashish; Yoon, Hwan Su

    2016-01-01

    The integration of foreign DNA into algal and plant plastid genomes is a rare event, with only a few known examples of horizontal gene transfer (HGT). Plasmids, which are well-studied drivers of HGT in prokaryotes, have been reported previously in red algae (Rhodophyta). However, the distribution of these mobile DNA elements and their sites of integration into the plastid (ptDNA), mitochondrial (mtDNA), and nuclear genomes of Rhodophyta remain unknown. Here we reconstructed the complex evolutionary history of plasmid-derived DNAs in red algae. Comparative analysis of 21 rhodophyte ptDNAs, including new genome data for 5 species, turned up 22 plasmid-derived open reading frames (ORFs) that showed syntenic and copy number variation among species, but were conserved within different individuals in three lineages. Several plasmid-derived homologs were found not only in ptDNA but also in mtDNA and in the nuclear genome of green plants, stramenopiles, and rhizarians. Phylogenetic and plasmid-derived ORF analyses showed that the majority of plasmid DNAs originated within red algae, whereas others were derived from cyanobacteria, other bacteria, and viruses. Our results elucidate the evolution of plasmid DNAs in red algae and suggest that they spread as parasitic genetic elements. This hypothesis is consistent with their sporadic distribution within Rhodophyta. PMID:27030297

  3. Resident enhanced repair: novel repair process action on plasmid DNA transformed into Escherichia coli K-12

    International Nuclear Information System (INIS)

    Strike, P.; Roberts, R.J.

    1982-01-01

    The survival of UV-irradiated DNA of plasmid NTP16 was monitored after its transformation into recipient cells containing an essentially homologous undamaged plasmid, pLV9. The presence of pLV9 resulted in a substantial increase in the fraction of damaged NTP16 molecules which survived in the recipient cells. This enhanced survival requires the host uvrA + and uvrB + gene products, but not the host recA + gene product. The requirement for both homologous DNA and the uvrA + gene products suggests that a novel repair process may act on plasmid DNA. Possible mechanisms for this process are considered

  4. Construction and confirmation of the plasmid of human mitochondrial DNA 4977 bp deletion induced by ionizing radiation

    International Nuclear Information System (INIS)

    Chen Xiaosui; Zhou Lijun; Wang Yuxiao; Qu Jia; Feng Jiangbing; Lu Xue; Chen Deqing; Liu Qingjie

    2006-01-01

    Objective: To construct a stable plasmid that spanning deleted human mitochondrial DNA (mtDNA) 4977 bp induced by ionizing radiation and another one for control DNA fragment, in order to use in the human mitochondrial genome study in the future. Methods: The peripheral blood, which had no mtDNA 4977 bp deletion found in previous study, was exposed to 10 Gy 60 Co γ-rays in vitro. The total cell DNA was extracted and PCR was carried out: a nest-PCR of three-round PCR was used for the mtDNA 4977 bp deletion and one- round regular PCR was used for the control ND1 gene. The PCR products were used for transfection by electroporation and the positive clones were obtained after screening. The plasmid DNA was isolated and sequenced after enzymatic digestion and purification. The sequence result was BLASTed with the human mitochondrial genome. Results: The sizes of PCR products for the flanked 4977 bp deletion and the ND1 gene were similar with those predicted according to GeneBank. The sequences for the positive clones were above 99 per cent homologous with the human mitochondrial genome after BLASTed. Conclusion: The plasmids for deleted human mtDNA 4977 bp and control DNA fragment have been constructed successfully, and they could be used in the quality and quantity studies on human mtDNA 4977 bp deletion. (authors)

  5. Treatment with Cefotaxime Affects Expression of Conjugation Associated Proteins and Conjugation Transfer Frequency of an IncI1 Plasmid in Escherichia coli

    DEFF Research Database (Denmark)

    Møller, Thea S B; Liu, Gang; Boysen, Anders

    2017-01-01

    research suggests that the effect of antibiotic treatment on plasmid conjugation frequencies, and hence the spread of resistance plasmids, may have been overestimated. We addressed the question by quantifying transfer proteins and conjugation frequencies of a blaCTX-M-1 encoding IncI1 resistance plasmid....... The frequency of plasmid conjugation, measured in an antibiotic free environment, increased significantly when the donor was pre-grown in broth containing CTX compared to growth without this drug, regardless of whether blaCTX-M-1 was located on the plasmid or in trans on the chromosome. The results shows...

  6. Radioautographic test for genetic cotton transformation by pCaVItoxneo hybrid plasmid

    International Nuclear Information System (INIS)

    Imamkhodjaeva, A.S.

    2006-01-01

    Full text: Search for novel technologies in biology, application of up-to-date methods in gene engineering, manipulation with the recombinant DNA, in particular, open opportunities for experiments with plants. To identify some DNA fragments in an organism's genome, radioautographic methods, such as dot- and blot-hybridization are frequently used. As a rule, genomic DNA is first isolated from the plant's organ. Its purification and subsequent manipulation is followed by hybridization with a probe labeled with radioactive components. The purified DNA, cDNA of RNA reverse transcription or a DNA fragment cloned in E-coli could serve as the probe. Radioautography shows homologically hybridized fragments. We have performed express dot-hybridization analysis on hybrid plasmid transformation of G.Hirsutum L. (108F) and G. Barbadense L. (C-6037) cotton sorts. pCaVItoxneo plasmid obtained on the basis of independently replicated plasmid-like DNA of the G.Hirsutum L. (pGHm2) cotton mitochondria was used (Yusupov T., 1994). There are hybrid two-domain gene of insectotoxin and enzymatically active kanamycine - phosphotransferase in the plasmid. The whole content is controlled by the plant promoter of cauliflower mosaic virus (19 S SFMV). The plasmid in question was added to the pollen sprouting medium followed by the transfer of the suspension on the pistil stigmas of the pre-prepared cotton flowers. The seed budding as the result of the experiment were analyzed by means of dot-hybridization method. DNA probes used for radioactive hybridization were labeled by method of Fainberg and Vagelstein (1990). To perform that DNA was dissolved in Tris-EDTA (10:1), containing 10mM of Tris HCl and 1mM EDTA, denaturated at 100 d eg C for 2 minutes with subsequent addition of oligonucleotide primers and annealing. DNA synthesis in the presence of 32 P labeled dATP and dCTP (Tashkent) was performed in the reaction mixture of potassium-phosphate buffer containing 67mM of MgCl 2 , 1 mg/ml of

  7. Protein Structure Initiative Material Repository: an open shared public resource of structural genomics plasmids for the biological community

    Science.gov (United States)

    Cormier, Catherine Y.; Mohr, Stephanie E.; Zuo, Dongmei; Hu, Yanhui; Rolfs, Andreas; Kramer, Jason; Taycher, Elena; Kelley, Fontina; Fiacco, Michael; Turnbull, Greggory; LaBaer, Joshua

    2010-01-01

    The Protein Structure Initiative Material Repository (PSI-MR; http://psimr.asu.edu) provides centralized storage and distribution for the protein expression plasmids created by PSI researchers. These plasmids are a resource that allows the research community to dissect the biological function of proteins whose structures have been identified by the PSI. The plasmid annotation, which includes the full length sequence, vector information and associated publications, is stored in a freely available, searchable database called DNASU (http://dnasu.asu.edu). Each PSI plasmid is also linked to a variety of additional resources, which facilitates cross-referencing of a particular plasmid to protein annotations and experimental data. Plasmid samples can be requested directly through the website. We have also developed a novel strategy to avoid the most common concern encountered when distributing plasmids namely, the complexity of material transfer agreement (MTA) processing and the resulting delays this causes. The Expedited Process MTA, in which we created a network of institutions that agree to the terms of transfer in advance of a material request, eliminates these delays. Our hope is that by creating a repository of expression-ready plasmids and expediting the process for receiving these plasmids, we will help accelerate the accessibility and pace of scientific discovery. PMID:19906724

  8. Plasmid ColE1 as a Molecular Vehicle for Cloning and Amplification of DNA

    Science.gov (United States)

    Hershfield, Vickers; Boyer, Herbert W.; Yanofsky, Charles; Lovett, Michael A.; Helinski, Donald R.

    1974-01-01

    DNA fragments obtained from EcoRI endonuclease digestion of bacteriophage ϕ80pt190 (trp+) and the plasmid ColE1 were covalently joined with polynucleotide ligase. Transformation of Escherichia coli trp- strains to tryptophan independence with the recombined DNA selected for reconstituted ColE1 plasmids containing the tryptophan operon and the ϕ80 immunity region. Similarly, an EcoRI endonuclease generated fragment of plasmid pSC105 DNA containing the genetic determinant of kanamycin resistance was inserted into the ColE1 plasmid and recovered in E. coli. The plasmids containing the trp operon (ColE1-trp) and the kanamycin resistance gene were maintained under logarithmic growth conditions at a level of 25-30 copies per cell and accumulate to the extent of several hundred copies per cell in the presence of chloramphenicol. Cells carrying the ColE1-trp plasmid determined the production of highly elevated levels of trp operon-specific mRNA and tryptophan biosynthetic enzymes. Images PMID:4610576

  9. Characterization of plasmids in a human clinical strain of Lactococcus garvieae.

    Directory of Open Access Journals (Sweden)

    Mónica Aguado-Urda

    Full Text Available The present work describes the molecular characterization of five circular plasmids found in the human clinical strain Lactococcus garvieae 21881. The plasmids were designated pGL1-pGL5, with molecular sizes of 4,536 bp, 4,572 bp, 12,948 bp, 14,006 bp and 68,798 bp, respectively. Based on detailed sequence analysis, some of these plasmids appear to be mosaics composed of DNA obtained by modular exchange between different species of lactic acid bacteria. Based on sequence data and the derived presence of certain genes and proteins, the plasmid pGL2 appears to replicate via a rolling-circle mechanism, while the other four plasmids appear to belong to the group of lactococcal theta-type replicons. The plasmids pGL1, pGL2 and pGL5 encode putative proteins related with bacteriocin synthesis and bacteriocin secretion and immunity. The plasmid pGL5 harbors genes (txn, orf5 and orf25 encoding proteins that could be considered putative virulence factors. The gene txn encodes a protein with an enzymatic domain corresponding to the family actin-ADP-ribosyltransferases toxins, which are known to play a key role in pathogenesis of a variety of bacterial pathogens. The genes orf5 and orf25 encode two putative surface proteins containing the cell wall-sorting motif LPXTG, with mucin-binding and collagen-binding protein domains, respectively. These proteins could be involved in the adherence of L. garvieae to mucus from the intestine, facilitating further interaction with intestinal epithelial cells and to collagenous tissues such as the collagen-rich heart valves. To our knowledge, this is the first report on the characterization of plasmids in a human clinical strain of this pathogen.

  10. Plasmid construction using recombination activity in the fission yeast Schizosaccharomyces pombe.

    Directory of Open Access Journals (Sweden)

    Ayako Chino

    Full Text Available BACKGROUND: Construction of plasmids is crucial in modern genetic manipulation. As of now, the common method for constructing plasmids is to digest specific DNA sequences with restriction enzymes and to ligate the resulting DNA fragments with DNA ligase. Another potent method to construct plasmids, known as gap-repair cloning (GRC, is commonly used in the budding yeast Saccharomyces cerevisiae. GRC makes use of the homologous recombination activity that occurs within the yeast cells. Due to its flexible design and efficiency, GRC has been frequently used for constructing plasmids with complex structures as well as genome-wide plasmid collections. Although there have been reports indicating GRC feasibility in the fission yeast Schizosaccharomyces pombe, this species is not commonly used for GRC as systematic studies of reporting GRC efficiency in S. pombe have not been performed till date. METHODOLOGY/PRINCIPAL FINDINGS: We investigated GRC efficiency in S. pombe in this study. We first showed that GRC was feasible in S. pombe by constructing a plasmid that contained the LEU2 auxotrophic marker gene in vivo and showed sufficient efficiency with short homology sequences (>25 bp. No preference was shown for the sequence length from the cut site in the vector plasmid. We next showed that plasmids could be constructed in a proper way using 3 DNA fragments with 70% efficiency without any specific selections being made. The GRC efficiency with 3 DNA fragments was dramatically increased >95% in lig4Delta mutant cell, where non-homologous end joining is deficient. Following this approach, we successfully constructed plasmid vectors with leu1+, ade6+, his5+, and lys1+ markers with the low-copy stable plasmid pDblet as a backbone by applying GRC in S. pombe. CONCLUSIONS/SIGNIFICANCE: We concluded that GRC was sufficiently feasible in S. pombe for genome-wide gene functional analysis as well as for regular plasmid construction. Plasmids with different

  11. Plasmid DNA damage induced by helium atmospheric pressure plasma jet

    Science.gov (United States)

    Han, Xu; Cantrell, William A.; Escobar, Erika E.; Ptasinska, Sylwia

    2014-03-01

    A helium atmospheric pressure plasma jet (APPJ) is applied to induce damage to aqueous plasmid DNA. The resulting fractions of the DNA conformers, which indicate intact molecules or DNA with single- or double-strand breaks, are determined using agarose gel electrophoresis. The DNA strand breaks increase with a decrease in the distance between the APPJ and DNA samples under two working conditions of the plasma source with different parameters of applied electric pulses. The damage level induced in the plasmid DNA is also enhanced with increased plasma irradiation time. The reactive species generated in the APPJ are characterized by optical emission spectra, and their roles in possible DNA damage processes occurring in an aqueous environment are also discussed.

  12. Identification and characterization of two novel bla(KLUC resistance genes through large-scale resistance plasmids sequencing.

    Directory of Open Access Journals (Sweden)

    Teng Xu

    Full Text Available Plasmids are important antibiotic resistance determinant carriers that can disseminate various drug resistance genes among species or genera. By using a high throughput sequencing approach, two groups of plasmids of Escherichia coli (named E1 and E2, each consisting of 160 clinical E. coli strains isolated from different periods of time were sequenced and analyzed. A total of 20 million reads were obtained and mapped onto the known resistance gene sequences. As a result, a total of 9 classes, including 36 types of antibiotic resistant genes, were identified. Among these genes, 25 and 27 single nucleotide polymorphisms (SNPs appeared, of which 9 and 12 SNPs are nonsynonymous substitutions in the E1 and E2 samples. It is interesting to find that a novel genotype of bla(KLUC, whose close relatives, bla(KLUC-1 and bla(KLUC-2, have been previously reported as carried on the Kluyvera cryocrescens chromosome and Enterobacter cloacae plasmid, was identified. It shares 99% and 98% amino acid identities with Kluc-1 and Kluc-2, respectively. Further PCR screening of 608 Enterobacteriaceae family isolates yielded a second variant (named bla(KLUC-4. It was interesting to find that Kluc-3 showed resistance to several cephalosporins including cefotaxime, whereas bla(KLUC-4 did not show any resistance to the antibiotics tested. This may be due to a positively charged residue, Arg, replaced by a neutral residue, Leu, at position 167, which is located within an omega-loop. This work represents large-scale studies on resistance gene distribution, diversification and genetic variation in pooled multi-drug resistance plasmids, and provides insight into the use of high throughput sequencing technology for microbial resistance gene detection.

  13. Integral parametrization of the Kinetics of Crosslink production in plasmid DNA as a function of 8-methoxypsoralen concentration

    Energy Technology Data Exchange (ETDEWEB)

    Vidania, R. de; Paramio, J. M.; Bauluz, C.

    1986-07-01

    In this paper we present results of crosslink production in pBR322 DNA along a wide range of 8-methoxypsoralen (8-MOP) concentration. Experimental data were obtained as DNA renaturation percentages, from the shift in hyperchromicity after a temperature-dependent denaturation-renaturation process. the experimental results showed a three-stage profile when represented as a function of the natural logarithms of 8-MOP concentration. an integral parametrization which allows a simultaneous fit of the three observed stages is presented here. the theoretical values of crosslink production determined from the fit are useful to asses the genotoxicity of psoralen-induced crosslinks in plasmid DNA. (Author) 24 refs.

  14. Integral parametrization of the Kinetics of Crosslink production in plasmid DNA as a function of 8-methoxypsoralen concentration

    International Nuclear Information System (INIS)

    Vidania, R. de; Paramio, J. M.; Bauluz, C.

    1986-01-01

    In this paper we present results of crosslink production in pBR322 DNA along a wide range of 8-methoxypsoralen (8-MOP) concentration. Experimental data were obtained as DNA renaturation percentages, from the shift in hyperchromicity after a temperature-dependent denaturation-renaturation process. the experimental results showed a three-stage profile when represented as a function of the natural logarithms of 8-MOP concentration. an integral parametrization which allows a simultaneous fit of the three observed stages is presented here. the theoretical values of crosslink production determined from the fit are useful to asses the genotoxicity of psoralen-induced crosslinks in plasmid DNA. (Author) 24 refs

  15. Transposition of Tn5096 from a temperature-sensitive transducible plasmid in Streptomyces spp.

    OpenAIRE

    McHenney, M A; Baltz, R H

    1991-01-01

    Transposon Tn5096 was inserted into a derivative of the temperature-sensitive plasmid pMT660 containing the bacteriophage FP43 pac site. The resulting plasmid, pRHB126, was transduced by FP43 into several Streptomyces species. Tn5096 transposed from pRHB126 into different sites in the genomes of Streptomyces ambofaciens, Streptomyces cinnamonensis, Streptomyces coelicolor A3(2), Streptomyces fradiae, Streptomyces griseofuscus, and Streptomyces thermotolerans.

  16. PLASMID PROFILES AND PHAGE TYPES OF SALMONELLA-TYPHIMURIUM ISOLATED FROM SUCCESSIVE FLOCKS OF CHICKENS ON 3 PARENT STOCK FARMS

    DEFF Research Database (Denmark)

    Baggesen, Dorte Lau; Olsen, J. E.; Bisgaard, M.

    1992-01-01

    Three-hundred-and-eighty-seven strains of Salmonella typhimurium obtained from successive generations of parent stock originating from three different rearing farms were characterized by phage typing and plasmid profiling. Seventy-six strains representing dominant types were selected for restrict......Three-hundred-and-eighty-seven strains of Salmonella typhimurium obtained from successive generations of parent stock originating from three different rearing farms were characterized by phage typing and plasmid profiling. Seventy-six strains representing dominant types were selected...

  17. Genomic analyses of the Chlamydia trachomatis core genome show an association between chromosomal genome, plasmid type and disease

    NARCIS (Netherlands)

    Versteeg, Bart; Bruisten, Sylvia M.; Pannekoek, Yvonne; Jolley, Keith A.; Maiden, Martin C. J.; van der Ende, Arie; Harrison, Odile B.

    2018-01-01

    Background: Chlamydia trachomatis (Ct) plasmid has been shown to encode genes essential for infection. We evaluated the population structure of Ct using whole-genome sequence data (WGS). In particular, the relationship between the Ct genome, plasmid and disease was investigated. Results: WGS data

  18. 3G vector-primer plasmid for constructing full-length-enriched cDNA libraries.

    Science.gov (United States)

    Zheng, Dong; Zhou, Yanna; Zhang, Zidong; Li, Zaiyu; Liu, Xuedong

    2008-09-01

    We designed a 3G vector-primer plasmid for the generation of full-length-enriched complementary DNA (cDNA) libraries. By employing the terminal transferase activity of reverse transcriptase and the modified strand replacement method, this plasmid (assembled with a polydT end and a deoxyguanosine [dG] end) combines priming full-length cDNA strand synthesis and directional cDNA cloning. As a result, the number of steps involved in cDNA library preparation is decreased while simplifying downstream gene manipulation, sequencing, and subcloning. The 3G vector-primer plasmid method yields fully represented plasmid primed libraries that are equivalent to those made by the SMART (switching mechanism at 5' end of RNA transcript) approach.

  19. Optimizing hyaluronidase dose and plasmid DNA delivery greatly improves gene electrotransfer efficiency in rat skeletal muscle

    DEFF Research Database (Denmark)

    Åkerström, Thorbjörn; Vedel, Kenneth; Needham Andersen, Josefine

    2015-01-01

    Transfection of rat skeletal muscle in vivo is a widely used research model. However, gene electrotransfer protocols have been developed for mice and yield variable results in rats. We investigated whether changes in hyaluronidase pre-treatment and plasmid DNA delivery can improve transfection...... with a homogenous distribution. We also show that transfection was stable over five weeks of regular exercise or inactivity. Our findings show that species-specific plasmid DNA delivery and hyaluronidase pre-treatment greatly improves transfection efficiency in rat skeletal muscle....... efficiency in rat skeletal muscle. We found that pre-treating the muscle with a hyaluronidase dose suitable for rats (0.56. U/g b.w.) prior to plasmid DNA injection increased transfection efficiency by >200% whereas timing of the pre-treatment did not affect efficiency. Uniformly distributing plasmid DNA...

  20. Co-evolution of genomes and plasmids within Chlamydia trachomatis and the emergence in Sweden of a new variant strain

    Directory of Open Access Journals (Sweden)

    Skilton Rachel J

    2009-05-01

    Full Text Available Abstract Background Chlamydia trachomatis is the most common cause of sexually transmitted infections globally and the leading cause of preventable blindness in the developing world. There are two biovariants of C. trachomatis: 'trachoma', causing ocular and genital tract infections, and the invasive 'lymphogranuloma venereum' strains. Recently, a new variant of the genital tract C. trachomatis emerged in Sweden. This variant escaped routine diagnostic tests because it carries a plasmid with a deletion. Failure to detect this strain has meant it has spread rapidly across the country provoking a worldwide alert. In addition to being a key diagnostic target, the plasmid has been linked to chlamydial virulence. Analysis of chlamydial plasmids and their cognate chromosomes was undertaken to provide insights into the evolutionary relationship between chromosome and plasmid. This is essential knowledge if the plasmid is to be continued to be relied on as a key diagnostic marker, and for an understanding of the evolution of Chlamydia trachomatis. Results The genomes of two new C. trachomatis strains were sequenced, together with plasmids from six C. trachomatis isolates, including the new variant strain from Sweden. The plasmid from the new Swedish variant has a 377 bp deletion in the first predicted coding sequence, abolishing the site used for PCR detection, resulting in negative diagnosis. In addition, the variant plasmid has a 44 bp duplication downstream of the deletion. The region containing the second predicted coding sequence is the most highly conserved region of the plasmids investigated. Phylogenetic analysis of the plasmids and chromosomes are fully congruent. Moreover this analysis also shows that ocular and genital strains diverged from a common C. trachomatis progenitor. Conclusion The evolutionary pathways of the chlamydial genome and plasmid imply that inheritance of the plasmid is tightly linked with its cognate chromosome. These data

  1. Evaluation of instrumental parameters for obtaining acceptable analytical results of the Dosimetry Laboratory of Chemistry of the Regional Center of Nuclear Sciences, CNEN-NE, Recife, Brazil

    International Nuclear Information System (INIS)

    Souza, V.L.B.; Figueiredo, M.D.C.; Cunha, M.S.

    2008-01-01

    Instrumental parameters need to be evaluated for obtaining acceptable analytical results for a specific instrument. The performance of the UV-VIS spectrophotometer can be verified for wavelengths and absorbances with appropriate materials (solutions of different concentrations of K 2 CrO 4 , for example). The aim of this work was to demonstrate the results of the procedures to control the quality of the measurements carried out in the laboratory in the last four years. The samples were analyzed in the spectrophotometer and control graphics were obtained for K 2 CrO 4 and Fe 3+ absorbance values. The variation in the results obtained for the stability of the spectrophotometer and for the control of its calibration did not exceed 2%. (author)

  2. Comparative Genomics of Rhodococcus equi Virulence Plasmids Indicates Host-Driven Evolution of the vap Pathogenicity Island.

    Science.gov (United States)

    MacArthur, Iain; Anastasi, Elisa; Alvarez, Sonsiray; Scortti, Mariela; Vázquez-Boland, José A

    2017-05-01

    The conjugative virulence plasmid is a key component of the Rhodococcus equi accessory genome essential for pathogenesis. Three host-associated virulence plasmid types have been identified the equine pVAPA and porcine pVAPB circular variants, and the linear pVAPN found in bovine (ruminant) isolates. We recently characterized the R. equi pangenome (Anastasi E, et al. 2016. Pangenome and phylogenomic analysis of the pathogenic actinobacterium Rhodococcus equi. Genome Biol Evol. 8:3140-3148.) and we report here the comparative analysis of the virulence plasmid genomes. Plasmids within each host-associated type were highly similar despite their diverse origins. Variation was accounted for by scattered single nucleotide polymorphisms and short nucleotide indels, while larger indels-mostly in the plasticity region near the vap pathogencity island (PAI)-defined plasmid genomic subtypes. Only one of the plasmids analyzed, of pVAPN type, was exceptionally divergent due to accumulation of indels in the housekeeping backbone. Each host-associated plasmid type carried a unique PAI differing in vap gene complement, suggesting animal host-specific evolution of the vap multigene family. Complete conservation of the vap PAI was observed within each host-associated plasmid type. Both diversity of host-associated plasmid types and clonality of specific chromosomal-plasmid genomic type combinations were observed within the same R. equi phylogenomic subclade. Our data indicate that the overall strong conservation of the R. equi host-associated virulence plasmids is the combined result of host-driven selection, lateral transfer between strains, and geographical spread due to international livestock exchanges. © The Author(s) 2017. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  3. Dichromatic laser radiation effects on DNA of Escherichia coli and plasmids

    Science.gov (United States)

    Martins, W. A.; Polignano, G. A. C.; Guimarães, O. R.; Geller, M.; Paoli, F.; Fonseca, A. S.

    2015-04-01

    Dichromatic and consecutive laser radiations have attracted increased attention for clinical applications as offering new tools for the treatment of dysfunctional tissues in situations where monochromatic radiation is not effective. This work evaluated the survival, filamentation and morphology of Escherichia coli cells, and the induction of DNA lesions, in plasmid DNA exposed to low-intensity consecutive dichromatic laser radiation. Exponential and stationary wild type and formamidopyrimidine DNA glycosylase/MutM protein deficient E. coli cultures were exposed to consecutive low-intensity dichromatic laser radiation (infrared laser immediately after red laser) to study the survival, filamentation and morphology of bacterial cells. Plasmid DNA samples were exposed to dichromatic radiation to study DNA lesions by electrophoretic profile. Dichromatic laser radiation affects the survival, filamentation and morphology of E. coli cultures depending on the growth phase and the functional repair mechanism of oxidizing lesions in DNA, but does not induce single/double strands breaks or alkali-labile DNA lesions. Results show that low-intensity consecutive dichromatic laser radiation induces biological effects that differ from those induced by monochromatic laser radiation, suggesting that other therapeutic effects could be obtained using dichromatic radiation.

  4. F-127-PEI co-delivering docetaxel and TFPI-2 plasmid for nasopharyngeal cancer therapy

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Tao, E-mail: taoliu18@126.com [Department of Otolaryngology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282 (China); Zhang, Xinyu [Department of Otolaryngology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282 (China); Ke, Bo [Jiangxi Key Laboratory of Hematological Oncology and Cell Biology, Jiangxi Provincial People' s Hospital, Nanchang 330006 (China); Wang, Yigang [School of Life Sciences, Zhejiang Sci-Tech University, Hangzhou 310018 (China); Wu, Xidong [Department of Pharmacology, Jiangxi Institute of Materia Medica,Nanchang 330029 (China); Jiang, Gang [Department of Otolaryngology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282 (China); Wu, Ting [Department of Light Chemical Engineering, Guangdong Polytechnic, Foshan 528041 (China); Nie, Guohui, E-mail: nghui@21cn.com [Department of Otolaryngological, Peking University Shenzhen Hospital, Shenzhen 518036 (China)

    2016-04-01

    The co-delivery of drug and gene has become the primary strategy in cancer therapy. However, to construct one safe co-delivering system with higher drug loading and gene transfection efficiency for cancer therapy is still challenging. Herein, a novel degradable nanocarriers were synthesized and characterized in this study, which was composed of polyethylenimine (PEI)-linked PEO–PPO–PEO (Pluronic F127), called F127-PEI. Then the nanocarrier was used for hydrophobic docetaxel (DOC) and functional gene (TFPI-2 plasmid) co-delivery to treat nasopharyngeal cancer (NPC). The results indicated that F127-PEI nanocarriers had higher DOC loading amount and possessed good gene delivery effect in vitro. For co-delivery analysis, the obtained F127-PEI/DOC/TFPI-2 complexes could induce a more significant apoptosis than DOC or TFPI-2 alone, and decreased invasive capacity of NPC HNE-1 cells more obviously. Moreover, the F127-PEI copolymer exhibited better blood compatibility and lower cytotoxicity compared to PEI-25k by the hemolysis and MTT assays, which suggests a promising potential for NPC therapy. - Highlights: • F127-PEI was synthesized and used for drug and gene co-delivery. • F127-PEI showed good delivery ability to docetaxel and TFPI-2 plasmid. • The co-loaded complexes showed synergistic effect to nasopharyngeal carcinoma. • F127-PEI showed better blood safety and lower cytotoxicity compared to PEI-25k.

  5. F-127-PEI co-delivering docetaxel and TFPI-2 plasmid for nasopharyngeal cancer therapy

    International Nuclear Information System (INIS)

    Liu, Tao; Zhang, Xinyu; Ke, Bo; Wang, Yigang; Wu, Xidong; Jiang, Gang; Wu, Ting; Nie, Guohui

    2016-01-01

    The co-delivery of drug and gene has become the primary strategy in cancer therapy. However, to construct one safe co-delivering system with higher drug loading and gene transfection efficiency for cancer therapy is still challenging. Herein, a novel degradable nanocarriers were synthesized and characterized in this study, which was composed of polyethylenimine (PEI)-linked PEO–PPO–PEO (Pluronic F127), called F127-PEI. Then the nanocarrier was used for hydrophobic docetaxel (DOC) and functional gene (TFPI-2 plasmid) co-delivery to treat nasopharyngeal cancer (NPC). The results indicated that F127-PEI nanocarriers had higher DOC loading amount and possessed good gene delivery effect in vitro. For co-delivery analysis, the obtained F127-PEI/DOC/TFPI-2 complexes could induce a more significant apoptosis than DOC or TFPI-2 alone, and decreased invasive capacity of NPC HNE-1 cells more obviously. Moreover, the F127-PEI copolymer exhibited better blood compatibility and lower cytotoxicity compared to PEI-25k by the hemolysis and MTT assays, which suggests a promising potential for NPC therapy. - Highlights: • F127-PEI was synthesized and used for drug and gene co-delivery. • F127-PEI showed good delivery ability to docetaxel and TFPI-2 plasmid. • The co-loaded complexes showed synergistic effect to nasopharyngeal carcinoma. • F127-PEI showed better blood safety and lower cytotoxicity compared to PEI-25k

  6. Characterization of a plasmid carrying cat, ermB and tetS genes in a foodborne Listeria monocytogenes strain and uptake of the plasmid by cariogenic Streptococcus mutans

    DEFF Research Database (Denmark)

    Li, Lili; Olsen, Rikke Heidemann; Shi, Lei

    2016-01-01

    A multi-drug resistant (MDR) Listeria monocytogenes isolate (serotype 1/2c) was recovered from a quick-frozen rice flour product collected from Langfang city in northern China. PCR screening identified the presence of cat, ermB and tetS genes. The plasmid profile of the strain showed the presence...... of an approximately 22.4-kb plasmid. Curing of this plasmid resulted in the loss of cat, ermB and tetS genes and increased susceptibility to several antibiotics, suggesting the involvement of the plasmid in multiple antibiotic resistances. Moreover, the plasmid was able to be uptaken by human oral pathogen...

  7. Plasmid transfer by conjugation in Xylella fastidiosa.

    Science.gov (United States)

    Recombination and horizontal gene transfer have been implicated in the adaption of Xylella fastidiosa (Xf) to infect a wide variety of different plant species. There is evidence that certain strains of Xf carry native plasmids equipped with transfer and mobilization genes, suggesting conjugation as ...

  8. Standardized Cloning and Curing of Plasmids

    DEFF Research Database (Denmark)

    Lauritsen, Ida; Kim, Se Hyeuk; Porse, Andreas

    2018-01-01

    and exchange of genetic parts in the Standard European Vectors Architecture (SEVA) vector system. Additionally, to facilitate rapid testing and iterative bioengineering using different vector designs, we provide a one-step protocol for a universal CRISPR-Cas9-based plasmid curing system (pFREE) and demonstrate...

  9. Plasmid mediated quinolone resistance in Enterobacteriaceae

    NARCIS (Netherlands)

    Veldman, K.T.; LS Klinisch Onderzoek Wagenaar

    2014-01-01

    This thesis describes the occurrence of Plasmid Mediated Quinolone Resistance (PMQR) in Salmonella and E. coli from The Netherlands and other European countries. Furthermore, the genetic background of these genes was characterized. Fluoroquinolones are widely used antibiotics in both human and

  10. Plasmid and chromosome segregation in prokaryotes

    DEFF Research Database (Denmark)

    Møller-Jensen, Jakob; Bugge Jensen, Rasmus; Gerdes, Kenn

    2000-01-01

    Recent major advances in the understanding of prokaryotic DNA segregation have been achieved by using fluorescence microscopy to visualize the localization of cellular components. Plasmids and bacterial chromosomes are partitioned in a highly dynamic fashion, suggesting the presence of a mitotic...

  11. Antimicrobial resistance and plasmid profiles of Aeromonas ...

    African Journals Online (AJOL)

    The purpose of this study was to investigate the presence of Aeromonas hydrophila at commonly used water collection points on the River Njoro and to determine the in-vitro antimicrobial susceptibility and plasmid profiles of isolates. In total, 126 samples were collected and 36.5% of them were positive for A. hydrophila.

  12. Antimicrobial resistance patterns and plasmid profiles of ...

    African Journals Online (AJOL)

    Objectives: To determine the frequency of resistance of Staphylococcus aureus to various antimicrobial agents, and the relationship between antimicrobial resistance of the isolates and carriage of plasmids. Design: A random sampling of milk and meat samples was carried out. Setting: Milk was collected from various dairy ...

  13. Contribution of plasmid-encoded peptidase S8 (PrtP) to adhesion and transit in the gut of Lactococcus lactis IBB477 strain.

    Science.gov (United States)

    Radziwill-Bienkowska, Joanna Maria; Robert, Véronique; Drabot, Karolina; Chain, Florian; Cherbuy, Claire; Langella, Philippe; Thomas, Muriel; Bardowski, Jacek Karol; Mercier-Bonin, Muriel; Kowalczyk, Magdalena

    2017-07-01

    The ability of Lactococcus lactis to adhere to the intestinal mucosa can potentially prolong the contact with the host, and therefore favour its persistence in the gut. In the present study, the contribution of plasmid-encoded factors to the adhesive and transit properties of the L. lactis subsp. cremoris IBB477 strain was investigated. Plasmid-cured derivatives as well as deletion mutants were obtained and analysed. Adhesion tests were performed using non-coated polystyrene plates, plates coated with mucin or fibronectin and mucus-secreting HT29-MTX intestinal epithelial cells. The results indicate that two plasmids, pIBB477a and b, are involved in adhesion of the IBB477 strain. One of the genes localised on plasmid pIBB477b (AJ89_14230), which encodes cell wall-associated peptidase S8 (PrtP), mediates adhesion of the IBB477 strain to bare, mucin- and fibronectin-coated polystyrene, as well as to HT29-MTX cells. Interactions between bacteria and mucus secreted by HT29-MTX cells were further investigated by fluorescent staining and confocal microscopy. Confocal images showed that IBB477 forms dense clusters embedded in secreted mucus. Finally, the ability of IBB477 strain and its ΔprtP deletion mutant to colonise the gastrointestinal tract of conventional C57Bl/6 mice was determined. Both strains were present in the gut for up to 72 h. In summary, adhesion and persistence of IBB477 were analysed by in vitro and in vivo approaches, respectively. Our studies revealed that plasmidic genes encoding cell surface proteins are more involved in the adhesion of IBB477 strain than in the ability to confer a selective advantage in the gut.

  14. Comparative metagenomic analysis of plasmid encoded functions in the human gut microbiome

    Directory of Open Access Journals (Sweden)

    Marchesi Julian R

    2010-01-01

    Full Text Available Abstract Background Little is known regarding the pool of mobile genetic elements associated with the human gut microbiome. In this study we employed the culture independent TRACA system to isolate novel plasmids from the human gut microbiota, and a comparative metagenomic analysis to investigate the distribution and relative abundance of functions encoded by these plasmids in the human gut microbiome. Results Novel plasmids were acquired from the human gut microbiome, and homologous nucleotide sequences with high identity (>90% to two plasmids (pTRACA10 and pTRACA22 were identified in the multiple human gut microbiomes analysed here. However, no homologous nucleotide sequences to these plasmids were identified in the murine gut or environmental metagenomes. Functions encoded by the plasmids pTRACA10 and pTRACA22 were found to be more prevalent in the human gut microbiome when compared to microbial communities from other environments. Among the most prevalent functions identified was a putative RelBE toxin-antitoxin (TA addiction module, and subsequent analysis revealed that this was most closely related to putative TA modules from gut associated bacteria belonging to the Firmicutes. A broad phylogenetic distribution of RelE toxin genes was observed in gut associated bacterial species (Firmicutes, Bacteroidetes, Actinobacteria and Proteobacteria, but no RelE homologues were identified in gut associated archaeal species. We also provide indirect evidence for the horizontal transfer of these genes between bacterial species belonging to disparate phylogenetic divisions, namely Gram negative Proteobacteria and Gram positive species from the Firmicutes division. Conclusions The application of a culture independent system to capture novel plasmids from the human gut mobile metagenome, coupled with subsequent comparative metagenomic analysis, highlighted the unexpected prevalence of plasmid encoded functions in the gut microbial ecosystem. In

  15. Review of the scientific results obtained at the research reactor-booster IBR-30 and the Program of investigations at the neutron source IREN

    CERN Document Server

    Furman, W

    2002-01-01

    Brief review of the main scientific results obtained at research reactor booster IBR-30 and its predecessor IBR and IBR-1 for the period 1960 - 2001 is presented. The thesis of the scientific program for the upgrade of IBR-30 resonance neutron source IREN are adduced

  16. Aircraft and ground vehicle friction correlation test results obtained under winter runway conditions during joint FAA/NASA Runway Friction Program

    Science.gov (United States)

    Yager, Thomas J.; Vogler, William A.; Baldasare, Paul

    1988-01-01

    Aircraft and ground vehicle friction data collected during the Joint FAA/NASA Runway Friction Program under winter runway conditions are discussed and test results are summarized. The relationship between the different ground vehicle friction measurements obtained on compacted snow- and ice-covered conditions is defined together with the correlation to aircraft tire friction performance under similar runway conditions.

  17. Simple method for identification of plasmid-coded proteins

    International Nuclear Information System (INIS)

    Sancar, A.; Hack, A.M.; Rupp, W.D.

    1979-01-01

    Proteins encoded by plasmid DNA are specifically labeled in uv-irradiated cells of Escherichia coli carrying recA and uvrA mutations because extensive degradation of the chromosome DNA occurs concurrently with amplification of plasmid DNA

  18. Most recent developments in combined radium and high voltage therapy of uterine tumours as shown by the results obtained at the municipal hospital Passau

    International Nuclear Information System (INIS)

    Moos, G.

    1987-01-01

    It was the aim of the study described here to review data on healing successes achieved in malignant changes of the uterus. The results obtained were analysed on the bais of tumour stage and, in particular, the radiation dose actually administered. It could be shown that the treatment results currently achieved at this clinic are in keeping with relevant patient data collected previously or by other centres. (orig./MG) [de

  19. Interobserver agreement of the injury diagnoses obtained by postmortem computed tomography of traffic fatality victims and a comparison with autopsy results

    DEFF Research Database (Denmark)

    Leth, Peter Mygind; Struckmann, Henrik; Lauritsen, Jens

    2013-01-01

    The present study investigated the interobserver variation between a radiologist and a forensic pathologist in 994 injury diagnoses obtained by postmortem computed tomography (CT) of 67 traffic fatality victims, and the results were compared with diagnoses obtained by autopsy. The injuries were...... system, but the pathologist diagnosed more organ injuries. We recommend the use of a radiologist as a consultant for the evaluation of postmortem CT images. Training in radiology should be included in forensic medicine postgraduate training. CT was superior to autopsy in detecting abnormal air...

  20. Bacterial mitosis: Partitioning protein ParA oscillates in spiral-shaped structures and positions plasmids at mid-cell

    DEFF Research Database (Denmark)

    Ebersbach, G.; Gerdes, Kenn

    2004-01-01

    The par2 locus of Escherichia coli plasmid pB171 encodes oscillating ATPase ParA, DNA binding protein ParB and two cis-acting DNA regions to which ParB binds (parC1 and parC2). Three independent techniques were used to investigate the subcellular localization of plasmids carrying par2. In cells......A-GFP oscillated in spiral-shaped structures. Amino acid substitutions in ParA simultaneously abolished ParA spiral formation, oscillation and either plasmid localization or plasmid separation at mid-cell. Therefore, our results suggest that ParA spirals position plasmids at the middle of the bacterial nucleoid...

  1. Comparative analysis of the results obtained by computer code ASTEC V2 and RELAP 5.3.2 for small leak ID 80 for VVER 1000

    International Nuclear Information System (INIS)

    Atanasova, B.; Grudev, P.

    2011-01-01

    The purpose of this report is to present the results obtained by simulation and subsequent analysis of emergency mode for small leak with ID 80 for WWER 1000/B320 - Kozloduy NPP Units 5 and 6. Calculations were performed with the ASTEC v2 computer code used for calculation of severe accident, which was designed by French and German groups - IRSN and GRS. Integral RELAP5 computer code is used as a reference for comparison of results. The analyzes are focused on the processes occurring in reactor internals phase of emergency mode with significant core damage. The main thermohydraulic parameters, start of reactor core degradation and subsequent fuel relocalization till reactor vessel failure are evaluated in the analysis. RELAP5 computer code is used as a reference code to compare the results obtained till early core degradation that occurs after core stripping and excising of fuel temperature above 1200 0 C

  2. Plasmid mediated enhancement of uv resistance in Streptococcus faecalis

    International Nuclear Information System (INIS)

    Miehl, R.; Miller, M.; Yasbin, R.E.

    1980-01-01

    A 38.5-Mdal plasmid of Streptococcus faecalis subdp. zymogenes has been shown to enhance survival following uv irradiation. In addition, the presence of this plasmid increases the mutation frequencies following uv irradiation and enhanced W-reactivation. The data presented indicate that S. faecalis has an inducible error-prone repair system and that the plasmid enhances these repair functions

  3. Isolation of a conjugative F-like plasmid from a multidrug-resistant Escherichia coli strain CM6 using tandem shock wave-mediated transformation.

    Science.gov (United States)

    Soto-Alonso, G; Cruz-Medina, J A; Caballero-Pérez, J; Arvizu-Hernández, I; Ávalos-Esparza, L M; Cruz-Hernández, A; Romero-Gómez, S; Rodríguez, A L; Pastrana-Martínez, X; Fernández, F; Loske, A M; Campos-Guillén, J

    2015-07-01

    Genetic characterization of plasmids from bacterial strains provides insight about multidrug resistance. Ten wild type Escherichia coli (E. coli) strains isolated from cow fecal samples were characterized by their antibiotic resistance profile, plasmid patterns and three different identification methods. From one of the strains, a fertility factor-like plasmid was replicated using tandem shock wave-mediated transformation. Underwater shock waves with a positive pressure peak of up to approximately 40 MPa, followed by a pressure trough of approximately -19 MPa were generated using an experimental piezoelectric shock wave source. Three different shock wave energies and a fixed delay of 750 μs were used to study the relationship between energy and transformation efficiency (TE), as well as the influence of shock wave energy on the integrity of the plasmid. Our results showed that the mean shock wave-mediated TE and the integrity of the large plasmid (~70 kb) were reduced significantly at the energy levels tested. The sequencing analysis of the plasmid revealed a high identity to the pHK17a plasmid, including the replication system, which was similar to the plasmid incompatibility group FII. It also showed that it carried an extended spectrum beta-lactamase gene, ctx-m-14. Furthermore, diverse genes for the conjugative mechanism were identified. Our results may be helpful in improving methodologies for conjugative plasmid transfer and directly selecting the most interesting plasmids from environmental samples. Copyright © 2015 Elsevier B.V. All rights reserved.

  4. Fermentation Results and Chemical Composition of Agricultural Distillates Obtained from Rye and Barley Grains and the Corresponding Malts as a Source of Amylolytic Enzymes and Starch.

    Science.gov (United States)

    Balcerek, Maria; Pielech-Przybylska, Katarzyna; Dziekońska-Kubczak, Urszula; Patelski, Piotr; Strąk, Ewelina

    2016-10-01

    The objective of this study was to determine the efficiency of rye and barley starch hydrolysis in mashing processes using cereal malts as a source of amylolytic enzymes and starch, and to establish the volatile profile of the obtained agricultural distillates. In addition, the effects of the pretreatment method of unmalted cereal grains on the physicochemical composition of the prepared mashes, fermentation results, and the composition of the obtained distillates were investigated. The raw materials used were unmalted rye and barley grains, as well as the corresponding malts. All experiments were first performed on a semi-technical scale, and then verified under industrial conditions in a Polish distillery. The fermentable sugars present in sweet mashes mostly consisted of maltose, followed by glucose and maltotriose. Pressure-thermal treatment of unmalted cereals, and especially rye grains, resulted in higher ethanol content in mashes in comparison with samples subjected to pressureless liberation of starch. All agricultural distillates originating from mashes containing rye and barley grains and the corresponding malts were characterized by low concentrations of undesirable compounds, such as acetaldehyde and methanol. The distillates obtained under industrial conditions contained lower concentrations of higher alcohols (apart from 1-propanol) than those obtained on a semi-technical scale.

  5. Fermentation Results and Chemical Composition of Agricultural Distillates Obtained from Rye and Barley Grains and the Corresponding Malts as a Source of Amylolytic Enzymes and Starch

    Directory of Open Access Journals (Sweden)

    Maria Balcerek

    2016-10-01

    Full Text Available The objective of this study was to determine the efficiency of rye and barley starch hydrolysis in mashing processes using cereal malts as a source of amylolytic enzymes and starch, and to establish the volatile profile of the obtained agricultural distillates. In addition, the effects of the pretreatment method of unmalted cereal grains on the physicochemical composition of the prepared mashes, fermentation results, and the composition of the obtained distillates were investigated. The raw materials used were unmalted rye and barley grains, as well as the corresponding malts. All experiments were first performed on a semi-technical scale, and then verified under industrial conditions in a Polish distillery. The fermentable sugars present in sweet mashes mostly consisted of maltose, followed by glucose and maltotriose. Pressure-thermal treatment of unmalted cereals, and especially rye grains, resulted in higher ethanol content in mashes in comparison with samples subjected to pressureless liberation of starch. All agricultural distillates originating from mashes containing rye and barley grains and the corresponding malts were characterized by low concentrations of undesirable compounds, such as acetaldehyde and methanol. The distillates obtained under industrial conditions contained lower concentrations of higher alcohols (apart from 1-propanol than those obtained on a semi-technical scale.

  6. Prevalence and characterization of plasmids carrying sulfonamide resistance genes among Escherichia coli from pigs, pig carcasses and human

    Directory of Open Access Journals (Sweden)

    Hammerum Anette M

    2010-07-01

    Full Text Available Abstract Background Sulfonamide resistance is very common in Escherichia coli. The aim of this study was to characterize plasmids carrying sulfonamide resistance genes (sul1, sul2 and sul3 in E. coli isolated from pigs and humans with a specific objective to assess the genetic diversity of plasmids involved in the mobility of sul genes. Methods A total of 501 E. coli isolates from pig feces, pig carcasses and human stools were tested for their susceptibility to selected antimicrobial. Multiplex PCR was conducted to detect the presence of three sul genes among the sulfonamide-resistant E. coli isolates. Fifty-seven sulfonamide-resistant E. coli were selected based on presence of sul resistance genes and subjected to conjugation and/or transformation experiments. S1 nuclease digestion followed by pulsed-field gel electrophoresis was used to visualize and determine the size of plasmids. Plasmids carrying sul genes were characterized by PCR-based replicon typing to allow a comparison of the types of sul genes, the reservoir and plasmid present. Results A total of 109/501 isolates exhibited sulfonamide resistance. The relative prevalences of sul genes from the three reservoirs (pigs, pig carcasses and humans were 65%, 45% and 12% for sul2, sul1, and sul3, respectively. Transfer of resistance through conjugation was observed in 42/57 isolates. Resistances to streptomycin, ampicillin and trimethoprim were co-transferred in most strains. Class 1 integrons were present in 80% of sul1-carrying plasmids and 100% of sul3-carrying plasmids, but only in 5% of sul2-carrying plasmids. The sul plasmids ranged from 33 to 160-kb in size and belonged to nine different incompatibility (Inc groups: FII, FIB, I1, FIA, B/O, FIC, N, HI1 and X1. IncFII was the dominant type in sul2-carrying plasmids (52%, while IncI1 was the most common type in sul1 and sul3-carrying plasmids (33% and 45%, respectively. Multireplicons were found associated with all three sul genes

  7. LEP and results obtained by DELPHI after four years of operation; LEP i wyniki uzyskane przez DELPHI po czterech latach dzialania

    Energy Technology Data Exchange (ETDEWEB)

    Blocki, J. [Warsaw Univ. (Poland). Inst. Fizyki Doswiadczalnej; Brueckman de Renstrom, P.; Budziak, A. [Institute of Nuclear Physics, Cracow (Poland)] [and others

    1993-10-01

    We characterize the most important problems of modern elementary particles physics, for the solution of which the LEP (Large Electron Positron) accelerator was built. We present the characteristics of this accelerator. The structure and properties of the DELPHI detector are described with special emphasis on the contribution of Polish groups. The most important results obtained so far in the LEP accelerator are discussed. (author). 12 refs, 17 figs, 1 tab.

  8. Carbon and nitrogen determination in Zr by photon or proton activation analysis. Comparison between the results obtained by this method and other analytical techniques

    International Nuclear Information System (INIS)

    Petit, J.; Gosset, J.; Engelmann, C.

    1977-01-01

    Carbon and nitrogen are determined by the following nuclear reactions: 12 C(γ,n) 11 C and 14 N(p,α) 11 C. The performances of the method and the main interferences are considered. The process developed for the separation of carbon-11 from zirconium is described and its efficiency evaluated. The results obtained are compared with those given by different laboratories using various analytical techniques [fr

  9. Sequence Analysis of the Cryptic Plasmid pMG101 from Rhodopseudomonas palustris and Construction of Stable Cloning Vectors

    Science.gov (United States)

    Inui, Masayuki; Roh, Jung Hyeob; Zahn, Kenneth; Yukawa, Hideaki

    2000-01-01

    A 15-kb cryptic plasmid was obtained from a natural isolate of Rhodopseudomonas palustris. The plasmid, designated pMG101, was able to replicate in R. palustris and in closely related strains of Bradyrhizobium japonicum and phototrophic Bradyrhizobium species. However, it was unable to replicate in the purple nonsulfur bacterium Rhodobacter sphaeroides and in Rhizobium species. The replication region of pMG101 was localized to a 3.0-kb SalI-XhoI fragment, and this fragment was stably maintained in R. palustris for over 100 generations in the absence of selection. The complete nucleotide sequence of this fragment revealed two open reading frames (ORFs), ORF1 and ORF2. The deduced amino acid sequence of ORF1 is similar to sequences of Par proteins, which mediate plasmid stability from certain plasmids, while ORF2 was identified as a putative rep gene, coding for an initiator of plasmid replication, based on homology with the Rep proteins of several other plasmids. The function of these sequences was studied by deletion mapping and gene disruptions of ORF1 and ORF2. pMG101-based Escherichia coli-R. palustris shuttle cloning vectors pMG103 and pMG105 were constructed and were stably maintained in R. palustris growing under nonselective conditions. The ability of plasmid pMG101 to replicate in R. palustris and its close phylogenetic relatives should enable broad application of these vectors within this group of α-proteobacteria. PMID:10618203

  10. Gene electrotransfer of plasmid antiangiogenic metargidin peptide (AMEP) in disseminated melanoma

    DEFF Research Database (Denmark)

    Spanggaard, Iben; Snoj, Marko; Cavalcanti, Andrea

    2013-01-01

    ). In each patient, two cutaneous lesions were identified (one treated and one control). At day 1 and day 8, plasmid AMEP was injected intratumorally followed by electrotransfer. Patients were monitored weekly until day 29, and at day 64. Local efficacy was assessed at day 29 by direct measurement...... lesions increased more than 20%. No response occurred in distant lesions. This first-in-man study on electrotransfer of plasmid AMEP into cutaneous melanoma shows that the procedure and drug are safe and that local transfection was obtained....

  11. Construction of recombinant ZNF230/GFP fused plasmids and their expression and cellular localization

    DEFF Research Database (Denmark)

    Xu, Wen-Ming; Zhang, Si-Zhong; Qiu, Wei-Min

    2004-01-01

    To use green fluorescent protein as a marker to study the localization of the fusion protein, the mutant full length cDNAs of human ZNF230 and mouse znf230 with their stop codon TGA changed to TGG were obtained by PCR amplification, and then cloned into pGEM-Teasy vector. After the double enzyme...... cutting, the mutated human and mouse ZNF230(znf230) were inserted into mammalian expression plasmid pEGFP-N1. Thus we constructed the plasmid with fusion gene of ZNF230 and green fluorescent protein(GFP). Then the Cos cell was transfected with the fused gene by liposome. Fluorescence microscopy showed...

  12. Formation of Escherichia coli Hfr strains by integrative suppression with the P group plasmid RP1.

    OpenAIRE

    Martin, R R; Thorlton, C L; Unger, L

    1981-01-01

    Hfr strains of Escherichia coli were obtained by integrative suppression of a dnaA(Ts) mutation by the Inc P-1 plasmid RP1 without prior creation of an unnatural homology between the plasmid and the E. coli chromosome. Unmodified RP1 mobilized the polarized transfer of the chromosome in a counterclock-wise direction from a distinct origin between 81 min (pyrE) and 82 min (dnaA) with pyrE as a leading marker. Inheritance of RP1-Hfr chromosomal and antibiotic resistance genes was due to recombi...

  13. Ecological and genetic determinants of plasmid distribution in Escherichia coli.

    Science.gov (United States)

    Medaney, Frances; Ellis, Richard J; Raymond, Ben

    2016-11-01

    Bacterial plasmids are important carriers of virulence and antibiotic resistance genes. Nevertheless, little is known of the determinants of plasmid distribution in bacterial populations. Here the factors affecting the diversity and distribution of the large plasmids of Escherichia coli were explored in cattle grazing on semi-natural grassland, a set of populations with low frequencies of antibiotic resistance genes. Critically, the population genetic structure of bacterial hosts was chararacterized. This revealed structured E. coli populations with high diversity between sites and individuals but low diversity within cattle hosts. Plasmid profiles, however, varied considerably within the same E. coli genotype. Both ecological and genetic factors affected plasmid distribution: plasmid profiles were affected by site, E. coli diversity, E. coli genotype and the presence of other large plasmids. Notably 3/26 E. coli serotypes accounted for half the observed plasmid-free isolates indicating that within species variation can substantially affect carriage of the major conjugative plasmids. The observed population structure suggest that most of the opportunities for within species plasmid transfer occur between different individuals of the same genotype and support recent experimental work indicating that plasmid-host coevolution, and epistatic interactions on fitness costs are likely to be important in determining occupancy. © 2016 The Authors. Environmental Microbiology published by Society for Applied Microbiology and John Wiley & Sons Ltd.

  14. An Enterobacter plasmid as a new genetic background for the transposon Tn1331

    Directory of Open Access Journals (Sweden)

    Alavi MR

    2011-11-01

    Full Text Available Mohammad R Alavi1,2, Vlado Antonic2, Adrien Ravizee1, Peter J Weina3, Mina Izadjoo1,2, Alexander Stojadinovic21Division of Wound Biology and Translational Research, Armed Forces Institute of Pathology and American Registry of Pathology, Washington DC, 2Combat Wound Initiative Program, Walter Reed Army Medical Center, Washington DC, 3The Walter Reed Army Institute of Research, Silver Spring, MD, USABackground: Genus Enterobacter includes important opportunistic nosocomial pathogens that could infect complex wounds. The presence of antibiotic resistance genes in these microorganisms represents a challenging clinical problem in the treatment of these wounds. In the authors’ screening of antibiotic-resistant bacteria from complex wounds, an Enterobacter species was isolated that harbors antibiotic-resistant plasmids conferring resistance to Escherichia coli. The aim of this study was to identify the resistance genes carried by one of these plasmids.Methods: The plasmids from the Enterobacter isolate were propagated in E. coli and one of the plasmids, designated as pR23, was sequenced by the Sanger method using fluorescent dye-terminator chemistry on a genetic analyzer. The assembled sequence was annotated by search of the GenBank database.Results: Plasmid pR23 is composed of the transposon Tn1331 and a backbone plasmid that is identical to the plasmid pPIGDM1 from Enterobacter agglomerans. The multidrug-resistance transposon Tn1331, which confers resistance to aminoglycoside and beta lactam antibiotics, has been previously isolated only from Klebsiella. The Enterobacter plasmid pPIGDM1, which carries a ColE1-like origin of replication and has no apparent selective marker, appears to provide a backbone for propagation of Tn1331 in Enterobacter. The recognition sequence of Tn1331 transposase for insertion into pPIGDM1 is the pentanucleotide TATTA, which occurs only once throughout the length of this plasmid.Conclusion: Transposition of Tn1331 into

  15. Increasing plasmid transformation efficiency of natural spizizen method in Bacillus Subtilis by a cell permeable peptide

    Directory of Open Access Journals (Sweden)

    Mehrdad Moosazadeh Moghaddam

    2013-01-01

    Full Text Available Introduction: Some of bacterial species are able to uptake DNA molecule from environment, the yield of this process depends on some conditions such as plasmid size and host type. In the case of Bacillus subtilis, DNA uptake has low efficacy. Using Spizizen minimal medium is common method in plasmid transformation into B. subtilis, but rate of this process is not suitable and noteworthy. The aim of this study was investigation of novel method for improvement of DNA transformation into B. subtilis based on CM11 cationic peptide as a membrane permeable agent.Materials and methods: In this study, for optimization of pWB980 plasmid transformation into B. subtilis, the CM11 cationic peptide was used. For this purpose, B. subtilis competent cell preparation in the present of different concentration of peptide was implemented by two methods. In the first method, after treatment of bacteria with different amount of peptide for 14h, plasmid was added. In the second method, several concentration of peptide with plasmid was exposed to bacteria simultaneously. Bacteria that uptake DNA were screened on LB agar medium containing kanamycin. The total transformed bacteria per microgram of DNA was calculated and compared with the control.Results: Plasmid transformation in best conditions was 6.5 folds higher than the control. This result was statistically significant (P value <0.001.Discussion and conclusion: This study showed that CM11 cationic peptide as a membrane permeable agent was able to increase plasmid transformation rate into B. subtilis. This property was useful for resolution of low transformation efficacy.

  16. Rapid and inexpensive method for isolating plasmid DNA

    International Nuclear Information System (INIS)

    Aljanabi, S. M.; Al-Awadi, S. J.; Al-Kazaz, A. A.; Baghdad Univ.

    1997-01-01

    A small-scale and economical method for isolating plasmid DNA from bacteria is described. The method provides DNA of suitable quality for most DNA manipulation techniques. This DNA can be used for restriction endonuclease digestion, southern blot hybridization, nick translation and end labeling of DNA probes, Polymerase Chain Reaction (PCR) -based techniques, transformation, DNA cycle-sequencing, and Chain-termination method for DNA sequencing. The entire procedure is adapted to 1.5 ml microfuge tubes and takes approximately 30 mins. The DNA isolated by this method has the same purity produced by CTAB and cesium chloride precipitation and purification procedures respectively. The two previous methods require many hours to obtain the final product and require the use of very expensive equipment as ultracentrifuge. This method is well suited for the isolation of plasmid DNA from a large number of bacterial samples and in a very short time and low cost in laboratories where chemicals, expensive equipment and finance are limited factors in conducting molecular research. (authors). 11refs. 11refs

  17. Effects of different replicons in conjugative plasmids on transformation efficiency, plasmid stability, gene expression and n-butanol biosynthesis in Clostridium tyrobutyricum

    Energy Technology Data Exchange (ETDEWEB)

    Yu, Mingrui; Du, Yinming; Jiang, Wenyan; Chang, Wei-Lun; Yang, Shang-Tian [Ohio State Univ., Columbus, OH (United States). William G. Lowrie Dept. of Chemical and Biomolecular Engineering; Tang, I-Ching [Bioprocessing Innovative Company, Dublin, OH (United States)

    2012-01-15

    Clostridium tyrobutyricum ATCC 25755 can produce butyric acid, acetic acid, and hydrogen as the main products from various carbon sources. In this study, C. tyrobutyricum was used as a host to produce n-butanol by expressing adhE2 gene under the control of a native thiolase promoter using four different conjugative plasmids (pMTL82151, 83151, 84151, and 85151) each with a different replicon (pBP1 from C. botulinum NCTC2916, pCB102 from C. butyricum, pCD6 from Clostridium difficile, and pIM13 from Bacillus subtilis). The effects of different replicons on transformation efficiency, plasmid stability, adhE2 expression and aldehyde/alcohol dehydrogenase activities, and butanol production by different mutants of C. tyrobutyricum were investigated. Among the four plasmids and replicons studied, pMTL82151 with pBP1 gave the highest transformation efficiency, plasmid stability, gene expression, and butanol biosynthesis. Butanol production from various substrates, including glucose, xylose, mannose, and mannitol were then investigated with the best mutant strain harboring adhE2 in pMTL82151. A high butanol titer of 20.5 g/L with 0.33 g/g yield and 0.32 g/L h productivity was obtained with mannitol as the substrate in batch fermentation with pH controlled at {proportional_to}6.0. (orig.)

  18. Comparison between two sampling methods by results obtained using petrographic techniques, specially developed for minerals of the Itataia uranium phosphate deposit, Ceara, Brazil

    International Nuclear Information System (INIS)

    Salas, H.T.; Murta, R.L.L.

    1985-01-01

    The results of comparison of two sampling methods applied to a gallery of the uranium-phosphate ore body of Itataia-Ceara State, Brazil, along 235 metres of mineralized zone, are presented. The results were obtained through petrographic techniques especially developed and applied to both samplings. In the first one it was studied hand samples from a systematically sampling made at intervals of 2 metres. After that, the estimated mineralogical composition studies were carried out. Some petrogenetic observations were for the first time verified. The second sampling was made at intervals of 20 metres and 570 tons of ore extracted and distributed in sections and a sample representing each section was studied after crushing at -65. Their mineralogy were quantified and the degree of liberation of apatite calculated. Based on the mineralogical data obtained it was possible to represent both samplings and to make the comparison of the main mineralogical groups (phosphates, carbonates and silicates). In spite of utilizing different methods and methodology and the kind of mineralization, stockwork, being quite irregular, the results were satisfactory. (Author) [pt

  19. Interactions Between the Cytomegalovirus Promoter and the Estrogen Response Element: Implications for Design of Estrogen-Responsive Reporter Plasmids

    OpenAIRE

    Derecka, K.; Wang, C.K.; Flint, A.P.F.

    2006-01-01

    We aimed to produce an estrogen-responsive reporter plasmid that would permit monitoring of estrogen receptor function in the uterus in vivo. The plasmid pBL-tk-CAT(+)ERE was induced by estrogen in bovine endometrial stromal cells. When the CAT gene was replaced by the secreted alkaline phosphatase SeAP, the resulting construct pBL-tk-SeAP(+)ERE remained estrogen responsive. However when the tk promoter was replaced by the cytomegalovirus (cmv) promoter, the resulting plasmid (pBL-cmv-SeAP(+)...

  20. Warm magnetic measurements of MCBCs: comparison between the results obtained with the Quadrupole Industrial Magnetic Measurement and the Corrector Industrial Magnetic Measurement systems

    CERN Document Server

    Remondino, Vittorio; Catalan-Lasheras, N

    2005-01-01

    MCBCs modules are first magnetically measured at Tesla using a second-generation Corrector Industrial Magnetic Measurement (CIMM). After assembly into the SSS quadrupoles, measurements are repeated at CERN using the Quadrupole Industrial Magnetic Measurement (QIMM) system. In this note, we compare the measurements provided by the two systems. In all the 18 cases examined, the correlation found is excellent. The consistency of the results obtained indicates that both systems are effective, that modules are correctly measured by the firm personnel and that magnetic characteristics of the modules do not change during their assembly in cold masses.

  1. Transformation of UV-hypersensitive Chinese hamster ovary cell mutants with UV-irradiated plasmids

    International Nuclear Information System (INIS)

    Nairn, R.S.; Humphrey, R.M.; Adair, G.M.

    1988-01-01

    Transfection of UV-hypersensitive, DNA repair-deficient Chinese hamster ovary (CHO) cell lines and parental, repair-proficient CHO cells with UV-irradiated pHaprt-1 or pSV2gpt plasmids resulted in different responses by recipient cell lines to UV damage in transfected DNA. Unlike results reported for human cells, UV irradiation of transfecting DNA did not stimulate genetic transformation of CHO recipient cells. In repair-deficient CHO cells, proportionally fewer transformants were produced with increasing UV damage than in repair-proficient cells in transfections with UV-irradiated hamster adenine phosphoribosyltransferase (APRT) gene contained in plasmid pHaprt-1. Transfection of CHO cells with UV-irradiated pSV2gpt resulted in neither decline in transformation frequencies in repair-deficient cell lines relative to repair-proficient cells nor stimulation of genetic transformation by UV damage in the plasmid. Blot hybridization analysis of DNA samples isolated from transformed cells showed no dramatic changes in copy number or arrangement of transfected plasmid DNA with increasing UV dose. The authors conclude responses of recipient cells to UV-damaged transfecting plasmids depend on type of recipient cell and characteristics of the genetic sequence used for transfection. (author)

  2. CARTOGRAPHIE DU PLASMIDE pSU100, PLASMIDE CRYPTIQUE DE LACTOBACILLUS CASEI

    Directory of Open Access Journals (Sweden)

    F BENSALAH

    2003-06-01

    Ce plasmide appelé pSU100 a été cloné dans le vecteur de transformation pUC18 au site EcoRI chez E. coli JM103. Les profils électrophorétiques de restriction obtenus par des digestions simples, doubles et triples sous l’action de 33 endonucléases, ont contribué à l’élaboration d’une carte de restriction de ce plasmide. Cinq sites uniques ont été identifiés, ainsi que d’autres sites doubles et multiples. Une étude préliminaire du rôle physiologique de ce plasmide a permis de déceler une résistance à la kanamycine.

  3. Comparison of visual field test results obtained through Humphrey matrix frequency doubling technology perimetry versus standard automated perimetry in healthy children

    Directory of Open Access Journals (Sweden)

    Sibel Kocabeyoglu

    2013-01-01

    Full Text Available Aims : The aim of this study was to compare the visual field test results in healthy children obtained via the Humphrey matrix 24-2 threshold program and standard automated perimetry (SAP using the Swedish interactive threshold algorithm (SITA-Standard 24-2 test. Materials and Methods: This prospective study included 55 healthy children without ocular or systemic disorders who underwent both SAP and frequency doubling technology (FDT perimetry visual field testing. Visual field test reliability indices, test duration, global indices (mean deviation [MD], and pattern standard deviation [PSD] were compared between the 2 tests using the Wilcoxon signed-rank test and paired t-test. The performance of the Humphrey field analyzer (HFA 24-2 SITA-standard and frequency-doubling technology Matrix 24-2 tests between genders were compared with Mann-Whitney U-test. Results: Fifty-five healthy children with a mean age of 12.2 ± 1.9 years (range from 8 years to 16 years were included in this prospective study. The test durations of SAP and FDT were similar (5.2 ± 0.5 and 5.1 ± 0.2 min, respectively, P = 0.651. MD and the PSD values obtained via FDT Matrix were significantly higher than those obtained via SAP (P < 0.001, and fixation losses and false negative errors were significantly less with SAP (P < 0.05. A weak positive correlation between the two tests in terms of MD (r = 0.352, P = 0.008 and PSD (r = 0.329, P = 0.014 was observed. Conclusion: Children were able to complete both the visual test algorithms successfully within 6 min. However, SAP testing appears to be associated with less depression of the visual field indices of healthy children. FDT Matrix and SAP should not be used interchangeably in the follow-up of children.

  4. Reversible entrapment of plasmid deoxyribonucleic acid on different chromatographic supports.

    Science.gov (United States)

    Gabor, Boštjan; Černigoj, Urh; Barut, Miloš; Štrancar, Aleš

    2013-10-11

    HPLC based analytical assay is a powerful technique that can be used to efficiently monitor plasmid DNA (pDNA) purity and quantity throughout the entire purification process. Anion exchange monolithic and non-porous particle based stationary phases were used to study the recovery of the different pDNA isoforms from the analytical column. Three differently sized pDNA molecules of 3.0kbp, 5.2kbp and 14.0kbp were used. Plasmid DNA was injected onto columns under the binding conditions and the separation of the isoforms took place by increasing the ionic strength of the elution buffer. While there was no substantial decrease of the recovered supercoiled and linear isoforms of the pDNA with the increase of the plasmid size and with the increase of the flow rate (recoveries in all cases larger than 75%), a pronounced decrease of the oc isoform recovery was observed. The entrapment of the oc pDNA isoform occurred under non-binding conditions as well. The partial oc isoform elution from the column could be achieved by decreasing the flow rate of the elution mobile phase. The results suggested a reversible entrapment of the oc isoform in the restrictions within the pores of the monolithic material as well as within the intra-particle space of the non-porous particles. This phenomenon was observed on both types of the stationary phase morphologies and could only be connected to the size of a void space through which the pDNA needs to migrate. A prediction of reversible pDNA entrapment was successfully estimated with the calculation of Peclet numbers, Pe, which defines the ratio between a convective and diffusive mass transport. Copyright © 2013 Elsevier B.V. All rights reserved.

  5. Determination of residual stresses in steel 20 comparison of the results obtained by the Barkhausen noise and X-ray diffraction

    International Nuclear Information System (INIS)

    Fernandez, L. M.; Herrera, V.; Suarez, J.C.; Merino, F.J.

    1997-01-01

    During a heat treatment (quenching, surface hardening, Thermochemical treatment) the metallic alloys undergo temperature variations and phase transformation. The resulting changes give rise to internal stresses in the piece. Phase transformation modifies the thermomechanical behavior of the material through the change of mechanical properties ( hardness, fatigue-behaviour, corrosion resistance, electric resistivity, etc.) The aim of this paper is residual stress measurements in specimens of steel-20 submitted to different heat treatment, by two non destructive methods: Barkhausen and X-ray diffraction. Brakhausen Effects utilizes the magnetic properties of ferromagnetic materials such as ferritic and perlitic steels and provides a practical tool for surface streets evaluation. X-ray streets analysis is based on the measurement of lattice strains in different directions of specimen The results obtained by two techniques are compared, avowing a good agreement

  6. Cholesterol-conjugated supramolecular assemblies of low generations polyamidoamine dendrimers for enhanced EGFP plasmid DNA transfection

    Energy Technology Data Exchange (ETDEWEB)

    Golkar, Nasim; Samani, Soliman Mohammadi; Tamaddon, Ali Mohammad, E-mail: amtamadon@gmail.com [Shiraz University of Medical Sciences, Department of Pharmaceutics, School of Pharmacy (Iran, Islamic Republic of)

    2016-05-15

    Aimed to prepare an enhanced gene delivery system with low cytotoxicity and high transfection efficiency, various cholesterol-conjugated derivates of low generation polyamidoamine (PAMAM) dendrimers were prepared. The conjugates were characterized by TNBS assay, FTIR, and {sup 1}H-NMR spectroscopy. Self-assembly of the dendrimer conjugates (G1-Chol, G2-Chol, and G3-Chol) was investigated by pyrene assay. Following formation of the complexes between enhanced green fluorescence protein plasmid and the dendrimer conjugates at various N (primary amine)/P (phosphate) mole ratios, plasmid condensation, biologic stability, cytotoxicity, and protein expression were investigated. The conjugates self-assembled into micellar dispersions with the critical micelle concentration values (<50 µg/ml) depending on the dendrimer generation and cholesterol/amine mole ratio. Cholesterol conjugation resulted in higher resistance of the condensed plasmid DNA in a competition assay with heparin sulfate. Also, the transfection efficiency was determined higher for the cholesterol conjugates than unmodified dendrimers in HepG2 cells, showing the highest for G2-Chol at 40 % degree of cholesterol modification (G2-Chol{sub 40 %}) among various dendrimer generations. Interestingly, such conjugate showed a complete protection of plasmid against serum nucleases. Our results confirmed that the cholesterol conjugation to PAMAM dendrimers of low generations bearing little cytotoxicity improves their several physicochemical and biological characteristics required for an enhanced delivery of plasmid DNA into cells.

  7. Direct identification of antibiotic resistance genes on single plasmid molecules using CRISPR/Cas9 in combination with optical DNA mapping

    Science.gov (United States)

    Müller, Vilhelm; Rajer, Fredrika; Frykholm, Karolin; Nyberg, Lena K.; Quaderi, Saair; Fritzsche, Joachim; Kristiansson, Erik; Ambjörnsson, Tobias; Sandegren, Linus; Westerlund, Fredrik

    2016-12-01

    Bacterial plasmids are extensively involved in the rapid global spread of antibiotic resistance. We here present an assay, based on optical DNA mapping of single plasmids in nanofluidic channels, which provides detailed information about the plasmids present in a bacterial isolate. In a single experiment, we obtain the number of different plasmids in the sample, the size of each plasmid, an optical barcode that can be used to identify and trace the plasmid of interest and information about which plasmid that carries a specific resistance gene. Gene identification is done using CRISPR/Cas9 loaded with a guide-RNA (gRNA) complementary to the gene of interest that linearizes the circular plasmids at a specific location that is identified using the optical DNA maps. We demonstrate the principle on clinically relevant extended spectrum beta-lactamase (ESBL) producing isolates. We discuss how the gRNA sequence can be varied to obtain the desired information. The gRNA can either be very specific to identify a homogeneous group of genes or general to detect several groups of genes at the same time. Finally, we demonstrate an example where we use a combination of two gRNA sequences to identify carbapenemase-encoding genes in two previously not characterized clinical bacterial samples.

  8. The porcine circovirus type 1 capsid gene promoter improves antigen expression and immunogenicity in a HIV-1 plasmid vaccine

    Directory of Open Access Journals (Sweden)

    Burger Marieta

    2011-02-01

    Full Text Available Abstract Background One of the promising avenues for development of vaccines against Human immunodeficiency virus type 1 (HIV-1 and other human pathogens is the use of plasmid-based DNA vaccines. However, relatively large doses of plasmid must be injected for a relatively weak response. We investigated whether genome elements from Porcine circovirus type 1 (PCV-1, an apathogenic small ssDNA-containing virus, had useful expression-enhancing properties that could allow dose-sparing in a plasmid vaccine. Results The linearised PCV-1 genome inserted 5' of the CMV promoter in the well-characterised HIV-1 plasmid vaccine pTHgrttnC increased expression of the polyantigen up to 2-fold, and elicited 3-fold higher CTL responses in mice at 10-fold lower doses than unmodified pTHgrttnC. The PCV-1 capsid gene promoter (Pcap alone was equally effective. Enhancing activity was traced to a putative composite host transcription factor binding site and a "Conserved Late Element" transcription-enhancing sequence previously unidentified in circoviruses. Conclusions We identified a novel PCV-1 genome-derived enhancer sequence that significantly increased antigen expression from plasmids in in vitro assays, and improved immunogenicity in mice of the HIV-1 subtype C vaccine plasmid, pTHgrttnC. This should allow significant dose sparing of, or increased responses to, this and other plasmid-based vaccines. We also report investigations of the potential of other circovirus-derived sequences to be similarly used.

  9. Comparison of visual field test results obtained through Humphrey matrix frequency doubling technology perimetry versus standard automated perimetry in healthy children.

    Science.gov (United States)

    Kocabeyoglu, Sibel; Uzun, Salih; Mocan, Mehmet Cem; Bozkurt, Banu; Irkec, Murat; Orhan, Mehmet

    2013-10-01

    The aim of this study was to compare the visual field test results in healthy children obtained via the Humphrey matrix 24-2 threshold program and standard automated perimetry (SAP) using the Swedish interactive threshold algorithm (SITA)-Standard 24-2 test. This prospective study included 55 healthy children without ocular or systemic disorders who underwent both SAP and frequency doubling technology (FDT) perimetry visual field testing. Visual field test reliability indices, test duration, global indices (mean deviation [MD], and pattern standard deviation [PSD]) were compared between the 2 tests using the Wilcoxon signed-rank test and paired t-test. The performance of the Humphrey field analyzer (HFA) 24-2 SITA-standard and frequency-doubling technology Matrix 24-2 tests between genders were compared with Mann-Whitney U-test. Fifty-five healthy children with a mean age of 12.2 ± 1.9 years (range from 8 years to 16 years) were included in this prospective study. The test durations of SAP and FDT were similar (5.2 ± 0.5 and 5.1 ± 0.2 min, respectively, P = 0.651). MD and the PSD values obtained via FDT Matrix were significantly higher than those obtained via SAP (P tests in terms of MD (r = 0.352, P = 0.008) and PSD (r = 0.329, P = 0.014) was observed. Children were able to complete both the visual test algorithms successfully within 6 min. However, SAP testing appears to be associated with less depression of the visual field indices of healthy children. FDT Matrix and SAP should not be used interchangeably in the follow-up of children.

  10. Comparative genomics of multidrug resistance-encoding IncA/C plasmids from commensal and pathogenic Escherichia coli from multiple animal sources.

    Science.gov (United States)

    Fernández-Alarcón, Claudia; Singer, Randall S; Johnson, Timothy J

    2011-01-01

    Incompatibility group A/C (IncA/C) plasmids have received recent attention for their broad host range and ability to confer resistance to multiple antimicrobial agents. Due to the potential spread of multidrug resistance (MDR) phenotypes from foodborne pathogens to human pathogens, the dissemination of these plasmids represents a public health risk. In this study, four animal-source IncA/C plasmids isolated from Escherichia coli were sequenced and analyzed, including isolates from commercial dairy cows, pigs and turkeys in the U.S. and Chile. These plasmids were initially selected because they either contained the floR and tetA genes encoding for florfenicol and tetracycline resistance, respectively, and/or the bla(CMY-2) gene encoding for extended spectrum β-lactamase resistance. Overall, sequence analysis revealed that each of the four plasmids retained a remarkably stable and conserved backbone sequence, with differences observed primarily within their accessory regions, which presumably have evolved via horizontal gene transfer events involving multiple modules. Comparison of these plasmids with other available IncA/C plasmid sequences further defined the core and accessory elements of these plasmids in E. coli and Salmonella. Our results suggest that the bla(CMY-2) plasmid lineage appears to have derived from an ancestral IncA/C plasmid type harboring floR-tetAR-strAB and Tn21-like accessory modules. Evidence is mounting that IncA/C plasmids are widespread among enteric bacteria of production animals and these emergent plasmids have flexibility in their acquisition of MDR-encoding modules, necessitating further study to understand the evolutionary mechanisms involved in their dissemination and stability in bacterial populations.

  11. The Use of a Combined Bioinformatics Approach to Locate Antibiotic Resistance Genes on Plasmids From Whole Genome Sequences of Salmonella enterica Serovars From Humans in Ghana

    Directory of Open Access Journals (Sweden)

    Egle Kudirkiene

    2018-05-01

    Full Text Available In the current study, we identified plasmids carrying antimicrobial resistance genes in draft whole genome sequences of 16 selected Salmonella enterica isolates representing six different serovars from humans in Ghana. The plasmids and the location of resistance genes in the genomes were predicted using a combination of PlasmidFinder, ResFinder, plasmidSPAdes and BLAST genomic analysis tools. Subsequently, S1-PFGE was employed for analysis of plasmid profiles. Whole genome sequencing confirmed the presence of antimicrobial resistance genes in Salmonella isolates showing multidrug resistance phenotypically. ESBL, either blaTEM52−B or blaCTX−M15 were present in two cephalosporin resistant isolates of S. Virchow and S. Poona, respectively. The systematic genome analysis revealed the presence of different plasmids in different serovars, with or without insertion of antimicrobial resistance genes. In S. Enteritidis, resistance genes were carried predominantly on plasmids of IncN type, in S. Typhimurium on plasmids of IncFII(S/IncFIB(S/IncQ1 type. In S. Virchow and in S. Poona, resistance genes were detected on plasmids of IncX1 and TrfA/IncHI2/IncHI2A type, respectively. The latter two plasmids were described for the first time in these serovars. The combination of genomic analytical tools allowed nearly full mapping of the resistance plasmids in all Salmonella strains analyzed. The results suggest that the improved analytical approach used in the current study may be used to identify plasmids that are specifically associated with resistance phenotypes in whole genome sequences. Such knowledge would allow the development of rapid multidrug resistance tracking tools in Salmonella populations using WGS.

  12. Comparative genomics of multidrug resistance-encoding IncA/C plasmids from commensal and pathogenic Escherichia coli from multiple animal sources.

    Directory of Open Access Journals (Sweden)

    Claudia Fernández-Alarcón

    Full Text Available Incompatibility group A/C (IncA/C plasmids have received recent attention for their broad host range and ability to confer resistance to multiple antimicrobial agents. Due to the potential spread of multidrug resistance (MDR phenotypes from foodborne pathogens to human pathogens, the dissemination of these plasmids represents a public health risk. In this study, four animal-source IncA/C plasmids isolated from Escherichia coli were sequenced and analyzed, including isolates from commercial dairy cows, pigs and turkeys in the U.S. and Chile. These plasmids were initially selected because they either contained the floR and tetA genes encoding for florfenicol and tetracycline resistance, respectively, and/or the bla(CMY-2 gene encoding for extended spectrum β-lactamase resistance. Overall, sequence analysis revealed that each of the four plasmids retained a remarkably stable and conserved backbone sequence, with differences observed primarily within their accessory regions, which presumably have evolved via horizontal gene transfer events involving multiple modules. Comparison of these plasmids with other available IncA/C plasmid sequences further defined the core and accessory elements of these plasmids in E. coli and Salmonella. Our results suggest that the bla(CMY-2 plasmid lineage appears to have derived from an ancestral IncA/C plasmid type harboring floR-tetAR-strAB and Tn21-like accessory modules. Evidence is mounting that IncA/C plasmids are widespread among enteric bacteria of production animals and these emergent plasmids have flexibility in their acquisition of MDR-encoding modules, necessitating further study to understand the evolutionary mechanisms involved in their dissemination and stability in bacterial populations.

  13. Plasmid Mediated Antibiotic and Heavy Metal Resistance in Bacillus Strains Isolated From Soils in Rize, Turkey

    Directory of Open Access Journals (Sweden)

    Elif SEVİM

    2015-09-01

    Full Text Available Fifteen Bacillus strains which were isolated from soil samples were examined for resistance to 17 different antibiotics (ampicillin, methicillin, erythromycin, norfloxacin, cephalotine, gentamycin, ciprofloxacin, streptomycin, tobramycin, chloramphenicol, trimethoprim-sulfamethoxazole, tetracycline, vancomycin, oxacilin, neomycin, kanamycin and, novabiocin and to 10 different heavy metals (copper, lead, cobalt, chrome, iron, mercury, zinc, nickel, manganese and, cadmium and for the presence of plasmid DNA. A total of eleven strains (67% were resistant to at least one antibiotic. The most common resistance was observed against methicillin and oxacillin. The most resistance strains were found as Bacillus sp. B3 and Bacillus sp. B11. High heavy metal resistance against copper, chromium, zinc, iron and nickel was detected, but mercury and cobalt resistance was not detected, except for 3 strains (B3, B11, and B12 which showed mercury resistance. It has been determined that seven Bacillus strains have plasmids. The isolated plasmids were transformed into the Bacillus subtilis W168 and it was shown that heavy metal and antibiotic resistance determinants were carried on these plasmids. These results showed that there was a correlation between plasmid content and resistance for both antibiotic and heavy metal resistance

  14. Ultrasound-targeted microbubble destruction enhances naked plasmid DNA transfection in rabbit Achilles tendons in vivo.

    Science.gov (United States)

    Qiu, L; Zhang, L; Wang, L; Jiang, Y; Luo, Y; Peng, Y; Lin, L

    2012-07-01

    The study was to investigate the probability of increasing the transfection of the gene in tendons by ultrasound-targeted microbubble destruction (UTMD), and to search for the most suitable transfection conditions. A mixture of microbubbles and enhanced green fluorescent protein (EGFP) plasmids was injected into rabbit Achilles tendons by different administration routes and the tendons were ultrasound pulse by different ultrasonic conditions in order to determine the most appropriate conditions. Then, the rabbits were divided into four groups: (1) ultrasound + microbubbles + plasmid; (2) ultrasound+ plasmid; (3) microbubble + plasmid; (4) plasmid only. EGFP expression in the tendons and other tissues, and the damage to tendon and paratenon were all observed. The results showed that EGFP expression in the tendon was higher by ultrasound pulse with 2 W cm(-2) of output intensity and a 20% duty cycle for 10 min. Local injection was determined to be the better administration route. Among the four groups, EGFP expression in Group 1 was higher than that in other groups. EGFP expression was highest on seventh day, then it gradually decrease over time, and lasted more than 56 days. EGFP expression was not found in other tissues. There was no obvious injury caused by UTMD. Under suitable conditions, it is feasible to use UTMD as a safe and effective gene transfection therapy for tendon injuries.

  15. Sequential acquisition of R-plasmids in vivo by Salmonella typhimurium.

    Science.gov (United States)

    Platt, D J; Sommerville, J S; Gribben, J

    1984-01-01

    Salmonella typhimurium, resistant only to trimethoprim and sulphamethoxazole, was isolated from the faeces and blood of a chronic alcoholic patient in acute renal failure. The isolates harboured an 18 Md non-conjugative plasmid. He was dialysed peritoneally and treated with ampicillin; four days later there was no clinical improvement and his peritoneal dialysis fluid (PDF) had become infected. Salm. typhimurium was isolated from faeces and PDF. Both isolates were additionally resistant to ampicillin and contained two plasmids (55 Md and 18 Md). Therapy was changed to chloramphenicol and gentamicin was added to the PDF. Two weeks later Salm. typhimurium was again isolated from PDF and faeces. The PDF isolate was unchanged but 4% of the colonies isolated from this faecal specimen were resistant to chloramphenicol and had acquired an additional 62 Md plasmid. From all PDF and faecal specimens two different strains of Escherichia coli and one strain of Klebsiella pneumoniae were isolated which contained plasmids indistinguishable, on the basis of molecular weight and transferable resistance markers, from those acquired by Salm. typhimurium. The transferability of these plasmids in vitro to E. coli K12 and to the patient's initial Salm. typhimurium was studied and the results discussed.

  16. Transformation of Saccharomyces cerevisiae with UV-irradiated single-stranded plasmid.

    Science.gov (United States)

    Zgaga, Z

    1991-08-01

    UV-irradiated single-stranded replicative plasmids were used to transform different yeast strains. The low doses of UV used in this study (10-75 J/m2) caused a significant decrease in the transforming efficiency of plasmid DNA in the Rad+ strain, while they had no effect on transformation with double-stranded plasmids of comparable size. Neither the rev3 mutation, nor the rad18 or rad52 mutations influenced the efficiency of transformation with irradiated single-stranded plasmid. However, it was found to be decreased in the double rev3 rad52 mutant. Extracellular irradiation of plasmid that contains both URA3 and LEU2 genes (psLU) gave rise to up to 5% Leu- transformants among selected Ura+ ones in the repair-proficient strain. Induction of Leu- transformants was dose-dependent and only partially depressed in the rev3 mutant. These results suggest that both mutagenic and recombinational repair processes operate on UV-damaged single-stranded DNA in yeast.

  17. Protein-Nanocrystal Conjugates Support a Single Filament Polymerization Model in R1 Plasmid Segregation

    Energy Technology Data Exchange (ETDEWEB)

    Choi, Charina L.; Claridge, Shelley A.; Garner, Ethan C.; Alivisatos, A. Paul; Mullins, R. Dyche

    2008-07-15

    To ensure inheritance by daughter cells, many low-copy number bacterial plasmids, including the R1 drug-resistance plasmid, encode their own DNA segregation systems. The par operon of plasmid R1 directs construction of a simple spindle structure that converts free energy of polymerization of an actin-like protein, ParM, into work required to move sister plasmids to opposite poles of rod-shaped cells. The structures of individual components have been solved, but little is known about the ultrastructure of the R1 spindle. To determine the number of ParM filaments in a minimal R1 spindle, we used DNA-gold nanocrystal conjugates as mimics of the R1 plasmid. Wefound that each end of a single polar ParM filament binds to a single ParR/parC-gold complex, consistent with the idea that ParM filaments bind in the hollow core of the ParR/parC ring complex. Our results further suggest that multifilament spindles observed in vivo are associated with clusters of plasmidssegregating as a unit.

  18. Analysis of plasmid genes by phylogenetic profiling and visualization of homology relationships using Blast2Network

    Directory of Open Access Journals (Sweden)

    Bazzicalupo Marco

    2008-12-01

    Full Text Available Abstract Background Phylogenetic methods are well-established bioinformatic tools for sequence analysis, allowing to describe the non-independencies of sequences because of their common ancestor. However, the evolutionary profiles of bacterial genes are often complicated by hidden paralogy and extensive and/or (multiple horizontal gene transfer (HGT events which make bifurcating trees often inappropriate. In this context, plasmid sequences are paradigms of network-like relationships characterizing the evolution of prokaryotes. Actually, they can be transferred among different organisms allowing the dissemination of novel functions, thus playing a pivotal role in prokaryotic evolution. However, the study of their evolutionary dynamics is complicated by the absence of universally shared genes, a prerequisite for phylogenetic analyses. Results To overcome such limitations we developed a bioinformatic package, named Blast2Network (B2N, allowing the automatic phylogenetic profiling and the visualization of homology relationships in a large number of plasmid sequences. The software was applied to the study of 47 completely sequenced plasmids coming from Escherichia, Salmonella and Shigella spps. Conclusion The tools implemented by B2N allow to describe and visualize in a new way some of the evolutionary features of plasmid molecules of Enterobacteriaceae; in particular it helped to shed some light on the complex history of Escherichia, Salmonella and Shigella plasmids and to focus on possible roles of unannotated proteins. The proposed methodology is general enough to be used for comparative genomic analyses of bacteria.

  19. Development of the safety evaluation system in the respects of organizational factors and workers' consciousness. Pt. 1. Study of validities of functions for necessary evaluation and results obtained

    International Nuclear Information System (INIS)

    Takano, Kenichi; Tsuge, Tadafumi; Hasegawa, Naoko; Hirose, Ayako; Sasou, Kunihide

    2002-01-01

    CRIEPI decided to develop the safety evaluation system to investigate the safety level of the industrial sites due to questionnaires of organizational climate, safety managements, and workers' safety consciousness to workers. This report describes the questionnaire survey to apply to the domestic nuclear power plant for using obtained results as a fundamental data in order to construct the safety evaluation system. This system will be used for promoting safety culture in organizations of nuclear power plants. The questionnaire survey was conducted to 14 nuclear power stations for understanding the present status relating to safety issues. This questionnaire involves 122 items classified into following three categories: (1) safety awareness and behavior of plant personnel; (2) safety management; (3) organizational climate, based on the model considering contributing factor groups to safety culture. Obtained results were analyzed by statistical method to prepare functions of evaluation. Additionally, by applying a multivariate analysis, it was possible to extract several crucial factors influencing safety performance and to find a comprehensive safety indicator representing total organizational safety level. Significant relations were identified between accident rates (both labor accidents and facility failures) and above comprehensive safety indicator. Next, 122 questionnaire items were classified into 20 major safety factors to grasp the safety profiles of each site. This profile is considered as indicating the features of each site and also indicating the direction of progress for improvement of safety situation in the site. These findings can be reflected in developing the safety evaluation system, by confirming the validity of the evaluation method and giving specific functions. (author)

  20. Lactation curves and economic results of Saanen goats fed increasing dietary energy levels obtained by the addition of calcium salts of fatty acids

    Directory of Open Access Journals (Sweden)

    Rodrigo de Souza

    2014-02-01

    Full Text Available The objective of this study was to evaluate the use of calcium salts of fatty acids (CSFA to increase the dietary energy levels for Saanen goats and their effects on the lactation curve, dry matter intake, body weight, and economic results of the goats. Twenty multiparous goats, weighing an average of 63.5±10.3 kg, were randomly assigned to one of four treatment groups, each receiving one of the following dietary energy levels: a control diet consisting of 2.6 Mcal of metabolizable energy per kg of dry matter (Mcal ME/kg DM or a test diet supplemented with CSFA (Lactoplus® to obtain 2.7, 2.8, or 2.9 Mcal ME/kg DM. Goats were housed in individual stalls and were fed and milked twice daily. The animals were evaluated until 180 days in milk by measuring dry matter intake and milk yield. These measurements were used to calculate feed efficiencies and the cost-benefit ratio of diet and lactation curves using Wood's nonlinear model. Increasing dietary energy levels showed no effect on body weight. Supplementation with CSFA did not limit dry matter intake; however, it changed the shape of the lactation curve by promoting a late peak lactation with a longer duration. Milk yields at 180 days in milk had a quadratic increase with a maximum energy level at 2.85 Mcal ME/kg DM. Increasing the dietary energy level for Saanen goats using CSFA changes their lactation curves, with the best milk production achieved with a 2.85 Mcal ME/kg DM diet; however, the greatest economic results were obtained with a 2.7 Mcal ME/kg DM diet.

  1. CAPRRESI: Chimera Assembly by Plasmid Recovery and Restriction Enzyme Site Insertion.

    Science.gov (United States)

    Santillán, Orlando; Ramírez-Romero, Miguel A; Dávila, Guillermo

    2017-06-25

    Here, we present chimera assembly by plasmid recovery and restriction enzyme site insertion (CAPRRESI). CAPRRESI benefits from many strengths of the original plasmid recovery method and introduces restriction enzyme digestion to ease DNA ligation reactions (required for chimera assembly). For this protocol, users clone wildtype genes into the same plasmid (pUC18 or pUC19). After the in silico selection of amino acid sequence regions where chimeras should be assembled, users obtain all the synonym DNA sequences that encode them. Ad hoc Perl scripts enable users to determine all synonym DNA sequences. After this step, another Perl script searches for restriction enzyme sites on all synonym DNA sequences. This in silico analysis is also performed using the ampicillin resistance gene (ampR) found on pUC18/19 plasmids. Users design oligonucleotides inside synonym regions to disrupt wildtype and ampR genes by PCR. After obtaining and purifying complementary DNA fragments, restriction enzyme digestion is accomplished. Chimera assembly is achieved by ligating appropriate complementary DNA fragments. pUC18/19 vectors are selected for CAPRRESI because they offer technical advantages, such as small size (2,686 base pairs), high copy number, advantageous sequencing reaction features, and commercial availability. The usage of restriction enzymes for chimera assembly eliminates the need for DNA polymerases yielding blunt-ended products. CAPRRESI is a fast and low-cost method for fusing protein-coding genes.

  2. Recent Advances in Solid Catalysts Obtained by Metalloporphyrins Immobilization on Layered Anionic Exchangers: A Short Review and Some New Catalytic Results

    Directory of Open Access Journals (Sweden)

    Shirley Nakagaki

    2016-02-01

    Full Text Available Layered materials are a very interesting class of compounds obtained by stacking of two-dimensional layers along the basal axis. A remarkable property of these materials is their capacity to interact with a variety of chemical species, irrespective of their charge (neutral, cationic or anionic. These species can be grafted onto the surface of the layered materials or intercalated between the layers, to expand or contract the interlayer distance. Metalloporphyrins, which are typically soluble oxidation catalysts, are examples of molecules that can interact with layered materials. This work presents a short review of the studies involving metalloporphyrin immobilization on two different anionic exchangers, Layered Double Hydroxides (LDHs and Layered Hydroxide Salts (LHSs, published over the past year. After immobilization of anionic porphyrins, the resulting solids behave as reusable catalysts for heterogeneous oxidation processes. Although a large number of publications involving metalloporphyrin immobilization on LDHs exist, only a few papers have dealt with LHSs as supports, so metalloporphyrins immobilized on LHSs represent a new and promising research field. This work also describes new results on an anionic manganese porphyrin (MnP immobilized on Mg/Al-LDH solids with different nominal Mg/Al molar ratios (2:1, 3:1 and 4:1 and intercalated with different anions (CO32− or NO3−. The influence of the support composition on the MnP immobilization rates and the catalytic performance of the resulting solid in cyclooctene oxidation reactions will be reported.

  3. Assessment of the variations in fat content in normal liver using a fast MR imaging method in comparison with results obtained by spectroscopic imaging

    International Nuclear Information System (INIS)

    Irwan, Roy; Edens, Mireille A.; Sijens, Paul E.

    2008-01-01

    A recently published Dixon-based MRI method for quantifying liver fat content using dual-echo breath-hold gradient echo imaging was validated by phantom experiments and compared with results of biopsy in two patients (Radiology 2005;237:1048-1055). We applied this method in ten healthy volunteers and compared the outcomes with the results of MR spectroscopy (MRS), the gold standard in quantifying liver fat content. Novel was the use of spectroscopic imaging yielding the variations in fat content across the liver rather than a single value obtained by single voxel MRS. Compared with the results of MRS, liver fat content according to MRI was too high in nine subjects (range 3.3-10.7% vs. 0.9-7.7%) and correct in one (21.1 vs. 21.3%). Furthermore, in one of the ten subjects the MRI fat content according to the Dixon-based MRI method was incorrect due to a (100-x) versus x percent lipid content mix-up. The second problem was fixed by a minor adjustment of the MRI algorithm. Despite systematic overestimation of liver fat contents by MRI, Spearman's correlation between the adjusted MRI liver fat contents with MRS was high (r = 0.927, P < 0.001). Even after correction of the algorithm, the problem remaining with the Dixon-based MRI method for the assessment of liver fat content,is that, at the lower end range, liver fat content is systematically overestimated by 4%. (orig.)

  4. Recent Advances in Solid Catalysts Obtained by Metalloporphyrins Immobilization on Layered Anionic Exchangers: A Short Review and Some New Catalytic Results.

    Science.gov (United States)

    Nakagaki, Shirley; Mantovani, Karen Mary; Machado, Guilherme Sippel; Castro, Kelly Aparecida Dias de Freitas; Wypych, Fernando

    2016-02-29

    Layered materials are a very interesting class of compounds obtained by stacking of two-dimensional layers along the basal axis. A remarkable property of these materials is their capacity to interact with a variety of chemical species, irrespective of their charge (neutral, cationic or anionic). These species can be grafted onto the surface of the layered materials or intercalated between the layers, to expand or contract the interlayer distance. Metalloporphyrins, which are typically soluble oxidation catalysts, are examples of molecules that can interact with layered materials. This work presents a short review of the studies involving metalloporphyrin immobilization on two different anionic exchangers, Layered Double Hydroxides (LDHs) and Layered Hydroxide Salts (LHSs), published over the past year. After immobilization of anionic porphyrins, the resulting solids behave as reusable catalysts for heterogeneous oxidation processes. Although a large number of publications involving metalloporphyrin immobilization on LDHs exist, only a few papers have dealt with LHSs as supports, so metalloporphyrins immobilized on LHSs represent a new and promising research field. This work also describes new results on an anionic manganese porphyrin (MnP) immobilized on Mg/Al-LDH solids with different nominal Mg/Al molar ratios (2:1, 3:1 and 4:1) and intercalated with different anions (CO₃(2-) or NO₃(-)). The influence of the support composition on the MnP immobilization rates and the catalytic performance of the resulting solid in cyclooctene oxidation reactions will be reported.

  5. Comparison of Hematologic and Biochemical Test Results in Blood Samples Obtained by Jugular Venipuncture Versus Nail Clip in Moluccan Cockatoos (Cacatua moluccensis).

    Science.gov (United States)

    Bennett, Tracy D; Lejnieks, Daniel V; Koepke, Hoyt; Grimson, Fiona; Szucs, Jennifer; Omaits, Kerri; Lane, Rosalie

    2015-12-01

    In birds, blood samples are often collected from the jugular, medial metatarsal, and basilic vein. Samples are sometimes collected by toe nail clip, but concerns to avoid drawing blood from the nail include pain after nail clips for blood collection, potential differences in complete blood count (CBC) results, and potential contamination with uric acid values. To compare differences in biochemical and hematologic values in blood samples obtained by jugular venipuncture versus toenail clip, blood samples were collected from Moluccan cockatoos (Cacatua moluccensis) (N = 23) and sent to a commercial laboratory for routine CBCs and serum biochemical analysis. Results showed good agreement between venipuncture and nail clip blood samples in red blood cell count, packed cell volume, heterophil count and percentage, lymphocyte count and percentage, aspartate aminotransferase, chloride, creatine phosphokinase, glucose, lactate dehydrogenase, total protein, and uric acid values. Constant bias was found in values of bile acids, cholesterol, and hemoglobin. Proportional bias toward higher values in the jugular sample were found in total white blood cell (WBC) count and inorganic phosphorus. Serum calcium plots revealed a proportional bias toward higher values in the toe nail blood when values were increased. Results suggest some differences in WBC count, bile acids, calcium, cholesterol, hemoglobin, and phosphorus values between blood samples collected by jugular venipuncture and samples collected by toe nail clip, but the differences are mostly minor and, with the possible exception of inorganic phosphorus and marginally elevated or very low WBC counts, are unlikely to affect the use or interpretation of the avian blood panel.

  6. A review of experimental results obtained in Au-Au collisions at the RHIC-BNL energies using BRAHMS experimental set-up

    International Nuclear Information System (INIS)

    Jipa, Alexandru

    2003-01-01

    Ultrarelativistic heavy ion collisions are useful tools to investigate highly excited dense nuclear matter. At sufficiently high temperature, nuclear matter is expected to undergo a phase transition to quark-gluon plasma. Therefore, new experiments have been proposed to investigate the behaviour of the highly excited dense nuclear matter. One of the experiments performed at the Relativistic Heavy Ion Collider from Brookhaven National Laboratory is the BRAHMS Experiment. In the three running years many experimental data have been obtained. These data permitted to establish significant experimental results on charged pion, charged kaon, proton and antiproton production in Au-Au collisions at 130 A GeV and 200 A GeV in CMS. The present work summarizes the main experimental results on charged particle multiplicities, rapidity distributions, momentum spectra, antiparticle to particle ratios and connects these results with some interesting physical quantities. Their dependencies on the available energies in the CMS in similar collisions are included. Interesting behaviours with the increase of this energy are reported. The most interesting are related to the behaviour of the antiproton to proton ratio on the collision centrality and on the available energy in the centre of mass system, the strangeness production, Coulomb repulsion and slope parameters of the momentum spectra. A few new steps in the knowledge of the behaviour of the highly excited and dense nuclear matter can be reported. (authors)

  7. Containment Evaluation under Severe Accidents (CESA): synthesis of the predictive calculations and analysis of the first experimental results obtained on the Civaux mock-up

    International Nuclear Information System (INIS)

    Granger, L.; Rieg, C.Y.; Touret, J.P.; Fleury, F.; Nahas, G.; Danisch, R.; Brusa, L.; Millard, A.; Laborderie, C.; Ulm, F.; Contri, P.; Schimmelpfennig, K.; Barre, F.; Firnhaber, M.; Gauvain, J; Coulon, N.; Dutton, L.M.C.; Tuson, A.

    2001-01-01

    In 1996, EDF decided to build a containment model at the scale 1:3, the MAEVA mock-up, in order to check and study the behaviour of a pre-stressed concrete containment vessel without a liner in terms of mechanical strength and leaktightness, for loadings corresponding to its design and beyond design conditions. In parallel with the construction and testing of the mock-up, a cost-shared R and D action supported by the European Union, the CESA project, is dealing with quantification of leak rates through concrete cracks and porosity, predictive calculations of the behaviour of the mock-up and analysis of the experimental results. In this paper, we propose a synthesis of the main theoretical and experimental results, obtained after 2.5 years. It should however be noted that, due to some unexpected delays in the experimental programme, quite natural with such a huge and unique experimental set-up, only the design-basis accident sequences, already performed, have been reported in this paper. The first results are nevertheless very interesting, both from a scientific and nuclear utility point of view

  8. Localization of Low Copy Number Plasmid pRC4 in Replicating Rod and Non-Replicating Cocci Cells of Rhodococcus erythropolis PR4.

    Directory of Open Access Journals (Sweden)

    Divya Singhi

    Full Text Available Rhodococcus are gram-positive bacteria, which can exist in two different shapes rod and cocci. A number of studies have been done in the past on replication and stability of small plasmids in this bacterium; however, there are no reports on spatial localization and segregation of these plasmids. In the present study, a low copy number plasmid pDS3 containing pRC4 replicon was visualized in growing cells of Rhodococcus erythropolis PR4 (NBRC100887 using P1 parS-ParB-GFP system. Cells were initially cocci and then became rod shaped in exponential phase. Cocci cells were found to be non-replicating as evident by the presence of single fluorescence focus corresponding to the plasmid and diffuse fluorescence of DnaB-GFP. Rod shaped cells contained plasmid either present as one fluorescent focus observed at the cell center or two foci localized at quarter positions. The results suggest that the plasmid is replicated at the cell center and then it goes to quarter position. In order to observe the localization of plasmid with respect to nucleoid, plasmid segregation was also studied in filaments where it was found to be replicated at the cell center in a nucleoid free region. To the best of our knowledge, this is the first report on segregation of small plasmids in R. erythropolis.

  9. Diversity and homogeneity among small plasmids of Aeromonas salmonicida subsp. salmonicida linked with geographical origin

    Directory of Open Access Journals (Sweden)

    Sabrina A Attéré

    2015-11-01

    Full Text Available Furunculosis, which is caused by Aeromonas salmonicida subsp. salmonicida, is a major salmonid disease in fish farms worldwide. Several plasmids found in this bacterium confer phenotypes such drug resistance and virulence. Small plasmids (pAsa1, pAsa2, pAsa3, and pAsal1 related to ColE1- and ColE2-type replicons are usually present in its normal plasmidome. In the present study, with the objective to investigate if these plasmids display particularities related to the origin of the isolates bearing them, a total of 153 isolates, including 78 new and 75 previously described, were analyzed for the presence of small plasmids by PCR and DNA restriction fragment profiling. A geographical dichotomy between Canadian and European isolates for their propensity to do not have pAsa3 or pAsal1 was found. In addition, the genotyping analysis led to the identification of two European isolates harboring an unusual pAsal1. An investigation by next-generation sequencing (NGS of these two isolates shed light on two pAsal1 variants (pAsal1C and pAsal1D. As with pAsal1B, another pAsal1 variant previously described, these two new variants bore a second insertion sequence (ISAS5 in addition to the usual ISAS11. The characterization of these variants suggested that they could predominate over the wild-type pAsal1 in stressful conditions such as growth at temperatures of 25°C and above. To obtain a comprehensive portrait of the mutational pressure on small plasmids, 26 isolates whose DNA had been sequenced by NGS were investigated. pAsa3 and pAsal1 were more prone to mutations than pAsa1 and pAsa2, especially in the mobA gene, which encodes a relaxase and a primase. Lastly, the average copy number of each plasmid per cell was assessed using raw sequencing data. A clear trend with respect to the relative proportion per cell of each plasmid was identified. Our large-scale study revealed a geographical dichotomy in small plasmid repertoire in addition to a clear trend

  10. Some Results on Sea Ice Rheology for the Seasonal Ice Zone, Obtained from the Deformation Field of Sea Ice Drift Pattern

    Science.gov (United States)

    Toyota, T.; Kimura, N.

    2017-12-01

    Sea ice rheology which relates sea ice stress to the large-scale deformation of the ice cover has been a big issue to numerical sea ice modelling. At present the treatment of internal stress within sea ice area is based mostly on the rheology formulated by Hibler (1979), where the whole sea ice area behaves like an isotropic and plastic matter under the ordinary stress with the yield curve given by an ellipse with an aspect ratio (e) of 2, irrespective of sea ice area and horizontal resolution of the model. However, this formulation was initially developed to reproduce the seasonal variation of the perennial ice in the Arctic Ocean. As for its applicability to the seasonal ice zones (SIZ), where various types of sea ice are present, it still needs validation from observational data. In this study, the validity of this rheology was examined for the Sea of Okhotsk ice, typical of the SIZ, based on the AMSR-derived ice drift pattern in comparison with the result obtained for the Beaufort Sea. To examine the dependence on a horizontal scale, the coastal radar data operated near the Hokkaido coast, Japan, were also used. Ice drift pattern was obtained by a maximum cross-correlation method with grid spacings of 37.5 km from the 89 GHz brightness temperature of AMSR-E for the entire Sea of Okhotsk and the Beaufort Sea and 1.3 km from the coastal radar for the near-shore Sea of Okhotsk. The validity of this rheology was investigated from a standpoint of work rate done by deformation field, following the theory of Rothrock (1975). In analysis, the relative rates of convergence were compared between theory and observation to check the shape of yield curve, and the strain ellipse at each grid cell was estimated to see the horizontal variation of deformation field. The result shows that the ellipse of e=1.7-2.0 as the yield curve represents the observed relative conversion rates well for all the ice areas. Since this result corresponds with the yield criterion by Tresca and

  11. Plasmid P1 replication: negative control by repeated DNA sequences.

    OpenAIRE

    Chattoraj, D; Cordes, K; Abeles, A

    1984-01-01

    The incompatibility locus, incA, of the unit-copy plasmid P1 is contained within a fragment that is essentially a set of nine 19-base-pair repeats. One or more copies of the fragment destabilizes the plasmid when present in trans. Here we show that extra copies of incA interfere with plasmid DNA replication and that a deletion of most of incA increases plasmid copy number. Thus, incA is not essential for replication but is required for its control. When cloned in a high-copy-number vector, pi...

  12. Plasmids foster diversification and adaptation of bacterial populations in soil.

    Science.gov (United States)

    Heuer, Holger; Smalla, Kornelia

    2012-11-01

    It is increasingly being recognized that the transfer of conjugative plasmids across species boundaries plays a vital role in the adaptability of bacterial populations in soil. There are specific driving forces and constraints of plasmid transfer within bacterial communities in soils. Plasmid-mediated genetic variation allows bacteria to respond rapidly with adaptive responses to challenges such as irregular antibiotic or metal concentrations, or opportunities such as the utilization of xenobiotic compounds. Cultivation-independent detection and capture of plasmids from soil bacteria, and complete sequencing have provided new insights into the role and ecology of plasmids. Broad host range plasmids such as those belonging to IncP-1 transfer a wealth of accessory functions which are carried by similar plasmid backbones. Plasmids with a narrower host range can be more specifically adapted to particular species and often transfer genes which complement chromosomally encoded functions. Plasmids seem to be an ancient and successful strategy to ensure survival of a soil population in spatial and temporal heterogeneous conditions with various environmental stresses or opportunities that occur irregularly or as a novel challenge in soil. © 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  13. Permissiveness of soil microbial communities towards broad host range plasmids

    DEFF Research Database (Denmark)

    Klümper, Uli

    . Plasmids are implicated in the rapid spread of antibiotic resistance and the emergence of multi-resistant pathogenic bacteria, making it crucial to be able to quantify, understand, and, ideally, control plasmid transfer in mixed microbial communities. The fate of plasmids in microbial communities...... of microbial communities may be directly interconnected through transfer of BHR plasmids at a so far unrecognized level. The developed method furthermore enabled me to explore how agronomic practices may affect gene transfer in soil microbial communities. I compared bacterial communities extracted from plots...

  14. Comparison of arbitrarily primed-polymerase chain reaction and plasmid profiles typing of Pseudomonas aeruginosa strains from burn patients and hospital environment

    International Nuclear Information System (INIS)

    Japoni, A.; Farshad, S.; Alborzi, A.; Kalani, M.; Mohamadzadegan, R.

    2007-01-01

    Objective was to identify the strengths and weakness of arbitrary primed-polymerase chain reaction (A P-PCR) and plasmid profiles for typing of Pseudomonas aeruginosa (P. aeruginosa) and tracking of source of infections. Seventy-four strains of P. aeruginosa were isolated from burn patients and hospital environment between January to April 2003 in Ghotbadden Burn Hospital, Shiraz, Iran. The strains were classified by photo Capt Mw program, similarity and clustering of strains were assessed using NTSYS-PC version 2.02K software. Based on 50% and 64.7% and 67.5% similarity on the plotted dendrogram, 38 plasmid profiles were classified into: 2, 3 and 5 clusters, respectively. Photo Capt Mw program categorized AP-PCR products to 47 different types of 6 to 12 bands between 0.376 to 3.7 kb. Based on dendrogram pattern 3 levels (62 %, 81% and 84.6%) of similarity were selected. Using these criteria 2, 5and 11 clusters were obtained, respectively. As compared with plasmid profiles, AP-PCR analysis protocol is rapid, reproducible and differentiated the isolates with higher discrimination power. These results suggest that during admission of patients in burn center a limited number of common strains cross-contaminate burn victims. However, transmissions of infection from hospital environment to patients also occur in the minority of the victims. To control cross-contamination of the patient wounds with antibiotics resistant isolates, strong disinfection of patients' bathroom after scrubbing of each patient wounds is mandatory. (author)

  15. On the influence of infiltration of radioactive beds by flushing fluid on γ-ray logging results obtained from uranium deposit No. 387

    International Nuclear Information System (INIS)

    Ren Bingxiang; Zhang Yuechun; Ai Shuyi.

    1985-01-01

    A large number of field logging data obtained during the course of exploration on uranium deposit No. 387 show when the radioactive beds are encountered by drill holes and the drilling continues the γ-ray intensity decreases. It is considered that the escape of emanation does not considerably influence the logging results. Therefore, based upon the experiment of immerseing ores in fluid and geological and hydrogeological data of the deposit, the hypotheses that the γ-ray intensity in drill hole is closely related with the infiltration of flushing fluid is suggested. The ore bodies are strictly controlled by faults. Most of the uranium and thorium are absorbed by pelitic-carbonaceous cements or fill the porous spaces in structural breccia and cataclasite. They are easy to dissolve in water. Moreover, the ore-bearing structures are the unique water aquifers which is under pressure. As the level of flushing fluid is higher than the pressure head so it continuously pours into the ore-bearing beds, resulting in the infiltration of U and Ra. Consequently, the radioactivity detected is low

  16. Answers to the questions about food irradiation. Concerning results of animal experiments in the specified integrated research. Data carrying a problem in human health were obtained?

    International Nuclear Information System (INIS)

    Ito, Hitoshi

    2007-01-01

    Experts of Food and Agriculture Organization (FAO)/ International Atomic Energy Agency (IAEA)/ World Health Organization (WHO) committee obtained their conclusion in 1980 that food irradiated with <10 kGy of radiation is safe for human health, which is now globally approved. However, in Japan, there have been still opposite opinions based on the doubt in the title on the safety of irradiated food. In this paper, the author answers those questions as he was a member to arrange the Research in the title for food irradiation. Described are data presentation and explanation about results of toxicity studies of diets added with irradiated materials of: weight reductions in rat ovary by irradiated potato (ip) in chronic studies, and in mouse testicle and ovary of F3 generation from the ancestor mice kept on diet with irradiated onion (io); bone malformation in mice by io; and reduction of body weight gain in female rats by ip and increase of mortality of male rats by ip. These are analyzed on the aspects of radiation dose-response, sustained tendency of results throughout the living period or generation, and apparent abnormality by other factors; and normal variation due to individual difference is pointed out to contribute to these findings. The safety test of irradiated food has been conducted valid not only in animal experiments but also other tests like genotoxicity and analysis of radiation-degraded products. (R.T.)

  17. Breaks in plasmid DNA strand induced by laser radiation at a wavelength of 193 nm

    International Nuclear Information System (INIS)

    Gurzadyan, G.G.; Shul'te Frolinde, D.

    1996-01-01

    DNA of plasmid pB322 irradiated with laser at a wavelength of 193 nm was treated with an extract containing proteins from E.coli K12 AB1157 (wild-type). The enzymes were found to produce single- and double-strand DNA breaks, which was interpreted as a transformation of a portion of cyclobutane pyrimidine dimers and (6-4) photoproducts into nonrepairable single-strand DNA breaks. The products resulted from ionization of DNA, in particular, single-strand breaks, transform to double-strand breaks. A comparison of these data with the data on survival of plasmid upon transformation of E.coli K12 AB1157 enables one to assess the biological significance of single- and double-strand breaks. The inactivation of the plasmid is mainly determined by the number of directly formed laser-induced single-strand breaks. 26 refs.; 2 figs

  18. Plasmid-Mediated Antibiotic Resistance and Virulence in Gram-negatives: the Klebsiella pneumoniae Paradigm.

    Science.gov (United States)

    Ramirez, Maria S; Traglia, German M; Lin, David L; Tran, Tung; Tolmasky, Marcelo E

    Plasmids harbor genes coding for specific functions including virulence factors and antibiotic resistance that permit bacteria to survive the hostile environment found in the host and resist treatment. Together with other genetic elements such as integrons and transposons, and using a variety of mechanisms, plasmids participate in the dissemination of these traits resulting in the virtual elimination of barriers among different kinds of bacteria. In this article we review the current information about physiology and role in virulence and antibiotic resistance of plasmids from the gram-negative opportunistic pathogen Klebsiella pneumoniae . This bacterium has acquired multidrug resistance and is the causative agent of serious communityand hospital-acquired infections. It is also included in the recently defined ESKAPE group of bacteria that cause most of US hospital infections.

  19. Characteristics of plasmids in multi-drug-resistant Enterobacteriaceae isolated during prospective surveillance of a newly opened hospital in Iraq.

    Directory of Open Access Journals (Sweden)

    Xiao-Zhe Huang

    Full Text Available BACKGROUND: Gram-negative multidrug-resistant (MDR bacteria are major causes of nosocomial infections, and antibiotic resistance in these organisms is often plasmid mediated. Data are scarce pertaining to molecular mechanisms of antibiotic resistance in resource constrained areas such as Iraq. METHODOLOGY/PRINCIPAL FINDINGS: In this study, all MDR Enterobacteriaceae (n = 38 and randomly selected non-MDR counterparts (n = 41 isolated from patients, healthcare workers and environmental surfaces in a newly opened hospital in Iraq were investigated to characterize plasmids found in these isolates and determine their contribution to antibiotic resistance. Our results demonstrated that MDR E. coli and K. pneumoniae isolates harbored significantly more (≥ 3 plasmids compared to their non-MDR counterparts, which carried ≤ 2 plasmids (p<0.01. Various large plasmids (~52 to 100 kb from representative isolates were confirmed to contain multiple resistance genes by DNA microarray analysis. Aminoglycoside (acc, aadA, aph, strA/B, and ksgA, β-lactam (bla(TEM1, bla(AMPC, bla(CTX-M-15, bla(OXA-1, bla(VIM-2 and bla(SHV, sulfamethoxazole/trimethoprim (sul/dfr, tetracycline (tet and chloramphenicol (cat resistance genes were detected on these plasmids. Additionally, multiple plasmids carrying multiple antibiotic resistance genes were found in the same host strain. Genetic transfer-associated genes were identified on the plasmids from both MDR and non-MDR isolates. Seven plasmid replicon types (FII, FIA, FIB, B/O, K, I1 and N were detected in the isolates, while globally disseminated IncA/C and IncHI1 plasmids were not detected in these isolates. CONCLUSIONS/SIGNIFICANCE: This is the first report of the characteristics of the plasmids found in Enterobacteriaceae isolated following the opening of a new hospital in Iraq. The information provided here furthers our understanding of the mechanisms of drug resistance in this specific region and their evolutionary

  20. Transfection of embryonated Muscovy duck eggs with a recombinant plasmid is suitable for rescue of infectious Muscovy duck parvovirus.

    Science.gov (United States)

    Wang, Jianye; Huang, Yu; Ling, Jueyi; Wang, Zhixiang; Zhu, Guoqiang

    2017-12-01

    For members of the family Parvoviridae, rescue of infectious virus from recombinant plasmid is usually done in cultured cells. In this study, the whole genome of the pathogenic Muscovy duck parvovirus (MDPV) strain YY was cloned into the pBluescript II (SK) vector, generating recombinant plasmid pYY. With the aid of a transfection reagent, pYY plasmid was inoculated into 11-day-old embryonated Muscovy duck eggs via the chorioallantoic membrane route, resulting in the successful rescue of infectious virus and death of the embryos. The rescued virus exhibited pathogenicity in Muscovy ducklings similar to that of its parental strain, as evaluated based on the mortality rate. The results demonstrate that plasmid transfection in embryonated Muscovy duck eggs is a convenient and efficacious method for rescue of infectious MDPV in comparison to transfection of primary cells, which is somewhat time-consuming and laborious.

  1. A comparison of two methods for prediction of response and rates of inbreeding in selected populations with the results obtained in two selection experiments

    Directory of Open Access Journals (Sweden)

    Verrier Etienne

    2005-05-01

    Full Text Available Abstract Selection programmes are mainly concerned with increasing genetic gain. However, short-term progress should not be obtained at the expense of the within-population genetic variability. Different prediction models for the evolution within a small population of the genetic mean of a selected trait, its genetic variance and its inbreeding have been developed but have mainly been validated through Monte Carlo simulation studies. The purpose of this study was to compare theoretical predictions to experimental results. Two deterministic methods were considered, both grounded on a polygenic additive model. Differences between theoretical predictions and experimental results arise from differences between the true and the assumed genetic model, and from mathematical simplifications applied in the prediction methods. Two sets of experimental lines of chickens were used in this study: the Dutch lines undergoing true truncation mass selection, the other lines (French undergoing mass selection with a restriction on the representation of the different families. This study confirmed, on an experimental basis, that modelling is an efficient approach to make useful predictions of the evolution of selected populations although the basic assumptions considered in the models (polygenic additive model, normality of the distribution, base population at the equilibrium, etc. are not met in reality. The two deterministic methods compared yielded results that were close to those observed in real data, especially when the selection scheme followed the rules of strict mass selection: for instance, both predictions overestimated the genetic gain in the French experiment, whereas both predictions were close to the observed values in the Dutch experiment.

  2. Obtaining land cover changes information from multitemporal analysis of Landsat-TM images: results from a case study in West African dryland

    Science.gov (United States)

    Nutini, F.; Boschetti, M.; Brivio, P. A.; Antoninetti, M.

    2012-04-01

    The Sahelian belt of West Africa is a semiarid region characterized by wide climate variations, which can in turn affect the livelihood of local populations particularly in rangeland areas, as happens during the dramatic food crisis in the 70-80s caused by rainfall scarcity. The monitoring of natural resources and rainfed agricultural activities, with the aim to provide information to support Sahelian food security action, needs the production of detailed thematic maps as emphasized by several scientific papers. In this framework, a study was conducted to develop a method to exploit time series of remote sensed satellite data to 1) provide reliable land cover (LC) map at local scale in a dry region and 2) obtain a LC change (LCC) map that contribute to identify the plausible causes of local environmental instability. Satellite images used for this work consist in a time series of Landsat Thematic Mapper (TM) (path row 195-50) acquired in the 2000 (6 scenes) and 2007 (9 scenes) from February (Dry season) to September (end of wet season). The study investigates the different contribution provided by spectra information of a single Landsat TM image and by time series of derived NDVI. Different tests have been conducted with different combination of data set (spectral and temporal)in order to identify the best approach to obtain a LC map in five classes of interest: Shrubland, Cultivated Land, Water body, Herbaceous vegetation and Bare soil. The best classification approach is exposed and applied on two years in the last decade. The comparison between this two LC results in land cover change map, that displays the changes of vegetation patterns that have been characterized the area. The discussed results show a largely stable dryland region, but locally characterized by hot-spot of decreasing in natural vegetation inside the rangelands and an increasing of cultivations along fossil valleys where human activities are slightly intense. The discussion shows that this hot

  3. Comparison of the identification results of Candida species obtained by BD Phoenix™ and Maldi-TOF (Bruker Microflex LT Biotyper 3.1).

    Science.gov (United States)

    Marucco, Andrea P; Minervini, Patricia; Snitman, Gabriela V; Sorge, Adriana; Guelfand, Liliana I; Moral, Laura López

    2018-02-05

    In patients with invasive fungal infections, the accurate and rapid identification of the genus Candida is of utmost importance since antimycotic sensitivity is closely related to the species. The aim of the present study was to compare the identification results of species of the genus Candida obtained by BD Phoenix™ (Becton Dickinson [BD]) and Maldi-TOF MS (Bruker Microflex LT Biotyper 3.1). A total of 192 isolates from the strain collection belonging to the Mycology Network of the Autonomous City of Buenos Aires, Argentina, were analyzed. The observed concordance was 95%. Only 10 strains (5%) were not correctly identified by the BD Phoenix™ system. The average identification time with the Yeast ID panels was 8h 22min. The BD Phoenix™ system proved to be a simple, reliable and effective method for identifying the main species of the genus Candida. Copyright © 2017 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  4. KRAS mutation testing in colorectal cancer: comparison of the results obtained using 3 different methods for the analysis of codons G12 and G13.

    Science.gov (United States)

    Bihl, Michel P; Hoeller, Sylvia; Andreozzi, Maria Carla; Foerster, Anja; Rufle, Alexander; Tornillo, Luigi; Terracciano, Luigi

    2012-03-01

    Targeting the epidermal growth factor receptor (EGFR) is a new therapeutic option for patients with metastatic colorectal or lung carcinoma. However, the therapy efficiency highly depends on the KRAS mutation status in the given tumour. Therefore a reliable and secure KRAS mutation testing is crucial. Here we investigated 100 colorectal carcinoma samples with known KRAS mutation status (62 mutated cases and 38 wild type cases) in a comparative manner with three different KRAS mutation testing techniques (Pyrosequencing, Dideoxysequencing and INFINITI) in order to test their reliability and sensitivity. For the large majority of samples (96/100, 96%), the KRAS mutation status obtained by all three methods was the same. Only two cases with clear discrepancies were observed. One case was reported as wild type by the INFINITI method while the two other methods detected a G13C mutation. In the second case the mutation could be detected by the Pyrosequencing and INFINITI method (15% and 15%), while no signal for mutation could be observed with the Dideoxysequencing method. Additional two unclear results were due to a detection of a G12V with the INFINITI method, which was below cut-off when repeated and which was not detectable by the other two methods and very weak signals in a G12V mutated case with the Dideoxy- and Pyroseqencing method compared to the INFINITI method, respectively. In summary all three methods are reliable and robust methods in detecting KRAS mutations. INFINITI, however seems to be slightly more sensitive compared to Dideoxy- and Pyrosequencing.

  5. Influence of the microstructure on the resulting 18R martensitic transformation of polycrystalline Cu−Al−Zn thin films obtained by sputtering and reactive annealing

    International Nuclear Information System (INIS)

    Domenichini, P.; Condó, A.M.; Soldera, F.; Sirena, M.; Haberkorn, N.

    2016-01-01

    We report the influence of the microstructure on the martensitic transformation in polycrystalline Cu−Zn−Al thin films with 18R structure. The films are grown in two steps. First, Cu−Al thin films are obtained by DC sputtering. Second, the Zn is introduced in the Cu−Al thin films by the annealing them together with a bulk Cu−Zn−Al reference. The crystalline structure of the films was analyzed by X-ray diffraction and transmission electron microscopy. The martensitic transformation temperature was measured by electrical transport using conventional four probe geometry. It was observed that temperatures above 973 K are necessary for zincification of the samples to occur. The resulting martensitic transformation and its hysteresis (barrier for the transformation) depend on the grain size, topology and films thickness. - Highlights: • Polycrystalline Cu−Al−Zn thin films with nanometric grain size are sintered. • Influence of thermal annealing process on the microstructure is analyzed. • Martensitic transformation of Cu−Al−Zn thin films is strongly affected by the microstructure.

  6. Influence of the microstructure on the resulting 18R martensitic transformation of polycrystalline Cu−Al−Zn thin films obtained by sputtering and reactive annealing

    Energy Technology Data Exchange (ETDEWEB)

    Domenichini, P. [Instituto Balseiro, Bustillo 9500, S. C. de Bariloche (Argentina); Condó, A.M. [Instituto Balseiro, Bustillo 9500, S. C. de Bariloche (Argentina); Centro Atómico Bariloche, Comisión Nacional de Energía Atómica, Av. Bustillo 9500, 8400 San Carlos de Bariloche (Argentina); Soldera, F. [Department of Materials Science & Engineering, Saarland University, D-66123 Saarbruecken (Germany); Sirena, M. [Instituto Balseiro, Bustillo 9500, S. C. de Bariloche (Argentina); Centro Atómico Bariloche, Comisión Nacional de Energía Atómica, Av. Bustillo 9500, 8400 San Carlos de Bariloche (Argentina); Haberkorn, N., E-mail: nhaberk@cab.cnea.gov.ar [Instituto Balseiro, Bustillo 9500, S. C. de Bariloche (Argentina); Centro Atómico Bariloche, Comisión Nacional de Energía Atómica, Av. Bustillo 9500, 8400 San Carlos de Bariloche (Argentina)

    2016-04-15

    We report the influence of the microstructure on the martensitic transformation in polycrystalline Cu−Zn−Al thin films with 18R structure. The films are grown in two steps. First, Cu−Al thin films are obtained by DC sputtering. Second, the Zn is introduced in the Cu−Al thin films by the annealing them together with a bulk Cu−Zn−Al reference. The crystalline structure of the films was analyzed by X-ray diffraction and transmission electron microscopy. The martensitic transformation temperature was measured by electrical transport using conventional four probe geometry. It was observed that temperatures above 973 K are necessary for zincification of the samples to occur. The resulting martensitic transformation and its hysteresis (barrier for the transformation) depend on the grain size, topology and films thickness. - Highlights: • Polycrystalline Cu−Al−Zn thin films with nanometric grain size are sintered. • Influence of thermal annealing process on the microstructure is analyzed. • Martensitic transformation of Cu−Al−Zn thin films is strongly affected by the microstructure.

  7. The Escompte - Marseille 2001 International Field Experiment: Ground Based and Lidar Results Obtained At St. Chamas By The Epfl Mobile Laboratory

    Science.gov (United States)

    Balin, I.; Jimenez, R.; Simeonov, V.; Ristori, P.; Navarette, M.; van den Bergh, H.; Calpini, B.

    The assessment of the air pollution problems in term of understanding of the non- linear chemical mechanisms, the transport or the meteorological processes, and the choice of the abatement strategies could be based on the air pollution models. Nowa- days, very few of these models were validated due to the lack of 3D measurements. The goal of the ESCOMPTE experiment was to provide such of 3D database in order to constrain the air pollution models. The EPFL-LPA mobile laboratory was part of the ESCOMPTE extensive network and was located on the northern side of the Berre Lake at St.Chamas. In this framework, measurements of the air pollutants (O3, SO2, NOx, polycyclic aromatic hydrocarbons, black carbon and particulate matter of less than 10 microns mean diameter) and meteorological parameters (wind, temperature, pressure and relative humidity) were continuously performed from June 10 to July 13, 2001. They were combined with ground based lidar observations for ozone and aerosol estimation from 100m above ground level up to the free troposphere at ca.7 km agl. This paper will present an overview of the results obtained and will highlight one of the intensive observation period (IOP) during which clean air conditions were initially observed followed by highly polluted air masses during the second half of the IOP.

  8. Broad host range plasmids can invade an unexpectedly diverse fraction of a soil bacterial community

    DEFF Research Database (Denmark)

    Klümper, Uli; Riber, Leise; Dechesne, Arnaud

    2014-01-01

    and Actinobacteria suggests that inter-Gram plasmid transfer of IncP-1 and IncPromA-type plasmids is a frequent phenomenon. While the plasmid receiving fractions of the community were both plasmid- and donor- dependent, we identified a core super-permissive fraction that could take up different plasmids from diverse...

  9. Initiation and termination of the bacteriophage phi X174 rolling circle DNA replication in vivo: packaging of plasmid single-stranded DNA into bacteriophage phi X174 coats

    NARCIS (Netherlands)

    van der Ende, A.; Teertstra, R.; Weisbeek, P. J.

    1982-01-01

    The bacteriophage phi X174 viral (+) origin when inserted in a plasmid can interact in vivo with the A protein produced by infecting phi X174 phages. A consequence of this interaction is packaging of single-stranded plasmid DNA into preformed phage coats resulting in infective particles (1). This

  10. Characterization and plasmid elimination of NDM-1-producing Acinetobacter calcoaceticus from China.

    Directory of Open Access Journals (Sweden)

    Yang Sun

    Full Text Available The presence of multidrug-resistant bacterial pathogens in the environment poses a serious threat to public health. The opportunistic Acinetobacter spp. are among the most prevalent causes of nosocomial infections. Here, we performed complete genome sequencing of the Acinetobacter calcoaceticus strain XM1570, which was originally cultivated from the sputum of a patient diagnosed with pneumonia in Xiamen in 2010. We identified carbapenem resistance associated gene bla(NDM-1 located on a 47.3-kb plasmid. Three methods--natural reproduction, sodium dodecyl sulfate treatment and nalidixic acid treatment--were used to eliminate the bla(NDM-1-encoding plasmid, which achieved elimination rates of 3.32% (10/301, 83.78% (278/332, and 84.17% (298/354, respectively. Plasmid elimination dramatically increased antibiotic sensitivity, reducing the minimum bacteriostatic concentration of meropenem from 256 µg/ml in the clinical strain to 0.125 µg/ml in the plasmid-eliminated strain. Conjugation transfer assays showed that the bla(NDM-1-containing plasmid could be transferred into Escherichia coli DH5α:pBR322 in vitro as well as in vivo in mice. The bla(NDM-1 genetic environment was in accordance with that of other bla(NDM-1 genes identified from India, Japan, and Hong-Kong. The multilocus sequence type of the isolate was identified as ST-70. Two novel genes encoding intrinsic OXA and ADC were identified and named as OXA-417 and ADC-72. The finding of bla(NDM-1 in species like A. calcoaceticus demonstrates the wide spread of this gene in gram-negative bacteria which is possible by conjugative plasmid transfer. The results of this study may help in the development of a treatment strategy for controlling NDM-1 bacterial infection and transmission.

  11. Diversity of Clostridium perfringens isolates from various sources and prevalence of conjugative plasmids.

    Science.gov (United States)

    Park, Miseon; Deck, Joanna; Foley, Steven L; Nayak, Rajesh; Songer, J Glenn; Seibel, Janice R; Khan, Saeed A; Rooney, Alejandro P; Hecht, David W; Rafii, Fatemeh

    2016-04-01

    Clostridium perfringens is an important pathogen, causing food poisoning and other mild to severe infections in humans and animals. Some strains of C. perfringens contain conjugative plasmids, which may carry antimicrobial resistance and toxin genes. We studied genomic and plasmid diversity of 145 C. perfringens type A strains isolated from soils, foods, chickens, clinical samples, and domestic animals (porcine, bovine and canine), from different geographic areas in the United States between 1994 and 2006, using multiple-locus variable-number tandem repeat analysis (MLVA) and/or pulsed-field gel electrophoresis (PFGE). MLVA detected the genetic diversity in a majority of the isolates. PFGE, using SmaI and KspI, confirmed the MLVA results but also detected differences among the strains that could not be differentiated by MLVA. All of the PFGE profiles of the strains were different, except for a few of the epidemiologically related strains, which were identical. The PFGE profiles of strains isolated from the same domestic animal species were clustered more closely with each other than with other strains. However, a variety of C. perfringens strains with distinct genetic backgrounds were found among the clinical isolates. Variation was also observed in the size and number of plasmids in the strains. Primers for the internal fragment of a conjugative tcpH gene of C. perfringens plasmid pCPF4969 amplified identical size fragments from a majority of strains tested; and this gene hybridized to the various-sized plasmids of these strains. The sequences of the PCR-amplified tcpH genes from 12 strains showed diversity among the tcpH genes. Regardless of the sources of the isolates, the genetic diversity of C. perfringens extended to the plasmids carrying conjugative genes. Published by Elsevier Ltd.

  12. Relationship between signal intensity of blood flow in the pulmonary artery obtained by magnetic resonance imaging and results of right cardiac catheterization in patients with pulmonary disease

    Energy Technology Data Exchange (ETDEWEB)

    Yuguchi, Yasutoshi; Nagao, Keiichi; Kouno, Norihiro; Tanabe, Nobuhiro; Okita, Shinya; Tojima, Hirokazu; Okada, Osamu; Kuriyama, Takayuki [Chiba Univ. (Japan). School of Medicine; Yamaguchi, Tetsuo

    1992-08-01

    Electrocardiogram-gated spin-echo magnetic resonance (MR) images of the chest were obtained in five normal controls and 35 patients with pulmonary disease (11 chronic obstructive pulmonary disease, 6 pulmonary thromboembolism, 5 primary pulmonary hypertension, 4 interstitial pulmonary disease, 4 pulmonary hypertension with disturbance of portal circulation, and 5 other diseases) who underwent right cardiac catheterization. In transverse images at the level of the right main pulmonary artery (rPA) and sagittal images at the level through the midsternal line and the spinal cord, the signal intensity of blood flow in the rPA was quantitatively evaluated, and the correlations with the MR signal intensity of intravascular flow and the parameters of hemodynamics were studied. In diastole MR images of both normal controls and patients mostly showed a significant signal and visible flow images. In systolic MR images, the mean values of hemodynamic parameters (mean pulmonary arterial pressure (mPAP), pulmonary arteriolar resistance (PAR), and cardiac index (CI)) were abnormal in patients with significant signal intensity of flow compared with those in patients without sufficient MR signal. The signal intensity was not correlated with mPAP; however, it significantly increased as PAR increased, and it increased as CI decreased both in diastole and in systole. Especially in systole, there was good correlation between the signal intensity in transverse MR images and CI and between signal intensity in sagittal MR images and PAR . These results suggest that the signal intensity of blood flow in the rPA on MR images can be used as an index of the severity of right heart failure associated with pulmonary disease. MR imaging is a useful modality to evaluate pulmonary circulation disturbance because of its ability to assess blood flow in the pulmonary artery noninvasively without interference from other structures such as bone and normal lung. (J.P.N.).

  13. Using some results about the Lie evolution of differential operators to obtain the Fokker-Planck equation for non-Hamiltonian dynamical systems of interest

    Science.gov (United States)

    Bianucci, Marco

    2018-05-01

    Finding the generalized Fokker-Planck Equation (FPE) for the reduced probability density function of a subpart of a given complex system is a classical issue of statistical mechanics. Zwanzig projection perturbation approach to this issue leads to the trouble of resumming a series of commutators of differential operators that we show to correspond to solving the Lie evolution of first order differential operators along the unperturbed Liouvillian of the dynamical system of interest. In this paper, we develop in a systematic way the procedure to formally solve this problem. In particular, here we show which the basic assumptions are, concerning the dynamical system of interest, necessary for the Lie evolution to be a group on the space of first order differential operators, and we obtain the coefficients of the so-evolved operators. It is thus demonstrated that if the Liouvillian of the system of interest is not a first order differential operator, in general, the FPE structure breaks down and the master equation contains all the power of the partial derivatives, up to infinity. Therefore, this work shed some light on the trouble of the ubiquitous emergence of both thermodynamics from microscopic systems and regular regression laws at macroscopic scales. However these results are very general and can be applied also in other contexts that are non-Hamiltonian as, for example, geophysical fluid dynamics, where important events, like El Niño, can be considered as large time scale phenomena emerging from the observation of few ocean degrees of freedom of a more complex system, including the interaction with the atmosphere.

  14. Plasmid Conjugation in E. coli and Drug Resistance | Igwe ...

    African Journals Online (AJOL)

    This study aimed at determining the antibiotics susceptibility pattern of E. coli isolates claimed to be multidrug resistance using disc diffusion method. It also determined the presence of transferable resistance plasmids through conjugation and evaluated the medical significance of plasmid encoding E. coli and drug ...

  15. Resistant plasmid profile analysis of multidrug resistant Escherichia ...

    African Journals Online (AJOL)

    Multiple drug resistance isolates causing UTI has seri- ous implications for the empiric therapy against patho- genic isolates and for the possible co-selection of antimicrobial resistant mediated by multi drug resistant plasmids21,22. E. coli from clinical isolates are known to harbour plasmids of different molecular sizes23.

  16. Plasmid-Mediated Antimicrobial Resistance in Staphylococci and Other Firmicutes.

    Science.gov (United States)

    Schwarz, Stefan; Shen, Jianzhong; Wendlandt, Sarah; Fessler, Andrea T; Wang, Yang; Kadlec, Kristina; Wu, Cong-Ming

    2014-12-01

    In staphylococci and other Firmicutes, resistance to numerous classes of antimicrobial agents, which are commonly used in human and veterinary medicine, is mediated by genes that are associated with mobile genetic elements. The gene products of some of these antimicrobial resistance genes confer resistance to only specific members of a certain class of antimicrobial agents, whereas others confer resistance to the entire class or even to members of different classes of antimicrobial agents. The resistance mechanisms specified by the resistance genes fall into any of three major categories: active efflux, enzymatic inactivation, and modification/replacement/protection of the target sites of the antimicrobial agents. Among the mobile genetic elements that carry such resistance genes, plasmids play an important role as carriers of primarily plasmid-borne resistance genes, but also as vectors for nonconjugative and conjugative transposons that harbor resistance genes. Plasmids can be exchanged by horizontal gene transfer between members of the same species but also between bacteria belonging to different species and genera. Plasmids are highly flexible elements, and various mechanisms exist by which plasmids can recombine, form cointegrates, or become integrated in part or in toto into the chromosomal DNA or into other plasmids. As such, plasmids play a key role in the dissemination of antimicrobial resistance genes within the gene pool to which staphylococci and other Firmicutes have access. This chapter is intended to provide an overview of the current knowledge of plasmid-mediated antimicrobial resistance in staphylococci and other Firmicutes.

  17. Functional analysis of three plasmids from Lactobacillus plantarum

    NARCIS (Netherlands)

    Kranenburg, R. van; Golic, N.; Bongers, R.; Leer, R.J.; Vos, W.M. de; Siezen, R.J.; Kleerebezem, M.

    2005-01-01

    Lactobacillus plantarum WCFS1 harbors three plasmids, pWCFS101, pWCFS102, and pWCFS103, with sizes of 1,917, 2,365, and 36,069 bp, respectively. The two smaller plasmids are of unknown function and contain replication genes that are likely to function via the rolling-circle replication mechanism.

  18. Production and pharmaceutical formulation of plasmid DNA vaccines

    NARCIS (Netherlands)

    van der Heijden, I.

    2013-01-01

    Research leading to the thesis ‘Production and pharmaceutical formulation of plasmid DNA vaccines‘ can be divided into two parts. The first part describes the development of a Good Manufacturing Practice (GMP) compliant plasmid DNA production process of pDNA vaccines for the treatment of Human

  19. Transfer of conjugative plasmids among bacteria under environmentally relevant conditions

    DEFF Research Database (Denmark)

    Musovic, Sanin

    Mobile genetiske elementer (f.eks. plasmider), der ofte bærer ekstra funktioner såsom antibiotikaresistens, eller kataboliske- og xenobiotiske nedbrydnings gener, antages at have en meget vigtigt evolutionær rolle for bakterier. I denne PhD afhandling undersøgte jeg størrelsen af plasmid overførs...

  20. Two novel conjugative plasmids from a single strain of Sulfolobus

    NARCIS (Netherlands)

    Erauso, G.; Stedman, K.M.; Werken, van de H.J.G.; Zillig, W.; Oost, van der J.

    2006-01-01

    Two conjugative plasmids (CPs) were isolated and characterized from the same 'Sulfolobus islandicus' strain, SOG2/4, The plasmids were separated from each other and transferred into Sulfolobus soltataricus. One has a high copy number and is not stable (pSOG1) whereas the other has a low copy number

  1. The technology of large-scale pharmaceutical plasmid purification ...

    African Journals Online (AJOL)

    Further test demonstrated that the pcDNAlacZ purified with CTAB and authoritative endotoxin-free plasmid Kit had the similar transfection efficiency in vivo and in vitro. CTAB can be used for plasmid purification; the main advantages of the DNAs purified with CTAB include the avoidance of animal-derived enzymes, toxic ...

  2. Radiation-chemical discussion on inverse dose-rate effect observed in radiation-induced strand breaks of plasmid DNA

    International Nuclear Information System (INIS)

    Masuda, Takahiro

    1994-01-01

    Experimental results of inverse dose-rate effect, so-called Kada Effects, which was published by Takakura and her coworkers on radiation-induced strand breaks of plasmid DNA in aerated aqueous solution, have been kinetically analyzed and discussed on the basis of radiation chemistry. the kinetic analysis indicates that there are two possible mechanisms; 1) equilibrium mixture of O 2 - and HO 2 is responsible for strand breaks of DNA, and 2) peroxyl radical produced from citrate is effective for the strand breaks. However, the detailed kinetic analysis revealed that the latter is improbable because unimolecular decay of the peroxyl radical must be assumed to be negligible for its participation despite fast decay of analogous organic peroxyl radicals. The analysis has also given 9.93±0.10 dm 3 mol -1 s -1 per nucleotide unit, which corresponds to 7.62 x 10 4 dm 3 mol -1 s -1 per DNA molecule, as the rate constant for the reaction of the equilibrium mixture with plasmid pBR 322 DNA. Furthermore the probability that the reaction of the mixture with a nucleotide unit of DNA leads to strand breaks was obtained to be 3.36 x 10 -3 for gamma-irradiated system and 1.98 x 10 -3 for beta-irradiated system, respectively. (author)

  3. Identification of IncA/C Plasmid Replication and Maintenance Genes and Development of a Plasmid Multilocus Sequence Typing Scheme.

    Science.gov (United States)

    Hancock, Steven J; Phan, Minh-Duy; Peters, Kate M; Forde, Brian M; Chong, Teik Min; Yin, Wai-Fong; Chan, Kok-Gan; Paterson, David L; Walsh, Timothy R; Beatson, Scott A; Schembri, Mark A

    2017-02-01

    Plasmids of incompatibility group A/C (IncA/C) are becoming increasingly prevalent within pathogenic Enterobacteriaceae They are associated with the dissemination of multiple clinically relevant resistance genes, including bla CMY and bla NDM Current typing methods for IncA/C plasmids offer limited resolution. In this study, we present the complete sequence of a bla NDM-1 -positive IncA/C plasmid, pMS6198A, isolated from a multidrug-resistant uropathogenic Escherichia coli strain. Hypersaturated transposon mutagenesis, coupled with transposon-directed insertion site sequencing (TraDIS), was employed to identify conserved genetic elements required for replication and maintenance of pMS6198A. Our analysis of TraDIS data identified roles for the replicon, including repA, a toxin-antitoxin system; two putative partitioning genes, parAB; and a putative gene, 053 Construction of mini-IncA/C plasmids and examination of their stability within E. coli confirmed that the region encompassing 053 contributes to the stable maintenance of IncA/C plasmids. Subsequently, the four major maintenance genes (repA, parAB, and 053) were used to construct a new plasmid multilocus sequence typing (PMLST) scheme for IncA/C plasmids. Application of this scheme to a database of 82 IncA/C plasmids identified 11 unique sequence types (STs), with two dominant STs. The majority of bla NDM -positive plasmids examined (15/17; 88%) fall into ST1, suggesting acquisition and subsequent expansion of this bla NDM -containing plasmid lineage. The IncA/C PMLST scheme represents a standardized tool to identify, track, and analyze the dissemination of important IncA/C plasmid lineages, particularly in the context of epidemiological studies. Copyright © 2017 American Society for Microbiology.

  4. Expansion of the IncX plasmid family for improved identification and typing of novel plasmids in drug-resistant Enterobacteriaceae

    DEFF Research Database (Denmark)

    Johnson, Timothy J.; Bielak, Eliza Maria; Fortini, Daniela

    2012-01-01

    and biofilm formation. Previous plasmid-based replicon typing procedures have indicated that the prevalence of IncX plasmids is low among members of the Enterobacteriaceae. However, examination of a number of IncX-like plasmid sequences and their occurrence in various organisms suggests that IncX plasmid...

  5. Plasmid Complement of Lactococcus lactis NCDO712 Reveals a Novel Pilus Gene Cluster.

    Science.gov (United States)

    Tarazanova, Mariya; Beerthuyzen, Marke; Siezen, Roland; Fernandez-Gutierrez, Marcela M; de Jong, Anne; van der Meulen, Sjoerd; Kok, Jan; Bachmann, Herwig

    2016-01-01

    Lactococcus lactis MG1363 is an important gram-positive model organism. It is a plasmid-free and phage-cured derivative of strain NCDO712. Plasmid-cured strains facilitate studies on molecular biological aspects, but many properties which make L. lactis an important organism in the dairy industry are plasmid encoded. We sequenced the total DNA of strain NCDO712 and, contrary to earlier reports, revealed that the strain carries 6 rather than 5 plasmids. A new 50-kb plasmid, designated pNZ712, encodes functional nisin immunity (nisCIP) and copper resistance (lcoRSABC). The copper resistance could be used as a marker for the conjugation of pNZ712 to L. lactis MG1614. A genome comparison with the plasmid cured daughter strain MG1363 showed that the number of single nucleotide polymorphisms that accumulated in the laboratory since the strains diverted more than 30 years ago is limited to 11 of which only 5 lead to amino acid changes. The 16-kb plasmid pSH74 was found to contain a novel 8-kb pilus gene cluster spaCB-spaA-srtC1-srtC2, which is predicted to encode a pilin tip protein SpaC, a pilus basal subunit SpaB, and a pilus backbone protein SpaA. The sortases SrtC1/SrtC2 are most likely involved in pilus polymerization while the chromosomally encoded SrtA could act to anchor the pilus to peptidoglycan in the cell wall. Overexpression of the pilus gene cluster from a multi-copy plasmid in L. lactis MG1363 resulted in cell chaining, aggregation, rapid sedimentation and increased conjugation efficiency of the cells. Electron microscopy showed that the over-expression of the pilus gene cluster leads to appendices on the cell surfaces. A deletion of the gene encoding the putative basal protein spaB, by truncating spaCB, led to more pilus-like structures on the cell surface, but cell aggregation and cell chaining were no longer observed. This is consistent with the prediction that spaB is involved in the anchoring of the pili to the cell.

  6. Host range of enterococcal vanA plasmids among Gram-positive intestinal bacteria

    DEFF Research Database (Denmark)

    Werner, Guido; Freitas, Ana R.; Coque, Teresa M.

    2010-01-01

    recipients. Transfer rates were calculated per donor and recipient. Transconjugants were confirmed by determining their phenotypic and genotypic properties. Stability of plasmids in the new host was assessed in long-term growth experiments. RESULTS: In total, 282 enterococcal matings and 73 inter...

  7. Plasmid-mediated UV-protection in Streptococcus lactis

    Energy Technology Data Exchange (ETDEWEB)

    Chopin, M.C.; Rouault, A. (Institut National de la Recherche Agronomique, Rennes (France). Lab. de Recherches de Technologie Laitiere); Moillo-Batt, A. (Institut National de la Sante et de la Recherche Medicale (INSERM), Hopital de Pontchaillon, 35 - Rennes (France))

    1985-02-01

    Streptococcus lactis strain IL594 contains 9 plasmids, designated pIL1 to pIL9. On the basis of protoplast-induced curing experiments the authors showed that derivatives containing pIL7 were resistant to UV-irradiation while derivatives lacking pIL7 were sensitive. The pIL7-determined UV-protection was confirmed by co-transfer of the plasmid and of the character into a plasmid-free derivative of S. lactis IL594. Moreover, prophage induction required higher UV-fluence in this derivative carrying pIL7 than in the plasmid-free strain. This is the first report of a plasmid-mediated UV-protection in group N streptococci.

  8. Plasmid-mediated UV-protection in Streptococcus lactis

    International Nuclear Information System (INIS)

    Chopin, M.-C.; Rouault, A.

    1985-01-01

    Streptococcus lactis strain IL594 contains 9 plasmids, designated pIL1 to pIL9. On the basis of protoplast-induced curing experiments the authors showed that derivatives containing pIL7 were resistant to UV-irradiation while derivatives lacking pIL7 were sensitive. The pIL7-determined UV-protection was confirmed by cotransfer of the plasmid and of the character into a plasmid-free derivative of S. lactis IL594. Moreover, prophage induction required higher UV-fluence in this derivative carrying pIL7 than in the plasmid-free strain. This is the first report of a plasmid-mediated UV-protection in group N streptococci. (orig.)

  9. An audit of the contribution to post-mortem examination diagnosis of individual analyte results obtained from biochemical analysis of the vitreous.

    Science.gov (United States)

    Mitchell, Rebecca; Charlwood, Cheryl; Thomas, Sunethra Devika; Bellis, Maria; Langlois, Neil E I

    2013-12-01

    Biochemical analysis of the vitreous humor from the eye is an accepted accessory test for post-mortem investigation of cause of death. Modern biochemical analyzers allow testing of a range of analytes from a sample. However, it is not clear which analytes should be requested in order to prevent unnecessary testing (and expense). The means and standard deviation of the values obtained from analysis of the vitreous humor for sodium, potassium, chloride, osmolality, glucose, ketones (β-hydroxybutyrate), creatinine, urea, calcium, lactate, and ammonia were calculated from which the contribution of each analyte was reviewed in the context of post-mortem findings and final cause of death. For sodium 32 cases were regarded as high (more than one standard deviation above the mean), from which 9 contributed to post-mortem diagnosis [drowning (4), heat related death (2), diabetic hyperglycemia (2), and dehydration (1)], but 25 low values (greater than one standard deviation below the mean) made no contribution. For chloride 29 high values contributed to 4 cases--3 drowning and 1 heat-related, but these were all previously identified by a high sodium level. There were 29 high and 35 low potassium values, none of which contributed to determining the final cause of death. Of 22 high values of creatinine, 12 contributed to a diagnosis of renal failure. From 32 high values of urea, 18 contributed to 16 cases of renal failure (2 associated with diabetic hyperglycemia), 1 heat-related death, and one case with dehydration. Osmolarity contributed to 12 cases (5 heat-related, 4 diabetes, 2 renal failure, and 1 dehydration) from 36 high values. There was no contribution from 32 high values and 19 low values of calcium and there was no contribution from 4 high and 2 low values of ammonia. There were 11 high values of glucose, which contributed to the diagnosis of 6 cases of diabetic hyperglycemia and 21 high ketone levels contributed to 8 cases: 4 diabetic ketosis, 3 hypothermia, 3

  10. Conjugative plasmid pAW63 brings new insights into the genesis of the Bacillus anthracis virulence plasmid pXO2 and of the Bacillus thuringiensis plasmid pBT9727

    Directory of Open Access Journals (Sweden)

    Mahillon Jacques

    2005-07-01

    Full Text Available Abstract Background Bacillus cereus, Bacillus anthracis and Bacillus thuringiensis belong to the genetically close-knit Bacillus cereus sensu lato group, a family of rod-shaped Gram-positive bacteria. pAW63 is the first conjugative plasmid from the B. cereus group to be completely sequenced. Results The 71,777 bp nucleotide sequence of pAW63 reveals a modular structure, including a 42 kb tra region encoding homologs of the Type IV secretion systems components VirB11, VirB4 and VirD4, as well as homologs of Gram-positive conjugation genes from Enterococcus, Lactococcus, Listeria, Streptococcus and Staphylococcus species. It also firmly establishes the existence of a common backbone between pAW63, pXO2 from Bacillus anthracis and pBT9727 from the pathogenic Bacillus thuringiensis serovar konkukian strain 97-27. The alignment of these three plasmids highlights the presence of well conserved segments, in contrast to distinct regions of high sequence plasticity. The study of their specific differences has provided a three-point reference framework that can be exploited to formulate solid hypotheses concerning the functionalities and the molecular evolution of these three closely related plasmids. This has provided insight into the chronology of their divergence, and led to the discovery of two Type II introns on pAW63, matching copies of the mobile element IS231L in different loci of pXO2 and pBT9727, and the identification on pXO2 of a 37 kb pathogenicity island (PAI containing the anthrax capsule genes. Conclusion The complete sequence determination of pAW63 has led to a functional map of the plasmid yielding insights into its conjugative apparatus, which includes T4SS-like components, as well as its resemblance to other large plasmids of Gram-positive bacteria. Of particular interest is the extensive homology shared between pAW63 and pXO2, the second virulence plasmid of B. anthracis, as well as pBT9727 from the pathogenic strain B. thuringiensis

  11. Functional characterization of replication and stability factors of an incompatibility group P-1 plasmid from Xylella fastidiosa.

    Science.gov (United States)

    Lee, Min Woo; Rogers, Elizabeth E; Stenger, Drake C

    2010-12-01

    Xylella fastidiosa strain riv11 harbors a 25-kbp plasmid (pXF-RIV11) belonging to the IncP-1 incompatibility group. Replication and stability factors of pXF-RIV11 were identified and used to construct plasmids able to replicate in X. fastidiosa and Escherichia coli. Replication in X. fastidiosa required a 1.4-kbp region from pXF-RIV11 containing a replication initiation gene (trfA) and the adjacent origin of DNA replication (oriV). Constructs containing trfA and oriV from pVEIS01, a related IncP-1 plasmid of the earthworm symbiont Verminephrobacter eiseniae, also were competent for replication in X. fastidiosa. Constructs derived from pXF-RIV11 but not pVEIS01 replicated in Agrobacterium tumefaciens, Xanthomonas campestris, and Pseudomonas syringae. Although plasmids bearing replication elements from pXF-RIV11 or pVEIS01 could be maintained in X. fastidiosa under antibiotic selection, removal of selection resulted in plasmid extinction after 3 weekly passages. Addition of a toxin-antitoxin addiction system (pemI/pemK) from pXF-RIV11 improved plasmid stability such that >80 to 90% of X. fastidiosa cells retained plasmid after 5 weekly passages in the absence of antibiotic selection. Expression of PemK in E. coli was toxic for cell growth, but toxicity was nullified by coexpression of PemI antitoxin. Deletion of N-terminal sequences of PemK containing the conserved motif RGD abolished toxicity. In vitro assays revealed a direct interaction of PemI with PemK, suggesting that antitoxin activity of PemI is mediated by toxin sequestration. IncP-1 plasmid replication and stability factors were added to an E. coli cloning vector to constitute a stable 6.0-kbp shuttle vector (pXF20-PEMIK) suitable for use in X. fastidiosa.

  12. Identification of novel Clostridium perfringens type E strains that carry an iota toxin plasmid with a functional enterotoxin gene.

    Directory of Open Access Journals (Sweden)

    Kazuaki Miyamoto

    Full Text Available Clostridium perfringens enterotoxin (CPE is a major virulence factor for human gastrointestinal diseases, such as food poisoning and antibiotic associated diarrhea. The CPE-encoding gene (cpe can be chromosomal or plasmid-borne. Recent development of conventional PCR cpe-genotyping assays makes it possible to identify cpe location (chromosomal or plasmid in type A isolates. Initial studies for developing cpe genotyping assays indicated that all cpe-positive strains isolated from sickened patients were typable by cpe-genotypes, but surveys of C. perfringens environmental strains or strains from feces of healthy people suggested that this assay might not be useful for some cpe-carrying type A isolates. In the current study, a pulsed-field gel electrophoresis Southern blot assay showed that four cpe-genotype untypable isolates carried their cpe gene on a plasmid of ∼65 kb. Complete sequence analysis of the ∼65 kb variant cpe-carrying plasmid revealed no intact IS elements and a disrupted cytosine methyltransferase (dcm gene. More importantly, this plasmid contains a conjugative transfer region, a variant cpe gene and variant iota toxin genes. The toxin genes encoded by this plasmid are expressed based upon the results of RT-PCR assays. The ∼65 kb plasmid is closely related to the pCPF4969 cpe plasmid of type A isolates. MLST analyses indicated these isolates belong to a unique cluster of C. perfringens. Overall, these isolates carrying a variant functional cpe gene and iota toxin genes represent unique type E strains.

  13. Plasmid Negative Regulation of CPAF Expression Is Pgp4 Independent and Restricted to Invasive Chlamydia trachomatis Biovars

    Directory of Open Access Journals (Sweden)

    Michael John Patton

    2018-01-01

    Full Text Available Chlamydia trachomatis is an obligate intracellular bacterial pathogen that causes blinding trachoma and sexually transmitted disease. C. trachomatis isolates are classified into 2 biovars—lymphogranuloma venereum (LGV and trachoma—which are distinguished biologically by their natural host cell infection tropism. LGV biovars infect macrophages and are invasive, whereas trachoma biovars infect oculo-urogenital epithelial cells and are noninvasive. The C. trachomatis plasmid is an important virulence factor in the pathogenesis of these infections. Central to its pathogenic role is the transcriptional regulatory function of the plasmid protein Pgp4, which regulates the expression of plasmid and chromosomal virulence genes. As many gene regulatory functions are post-transcriptional, we employed a comparative proteomic study of cells infected with plasmid-cured C. trachomatis serovars A and D (trachoma biovar, a L2 serovar (LGV biovar, and the L2 serovar transformed with a plasmid containing a nonsense mutation in pgp4 to more completely elucidate the effects of the plasmid on chlamydial infection biology. Our results show that the Pgp4-dependent elevations in the levels of Pgp3 and a conserved core set of chromosomally encoded proteins are remarkably similar for serovars within both C. trachomatis biovars. Conversely, we found a plasmid-dependent, Pgp4-independent, negative regulation in the expression of the chlamydial protease-like activity factor (CPAF for the L2 serovar but not the A and D serovars. The molecular mechanism of plasmid-dependent negative regulation of CPAF expression in the LGV serovar is not understood but is likely important to understanding its macrophage infection tropism and invasive infection nature.

  14. Enterococcus faecalis Sex Pheromone cCF10 Enhances Conjugative Plasmid Transfer In Vivo.

    Science.gov (United States)

    Hirt, Helmut; Greenwood-Quaintance, Kerryl E; Karau, Melissa J; Till, Lisa M; Kashyap, Purna C; Patel, Robin; Dunny, Gary M

    2018-02-13

    Cell-cell communication mediated by peptide pheromones (cCF10 [CF]) is essential for high-frequency plasmid transfer in vitro in Enterococcus faecalis To examine the role of pheromone signaling in vivo , we established either a CF-producing (CF+) recipient or a recipient producing a biologically inactive variant of CF (CF- recipient) in a germfree mouse model 3 days before donor inoculation and determined transfer frequencies of the pheromone-inducible plasmid pCF10. Plasmid transfer was detected in the upper and middle sections of the intestinal tract 5 h after donor inoculation and was highly efficient in the absence of antibiotic selection. The transconjugant/donor ratio reached a maximum level approaching 1 on day 4 in the upper intestinal tract. Plasmid transfer was significantly lower with the CF- recipient. While rescue of the CF- mating defect by coculture with CF+ recipients is easily accomplished in vitro , no extracellular complementation occurred in vivo This suggests that most pheromone signaling in the gut occurs between recipient and donor cells in very close proximity. Plasmid-bearing cells (donors plus transconjugants) steadily increased in the population from 0.1% after donor inoculation to about 10% at the conclusion of the experiments. This suggests a selective advantage of pCF10 carriage distinct from antibiotic resistance or bacteriocin production. Our results demonstrate that pheromone signaling is required for efficient pCF10 transfer in vivo In the absence of CF+ recipients, a low level of transfer to CF- recipients occurred in the gut. This may result from low-level host-mediated induction of the donors in the gastrointestinal (GI) tract, similar to that previously observed in serum. IMPORTANCE Horizontal gene transfer is a major factor in the biology of Enterococcus faecalis , an important nosocomial pathogen. Previous studies showing efficient conjugative plasmid transfer in the gastrointestinal (GI) tracts of experimental animals did

  15. A comparison of two methods for prediction of response and rates of inbreeding in selected populations with the results obtained in two selection experiments

    NARCIS (Netherlands)

    Loywyck, V.; Bijma, P.; Pinard-van der Laan, M.H.; Arendonk, van J.A.M.; Verrier, E.

    2005-01-01

    Selection programmes are mainly concerned with increasing genetic gain. However, short-term progress should not be obtained at the expense of the within-population genetic variability. Different prediction models for the evolution within a small population of the genetic mean of a selected trait,

  16. Tissue-specific Calibration of Real-time PCR Facilitates Absolute Quantification of Plasmid DNA in Biodistribution Studies

    Directory of Open Access Journals (Sweden)

    Joan K Ho

    2016-01-01

    Full Text Available Analysis of the tissue distribution of plasmid DNA after administration of nonviral gene delivery systems is best accomplished using quantitative real-time polymerase chain reaction (qPCR, although published strategies do not allow determination of the absolute mass of plasmid delivered to different tissues. Generally, data is expressed as the mass of plasmid relative to the mass of genomic DNA (gDNA in the sample. This strategy is adequate for comparisons of efficiency of delivery to a single site but it does not allow direct comparison of delivery to multiple tissues, as the mass of gDNA extracted per unit mass of each tissue is different. We show here that by constructing qPCR standard curves for each tissue it is possible to determine the dose of intact plasmid remaining in each tissue, which is a more useful parameter when comparing the fates of different formulations of DNA. We exemplify the use of this tissue-specific qPCR method by comparing the delivery of naked DNA, cationic DNA complexes, and neutral PEGylated DNA complexes after intramuscular injection. Generally, larger masses of intact plasmid were present 24 hours after injection of DNA complexes, and neutral complexes resulted in delivery of a larger mass of intact plasmid to the spleen.

  17. The cost of copy number in a selfish genetic element: the 2-μm plasmid of Saccharomyces cerevisiae.

    Science.gov (United States)

    Harrison, Ellie; Koufopanou, V; Burt, A; MacLean, R C

    2012-11-01

    Many autonomously replicating genetic elements exist as multiple copies within the cell. The copy number of these elements is often assumed to have important fitness consequences for both element and host, yet the forces shaping its evolution are not well understood. The 2 μm is a multicopy plasmid of Saccharomyces yeasts, encoding just four genes that are solely involved in plasmid replication. One simple model for the fitness relationship between yeasts and 2 μm is that plasmid copy number evolves as a trade-off between selection for increased vertical transmission, favouring high copy number, and selection for decreased virulence, favouring low copy number. To test this model, we experimentally manipulated the copy number of the plasmid and directly measured the fitness cost, in terms of growth rate reduction, associated with high plasmid copy number. We find that the fitness burden imposed by the 2 μm increases with plasmid copy number, such that each copy imposes a fitness burden of 0.17% (± 0.008%), greatly exceeding the cost expected for it to be stably maintained in yeast populations. Our results demonstrate the crucial importance of copy number in the evolution of yeast per 2 μm associations and pave the way for future studies examining how selection can shape the cost of multicopy elements. © 2012 The Authors. Journal of Evolutionary Biology © 2012 European Society For Evolutionary Biology.

  18. Virus-sized self-assembling lamellar complexes between plasmid DNA and cationic micelles promote gene transfer

    Science.gov (United States)

    Pitard, Bruno; Aguerre, Olivier; Airiau, Marc; Lachagès, Anne-Marie; Boukhnikachvili, Tsiala; Byk, Gérardo; Dubertret, Catherine; Herviou, Christian; Scherman, Daniel; Mayaux, Jean-François; Crouzet, Joël

    1997-01-01

    Gene therapy is based on the vectorization of genes to target cells and their subsequent expression. Cationic amphiphile-mediated delivery of plasmid DNA is the nonviral gene transfer method most often used. We examined the supramolecular structure of lipopolyamine/plasmid DNA complexes under various condensing conditions. Plasmid DNA complexation with lipopolyamine micelles whose mean diameter was 5 nm revealed three domains, depending on the lipopolyamine/plasmid DNA ratio. These domains respectively corresponded to negatively, neutrally, and positively charged complexes. Transmission electron microscopy and x-ray scattering experiments on complexes originating from these three domains showed that although their morphology depends on the lipopolyamine/plasmid DNA ratio, their particle structure consists of ordered domains characterized by even spacing of 80 Å, irrespective of the lipid/DNA ratio. The most active lipopolyamine/DNA complexes for gene transfer were positively charged. They were characterized by fully condensed DNA inside spherical particles (diameter: 50 nm) sandwiched between lipid bilayers. These results show that supercoiled plasmid DNA is able to transform lipopolyamine micelles into a supramolecular organization characterized by ordered lamellar domains. PMID:9405626

  19. High Prevalence of Plasmid-Mediated Quinolone Resistance and IncQ Plasmids Carrying qnrS2 Gene in Bacteria from Rivers near Hospitals and Aquaculture in China.

    Directory of Open Access Journals (Sweden)

    Yanping Wen

    Full Text Available Effluents from hospital and aquaculture are considered important sources of quinolone resistance. However, little information is available on the impact of this effluent on nearby rivers. In this study, 188 ciprofloxacin-resistant bacterial isolates obtained from rivers near hospitals and aquaculture were screened for plasmid-mediated quinolone resistance (PMQR genes. Species identification, antibiotic susceptibility testing, and PMQR gene transferability assessment were conducted for PMQR-positive bacteria. Representative qnrS2-encoding plasmids were subsequently sequenced using a primer-walking approach. In total, 44 isolates (23.4% were positive for qnr genes (16 qnrB2, 3 qnrS1, and 25 qnrS2 and 32 isolates (17.0% were positive for aac(6'-Ib-cr. Other PMQR genes were not detected. The qnrB2 and aac(6'-Ib-cr genes had a higher prevalence in aquaculture samples than in hospital samples, and were significantly associated with Enterobacteriaceae (p < 0.05. In contrast, the prevalence of qnrS2 was not site-related, but was significantly associated with Aeromonas spp. (p < 0.05. All PMQR isolates were resistant to three or more classes of antibiotics. Eleven qnrS2-harboring plasmids from Aeromonas spp., including a novel conjugative plasmid pHP18, were selected for sequencing. These plasmids were small in size (6,388-16,197 bp and belonged to the IncQ or IncU plasmid family, with qnrS2 being part of a mobile insertion cassette. Taken together, our findings suggest that aquaculture is a possible source for aac(6'-Ib-cr and qnrB2 dissemination, and demonstrate the ubiquity of qnrS2 in aquatic environments. Finally, Aeromonas spp. served as vectors for qnrS2 with the help of IncQ-type plasmids.

  20. Interplay of a non-conjugative integrative element and a conjugative plasmid in the spread of antibiotic resistance via suicidal plasmid transfer from an aquaculture Vibrio isolate.

    Science.gov (United States)

    Nonaka, Lisa; Yamamoto, Tatsuya; Maruyama, Fumito; Hirose, Yuu; Onishi, Yuki; Kobayashi, Takeshi; Suzuki, Satoru; Nomura, Nobuhiko; Masuda, Michiaki; Yano, Hirokazu

    2018-01-01

    The capture of antimicrobial resistance genes (ARGs) by mobile genetic elements (MGEs) plays a critical role in resistance acquisition for human-associated bacteria. Although aquaculture environments are recognized as important reservoirs of ARGs, intra- and intercellular mobility of MGEs discovered in marine organisms is poorly characterized. Here, we show a new pattern of interspecies ARGs transfer involving a 'non-conjugative' integrative element. To identify active MGEs in a Vibrio ponticus isolate, we conducted whole-genome sequencing of a transconjugant obtained by mating between Escherichia coli and Vibrio ponticus. This revealed integration of a plasmid (designated pSEA1) into the chromosome, consisting of a self-transmissible plasmid backbone of the MOBH group, ARGs, and a 13.8-kb integrative element Tn6283. Molecular genetics analysis suggested a two-step gene transfer model. First, Tn6283 integrates into the recipient chromosome during suicidal plasmid transfer, followed by homologous recombination between the Tn6283 copy in the chromosome and that in the newly transferred pSEA1. Tn6283 is unusual among integrative elements in that it apparently does not encode transfer function and its excision barely generates unoccupied donor sites. Thus, its movement is analogous to the transposition of insertion sequences rather than to that of canonical integrative and conjugative elements. Overall, this study reveals the presence of a previously unrecognized type of MGE in a marine organism, highlighting diversity in the mode of interspecies gene transfer.

  1. A comparison between the Structural Results obtained by X-ray Single Crystal Data and by Neutron Powder Data for BaVs/sb3/

    International Nuclear Information System (INIS)

    Marezio, M.

    1986-01-01

    The structure of BaVs/sb3/, as refined from X-ray single-crystal data to an R factor of 0.011, is compared to the structure of the same compound obtained from neutron powder data (Rsb(ro) = 6.82, Rsb(psilon) = 4.09). As expected, the X-ray standard deviations of the positional and thermal parameters are smaller than the corresponding neutron standard deviations. However, the dynamical disorder deduced from the anomalously large thermal vibrations of the vanadium atoms obtained from the X-ray data is also evidenced by the neutron refinement. On the other hand, the neutron standard deviations of the lattice parameters are smaller than the X-ray counterparts

  2. Quantifying and resolving multiple vector transformants in S. cerevisiae plasmid libraries

    Directory of Open Access Journals (Sweden)

    Gray Elizabeth C

    2009-11-01

    Full Text Available Abstract Background In addition to providing the molecular machinery for transcription and translation, recombinant microbial expression hosts maintain the critical genotype-phenotype link that is essential for high throughput screening and recovery of proteins encoded by plasmid libraries. It is known that Escherichia coli cells can be simultaneously transformed with multiple unique plasmids and thusly complicate recombinant library screening experiments. As a result of their potential to yield misleading results, bacterial multiple vector transformants have been thoroughly characterized in previous model studies. In contrast to bacterial systems, there is little quantitative information available regarding multiple vector transformants in yeast. Saccharomyces cerevisiae is the most widely used eukaryotic platform for cell surface display, combinatorial protein engineering, and other recombinant library screens. In order to characterize the extent and nature of multiple vector transformants in this important host, plasmid-born gene libraries constructed by yeast homologous recombination were analyzed by DNA sequencing. Results It was found that up to 90% of clones in yeast homologous recombination libraries may be multiple vector transformants, that on average these clones bear four or more unique mutant genes, and that these multiple vector cells persist as a significant proportion of library populations for greater than 24 hours during liquid outgrowth. Both vector concentration and vector to insert ratio influenced the library proportion of multiple vector transformants, but their population frequency was independent of transformation efficiency. Interestingly, the average number of plasmids born by multiple vector transformants did not vary with their library population proportion. Conclusion These results highlight the potential for multiple vector transformants to dominate yeast libraries constructed by homologous recombination. The

  3. Antibiotic resistance plasmids of Staphylococcus aureus and their clinical importance

    International Nuclear Information System (INIS)

    Lacey, R.W.

    1975-01-01

    A variety of plasmids were isolated physically, and most antibiotic resistance is thought to be plasmid mediated. A number of characters (e.g., resistance to erythromycin or methicillin, and production of pigment) are determined by genes that do not give clear indications of either plasmid or chromosomal location. Although the formation of a particular plasmid is probably, even in bacterial terms, a very rare event, once formed such an element can spread rapidly among the bacterial population. The spectacular increase in the incidence of penicillinase-producing hospital strains in the late 1940's could have been due in part to this process. Evidence is stronger, however, for the intercell transfer of recently isolated plasmids coding for resistance to fusidic acid (and penicillinase production), or for neomycin, or for tetracycline resistance. Study of bacterial plasmids can resolve fundamental biochemical problems, and give some insight into the life of the cell at the molecular level. But the immediate application of the study of staphylococcal plasmids may be directed towards improving the effectiveness of antibiotic therapy. The most important aspect of future anti-staphylococcal chemotherapy should thus be the limitation of the use of antibiotics, particularly for application to the skin and nose. (U.S.)

  4. Plasmid-associated sensitivity of Bacillus thuringiensis to UV light

    International Nuclear Information System (INIS)

    Benoit, T.G.; Wilson, G.R.; Bull, D.L.; Aronson, A.I.

    1990-01-01

    Spores and vegetative cells of Bacillus thuringiensis were more sensitive to UV light than were spores or cells of plasmid-cured B. thuringiensis strains or of the closely related Bacillus cereus. Introduction of B. thuringiensis plasmids into B. cereus by cell mating increased the UV sensitivity of the cells and spores. Protoxins encoded by one or more B. thuringiensis plasmids were not involved in spore sensitivity, since a B. thuringiensis strain conditional for protoxin accumulation was equally sensitive at the permissive and nonpermissive temperatures. In addition, introduction of either a cloned protoxin gene, the cloning vector, or another plasmid not containing a protoxin gene into a plasmid-cured strain of B. thuringiensis all increased the UV sensitivity of the spores. Although the variety of small, acid-soluble proteins was the same in the spores of all strains examined, the quantity of dipicolinic acid was about twice as high in the plasmid-containing strains, and this may account for the differences in UV sensitivity of the spores. The cells of some strains harboring only B. thuringiensis plasmids were much more sensitive than cells of any of the other strains, and the differences were much greater than observed with spores

  5. Distribution of Plasmids in Distinct Leptospira Pathogenic Species.

    Science.gov (United States)

    Wang, Yanzhuo; Zhuang, Xuran; Zhong, Yi; Zhang, Cuicai; Zhang, Yan; Zeng, Lingbing; Zhu, Yongzhang; He, Ping; Dong, Ke; Pal, Utpal; Guo, Xiaokui; Qin, Jinhong

    2015-11-01

    Leptospirosis, caused by pathogenic Leptospira, is a worldwide zoonotic infection. The genus Leptospira includes at least 21 species clustered into three groups--pathogens, non-pathogens, and intermediates--based on 16S rRNA phylogeny. Research on Leptospira is difficult due to slow growth and poor transformability of the pathogens. Recent identification of extrachromosomal elements besides the two chromosomes in L. interrogans has provided new insight into genome complexity of the genus Leptospira. The large size, low copy number, and high similarity of the sequence of these extrachromosomal elements with the chromosomes present challenges in isolating and detecting them without careful genome assembly. In this study, two extrachromosomal elements were identified in L. borgpetersenii serovar Ballum strain 56604 through whole genome assembly combined with S1 nuclease digestion following pulsed-field gel electrophoresis (S1-PFGE) analysis. Further, extrachromosomal elements in additional 15 Chinese epidemic strains of Leptospira, comprising L. borgpetersenii, L. weilii, and L. interrogans, were successfully separated and identified, independent of genome sequence data. Southern blot hybridization with extrachromosomal element-specific probes, designated as lcp1, lcp2 and lcp3-rep, further confirmed their occurrences as extrachromosomal elements. In total, 24 plasmids were detected in 13 out of 15 tested strains, among which 11 can hybridize with the lcp1-rep probe and 11 with the lcp2-rep probe, whereas two can hybridize with the lcp3-rep probe. None of them are likely to be species-specific. Blastp search of the lcp1, lcp2, and lcp3-rep genes with a nonredundant protein database of Leptospira species genomes showed that their homologous sequences are widely distributed among clades of pathogens but not non-pathogens or intermediates. These results suggest that the plasmids are widely distributed in Leptospira species, and further elucidation of their biological

  6. Calculation of multigroup constants in WIMS format with programs fedgroup and flange and comparison of the results obtained using different evaluated libraries

    International Nuclear Information System (INIS)

    Trkov, A.; Budnar, M.; Copic, M.; Perdan, A.; Ravnik, M.

    1982-01-01

    Multigroup constants for 1-H-1, 92-U-235, and 92-U-238 have been calculated. Averaged cross-sections and other constants have been prepared in the WIMS 69-group format. Comparison has been made between group constants obtained with several evaluated libraries (KEDAK-3 1975, 1979, ENDF/B-4, ENDF/B-5) and the WIMS-D library. Observed differences are most pronounced in the resonance and fast region. From test runs on fuel cell with the WIMS program it can be deduced that these differences affect the fewgroup constants significantly. (author)

  7. Quantifying and resolving multiple vector transformants in S. cerevisiae plasmid libraries.

    Science.gov (United States)

    Scanlon, Thomas C; Gray, Elizabeth C; Griswold, Karl E

    2009-11-20

    In addition to providing the molecular machinery for transcription and translation, recombinant microbial expression hosts maintain the critical genotype-phenotype link that is essential for high throughput screening and recovery of proteins encoded by plasmid libraries. It is known that Escherichia coli cells can be simultaneously transformed with multiple unique plasmids and thusly complicate recombinant library screening experiments. As a result of their potential to yield misleading results, bacterial multiple vector transformants have been thoroughly characterized in previous model studies. In contrast to bacterial systems, there is little quantitative information available regarding multiple vector transformants in yeast. Saccharomyces cerevisiae is the most widely used eukaryotic platform for cell surface display, combinatorial protein engineering, and other recombinant library screens. In order to characterize the extent and nature of multiple vector transformants in this important host, plasmid-born gene libraries constructed by yeast homologous recombination were analyzed by DNA sequencing. It was found that up to 90% of clones in yeast homologous recombination libraries may be multiple vector transformants, that on average these clones bear four or more unique mutant genes, and that these multiple vector cells persist as a significant proportion of library populations for greater than 24 hours during liquid outgrowth. Both vector concentration and vector to insert ratio influenced the library proportion of multiple vector transformants, but their population frequency was independent of transformation efficiency. Interestingly, the average number of plasmids born by multiple vector transformants did not vary with their library population proportion. These results highlight the potential for multiple vector transformants to dominate yeast libraries constructed by homologous recombination. The previously unrecognized prevalence and persistence of multiply

  8. Results of regular study on radionuclidic purity of sup(99m)Tc obtained from 99Mo-sup(99m)Tc generators

    International Nuclear Information System (INIS)

    Vlcek, J.; Rohacek, J.; Husak, V.

    1979-01-01

    A total of 39 sup(99m)Tc eluates obtained from 9 99 Mo-sup(99m)Tc generators delivered by The Radiochemical Centre Amersham during one year was studied with regard to their radionuclide purity. Using a Ge(Li) spectrometer the contaminants 60 Co, 103 Ru, 131 I, 134 Cs, 140 La and 188 Re were found in sup(99m)Tc-eluates with average levels ranging from 2.9 x 10 -3 to 2.8 x 10 -1 per cent of sup(99m)Tc activity. The additional total body absorbed dose caused by these contaminants, as calculated from their average content in sup(99m)Tc eluates, was less than 1% of the dose due to sup(99m)Tc pertechnetate. (orig.) 891 MG/orig. 892 CS [de

  9. From global to local statistical shape priors novel methods to obtain accurate reconstruction results with a limited amount of training shapes

    CERN Document Server

    Last, Carsten

    2017-01-01

    This book proposes a new approach to handle the problem of limited training data. Common approaches to cope with this problem are to model the shape variability independently across predefined segments or to allow artificial shape variations that cannot be explained through the training data, both of which have their drawbacks. The approach presented uses a local shape prior in each element of the underlying data domain and couples all local shape priors via smoothness constraints. The book provides a sound mathematical foundation in order to embed this new shape prior formulation into the well-known variational image segmentation framework. The new segmentation approach so obtained allows accurate reconstruction of even complex object classes with only a few training shapes at hand.

  10. Comparison of ATLAS Tilecal MODULE No 8 high-precision metrology measurement results obtained by laser (JINR) and photogrammetric (CERN) methods

    CERN Document Server

    Batusov, V; Gayde, J C; Khubua, J I; Lasseur, C; Lyablin, M V; Miralles-Verge, L; Nessi, Marzio; Rusakovitch, N A; Sissakian, A N; Topilin, N D

    2002-01-01

    The high-precision assembly of large experimental set-ups is of a principal necessity for the successful execution of the forthcoming LHC research programme in the TeV-beams. The creation of an adequate survey and control metrology method is an essential part of the detector construction scenario. This work contains the dimension measurement data for ATLAS hadron calorimeter MODULE No. 8 (6 m, 22 tons) which were obtained by laser and by photogrammetry methods. The comparative data analysis demonstrates the measurements agreement within +or-70 mu m. It means, these two clearly independent methods can be combined and lead to the rise of a new-generation engineering culture: high-precision metrology when precision assembling of large scale massive objects. (3 refs).

  11. Comparison of ATLAS tilecal module No. 8 high-precision metrology measurement results obtained by laser (JINR) and photogrammetric (CERN) methods

    International Nuclear Information System (INIS)

    Batusov, V.; Budagov, Yu.; Gayde, J.C.

    2002-01-01

    The high-precision assembly of large experimental set-ups is of a principal necessity for the successful execution of the forthcoming LHC research programme in the TeV-beams. The creation of an adequate survey and control metrology method is an essential part of the detector construction scenario. This work contains the dimension measurement data for ATLAS hadron calorimeter MODULE No. 8 (6 m, 22 tons) which were obtained by laser and by photogrammetry methods. The comparative data analysis demonstrates the measurements agreement within ± 70 μm. It means, these two clearly independent methods can be combined and lead to the rise of a new-generation engineering culture: high-precision metrology when precision assembling of large scale massive objects

  12. Dealing with the evolutionary downside of CRISPR immunity: bacteria and beneficial plasmids.

    Directory of Open Access Journals (Sweden)

    Wenyan Jiang

    Full Text Available The immune systems that protect organisms from infectious agents invariably have a cost for the host. In bacteria and archaea CRISPR-Cas loci can serve as adaptive immune systems that protect these microbes from infectiously transmitted DNAs. When those DNAs are borne by lytic viruses (phages, this protection can provide a considerable advantage. CRISPR-Cas immunity can also prevent cells from acquiring plasmids and free DNA bearing genes that increase their fitness. Here, we use a combination of experiments and mathematical-computer simulation models to explore this downside of CRISPR-Cas immunity and its implications for the maintenance of CRISPR-Cas loci in microbial populations. We analyzed the conjugational transfer of the staphylococcal plasmid pG0400 into Staphylococcus epidermidis RP62a recipients that bear a CRISPR-Cas locus targeting this plasmid. Contrary to what is anticipated for lytic phages, which evade CRISPR by mutations in the target region, the evasion of CRISPR immunity by plasmids occurs at the level of the host through loss of functional CRISPR-Cas immunity. The results of our experiments and models indicate that more than 10(-4 of the cells in CRISPR-Cas positive populations are defective or deleted for the CRISPR-Cas region and thereby able to receive and carry the plasmid. Most intriguingly, the loss of CRISPR function even by large deletions can have little or no fitness cost in vitro. These theoretical and experimental results can account for the considerable variation in the existence, number and function of CRISPR-Cas loci within and between bacterial species. We postulate that as a consequence of the opposing positive and negative selection for immunity, CRISPR-Cas systems are in a continuous state of flux. They are lost when they bear immunity to laterally transferred beneficial genes, re-acquired by horizontal gene transfer, and ascend in environments where phage are a major source of mortality.

  13. Enterococcus faecalis Sex Pheromone cCF10 Enhances Conjugative Plasmid Transfer In Vivo

    Directory of Open Access Journals (Sweden)

    Helmut Hirt

    2018-02-01

    Full Text Available Cell-cell communication mediated by peptide pheromones (cCF10 [CF] is essential for high-frequency plasmid transfer in vitro in Enterococcus faecalis. To examine the role of pheromone signaling in vivo, we established either a CF-producing (CF+ recipient or a recipient producing a biologically inactive variant of CF (CF− recipient in a germfree mouse model 3 days before donor inoculation and determined transfer frequencies of the pheromone-inducible plasmid pCF10. Plasmid transfer was detected in the upper and middle sections of the intestinal tract 5 h after donor inoculation and was highly efficient in the absence of antibiotic selection. The transconjugant/donor ratio reached a maximum level approaching 1 on day 4 in the upper intestinal tract. Plasmid transfer was significantly lower with the CF− recipient. While rescue of the CF− mating defect by coculture with CF+ recipients is easily accomplished in vitro, no extracellular complementation occurred in vivo. This suggests that most pheromone signaling in the gut occurs between recipient and donor cells in very close proximity. Plasmid-bearing cells (donors plus transconjugants steadily increased in the population from 0.1% after donor inoculation to about 10% at the conclusion of the experiments. This suggests a selective advantage of pCF10 carriage distinct from antibiotic resistance or bacteriocin production. Our results demonstrate that pheromone signaling is required for efficient pCF10 transfer in vivo. In the absence of CF+ recipients, a low level of transfer to CF− recipients occurred in the gut. This may result from low-level host-mediated induction of the donors in the gastrointestinal (GI tract, similar to that previously observed in serum.

  14. Characterization of Virulence Plasmids and Serotyping of Rhodococcus equi Isolates from Submaxillary Lymph Nodes of Pigs in Hungary

    OpenAIRE

    Makrai, László; Takayama, Saki; Dénes, Béla; Hajtós, István; Sasaki, Yukako; Kakuda, Tsutomu; Tsubaki, Shiro; Major, Andrea; Fodor, László; Varga, János; Takai, Shinji

    2005-01-01

    The plasmid types and serotypes of 164 Rhodococcus equi strains obtained from submaxillary lymph nodes of swine from different piggeries in 28 villages and towns located throughout the country were examined. The strains were tested by PCR for the presence of 15- to 17-kDa virulence-associated protein antigen (VapA) and 20-kDa virulence-associated protein antigen (VapB) genes. Plasmid DNAs were isolated and analyzed by digestion with restriction endonucleases to estimate size and compare their...

  15. results of Breeding Tomato Programme (lycopesicon esculentum mill) for the obtaining of new Tolerant varieties to low inputs hidric through the employment of the Radioinduced mutation

    International Nuclear Information System (INIS)

    Gonzalez, Maria C.; Caballero, Alberto; Mukandama, Jean Pierre; Mohamed Ali, Mansoor; Suarez, Lorenzo; Fuentes, Jorge L

    1999-01-01

    Induced mutation have been mostly used in the improvement of different crops. This papers shows the main results of Breeding Tomato Programme. It was possible to select some promising lines with a good behavior under hidric stress

  16. Impact of co-carriage of IncA/C plasmids with additional plasmids on the transfer of antimicrobial resistance in Salmonella enterica isolates.

    Science.gov (United States)

    Han, Jing; Pendleton, Sean J; Deck, Joanna; Singh, Ruby; Gilbert, Jeffrey; Johnson, Timothy J; Sanad, Yasser M; Nayak, Rajesh; Foley, Steven L

    2018-04-20

    Antimicrobial resistance in Salmonella enterica is often plasmid encoded. A key resistance plasmid group is the incompatibility group (Inc) A/C plasmids that often carry multiple resistance determinants. Previous studies showed that IncA/C plasmids were often co-located with other plasmids. The current study was undertaken to evaluate the impact of plasmid co-carriage on antimicrobial resistance and plasmid transfer. A total of 1267 Salmonella isolates, representing multiple serotypes and sources were previously subjected to susceptibility testing and 251 isolates with resistance to at least 5 antimicrobial agents were identified for further study. Each isolate was subjected to PCR-based replicon typing, and those with IncA/C plasmids were selected for plasmid isolation, PCR-based mapping of IncA/C plasmid backbone genes, and conjugation assays to evaluate resistance plasmid transferability. Of the 87 identified IncA/C positive isolates, approximately 75% carried a plasmid with another identified replicon type, with the most common being I1 (39%), FIA, FIIA, FIB and HI2 (each 15%). PCR-based mapping indicated significant diversity in IncA/C backbone content, especially in regions encoding transfer-associated and hypothetical proteins. Conjugation experiments showed that nearly 68% of the isolates transferred resistance plasmids, with 90% containing additional identified plasmids or larger (>50 kb) non-typeable plasmids. The majority of IncA/C-positive strains were able to conjugally transfer antimicrobial resistance to the recipient, encoded by IncA/C and/or co-carried plasmids. These findings highlight the importance of co-located plasmids for resistance dissemination either by directly transferring resistance genes or by potentially providing the needed conjugation machinery for IncA/C plasmid transfer. Copyright © 2018. Published by Elsevier B.V.

  17. Final results of the cadmium and spectral ratios obtained inside of the fuel rod positioned in the central position of the IPEN/MB-01 nuclear reactor

    Energy Technology Data Exchange (ETDEWEB)

    Bitelli, Ulysses d' Utra; Mura, Luiz Ernesto C.; Santos, Diogo Feliciano dos, E-mail: ubitelli@ipen.br [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil); Lambiasi, Beatriz G.N. [Centro Tecnologico da Marinha em Sao Paulo (CTMSP), SP (Brazil)

    2015-07-01

    The spectral ratios are very important to determine some nuclear reactors parameters such as reaction rates, fuel lifetime, etc and some safety operational conditions. This study aims to determine the spectral ratios in 2 (two) spatial positions located inside the core of the Nuclear Reactor IPEN/MB-01. These places are at the central position of the nuclear reactor core in an asymptotic neutron flux region. The experiment consists in inserting different activation foil detectors inside an experimental fuel rod. The experimental rod is assembled at the central position of the reactor core. Activation neutron foil detectors of different elements such as {sup 197}Au, {sup 238}U, {sup 45}Sc, {sup 58}Ni, {sup 24}Mg, {sup 47}Ti and {sup 115m}In were used to cover a large range of neutron spectrum. Saturation activity per target nucleus was obtained by gamma spectrometry using a HPGe system. The experimental cadmium ratios compared with values computed by MCNP-4C code show good agreement. (author)

  18. Results and interpretation of groundwater data obtained from multiport-instrumented coreholes (GW-131 through GS-135), fiscal years 1990 and 1991

    International Nuclear Information System (INIS)

    Dreier, R.B.; Early, T.O.; King, H.L.

    1993-01-01

    With the increased emphasis by Department of Energy personnel on assessing the environmental impact of past waste disposal practices at all of its facilities, there has been an associated increase in characterization activities that focus on delineating site-specific groundwater flow regimes and contaminant migration pathways. At the Oak Ridge Y-12 Plant, the complex geologic and hydrologic relationships require a more detailed understanding of the three-dimensional properties of groundwater flow regimes than can be obtained by conventional monitoring activities. Thus, as part of groundwater characterization activities conducted by the Environmental Surveillance Section staff of the Y-12 Plant Environmental Management Department, five existing deep core holes were instrumented with multiport monitoring systems to provide greatly enhanced resolution of the hydraulic and hydrochemical properties of the groundwater system within Bear Creek Valley. With a multiport system, it is possible to measure hydraulic head and hydraulic conductivity and collect water samples from multiple levels within a single borehole. In this report, multiport data collected during fiscal years (FYs) 1990 and 1991 are summarized

  19. Results obtained in a spectrometry using junction detectors (1962); Resultats en spectrometrie {alpha} a l'aide de ''detecteurs jonction'' (1962)

    Energy Technology Data Exchange (ETDEWEB)

    Beaumevieille, H; Fontenille, J; Lottgequeue, J P; Longequeue, N; Rabour, C [Commissariat a l' Energie Atomique, Grenoble (France). Centre d' Etudes Nucleaires

    1962-07-01

    Junction detectors have been used for studying {alpha} spectra obtained by nuclear reactions (p, {alpha}), the protons being produced by an electrostatic accelerator (E{sub p} varying from 100 to 600 keV). Difficulties have been encountered arising from parasitic noise from the accelerator and from the distance (60 metres) between detectors and electronic equipment. Resolution of the counter has been found to be 0.6 per cent with {alpha} from {sup 210}Po and 1.2 pour cent with {alpha} (5.8 MeV) from the reaction {sup 11}B (p, {alpha}{sub 0}). (authors) [French] Des detecteurs a jonction ont ete utilises pour l'etude de spectres {alpha} obtenus par reactions nucleaires (p, {alpha}), les protons etant acceleres par un accelerateur electrostatique (E{sub p} variant de 100 a 600 keV). Des difficultes ont ete recontrees dues aux parasites provenant de l'accelerateur et a la distance des cables (60 metres) entre detecteurs et appareils de mesure. Le pouvoir de resolution de l'ensemble a ete de 0, 6 pour cent avec les {alpha} du {sup 210}Po et de 1,2 pour cent avec les {alpha} (5,8 MeV) de la reaction {sup 11}B (p, {alpha}{sub 0}). (auteurs)

  20. KPC-mediated resistance in Klebsiella pneumoniae in two hospitals in Padua, Italy, June 2009-December 2011: massive spreading of a KPC-3-encoding plasmid and involvement of non-intensive care units

    Directory of Open Access Journals (Sweden)

    Richter Sara N

    2012-07-01

    Full Text Available Abstract Background Klebsiella pneumoniae carbapenemases (KPCs producing bacteria have emerged as a cause of multidrug-resistant nosocomial infections worldwide. KPCs are plasmid-encoded enzymes capable of hydrolysing a broad spectrum of beta-lactams, including carbapenems and monobactams, therefore worryingly limiting antimicrobial treatment options. Analysis of circulating bacterial strains and KPC alleles may help understanding the route of KPC dissemination and therefore help containing the infection. Methods KPC-producing Klebsiella pneumoniae dissemination in two 1580- and 300- bed hospitals in Padua, Italy, from initial outbreak in 2009 to late 2011 was analysed. Molecular and clinical epidemiology, including bacterial strains, KPC-encoding plasmid sequences and associated resistance genes, involved hospital wards and relocation of patients were described. Routine antimicrobial susceptibility testing and MIC of carbapenems on clinical isolates were performed. Detection of resistance genes was obtained by PCR and sequencing. MLST, PFGE and ERIC were used for molecular genotyping. Plasmid analysis was obtained by digestion with restriction enzymes and deep sequencing. Results KPC-positive clinical samples were isolated from nearly 200 patients. In the initial outbreak intensive care units were almost exclusively involved, while medical, surgical and long-term wards were successively massively concerned. Analysis of KPC alleles, plasmids and bacterial sequence types (STs indicated that during the initial outbreak KPC-3 in ST258 and KPC-2 in ST147 were each confined in one of the two surveilled hospitals. While KPC-2 dissemination was effectively contained, KPC-3 in ST258 cross-spreading was observed. The simultaneous presence of two carbapenemases, VIM-1 and KPC-2, in the same isolate was also observed in three patients. Total sequencing of plasmid content of two KPC-3 strains showed novel association of resistance plasmids. Conclusions The

  1. Comparative assessment of plasmid DNA delivery by encapsulation ...

    African Journals Online (AJOL)

    Tropical Journal of Pharmaceutical Research January 2018; 17 (1): 1-10 ... Purpose: To compare the gene delivery effectiveness of plasmid DNA (pDNA) ..... Intramuscular delivery of DNA ... copolymeric system for gene delivery in complete.

  2. The technology of large-scale pharmaceutical plasmid purification ...

    African Journals Online (AJOL)

    STORAGESEVER

    2010-01-04

    Jan 4, 2010 ... DNA vaccine, the cost of purification must be decreased. Although commonly .... Three mice were killed every 4 days interval. Tissues of heart, liver, .... Now, methods such as chromatography had good prospects in plasmid ...

  3. Mechanisms of Evolution in High-Consequence Drug Resistance Plasmids

    Directory of Open Access Journals (Sweden)

    Susu He

    2016-12-01

    Full Text Available The dissemination of resistance among bacteria has been facilitated by the fact that resistance genes are usually located on a diverse and evolving set of transmissible plasmids. However, the mechanisms generating diversity and enabling adaptation within highly successful resistance plasmids have remained obscure, despite their profound clinical significance. To understand these mechanisms, we have performed a detailed analysis of the mobilome (the entire mobile genetic element content of a set of previously sequenced carbapenemase-producing Enterobacteriaceae (CPE from the National Institutes of Health Clinical Center. This analysis revealed that plasmid reorganizations occurring in the natural context of colonization of human hosts were overwhelmingly driven by genetic rearrangements carried out by replicative transposons working in concert with the process of homologous recombination. A more complete understanding of the molecular mechanisms and evolutionary forces driving rearrangements in resistance plasmids may lead to fundamentally new strategies to address the problem of antibiotic resistance.

  4. Preliminary interpretations of geologic results obtained from boreholes UE25a-4, -5, -6, and -7, Yucca Mountain, Nevada Test Site

    International Nuclear Information System (INIS)

    Spengler, R.W.; Rosenbaum, J.G.

    1980-01-01

    Since 1978, the USGS (US Geological Survey) has been providing technical assistance in characterizing suitable rock masses at or contiguous to the NTS (Nevada Test Site) for long-term storage of high-level nuclear waste. Current efforts have been focused on investigating Yucca Mountain, a volcanic highland situated along the western boundary of NTS in southern Nevada. Detailed stratigraphic and structural studies have been in progress along a northeastern segment of the highland in a wedge-shaped area bounded by Basin and Range faults, most of which trend north-northeast. A series of four locally steep-walled, nearly parallel, linear washes transect the northeastern half of the area of interest and display trends similar to major faults to the northeast. Prior to the present study, drill hole UE25a-1, located about 1600 feet southeast of the edge of the area of interest, was cored to a depth of 2500 feet. Subsurface information derived from the upper 500 feet of this drill hole is included in this report to compare with recently acquired data. Surface electrical surveys have been conducted by both the University of Utah and the USGS perpendicular to the trend of the washes in an attempt to better understand factors that have influenced the present drainage pattern. Preliminary data of both pole-dipole and dipole-dipole resistivity/IP electrical methods indicate numerous vertical and horizontal discontinuities between adjacent resistive bodies that strongly suggest a broad zone of faulting, fracturing, and (or) brecciation. To verify the existence of structural discontinuities suggested by the linear washes and electrical anomalies, a drilling program was initiated in June 1979, to obtain geologic information within the southernmost of four northwest-trending washes

  5. Preliminary Results Obtained from Flight Test of a Rocket Model Having the Tail Only of the Grumman XF10F Airplane Configuration, TED No. NACA DE 354

    Science.gov (United States)

    Gardner, William N.; Edmondson, James L.

    1950-01-01

    A flight test was made to determine the servoplane effectiveness and stability characteristics of the free-floating horizontal stabilizer to be used on the XF10F airplane. The results of this test indicate that servoplane effectiveness is practically constant through the speed range up to a Mach number of 1.15, and the stabilizer static stability is satisfactory. A loss of damping occurs over a narrow Mach number range near M = 1.0, resulting in dynamic instability of the stabilizer in this narrow range. Above M = 1.0 there is a gradual positive trim change of the stabilizer.

  6. A comparison of texture results obtained using precession electron diffraction and neutron diffraction methods at diminishing length scales in ordered bimetallic nanolamellar composites

    International Nuclear Information System (INIS)

    Carpenter, J.S.; Liu, X.; Darbal, A.; Nuhfer, N.T.; McCabe, R.J.; Vogel, S.C.; LeDonne, J.E.; Rollett, A.D.; Barmak, K.; Beyerlein, I.J.; Mara, N.A.

    2012-01-01

    Precession electron diffraction (PED) is used to acquire orientation information in Cu–Nb nanolamellar composites fabricated by accumulative roll bonding (ARB). The resulting maps quantify the grain size, shape, orientation distributions and interface planes in the vicinity of nanometer-thick deformation twins. The PED-based texture results compare favorably with bulk textures provided by neutron diffraction measurements, indicating uniformity in the ARB Cu–Nb texture. Additionally, {1 1 2} Cu ||{1 1 2} Nb interfaces are present, suggesting that ARB techniques can lead to stable interfaces with a special crystallography.

  7. A comparison of the impact of screen-positive results obtained from ultrasound and biochemical screening for Down syndrome in the first trimester : a pilot study

    NARCIS (Netherlands)

    Weinans, M.J.; Kooij, L.; Muller, M.A.; Bilardo, K.M.; van Lith, J.M.; Tymstra, T.

    2004-01-01

    OBJECTIVE: To compare the experiences of women who received a screen-positive test result for Down syndrome after nuchal translucency screening or after biochemical screening in the first trimester of pregnancy in the Netherlands. METHOD: Semi-quantitative questionnaires were sent to 40 women with a

  8. Comparison of stapes prostheses: a retrospective analysis of individual audiometric results obtained after stapedotomy by implantation of a gold and a teflon piston

    NARCIS (Netherlands)

    de Bruijn, A. J.; Tange, R. A.; Dreschler, W. A.

    1999-01-01

    PURPOSE: Evaluation of hearing results after implantation of a Teflon piston (type Causse; Xomed Surgical Products, Jacksonville, FL) and of a pure gold piston (K-piston; Heinz Kurz GmbH Medizintechnik, Dusslingen, Germany), both with a shaft diameter of 0.4 mm in cases of otosclerosis requiring

  9. A retrospective study of the hearing results obtained after stapedotomy by the implantation of two Teflon pistons with a different diameter

    NARCIS (Netherlands)

    Grolman, W.; Tange, R. A.; de Bruijn, A. J.; Hart, A. A.; Schouwenburg, P. F.

    1997-01-01

    This study reports the evaluation of hearing results after implantation of a Teflon piston of a different diameter in cases of otosclerosis requiring stapedotomy. By random selection, a Teflon piston with a shaft diameter of 0.3 mm was inserted in 34 cases and a piston with a shaft diameter of 0.4

  10. Compatibility study of the different pole figures in one and the same specimen. Comparison between the results obtained by neutron and by X-ray diffraction

    International Nuclear Information System (INIS)

    Pernot, M.; Penelle, R.; Dervin, P.

    1981-01-01

    Development of a calculation to show directly the compatibility of the different pole figures in the same specimen is exposed. A review of the method by serial development is given. Experimental methods are described. The position of the anisotropy axes is determined. Compatibilities resulting from X-ray and from neutron diffraction is analysed [fr

  11. Isolation and properties of plasmids from Deinococcus radiodurans Sark

    International Nuclear Information System (INIS)

    Sjarief, S.H.; Kikuchi, Masahiro; Kurita, Hiromi; Kitayama, Shigeru; Watanabe, Hiroshi.

    1990-05-01

    Radioresistant bacterium, Deinococcus radiodurans, can repair completely almost all of DNA damages including double strand breaks induced by gamma-rays up to about 5 kGy. In order to reveal the repair mechanism, it is necessary to develop a cloning vector available for the genetic analysis. We tried to isolate plasmids from D.radiodurans Sark strain. In the present paper the isolation and properties of plasmids were described. (author)

  12. Mechanisms of Evolution in High-Consequence Drug Resistance Plasmids.

    Science.gov (United States)

    He, Susu; Chandler, Michael; Varani, Alessandro M; Hickman, Alison B; Dekker, John P; Dyda, Fred

    2016-12-06

    The dissemination of resistance among bacteria has been facilitated by the fact that resistance genes are usually located on a diverse and evolving set of transmissible plasmids. However, the mechanisms generating diversity and enabling adaptation within highly successful resistance plasmids have remained obscure, despite their profound clinical significance. To understand these mechanisms, we have performed a detailed analysis of the mobilome (the entire mobile genetic element content) of a set of previously sequenced carbapenemase-producing Enterobacteriaceae (CPE) from the National Institutes of Health Clinical Center. This analysis revealed that plasmid reorganizations occurring in the natural context of colonization of human hosts were overwhelmingly driven by genetic rearrangements carried out by replicative transposons working in concert with the process of homologous recombination. A more complete understanding of the molecular mechanisms and evolutionary forces driving rearrangements in resistance plasmids may lead to fundamentally new strategies to address the problem of antibiotic resistance. The spread of antibiotic resistance among Gram-negative bacteria is a serious public health threat, as it can critically limit the types of drugs that can be used to treat infected patients. In particular, carbapenem-resistant members of the Enterobacteriaceae family are responsible for a significant and growing burden of morbidity and mortality. Here, we report on the mechanisms underlying the evolution of several plasmids carried by previously sequenced clinical Enterobacteriaceae isolates from the National Institutes of Health Clinical Center (NIH CC). Our ability to track genetic rearrangements that occurred within resistance plasmids was dependent on accurate annotation of the mobile genetic elements within the plasmids, which was greatly aided by access to long-read DNA sequencing data and knowledge of their mechanisms. Mobile genetic elements such as

  13. Assessing representativeness of sampling methods for reaching men who have sex with men: a direct comparison of results obtained from convenience and probability samples.

    Science.gov (United States)

    Schwarcz, Sandra; Spindler, Hilary; Scheer, Susan; Valleroy, Linda; Lansky, Amy

    2007-07-01

    Convenience samples are used to determine HIV-related behaviors among men who have sex with men (MSM) without measuring the extent to which the results are representative of the broader MSM population. We compared results from a cross-sectional survey of MSM recruited from gay bars between June and October 2001 to a random digit dial telephone survey conducted between June 2002 and January 2003. The men in the probability sample were older, better educated, and had higher incomes than men in the convenience sample, the convenience sample enrolled more employed men and men of color. Substance use around the time of sex was higher in the convenience sample but other sexual behaviors were similar. HIV testing was common among men in both samples. Periodic validation, through comparison of data collected by different sampling methods, may be useful when relying on survey data for program and policy development.

  14. Development of integrated system of spatial coordinates for inputting cartographic information obtained as result of scientific researches at exclusion zone area

    International Nuclear Information System (INIS)

    Mazur, A.B.; Ermolenko, A.I.; Shaptala, D.V.; Postil, S.D.; Yachmenev, V.V.

    1999-01-01

    Integrated system of spatial coordinates was developed for modeling of diverse processes occurring in the exclusion zone. Technology of joint use of CAD and geoinformation system was worked out,which resulted in the 'Ukryttia' object fixing to the topographic base in accordance with geographic coordinates, editing of nomenclative sheets of the area adjacent to the ChNPP and renovation of its relief, as well as conversion of the files made in AutoCad to Arc View format files was performed

  15. Technique to Obtain a Predictable Aesthetic Result through Appropriate Placement of the Prosthesis/Soft Tissue Junction in the Edentulous Patient with a Gingival Smile.

    Science.gov (United States)

    Demurashvili, Georgy; Davarpanah, Keyvan; Szmukler-Moncler, Serge; Davarpanah, Mithridade; Raux, Didier; Capelle-Ouadah, Nedjoua; Rajzbaum, Philippe

    2015-10-01

    Treating the edentulous patient with a gingival smile requires securing the prosthesis/soft tissue junction (PSTJ) under the upper lip. To present a simple method that helps achieve a predictable aesthetic result when alveoplasty of the anterior maxilla is needed to place implants apical to the presurgical position of the alveolar ridge. The maximum smile line of the patient is recorded and carved on a thin silicone bite impression as a soft tissue landmark. During the three-dimensional radiographic examination, the patient wears the silicone guide loaded with radiopaque markers. The NobelClinician® software is then used to bring the hard and soft tissue landmarks together in a single reading. Using the software, a line is drawn 5 mm apical to the smile line; it dictates the position of the crestal ridge to be reached following the alveoplasty. Subsequently, the simulated implant position and the simulated residual bone height following alveoplasty can be simultaneously evaluated on each transverse section. An alveoplasty of the anterior maxilla was performed as simulated on the software, and implants were placed accordingly. The PSTJ was always under the upper lip, even during maximum smile events. The aesthetic result was, therefore, fully satisfactory. This simple method permits the placement of the PSTJ under the upper lip with a predictable outcome; it ensures a reliable aesthetic result for the edentulous patient with a gingival smile. © 2013 Wiley Periodicals, Inc.

  16. A feasibility study of prepubertal and over mature aged local goat in relation to results of In Vitro growth culture to obtain additional M-II oocyte resources

    Science.gov (United States)

    Ciptadi, Gatot; Ihsan, M. Nur; Rahayu, Sri; Widjaja, D. H. K.; Mudawamah, Mudawamah

    2017-11-01

    The aims of this research are to study the potential source of mature (M-II) oocytes of domestic animals using follicles isolated from prepubertal and over mature aged Indonesian local goats, resulting from an in vitro growth (IVG) method. This method of IVG could provide a new source of M-II oocytes for embryo production. In Indonesia, a very limited number of a good quality oocytes are available for research purposes, as there is a limited number of reproductive females slaughtered, which is dominated by prepubertal and old mature aged animals. IVG culture systems could be improved as an alternative method to provide a new source of a good quality oocytes for in vitro maturation of M-II oocytes. From a number of prepubertal and mature aged goats slaughtered in a local abattoir, the small oocytes in the preantral follicles were cultured in vitro to normal oocyte growth. The methods used in this research are experimental. Follicles were isolated, cultured in vitro for 14 days individually using a sticky medium containing 4% (w/v) polyvinylpyrrolidone in TCM 199 10% Fetal Bovine Serum supplemented with Follicle Stimulating Hormone, which was then evaluated for their follicle development and oocyte quality. The research results showed that a minimum follicle size and oocyte diameter is needed (>100 um) for early evaluation of maturation to be achieved, meanwhile oocytes recovered from IVG after being cultured in vitro for maturation resulted in a very low rate of maturation. However, in the future, IVG of the preantral follicles of Indonesian local goat could be considered as an alternative source of oocytes for both research purposes and embryo production in vitro.

  17. Contribution to a summary of the results obtained in the field of geology on the Oklo natural reactors and their environment

    International Nuclear Information System (INIS)

    Weber, F.

    1978-01-01

    The very special characteristics of the ore in the reaction zones may be attributed to the effects of the nuclear reactions themselves: de-structuration of the gangue by neutronic effects and hydrothermal leaching due to the generation of a true thermal syphon by the nuclear reactions. This mechanism contributes to the propagation of reactions from a number of initial reaction sites. The latter were initiated in certain parts of the deposit, enriched as a result of a tectonic episode, by oxidizing currents in the shear troughs. These enriched zones occur as anomalies within a pre-existent deposit, the formation of which still poses many questions of a metallogenic nature. (author)

  18. A nonparametric statistical method for determination of a confidence interval for the mean of a set of results obtained in a laboratory intercomparison

    International Nuclear Information System (INIS)

    Veglia, A.

    1981-08-01

    In cases where sets of data are obviously not normally distributed, the application of a nonparametric method for the estimation of a confidence interval for the mean seems to be more suitable than some other methods because such a method requires few assumptions about the population of data. A two-step statistical method is proposed which can be applied to any set of analytical results: elimination of outliers by a nonparametric method based on Tchebycheff's inequality, and determination of a confidence interval for the mean by a non-parametric method based on binominal distribution. The method is appropriate only for samples of size n>=10

  19. A cross-cultural study on noise problems: Comparison of the results obtained in Japan, West Germany, the U.S.A., China and Turkey

    Science.gov (United States)

    Namba, S.; Kuwano, S.; Schick, A.; Açlar, A.; Florentine, M.; Da Rui, Zheng

    1991-12-01

    Neighborhood noise has become a serious problem in many countries, and it is difficult to regulate by physical means alone. A cross-cultural survey was conducted in five countries, Japan, West Germany, the United States, China and Turkey, among residents of apartment houses in order to examine the cultural differences involved. Interesting differences were found in preferred countermeasures, highly annoying sounds, attitudes against noise, expectations for laws, etc. The use of loudspeakers in public places was accepted in all five countries, provided that they were used for conveying necessary information. The results suggest that it is important to take cultural or social backgrounds into consideration in order to find suitable countermeasures.

  20. Reducing the item number to obtain the same-length self-assessment scales: a systematic approach using result of graphical loglinear rasch models

    DEFF Research Database (Denmark)

    Nielsen, Tine; Kreiner, Svend

    2011-01-01

    The Revised Danish Learning Styles Inventory (R-D-LSI) (Nielsen 2005), which is an adaptation of Sternberg- Wagner Thinking Styles Inventory (Sternberg, 1997), comprises 14 subscales, each measuring a separate learning style. Of these 14 subscales, 9 are eight items long and 5 are seven items long...... Inventory (D-SA-LSI) comprising 14 subscales each with an item length of seven. The systematic approach to item reduction based on results of GLLRM will be presented and exemplified by its application to the R-D-LSI....

  1. On the Evaluation of Computational Results Obtained from Solving System of linear Equations With matlab The Dual affine Scalling interior Point

    International Nuclear Information System (INIS)

    Murfi, Hendri; Basaruddin, T.

    2001-01-01

    The interior point method for linear programming has gained extraordinary interest as an alternative to simplex method since Karmarkar presented a polynomial-time algorithm for linear programming based on interior point method. In implementation of the algorithm of this method, there are two important things that have impact heavily to performance of the algorithm; they are data structure and used method to solve linear equation system in the algorithm. This paper describes about solving linear equation system in variants of the algorithm called dual-affine scaling algorithm. Next, we evaluate experimentally results of some used methods, either direct method or iterative method. The experimental evaluation used Matlab

  2. Contribution to an understanding of the action of gamma radiation on granular starch - interpretation of results obtained using the enzymatic and chromatographic method

    International Nuclear Information System (INIS)

    Robin, J.P.; Tollier, M.Th.; Guilbot, A.

    1978-01-01

    Besides compounds of low molecular mass, the gamma irradiation of granular starch produces radiodextrins with a mass lying between that of the low-molecular-mass compounds and that of the amylose and amylopectin macromolecules from which they derive. The authors present the main results relating to characterization of the radiodextrins of strongly irradiated relating to characterization of the radiodextrins of strongly irradiated normal and waxy maize starches. The method of study - both enzymatic and chromatographic - is the one that has been in use for some ten years for studying the fine structure of α-1.4-1.6 glucanes. An attempt is made to interpret and integrate the results in the light of new data derived from a study of the controlled acid hydrolysis of starch. In particular, the following hypothesis is advanced: the 'hydrolytic' effect of irradiation is, at the qualitative level, independent of the nature of the starch and similar to the effect produced by acid hydrolysis; as with acid hydrolysis, the breaking of the covalent bonds is a function of the internal strucutre of the grain and especially of its amorphous/crystalline organization; the zones of an amorphous character are more easily degraded; on the other hand, the 'crystalline' zones are better protected; in fact, the 'hydrolytic' effect of irradiation is not homogeneous and does not conform to a statistical pattern. (author)

  3. The contribution of an asthma diagnostic consultation service in obtaining an accurate asthma diagnosis for primary care patients: results of a real-life study.

    Science.gov (United States)

    Gillis, R M E; van Litsenburg, W; van Balkom, R H; Muris, J W; Smeenk, F W

    2017-05-19

    Previous studies showed that general practitioners have problems in diagnosing asthma accurately, resulting in both under and overdiagnosis. To support general practitioners in their diagnostic process, an asthma diagnostic consultation service was set up. We evaluated the performance of this asthma diagnostic consultation service by analysing the (dis)concordance between the general practitioners working hypotheses and the asthma diagnostic consultation service diagnoses and possible consequences this had on the patients' pharmacotherapy. In total 659 patients were included in this study. At this service the patients' medical history was taken and a physical examination and a histamine challenge test were carried out. We compared the general practitioners working hypotheses with the asthma diagnostic consultation service diagnoses and the change in medication that was incurred. In 52% (n = 340) an asthma diagnosis was excluded. The diagnosis was confirmed in 42% (n = 275). Furthermore, chronic rhinitis was diagnosed in 40% (n = 261) of the patients whereas this was noted in 25% (n = 163) by their general practitioner. The adjusted diagnosis resulted in a change of medication for more than half of all patients. In 10% (n = 63) medication was started because of a new asthma diagnosis. The 'one-stop-shop' principle was met with 53% of patients and 91% (n = 599) were referred back to their general practitioner, mostly within 6 months. Only 6% (n = 41) remained under control of the asthma diagnostic consultation service because of severe unstable asthma. In conclusion, the asthma diagnostic consultation service helped general practitioners significantly in setting accurate diagnoses for their patients with an asthma hypothesis. This may contribute to diminish the problem of over and underdiagnosis and may result in more appropriate treatment regimens. SERVICE HELPS GENERAL PRACTITIONERS MAKE ACCURATE DIAGNOSES: A consultation service can

  4. Plasmid-cured Chlamydia caviae activates TLR2-dependent signaling and retains virulence in the guinea pig model of genital tract infection.

    Science.gov (United States)

    Frazer, Lauren C; Darville, Toni; Chandra-Kuntal, Kumar; Andrews, Charles W; Zurenski, Matthew; Mintus, Margaret; AbdelRahman, Yasser M; Belland, Robert J; Ingalls, Robin R; O'Connell, Catherine M

    2012-01-01

    Loss of the conserved "cryptic" plasmid from C. trachomatis and C. muridarum is pleiotropic, resulting in reduced innate inflammatory activation via TLR2, glycogen accumulation and infectivity. The more genetically distant C. caviae GPIC is a natural pathogen of guinea pigs and induces upper genital tract pathology when inoculated intravaginally, modeling human disease. To examine the contribution of pCpGP1 to C. caviae pathogenesis, a cured derivative of GPIC, strain CC13, was derived and evaluated in vitro and in vivo. Transcriptional profiling of CC13 revealed only partial conservation of previously identified plasmid-responsive chromosomal loci (PRCL) in C. caviae. However, 2-deoxyglucose (2DG) treatment of GPIC and CC13 resulted in reduced transcription of all identified PRCL, including glgA, indicating the presence of a plasmid-independent glucose response in this species. In contrast to plasmid-cured C. muridarum and C. trachomatis, plasmid-cured C. caviae strain CC13 signaled via TLR2 in vitro and elicited cytokine production in vivo similar to wild-type C. caviae. Furthermore, inflammatory pathology induced by infection of guinea pigs with CC13 was similar to that induced by GPIC, although we observed more rapid resolution of CC13 infection in estrogen-treated guinea pigs. These data indicate that either the plasmid is not involved in expression or regulation of virulence in C. caviae or that redundant effectors prevent these phenotypic changes from being observed in C. caviae plasmid-cured strains.

  5. Plasmid-cured Chlamydia caviae activates TLR2-dependent signaling and retains virulence in the guinea pig model of genital tract infection.

    Directory of Open Access Journals (Sweden)

    Lauren C Frazer

    Full Text Available Loss of the conserved "cryptic" plasmid from C. trachomatis and C. muridarum is pleiotropic, resulting in reduced innate inflammatory activation via TLR2, glycogen accumulation and infectivity. The more genetically distant C. caviae GPIC is a natural pathogen of guinea pigs and induces upper genital tract pathology when inoculated intravaginally, modeling human disease. To examine the contribution of pCpGP1 to C. caviae pathogenesis, a cured derivative of GPIC, strain CC13, was derived and evaluated in vitro and in vivo. Transcriptional profiling of CC13 revealed only partial conservation of previously identified plasmid-responsive chromosomal loci (PRCL in C. caviae. However, 2-deoxyglucose (2DG treatment of GPIC and CC13 resulted in reduced transcription of all identified PRCL, including glgA, indicating the presence of a plasmid-independent glucose response in this species. In contrast to plasmid-cured C. muridarum and C. trachomatis, plasmid-cured C. caviae strain CC13 signaled via TLR2 in vitro and elicited cytokine production in vivo similar to wild-type C. caviae. Furthermore, inflammatory pathology induced by infection of guinea pigs with CC13 was similar to that induced by GPIC, although we observed more rapid resolution of CC13 infection in estrogen-treated guinea pigs. These data indicate that either the plasmid is not involved in expression or regulation of virulence in C. caviae or that redundant effectors prevent these phenotypic changes from being observed in C. caviae plasmid-cured strains.

  6. An overview of experimental results obtained under the prestressed concrete nuclear pressure vessel development program at the Oak Ridge National Laboratory

    International Nuclear Information System (INIS)

    Naus, D.J.

    1979-01-01

    Under the Prestressed Concrete Nuclear Pressure Development Program at the Oak Ridge National Laboratory, various aspects of Prestressed Concrete Pressure Vessels (PCPVs) are investigated with respect to reliability, structural performance, constructability, and economy. These investigations are conducted under the High-Temperature Gas-Cooled Reactor (HTGR) Program and the Gas-Cooled Fast Reactor (GCFR) Program. The objectives are to: (1) provide technical support to ongoing PCPV design activities, (2) contribute to the overall technological data base, and (3) provide independent review and evaluations. Specific areas of interest at present include finite-element analysis development, materials and structural behaviour tests, instrumentation evaluations and development, and structural model tests. The following provides an overview of both the HTGR and GCFR PCPV activities and a summary of recent experimental results

  7. Correlation of results obtained by in-vivo optical spectroscopy with measured blood oxygen saturation using a positive linear regression fit

    Science.gov (United States)

    McCormick, Patrick W.; Lewis, Gary D.; Dujovny, Manuel; Ausman, James I.; Stewart, Mick; Widman, Ronald A.

    1992-05-01

    Near infrared light generated by specialized instrumentation was passed through artificially oxygenated human blood during simultaneous sampling by a co-oximeter. Characteristic absorption spectra were analyzed to calculate the ratio of oxygenated to reduced hemoglobin. A positive linear regression fit between diffuse transmission oximetry and measured blood oxygenation over the range 23% to 99% (r2 equals .98, p signal was observed in the patient over time. The procedure was able to be performed clinically without difficulty; rSO2 values recorded continuously demonstrate the usefulness of the technique. Using the same instrumentation, arterial input and cerebral response functions, generated by IV tracer bolus, were deconvoluted to measure mean cerebral transit time. Date collected over time provided a sensitive index of changes in cerebral blood flow as a result of therapeutic maneuvers.

  8. Can Good Infection Control Be Obtained in One-stage Exchange of the Infected TKA to a Rotating Hinge Design? 10-year Results.

    Science.gov (United States)

    Zahar, Akos; Kendoff, Daniel O; Klatte, Till O; Gehrke, Thorsten A

    2016-01-01

    Prosthetic joint infection (PJI) occurs in 1% to 2% of total knee arthroplasties (TKAs). Although two-stage exchange is the preferred management method of patients with chronic PJI in TKA in North America, one-stage exchange is an alternative treatment method, but long-term studies of this approach have not been conducted. We reviewed our minimum 9-year results of 70 patients who underwent one-stage exchange arthroplasty with a rotating hinge design to determine: (1) What was the proportion of patients free of infection? (2) What was the patient rate of survival free of any reoperation? (3) What were the clinical outcomes as measured by Hospital for Special Surgery scores? (4) What proportion of patients developed radiographic evidence of loosening? All one-stage revision TKAs for infection between January 1 and December 31, 2002, with a minimum 9-year followup (mean, 10 years; range, 9-11 years), in which patients had been seen within the last 1 year, were included in this retrospective review. During that period, 11 patients with infected TKAs were treated with other approaches (including two-stage approaches in eight); the general indication for one-stage revision was the diagnosis of PJI with a known causative organism. Exclusion criteria were culture-negative preoperative aspiration, known allergy to local antibiotics or bone cement, or cases in which radical débridement was impossible as a result of the involvement of important anatomical structures. Eighty-one patients with PJI were seen during this period; 70 underwent one-stage exchange using our strict protocol and were reimplanted with a rotating hinge TKA. Eleven patients (15.7%) were lost to followup. Hospital for Special Surgery scores were recorded and all radiographs were evaluated for prosthetic loosening. Failure was defined as revision surgery for infection or any other cause. Our 10-year infection-free survival was 93% (mean, 4.1; 95% confidence interval [CI], 89%-96%; p exchange techniques for

  9. Results of the new processing of images obtained from the surface of Venus in a TV experiment onboard the VENERA-9 lander (1975)

    Science.gov (United States)

    Ksanfomality, L. V.

    2012-09-01

    Data on the results of the analysis of the content of re-processed panorama of the VENERA-9 lander are presented. The panorama was transmitted historically for the first time from the surface of Venus in 1975. The low noise of the VENERA-9 data allowed allocating a large object of an unusual regular structure. Earlier, its fuzzy image was repeatedly cited in the literature being interpreted as a "strange stone". The complex shape and its other features suggest that the object may be a real habitant of the planet. It is not excluded that another similar object observed was damaged during the VENERA-9 landing. From the evidence of its movement and position of some other similar objects it is concluded that because of the limited energy capacity, the physical action of the Venusian fauna may be much slower than that of the Earth fauna. Another question considered is what sources of energy could be used by life in the conditions of the high temperature oxygenless atmosphere of the planet. It is natural to assume that, like on Earth, the Venusian fauna is heterotrophic and should be based on hypothetical flora, using photosynthesis (based on an unknown high temperature biophysical mechanism).

  10. Quantity and quality of black carbon molecular markers as obtained by two chromatographic methods (GC-FID and HPLC-DAD) - How do results compare?

    Science.gov (United States)

    Schneider, M. P. W.; Smittenberg, R. H.; Dittmar, T.; Schmidt, M. W. I.

    2009-04-01

    Chars produced by wildfires are an important source of black carbon (BC) in the environment. After their deposition on the soil surface they can be distributed into rivers, marine waters and sediments. The analysis of benzenepolycarboxylic acids (BPCAs) as a quantitative measure for black carbon (BC) in soil and sediment samples is a well-established method (Glaser et al., 1998; Brodowski et al., 2005). Briefly, the nitric acid oxidation of fused aromatic ring systems in BC forms eight molecular markers (BPCAs), which can be assigned to BC, and which subsequently can be quantified by GC-FID (gas chromatography with flame ionization detector). Recently, this method was modified for the quantification of BC in seawater samples using HPLC-DAD (High performance liquid chromatography with diode array detector) for the determination of individual BPCAs (Dittmar, 2008). A direct comparison of both analytical techniques is lacking but would be important for future data comparison aimed at the calculation of global BC budgets. Here we present a systematic comparison of the two BPCA quantification methods. We prepared chars under well-defined laboratory conditions. In order to cover a broad spectrum of char properties we used two sources of biomass and a wide range of pyrolysis temperatures. Chestnut hardwood chips (Castanea sativa) and rice straw (Oryza sativa) were pyrolysed at temperatures between 200 and 1000°C under a constant N2 stream. The maximum temperatures were held constant for 5 hours (Hammes et al., 2006). The BC contents of the chars have been analysed using the BPCA extraction method followed by either GC-FID or HPLC-DAD quantification. Preliminary results suggest that both methods yield similar total quantities of BPCA, and also the proportions of the individual markers are similar. Ongoing experiments will allow for a more detailed comparison of the two methods. The BPCA composition of chars formed at different temperatures and from different precursor

  11. Colorectal endoscopic submucosal dissection (ESD) in the West - when can satisfactory results be obtained? A single-operator learning curve analysis.

    Science.gov (United States)

    Spychalski, Michał; Skulimowski, Aleksander; Dziki, Adam; Saito, Yutaka

    2017-12-01

    Up to date we lack a detailed description of the colorectal endoscopic submucosal dissection (ESD) learning curve, that would represent the experience of the Western center. The aim of this study was to define the critical points of the learning curve and to draw up lesions qualification guidelines tailored to the endoscopists experience. We have carried out a single center prospective study. Between June 2013 and December 2016, 228 primary colorectal lesions were managed by ESD procedure. In order to create a learning curve model and to carry out the analysis the cases were divided into six periods, each consisting of 38 cases. The overall en bloc resection rate was 79.39%. The lowest en bloc resection rate (52.36%) was observed in the first period. After completing 76 procedures, the resection rate surged to 86% and it was accompanied by the significant increase in the mean procedure speed of ≥9 cm 2 /h. Lesions localization and diameter had a signification impact on the outcomes. After 76 procedures, en bloc resection rate of 90.9 and 90.67% were achieved for the left side of colon and rectum, respectively. In the right side of colon statistically significant lower resection rate of 67.57% was observed. We have proved that in the setting of the Western center, colorectal ESD can yield excellent results. It seems that the key to the success during the learning period is 'tailoring' lesions qualification guidelines to the experience of the endoscopist, as lesions diameter and localization highly influence the outcomes.

  12. Is the resulting phenotype of an embryo with balanced X-autosome translocation, obtained by means of preimplantation genetic diagnosis, linked to the X inactivation pattern?

    Science.gov (United States)

    Ferfouri, Fatma; Bernicot, Izabel; Schneider, Anouck; Haquet, Emmanuelle; Hédon, Bernard; Anahory, Tal

    2016-04-01

    To examine if a balanced female embryo with X-autosome translocation could, during its subsequent development, express an abnormal phenotype. Preimplantation genetic diagnosis (PGD) analysis on two female carriers with maternal inherited X-autosome translocations. Infertility center and genetic laboratory in a public hospital. Two female patients carriers undergoing PGD for a balanced X-autosome translocations: patient 1 with 46,X,t(X;2)(q27;p15) and patient 2 with 46,X,t(X;22)(q28;q12.3). PGD for balanced X-autosome translocations. PGD outcomes, fluorescence in situ hybridization in biopsied embryos and meiotic segregation patterns analysis of embryos providing from X-autosome translocation carriers. Controlled ovarian stimulation facilitated retrieval of a correct number of oocytes. One balanced embryo per patient was transferred and one developed, but the patient miscarried after 6 weeks of amenorrhea. In X-autosome translocation carriers, balanced Y-bearing embryos are most often phenotypically normal and viable. An ambiguous phenotype exists in balanced X-bearing embryos owing to the X inactivation mechanism. In 46,XX embryos issued from an alternate segregation, der(X) may be inactivated and partially spread transcriptional silencing into a translocated autosomal segment. Thus, the structural unbalanced genotype could be turned into a viable functional balanced one. It is relevant that a discontinuous silencing is observed with a partial and unpredictable inactivation of autosomal regions. Consequently, the resulting phenotype remains a mystery and is considered to be at risk of being an abnormal phenotype in the field of PGD. It is necessary to be cautious regarding to PGD management for this type of translocation, particularly in transferred female embryos. Copyright © 2016 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  13. Discordant results obtained with Francisella tularensis during in vitro and in vivo immunological studies are attributable to compromised bacterial structural integrity.

    Directory of Open Access Journals (Sweden)

    Anju Singh

    Full Text Available Francisella tularensis (Ft is a highly infectious intracellular pathogen and the causative agent of tularemia. Because Ft can be dispersed via small droplet-aerosols and has a very low infectious dose it is characterized as a category A Select Agent of biological warfare. Respiratory infection with the attenuated Live Vaccine Strain (LVS and the highly virulent SchuS4 strain of Ft engenders intense peribronchiolar and perivascular inflammation, but fails to elicit select pro-inflammatory mediators (e.g., TNF, IL-1β, IL-6, IL-12, and IFN-γ within the first ~72 h. This in vivo finding is discordant with the principally TH1-oriented response to Ft frequently observed in cell-based studies wherein the aforementioned cytokines are produced. An often overlooked confounding factor in the interpretation of experimental results is the influence of environmental cues on the bacterium's capacity to elicit certain host responses. Herein, we reveal that adaptation of Ft to its mammalian host imparts an inability to elicit select pro-inflammatory mediators throughout the course of infection. Furthermore, in vitro findings that non-host adapted Ft elicits such a response from host cells reflect aberrant recognition of the DNA of structurally-compromised bacteria by AIM2-dependent and -independent host cell cytosolic DNA sensors. Growth of Ft in Muller-Hinton Broth or on Muller-Hinton-based chocolate agar plates or genetic mutation of Ft was found to compromise the structural integrity of the bacterium thus rendering it capable of aberrantly eliciting pro-inflammatory mediators (e.g., TNF, IL-1β, IL-6, IL-12, and IFN-γ. Our studies highlight the profound impact of different growth conditions on host cell response to infection and demonstrate that not all in vitro-derived findings may be relevant to tularemia pathogenesis in the mammalian host. Rational development of a vaccine and immunotherapeutics can only proceed from a foundation of knowledge based upon

  14. Results from Mobile Lab Measurements Obtained in the Barnett Shale with Emphasis on Methane and Gaseous Mercury Emissions (Fort Worth, TX)

    Science.gov (United States)

    Laine, P. L.; Lan, X.; Anderson, D.; Talbot, R. W.

    2013-12-01

    Our work is part of a comprehensive analysis conducted through a collaboration of ground based measurements and airborne measurements with several research groups in order to gain a better understanding of methane and mercury emissions in the Barnett Shale. It's a vast rock formation that sits in the 5,000 square miles surrounding the Fort Worth area. To get the gas to market requires an underground highway of pipelines and compression stations. Texas state records show that since 2000 the number of gas compressors in the Barnett Shale has tripled (from a few hundred to 1,300), and they're ever infringing on populated areas. Recent preliminary data reported by Pétron et al. and Tollefson et al. (from the natural-gas operations in Denver-Julesburg Basin) point to CH4 loss from the process of 4-8%, not including additional losses in the pipeline and distribution system. Additionally, Howarth et al. have conducted a comprehensive analysis of greenhouse gases (methane, in particular) emitted from shale gas as a result of hydraulic fracturing and they estimate up to 8% of all natural gas mined from shale formations leaks to the atmosphere. Not only is this cause for alarm due to the global warming potential of methane, but we would expect similar losses of additional (potentially harmful) gases, i.e., atmospheric Hg, from the extraction systems. These preliminary findings are higher than the current U.S. Environment Protection Agency (EPA) leakage estimate of 2.3 percent. Our strategy employs the use of our mobile laboratory, a four door Chevrolet Silverado pickup truck with a camper shell, outfitted with trace gas instrumentation including a Picarro G2132i and a Tekran 2537 mercury analyzer. The Picarro cavity ring down instrument has high precision and accuracy H2O, CO2, CH4, and 13δC in CH4 and CO2 with very little drift due to precise temperature and pressure controls. The Tekran mercury analyzer allows for accurate total gaseous mercury measurements via

  15. Integration of replication-defective R68.45-like plasmids into the Pseudomonas aeruginosa chromosome.

    Science.gov (United States)

    Reimmann, C; Rella, M; Haas, D

    1988-06-01

    R68.45 and other similar broad-host-range (IncP) plasmids carrying a tandem repeat of the 2.1 kb insertion element IS21 mobilize the chromosome of many different Gram-negative bacteria. To analyse the structure of R68.45-chromosome cointegrates, whose involvement in the mobilization process had been postulated previously, we selected for the stable integration of R68.45-like plasmids into the Pseudomonas aeruginosa chromosome. Two plasmids were chosen: pME28, a transfer-deficient, mobilizable RP1 derivative with an inactive replication control (trfA) gene, and pME487, an R68.45 derivative with a trfA(ts) mutation causing temperature-sensitive replication. Chromosomally integrated pME28 and pME487 were found to be flanked by single IS21 elements. This structure is in agreement with a 'cut-and-paste' mode of R68.45 transposition. pME28 and pME487 showed a low specificity of insertion but rarely (less than 0.1%) induced auxotrophic mutations. Hfr (high-frequency-of-recombination) donors of P. aeruginosa could be obtained by chromosomal integration of pME487 or pME28; in the latter case, the transfer functions lacking from pME28 had to be provided in trans on an autonomous plasmid. Hfr donors gave higher conjugational linkage and transferred longer stretches of the P. aeruginosa chromosome than did R68.45 donors. This suggests that the integration of R68.45 into the donor chromosome is short-lived in P. aeruginosa.

  16. Genetic islands in pome fruit pathogenic and nonpathogenic Erwinia species and related plasmids

    Directory of Open Access Journals (Sweden)

    Pablo eLlop

    2015-08-01

    Full Text Available New pathogenic bacteria species belonging to the genus Erwinia associated with pome fruit trees (Erwinia pyrifoliae, E. piriflorinigrans, E. uzenensis have been increasingly described in the last years, and comparative analyses have found that all these species share several genetic characteristics. Studies at different level (whole genome comparison, virulence genes, plasmid content, etc. show a high intraspecies homogeneity (i.e. among E. amylovora strains and also abundant similarities appear between the different Erwinia species: presence of plasmids of similar size in the pathogenic species; high similarity in several genes associated with exopolysaccharide production and hence, with virulence, as well as in some other genes, in the chromosomes. Many genetic similarities have been observed also among some of the plasmids (and genomes from the pathogenic species and E. tasmaniensis or E. billingiae, two epiphytic species on the same hosts. The amount of genetic material shared in this genus varies from individual genes to clusters, genomic islands and genetic material that even may constitute a whole plasmid. Recent research on evolution of erwinias point out the horizontal transfer acquisition of some genomic islands that were subsequently lost in some species and several pathogenic traits that are still present. How this common material has been obtained and is efficiently maintained in different species belonging to the same genus sharing a common ecological niche provides an idea of the origin and evolution of the pathogenic Erwinia and the interaction with nonpathogenic species present in the same niche, and the role of the genes that are conserved in all of them.

  17. Circulation of a multiresistant, conjugative, IncA/C plasmid within the nosocomial Providencia stuartii population in the Athens area.

    Science.gov (United States)

    Giakkoupi, Panagiota; Tryfinopoulou, Kyriaki; Polemis, Michalis; Pappa, Olga; Miriagou, Vivi; Vatopoulos, Alkiviadis

    2015-05-01

    The objective of the study is to report a multidrug-resistant outbreak of Providencia stuartii that occurred in inpatients in the Athens area in 2012 resulting from a very successful transmissible A/C multidrug-resistant plasmid. Thirteen multidrug-resistant P. stuartii clinical isolates from 5 hospitals were studied. Molecular typing was performed by pulsed-field gel electrophoresis. Antibiotic resistance genes and their genetic surround were detected by PCR and sequencing. Plasmid analysis included conjugation experiments using liquid cultures, sizing by S1 digestion, and incompatibility replicon typing by PCR. Isolates were grouped into 2 distinct clonal types A and B, exhibiting similarity less than 70%. Isolates of type A were recovered from patients hospitalized in 4 different hospitals with no obvious epidemiological linkage, while isolates of type B were recovered from patients treated in a single hospital. Both clonal types harbored a conjugative plasmid of 130 bp and IncA/C replicon type carrying 5 β-lactamase genes bla(SHV-5), bla(VEB-1), bla(VIM-1), bla(OXA-10), and bla(TEM-1) and aminoglycosides resistant determinants. All β-lactamase genes were included in stable structures as IS26, IS1999, and In-e541. The current plasmid seemed to have many common determinants with previously reported plasmids derived from P. stuartii and Proteus mirabilis clinical isolates and exhibited the ability to circulate in nosocomial bacterial populations. Copyright © 2015 Elsevier Inc. All rights reserved.

  18. Characterization of Shigella Strains by Plasmid Profile Analysis and Antibiotic Susceptibility Patterns in a Pediatric Hospital in Ahvaz

    Directory of Open Access Journals (Sweden)

    Amin Sakhaei

    2015-11-01

    Full Text Available Background: High incidences of dysentery and diarrhea were reported in a pediatric hospital in Ahvaz, Iran during March to April, 2013. Objectives: A cross-sectional study was therefore undertaken to identify the causative agents. Patients and Methods: A total of 230 diarrhea samples were collected from the patients and analyzed by routine bacteriological methods. Bacterial identification, serological assay, antimicrobial susceptibility testing, extended spectrum β-lactamases (ESBLs screening and plasmid profile analysis were performed according to the standard guidelines. Results: A total of 70 Shigella strains including %70 (n = 49 S. sonnei and 30% (n = 21 S. flexneri were isolated from diarrhea samples. Most of the Shigella isolates showed high degrees of resistance to ampicillin, ulafamethoxazole- trimethoprime and cefexim. Concurrent resistance to sulafametoxazole- trimethoprime and ampicillin was the most common resistance pattern. Overall, 11.4% of Shigella isolates showed the ESBL producer criteria. The plasmid profile patterns of all the strains were determined by a modified alkaline lysis method. By plasmid profile analysis 23 genotypes were identified among all the isolates, 14 and 9 genotypes among the S. sonnei and S. Flexneri respectively. S. sonnei and S. flexneri isolates demonstrated unique plasmid profiles. Conclusions: These data demonstrated that S. sonnei strains are the main cause of shigellosis as the prevalent Shigella serotype in Iran. We also found that the antibiotic resistance rates are increasing among Shigella strains. Plasmid profile analysis is more reliable than antibiotic susceptibility patterns in epidemiologic studies.

  19. Results obtained during wall breaching research

    CSIR Research Space (South Africa)

    Hattingh, S

    2008-11-01

    Full Text Available To understand the physics of what is happening inside the wall directly after the detonation and the application of this knowledge in the improvement of the charge Measure the shock/stress waves in the masonry material and then in the wall as a whole... to maximise the effect of the charges on the walls and to broaden the knowledge of the physics of shock and stress waves. The thickness and characteristics of walls are not usually known in an operation. The effect of the charges on real buildings is still...

  20. Characterization of replication and conjugation of plasmid pWTY27 from a widely distributed Streptomyces species

    Directory of Open Access Journals (Sweden)

    Wang Tao

    2012-11-01

    Full Text Available Abstract Background Streptomyces species are widely distributed in natural habitats, such as soils, lakes, plants and some extreme environments. Replication loci of several Streptomyces theta-type plasmids have been reported, but are not characterized in details. Conjugation loci of some Streptomyces rolling-circle-type plasmids are identified and mechanism of conjugal transferring are described. Results We report the detection of a widely distributed Streptomyces strain Y27 and its indigenous plasmid pWTY27 from fourteen plants and four soil samples cross China by both culturing and nonculturing methods. The complete nucleotide sequence of pWTY27 consisted of 14,288 bp. A basic locus for plasmid replication comprised repAB genes and an adjacent iteron sequence, to a long inverted-repeat (ca. 105 bp of which the RepA protein bound specifically in vitro, suggesting that RepA may recognize a second structure (e.g. a long stem-loop of the iteron DNA. A plasmid containing the locus propagated in linear mode when the telomeres of a linear plasmid were attached, indicating a bi-directional replication mode for pWTY27. As for rolling-circle plasmids, a single traA gene and a clt sequence (covering 16 bp within traA and its adjacent 159 bp on pWTY27 were required for plasmid transfer. TraA recognized and bound specifically to the two regions of the clt sequence, one containing all the four DC1 of 7 bp (TGACACC and one DC2 (CCCGCCC and most of IC1, and another covering two DC2 and part of IC1, suggesting formation of a high-ordered DNA-protein complex. Conclusions This work (i isolates a widespread Streptomyces strain Y27 and sequences its indigenous theta-type plasmid pWTY27; (ii identifies the replication and conjugation loci of pWTY27 and; (iii characterizes the binding sequences of the RepA and TraA proteins.

  1. Characterization of replication and conjugation of plasmid pWTY27 from a widely distributed Streptomyces species

    Science.gov (United States)

    2012-01-01

    Background Streptomyces species are widely distributed in natural habitats, such as soils, lakes, plants and some extreme environments. Replication loci of several Streptomyces theta-type plasmids have been reported, but are not characterized in details. Conjugation loci of some Streptomyces rolling-circle-type plasmids are identified and mechanism of conjugal transferring are described. Results We report the detection of a widely distributed Streptomyces strain Y27 and its indigenous plasmid pWTY27 from fourteen plants and four soil samples cross China by both culturing and nonculturing methods. The complete nucleotide sequence of pWTY27 consisted of 14,288 bp. A basic locus for plasmid replication comprised repAB genes and an adjacent iteron sequence, to a long inverted-repeat (ca. 105 bp) of which the RepA protein bound specifically in vitro, suggesting that RepA may recognize a second structure (e.g. a long stem-loop) of the iteron DNA. A plasmid containing the locus propagated in linear mode when the telomeres of a linear plasmid were attached, indicating a bi-directional replication mode for pWTY27. As for rolling-circle plasmids, a single traA gene and a clt sequence (covering 16 bp within traA and its adjacent 159 bp) on pWTY27 were required for plasmid transfer. TraA recognized and bound specifically to the two regions of the clt sequence, one containing all the four DC1 of 7 bp (TGACACC) and one DC2 (CCCGCCC) and most of IC1, and another covering two DC2 and part of IC1, suggesting formation of a high-ordered DNA-protein complex. Conclusions This work (i) isolates a widespread Streptomyces strain Y27 and sequences its indigenous theta-type plasmid pWTY27; (ii) identifies the replication and conjugation loci of pWTY27 and; (iii) characterizes the binding sequences of the RepA and TraA proteins. PMID:23134842

  2. Sorbitol-Fermenting Enterohemorrhagic Escherichia coli O157:H- Isolates from Czech Patients with Novel Plasmid Composition Not Previously Seen in German Isolates.

    Science.gov (United States)

    Bauwens, Andreas; Marejková, Monika; Middendorf-Bauchart, Barbara; Prager, Rita; Kossow, Annelene; Zhang, Wenlan; Karch, Helge; Mellmann, Alexander; Bielaszewska, Martina

    2017-12-01

    Sorbitol-fermenting (SF) enterohemorrhagic Escherichia coli (EHEC) O157:H - strains, first identified in Germany, have emerged as important pathogens throughout Europe. Besides chromosomally encoded Shiga toxin 2a (the major virulence factor), several putative virulence loci, including the hly , etp , and sfp operons, encoding EHEC hemolysin, type II secretion system proteins, and Sfp fimbriae, respectively, are located on the 121-kb plasmid pSFO157 in German strains. Here we report novel SF EHEC O157:H - strains isolated from patients in the Czech Republic. These strains share the core genomes and chromosomal virulence loci encoding toxins ( stx 2a and the cdtV -ABC operon) and adhesins ( eae -γ, efa1 , lpfA O157OI-141 , and lpfA O157OI-154 ) with German strains but differ essentially in their plasmids. In contrast to all previously detected SF EHEC O157:H - strains, the Czech strains carry two plasmids, of 79 kb and 86 kb. The 79-kb plasmid harbors the sfp operon, but neither of the plasmids contains the hly and etp operons. Sequence analyses demonstrated that the 79-kb plasmid (pSFO157 258/98-1) evolved from pSFO157 of German strains by deletion of a 41,534-bp region via homologous recombination, resulting in loss of the hly and etp operons. The 86-kb plasmid (pSFO157 258/98-2) displays 98% sequence similarity to a 92.7-kb plasmid of an extraintestinal pathogenic E. coli bloodstream isolate. Our finding of this novel plasmid composition in SF EHEC O157:H - strains extends the evolutionary history of EHEC O157 plasmids. Moreover, the unique molecular plasmid characteristics permit the identification of such strains, thereby facilitating further investigations of their geographic distribution, clinical significance, and epidemiology. IMPORTANCE Since their first identification in Germany in 1989, sorbitol-fermenting enterohemorrhagic Escherichia coli O157:H - (nonmotile) strains have emerged as important causes of the life-threatening disease hemolytic

  3. Sorbitol-Fermenting Enterohemorrhagic Escherichia coli O157:H− Isolates from Czech Patients with Novel Plasmid Composition Not Previously Seen in German Isolates

    Science.gov (United States)

    Bauwens, Andreas; Marejková, Monika; Middendorf-Bauchart, Barbara; Prager, Rita; Kossow, Annelene; Zhang, Wenlan; Karch, Helge

    2017-01-01

    ABSTRACT Sorbitol-fermenting (SF) enterohemorrhagic Escherichia coli (EHEC) O157:H− strains, first identified in Germany, have emerged as important pathogens throughout Europe. Besides chromosomally encoded Shiga toxin 2a (the major virulence factor), several putative virulence loci, including the hly, etp, and sfp operons, encoding EHEC hemolysin, type II secretion system proteins, and Sfp fimbriae, respectively, are located on the 121-kb plasmid pSFO157 in German strains. Here we report novel SF EHEC O157:H− strains isolated from patients in the Czech Republic. These strains share the core genomes and chromosomal virulence loci encoding toxins (stx2a and the cdtV-ABC operon) and adhesins (eae-γ, efa1, lpfAO157OI-141, and lpfAO157OI-154) with German strains but differ essentially in their plasmids. In contrast to all previously detected SF EHEC O157:H− strains, the Czech strains carry two plasmids, of 79 kb and 86 kb. The 79-kb plasmid harbors the sfp operon, but neither of the plasmids contains the hly and etp operons. Sequence analyses demonstrated that the 79-kb plasmid (pSFO157 258/98-1) evolved from pSFO157 of German strains by deletion of a 41,534-bp region via homologous recombination, resulting in loss of the hly and etp operons. The 86-kb plasmid (pSFO157 258/98-2) displays 98% sequence similarity to a 92.7-kb plasmid of an extraintestinal pathogenic E. coli bloodstream isolate. Our finding of this novel plasmid composition in SF EHEC O157:H− strains extends the evolutionary history of EHEC O157 plasmids. Moreover, the unique molecular plasmid characteristics permit the identification of such strains, thereby facilitating further investigations of their geographic distribution, clinical significance, and epidemiology. IMPORTANCE Since their first identification in Germany in 1989, sorbitol-fermenting enterohemorrhagic Escherichia coli O157:H− (nonmotile) strains have emerged as important causes of the life-threatening disease hemolytic

  4. THE ENDOGENOUS BACILLUS-SUBTILIS (NATTO) PLASMIDS PTA1015 AND PTA1040 CONTAIN SIGNAL PEPTIDASE-ENCODING GENES - IDENTIFICATION OF A NEW STRUCTURAL MODULE ON CRYPTIC PLASMIDS

    NARCIS (Netherlands)

    MEIJER, WJJ; DEJONG, A; BEA, G; WISMAN, A; TJALSMA, H; VENEMA, G; BRON, S; MAARTEN, J; VANDIJL, JM

    Various strains of Bacillus subtilis (natto) contain small cryptic plasmids that replicate via the rolling-circle mechanism. Like plasmids from other Gram-positive bacteria, these plasmids are composed of several distinct structural modules. A new structural module was identified on the B. subtilis

  5. Dissemination of NDM-1-Producing Enterobacteriaceae Mediated by the IncX3-Type Plasmid

    Science.gov (United States)

    Fu, Ying; Du, Xiaoxing; Shen, Yuqin; Yu, Yunsong

    2015-01-01

    The emergence and spread of NDM-1-producing Enterobacteriaceae have resulted in a worldwide public health risk that has affected some provinces of China. China is an exceptionally large country, and there is a crucial need to investigate the epidemic of bla NDM-1-positive Enterobacteriaceae in our province. A total of 186 carbapenem-resistant Enterobacteriaceae isolates (CRE) were collected in a grade-3 hospital in Zhejiang province. Carbapenem-resistant genes, including bla KPC, bla IMP, bla VIM, bla OXA-48 and bla NDM-1 were screened and sequenced. Ninety isolates were identified as harboring the bla KPC-2 genes, and five bla NDM-1-positive isolates were uncovered. XbaI-PFGE revealed that three bla NDM-1-positive K. pneumoniae isolates belonged to two different clones. S1-PFGE and southern blot suggested that the bla NDM-1 genes were located on IncX3-type plasmids with two different sizes ranging from 33.3 to 54.7 kb (n=4) and 104.5 to 138.9 kb (n=1), respectively, all of which could easily transfer to Escherichia coli by conjugation and electrotransformation. The high-throughput sequencing of two plasmids was performed leading to the identification of a smaller 54-kb plasmid, which had high sequence similarity with a previously reported pCFNDM-CN, and a larger plasmid in which only a 7.8-kb sequence of a common gene environment around bla NDM-1 (bla NDM-1-trpF- dsbC-cutA1-groEL-ΔInsE,) was detected. PCR mapping and sequencing demonstrated that four smaller bla NDM-1 plasmids contained a common gene environment around bla NDM-1 (IS5-bla NDM-1-trpF- dsbC-cutA1-groEL). We monitored the CRE epidemic in our hospital and determined that KPC-2 carbapenemase was a major risk to patient health and the IncX3-type plasmid played a vital role in the spread of the bla NDM-1 gene among the CRE. PMID:26047502

  6. Multilocus sequence typing of IncN plasmids

    DEFF Research Database (Denmark)

    García-Fernández, Aurora; Villa, Laura; Moodley, Arshnee

    2011-01-01

    that spread and persistence of this particular IncN-carrying blaVIM-1 lineage in Greece. CONCLUSIONS: This study proposes the use of pMLST as a suitable and rapid method for identification of IncN epidemic plasmid lineages. The recent spread of blaCTX-M-1 among humans and animals seems to be associated......OBJECTIVES: Incompatibility group N (IncN) plasmids have been associated with the dissemination of antimicrobial resistance and are a major vehicle for the spread of blaVIM-1 in humans and blaCTX-M-1 in animals. A plasmid multilocus sequence typing (pMLST) scheme was developed for rapid...... in different countries from both animals and humans belonged to ST1, suggesting dissemination of an epidemic plasmid through the food chain. Fifteen of 17 plasmids carrying blaVIM-1 from Klebsiella pneumoniae and Escherichia coli, isolated during a 5year period in Greece were assigned to ST10, suggesting...

  7. Use of sperm plasmid DNA lipofection combined with REMI (restriction enzyme-mediated insertion) for production of transgenic chickens expressing eGFP (enhanced green fluorescent protein) or human follicle-stimulating hormone.

    Science.gov (United States)

    Harel-Markowitz, Eliane; Gurevich, Michael; Shore, Laurence S; Katz, Adi; Stram, Yehuda; Shemesh, Mordechai

    2009-05-01

    Linearized p-eGFP (plasmid-enhanced green fluorescent protein) or p-hFSH (plasmid human FSH) sequences with the corresponding restriction enzyme were lipofected into sperm genomic DNA. Sperm transfected with p-eGFP were used for artificial insemination in hens, and in 17 out of 19 of the resultant chicks, the exogenous DNA was detected in their lymphocytes as determined by PCR and expressed in tissues as determined by (a) PCR, (b) specific emission of green fluorescence by the eGFP, and (c) Southern blot analysis. A complete homology was found between the Aequorea Victoria eGFP DNA and a 313-bp PCR product of extracted DNA from chick blood cells. Following insemination with sperm lipofected with p-hFSH, transgenic offspring were obtained for two generations as determined by detection of the transgene for human FSH (PCR) and expression of the gene (RT-PCR and quantitative real-time PCR) and the presence of the protein in blood (radioimmunoassay). Data demonstrate that lipofection of plasmid DNA with restriction enzyme is a highly efficient method for the production of transfected sperm to produce transgenic offspring by direct artificial insemination.

  8. Intrathecal injection of naked plasmid DNA provides long-term expression of secreted proteins.

    Science.gov (United States)

    Hughes, Travis S; Langer, Stephen J; Johnson, Kirk W; Chavez, Raymond A; Watkins, Linda R; Milligan, Erin D; Leinwand, Leslie A

    2009-01-01

    Therapeutic benefit has been reported to result from intrathecal (i.t.) injection of transgene vectors, including naked DNA. However, most studies using naked DNA have measured only the transgene expression of intracellular proteins. Here we demonstrate that i.t. injection of naked DNA can result in long-term expression of secreted proteins. Plasmids expressing either secreted alkaline phosphatase (SEAP) or human interleukin-10 (hIL-10) were injected into the i.t. space in rats, and transgene products were repeatedly measured in the cerebrospinal fluid (CSF). Both SEAP and hIL-10 were maximal at 1 and 2 days after the injection and still detectable at 4 months. The utilization of a plasmid having two features that are hypothesized to increase gene expression (matrix attachment regions (MARs) and lack of CpG dinucleotides) resulted in a significant increase in gene expression. Reinjection of SEAP or hIL-10 plasmids after 4 months significantly increased protein levels at 1 and 14 days after the reinjection. SEAP was uniformly distributed between the DNA delivery site (approximately vertebral level T13) and the lumbar puncture site (L5/L6 inter-vertebral space), was reduced at the cisterna magna, and was detectable, though at much lower levels, in serum. These data suggest that naked DNA has the potential to be used as a therapeutic tool for applications that require long-term release of transgenes into the CSF.

  9. Enhanced extraction and purification of plasmid DNA from escherichia coli by applying a hybrid magnetic nanocomposite

    Energy Technology Data Exchange (ETDEWEB)

    Silva, R.J. da; Chavez-Guajardo, A.E.; Medina-llamas, J.C.; Alcaraz-Espinoza, J.J.; Melo, C.P. de [Universidade Federal de Pernambuco (UFPE), PE (Brazil)

    2016-07-01

    Full text: Plasmid DNA (pDNA), a special kind of nucleic acid usually found in bacteria, is a small molecule physically distinct from chromosomal DNA that can replicate independently. This genetic material has been used in a wide set of biotechnological methodologies, such as genetic engineering, production of recombinant drugs and gene therapy, among others. In all these applications, the extraction and purification of pDNA appears as a crucial step. In this work, we describe the synthesis of a polyaniline and maghemite (PANI/?-Fe2O3) magnetic nanocomposite (MNC) and its use in a new Escherichia coli (E. coli) pDNA extraction and purification protocol. We have used transmission electron microscopy (TEM), UV-Vis spectroscopy, infrared spectroscopy (FTIR), X-ray diffraction (XRD), dynamic light scattering (DLS) and magnetic measurements to characterize the MNC, which was synthesized through an emulsion polymerization method. The yield, purity and quality of the pDNA extracted by using our proposed MNC protocol were evaluated through UV-Vis, agarose gel electrophoreses and PCR techniques, respectively. After comparing our results to those obtained by use of a commercial kit (Promega Wizard Plus SV Minipreps), we suggest that the novel protocol here proposed appears as a competitive alternative methodology. Not only the purification step can be completed within only 10 min, but the high adsorption capacity of the MNC results in pDNA yields that are almost twice the best values obtained by using the commercial kit. Hence, this new MNC methodology can be of general interest and find widespread use in different types of biomedical applications. (author)

  10. F14:A-:B- and IncX4 Inc group cfr-positive plasmids circulating in Escherichia coli of animal origin in Northeast China.

    Science.gov (United States)

    Wang, Xiumei; Zhu, Yao; Hua, Xin; Chen, Fuguang; Wang, Changzhen; Zhang, Yanhe; Liu, Siguo; Zhang, Wanjiang

    2018-04-01

    The objective of this study was to investigate the prevalence of the cfr gene in Escherichia coli isolates from domestic animals in Northeast China and to characterize the cfr-containing plasmids. Between June 2015 and April 2016, 370 E. coli isolates were collected from pigs, chickens, and dairy cows in Northeast China. Among these, 111 were florfenicol resistant, including 109 isolates carrying the floR gene and 6 positives for cfr. The prevalence of cfr in E. coli isolates from the four northeast provinces in China was 1.6% (6/370), which was higher than that previously reported (0.08% and 0.5%). All six cfr-containing E. coli isolates were highly resistant to florfenicol (100%), cefotaxime (100%), and fosfomycin (100%). Complete sequence analysis of two cfr-carrying plasmids revealed high homology of the IncX4-type pEC14cfr plasmid with two other cfr-harboring plasmids, pSD11 and pGXEC6, found in swine E. coli isolates from southern China. pEC14cfr-like plasmids have been isolated in five provinces in southern and northern China. The isolation sites were up to 2700 kilometers apart, implying that pEC14cfr-like plasmids are likely to be national epidemic cfr-carrying plasmids that mediate the dissemination of cfr in China. Moreover, the genetic structure (IS26-IS26-cfr-rec-pre/mob-ramA-IS26) of the second cfr-carrying plasmid, IncF14:A-:B- pEC295cfr, represents a novel genetic environment for cfr identified for the first time in the present study. Sequence homology analysis indicated that the cfr-carrying element was most likely introduced into a cfr-negative pEC12 plasmid backbone, which evolved into the cfr-carrying vector, pEC295cfr. Moreover, isolation of the IncF14:A-:B- pEC295cfr plasmid harboring cfr suggests that IncFII plasmids maybe have become additional effective vehicles for cfr dissemination. These results highlight the importance of surveying the prevalence of IncX4 and IncFII plasmids in gram-negative bacteria, especially in swine E. coli

  11. Effect of TiO2 on conjugative transfer of RP4 plasmid

    International Nuclear Information System (INIS)

    Qian Di; Zhang Buchang; Yang Dong; Chen Zhaoli; Jin Min; Qiu Zhigang; Li Junwen

    2013-01-01

    Objective: To explore the effect and law of nano-titanium dioxide on the conjugative transfer of RP4 plasmid. Methods: Mating was conducted between Escherichia coli HB101 (RP4) and E. coli K12Rif in saline without stirring under certain conditions and the donor per recipient ratio was 1:1 constantly. The selective LB agar medium plates containing appropriate antibiotics were used to count the number of transconjugants and the conjugative transfer frequency. Results: Nano-titanium dioxide could promote the conjugative transfer of RP4. The nano-titanium dioxide concentration, bacterial concentration, mating temperature and mating time could affect the conjugative transfer of RP4. Conclusion: Nano-titanium dioxide can promote plasmid conjugal transfer in the liquid phase under certain conditions, which may pose a potential hazard to environmental and human health. (authors)

  12. Effect of 8-MOP plus treatment on survival and repair of plasmid pBR322

    International Nuclear Information System (INIS)

    Bauluz, C.; Vidania, R.

    1992-01-01

    We have studied the lethality produced in pBR322 DNA after PUVA treatment (8-MOP+UVA). As recipients, we used a collection of E. coli strains differing in their repair capacities and analysed the involvement of several DNA repair pathways in the removal of plasmid lesions. We have also studied the effect of UVA radiation alone, in order to determine more precisely the effect attributable only to psoralen molecules. Results showed a strong lethal effect derived from PUVA treatment; however, some plasmid recovery was achieved in bacterial hosts proficient in Excision repair and SOS repair. another repair pathway, only detectable at high density of lesions, appeared to be relevant for the removal of 8-MOP:DNA adducts. (author)

  13. Human Intestinal Cells Modulate Conjugational Transfer of Multidrug Resistance Plasmids between Clinical Escherichia coli Isolates

    DEFF Research Database (Denmark)

    Machado, Ana Manuel; Sommer, Morten

    2014-01-01

    Bacterial conjugation in the human gut microbiota is believed to play a major role in the dissemination of antibiotic resistance genes and virulence plasmids. However, the modulation of bacterial conjugation by the human host remains poorly understood and there is a need for controlled systems...... to study this process. We established an in vitro co-culture system to study the interaction between human intestinal cells and bacteria. We show that the conjugation efficiency of a plasmid encoding an extended spectrum beta-lactamase is reduced when clinical isolates of Escherichia coli are co...... of the intestinal cells exposed to bacteria leading to a two-fold reduction in conjugation efficiency. These results show that human gut epithelial cells can modulate bacterial conjugation and may have relevance to gene exchange in the gut....

  14. A binary plasmid system for shuffling combinatorial antibody libraries.

    Science.gov (United States)

    Collet, T A; Roben, P; O'Kennedy, R; Barbas, C F; Burton, D R; Lerner, R A

    1992-11-01

    We have used a binary system of replicon-compatible plasmids to test the potential for promiscuous recombination of heavy and light chains within sets of human Fab fragments isolated from combinatorial antibody libraries. Antibody molecules showed a surprising amount of promiscuity in that a particular heavy chain could recombine with multiple light chains with retention of binding to a protein antigen. The degree to which a given heavy chain productively paired with any light chain to bind antigen varied from 43% to 100% and depended strongly on the heavy-chain sequence. Such productive crosses resulted in a set of Fab fragments of similar apparent binding constants, which seemed to differ mainly in the amount of active Fab fragment produced in the bacterial cell. The dominance of the heavy chain in the antibody-antigen interaction was further explored in a set of directed crosses, in which heavy and light chains derived from antigen-specific clones were crossed with nonrelated heavy and light chains. In these crosses, an Fab fragment retained antigen binding only if it contained a heavy chain from an antigen-specific clone. In no case did the light chain confer detectable affinity when paired with indifferent heavy chains. The surprising promiscuity of heavy chains has ramifications for the evaluation of the diversity of combinatorial libraries made against protein antigens and should allow the combination of one such promiscuous heavy chain with an engineered light chain to form an Fab fragment carrying synthetic cofactors to assist in antibody catalysis.

  15. Development of electrochemical reporter assay using HeLa cells transfected with vector plasmids encoding various responsive elements

    Energy Technology Data Exchange (ETDEWEB)

    Shiku, Hitoshi, E-mail: shiku@bioinfo.che.tohoku.ac.jp [Graduate School of Environmental Studies, Tohoku University, 6-6-11-604 Aramaki-Aoba, Sendai 980-8579 (Japan); Takeda, Michiaki; Murata, Tatsuya [Graduate School of Environmental Studies, Tohoku University, 6-6-11-604 Aramaki-Aoba, Sendai 980-8579 (Japan); Akiba, Uichi; Hamada, Fumio [Graduate School of Engineering and Resource Science, Akita University, 1-1 Tegata gakuen-machi, Akita 010-8502 (Japan); Matsue, Tomokazu, E-mail: matsue@bioinfo.che.tohoku.ac.jp [Graduate School of Environmental Studies, Tohoku University, 6-6-11-604 Aramaki-Aoba, Sendai 980-8579 (Japan)

    2009-04-27

    Electrochemical assay using HeLa cell lines transfected with various plasmid vectors encoding SEAP (secreted alkaline phosphatase) as the reporter has been performed by using SECM (scanning electrochemical microscopy). The plasmid vector contains different responsive elements that include GRE (glucocorticoid response elements), CRE (cAMP responsive elements), or {kappa}B (binding site for NF{kappa}B (nuclear factor kappa B)) upstream of the SEAP sequence. The transfected HeLa cells were patterned on a culture dish in a 4 x 4 array of circles of diameter 300 {mu}m by using the PDMS (poly(dimethylsiloxane)) stencil technique. The cellular array was first exposed to 100 ng mL{sup -1} dexamethasone, 10 ng mL{sup -1} forskolin, or 100 ng mL{sup -1} TNF-{alpha} (tumor necrosis factor {alpha}) after which it was further cultured in an RPMI culture medium for 6 h. After incubation, the cellular array was soaked in a measuring solution containing 4.7 mM PAPP (p-aminophenylphosphate) at pH 9.5, following which electrochemical measurements were performed immediately within 40 min. The SECM method allows parallel evaluation of different cell lines transfected with pGRE-SEAP, pCRE-SEAP, and pNF{kappa}B-SEAP patterned on the same solid support for detection of the oxidation current of PAP (p-aminophenol) flux produced from only 300 HeLa cells in each stencil pattern. The results of the SECM method were highly sensitive as compared to those obtained from the conventional CL (chemiluminescence) protocol with at least 5 x 10{sup 4} cells per well.

  16. Microarray-based analysis of IncA/C plasmid-associated genes from multidrug-resistant Salmonella enterica.

    Science.gov (United States)

    Lindsey, Rebecca L; Frye, Jonathan G; Fedorka-Cray, Paula J; Meinersmann, Richard J

    2011-10-01

    In the family Enterobacteriaceae, plasmids have been classified according to 27 incompatibility (Inc) or replicon types that are based on the inability of different plasmids with the same replication mechanism to coexist in the same cell. Certain replicon types such as IncA/C are associated with multidrug resistance (MDR). We developed a microarray that contains 286 unique 70-mer oligonucleotide probes based on sequences from five IncA/C plasmids: pYR1 (Yersinia ruckeri), pPIP1202 (Yersinia pestis), pP99-018 (Photobacterium damselae), pSN254 (Salmonella enterica serovar Newport), and pP91278 (Photobacterium damselae). DNA from 59 Salmonella enterica isolates was hybridized to the microarray and analyzed for the presence or absence of genes. These isolates represented 17 serovars from 14 different animal hosts and from different geographical regions in the United States. Qualitative cluster analysis was performed using CLUSTER 3.0 to group microarray hybridization results. We found that IncA/C plasmids occurred in two lineages distinguished by a major insertion-deletion (indel) region that contains genes encoding mostly hypothetical proteins. The most variable genes were represented by transposon-associated genes as well as four antimicrobial resistance genes (aphA, merP, merA, and aadA). Sixteen mercury resistance genes were identified and highly conserved, suggesting that mercury ion-related exposure is a stronger pressure than anticipated. We used these data to construct a core IncA/C genome and an accessory genome. The results of our studies suggest that the transfer of antimicrobial resistance determinants by transfer of IncA/C plasmids is somewhat less common than exchange within the plasmids orchestrated by transposable elements, such as transposons, integrating and conjugative elements (ICEs), and insertion sequence common regions (ISCRs), and thus pose less opportunity for exchange of antimicrobial resistance.

  17. Origin and Evolution of Nitrogen Fixation Genes on Symbiosis Islands and Plasmid in Bradyrhizobium

    Science.gov (United States)

    Okubo, Takashi; Piromyou, Pongdet; Tittabutr, Panlada; Teaumroong, Neung; Minamisawa, Kiwamu

    2016-01-01

    The nitrogen fixation (nif) genes of nodule-forming Bradyrhizobium strains are generally located on symbiosis islands or symbiosis plasmids, suggesting that these genes have been transferred laterally. The nif genes of rhizobial and non-rhizobial Bradyrhizobium strains were compared in order to infer the evolutionary histories of nif genes. Based on all codon positions, the phylogenetic tree of concatenated nifD and nifK sequences showed that nifDK on symbiosis islands formed a different clade from nifDK on non-symbiotic loci (located outside of symbiosis islands and plasmids) with elongated branches; however, these genes were located in close proximity, when only the 1st and 2nd codon positions were analyzed. The guanine (G) and cytosine (C) content of the 3rd codon position of nifDK on symbiosis islands was lower than that on non-symbiotic loci. These results suggest that nif genes on symbiosis islands were derived from the non-symbiotic loci of Bradyrhizobium or closely related strains and have evolved toward a lower GC content with a higher substitution rate than the ancestral state. Meanwhile, nifDK on symbiosis plasmids clustered with nifDK on non-symbiotic loci in the tree representing all codon positions, and the GC content of symbiotic and non-symbiotic loci were similar. These results suggest that nif genes on symbiosis plasmids were derived from the non-symbiotic loci of Bradyrhizobium and have evolved with a similar evolutionary pattern and rate as the ancestral state. PMID:27431195

  18. Genetic characterization of plasmid pRJ5 of Staphylococcus aureus compared to plasmid pE194

    International Nuclear Information System (INIS)

    Oliveira, S.S. de; Freire Bastos, M.C. de

    1993-01-01

    The pRJ5, a naturally occurring constitutive macrolide, lincosamide and streptogramin B (MLS) resistance plasmid of Staphylococcus aureus, was compared to pE194, a plasmid that confers the inducible phenotype. pRJ5 was stable in all strains of S. aureus tested, even under growth at 43 O C, which distinguished it from pE194 which was shown to be thermo-sensitive for replication. pRJ5, like pE194, was highly unstable in Bacillus subtilis when the cells were grown in nonselective conditions. Multimeric forms of pRJ5 DNA were detected in the few cells of B. subtilis that retained this plasmid. pE194 was transduced by phages φ 11 and φ 443 at frequencies 400 and 20-fold higher, respectively, than pRJ5. Both plasmids were co-transduced with the plasmid pRJ4. pRJ5 was shown to be compatible with pE194. Therefore they belong to distinct Inc groups. Hybridization studies revealed that pRJ5 shares a 1.35 kb region of homology to pE194, which is limited to the erm gene, conferring MLS resistance. (author)

  19. Acquisition of Carbapenem Resistance by Plasmid-Encoded-AmpC-Expressing Escherichia coli.

    Science.gov (United States)

    van Boxtel, Ria; Wattel, Agnes A; Arenas, Jesús; Goessens, Wil H F; Tommassen, Jan

    2017-01-01

    Although AmpC β-lactamases can barely degrade carbapenems, if at all, they can sequester them and prevent them from reaching their targets. Thus, carbapenem resistance in Escherichia coli and other Enterobacteriaceae can result from AmpC production and simultaneous reduction of antibiotic influx into the periplasm by mutations in the porin genes. Here we investigated the route and genetic mechanisms of acquisition of carbapenem resistance in a clinical E. coli isolate carrying bla CMY-2 on a plasmid by selecting for mutants that are resistant to increasing concentrations of meropenem. In the first step, the expression of OmpC, the only porin produced in the strain under laboratory conditions, was lost, leading to reduced susceptibility to meropenem. In the second step, the expression of the CMY-2 β-lactamase was upregulated, leading to resistance to meropenem. The loss of OmpC was due to the insertion of an IS1 element into the ompC gene or to frameshift mutations and premature stop codons in this gene. The bla CMY-2 gene was found to be located on an IncIγ plasmid, and overproduction of the CMY-2 enzyme resulted from an increased plasmid copy number due to a nucleotide substitution in the inc gene. The clinical relevance of these genetic mechanisms became evident from the analysis of previously isolated carbapenem-resistant clinical isolates, which appeared to carry similar mutations. Copyright © 2016 American Society for Microbiology.

  20. CIEMAT Interlaboratories Comparison of the Results obtained in the Proficiency Test Run by IAEA; Comparacion Interlaboratorios del CIEMAT de los Resultados Obtenidos en la Prueba de Capacitacion de Analisis de Transuranicos en Cenizas propocionadas por el OIEA

    Energy Technology Data Exchange (ETDEWEB)

    Gasco, C.; Anton, M. P.; Alvarez, A.; Navarro, N.; Meral, J.; Gonzalez, A.; Higueras Lafaja, E. [Ciemat. Madrid (Spain)

    2000-07-01

    This report contains the results obtained by two different laboratories from CIEMAT after participating in the Proficiency Test organised by IAEA (International Atomic Energy Agency) in 1999. This test involves the analysis of fly ashes containing natural radionuclides and different amounts of added transuranics. The extraction techniques, counting methods and results obtained are detailed. This type of test are used for the labs to achieve their accreditation and check the reliability of the procedures routinely employed. (Author) 4 refs.

  1. Mitochondrial pAL2-1 plasmid homologs are senescence factors in Podospora anserina independent of intrinsic senescence

    NARCIS (Netherlands)

    Diepeningen, van A.D.; Debets, A.J.M.; Slakhorst-Wandel, S.M.; Hoekstra, R.F.

    2008-01-01

    Since the first description of a linear mitochondrial plasmid in Podospora anserina, pAL2-1, and homologous plasmids have gone from being considered beneficial longevity plasmids, via neutral genetic elements, toward mutator plasmids causing senescence. The plasmid has an invertron structure, with

  2. Mitochondrial pAL2-1 plasmid homologs are senescence factors in Podospora anserina independent of intrinsic senescence

    NARCIS (Netherlands)

    van Diepeningen, Anne D; Debets, Alfons J M; Slakhorst, S Marijke; Hoekstra, Rolf F

    Since the first description of a linear mitochondrial plasmid in Podospora anserina, pAL2-1, and homologous plasmids have gone from being considered beneficial longevity plasmids, via neutral genetic elements, toward mutator plasmids causing senescence. The plasmid has an invertron structure, with

  3. Recombinogenic engineering of conjugative plasmids with fluorescent marker cassettes

    DEFF Research Database (Denmark)

    Reisner, A.; Molin, Søren; Zechner, E.L.

    2002-01-01

    An efficient approach for the insertion of fluorescent marker genes with sequence specificity into conjugative plasmids in Escherichia coli is described. For this purpose, homologous recombination of linear double-stranded targeting DNA was mediated by the bacteriophage lambda recombination...... resistance genes and fluorescent markers. The choice of 5' non-homologous extensions in primer pairs used for amplifying the marker cassettes determines the site specificity of the targeting DNA. This methodology is applicable to the modification of all plasmids that replicate in E coli and is not restricted...

  4. Novel assay to measure the plasmid mobilizing potential of mixed microbial communities

    DEFF Research Database (Denmark)

    Klümper, Uli; Droumpali, Ariadni; Dechesne, Arnaud

    2014-01-01

    Mobilizable plasmids lack necessary genes for complete conjugation and are therefore non-self-transmissible. Instead, they rely on the conjugation system of conjugal plasmids to be horizontally transferred to new recipients. While community permissiveness, the fraction of a mixed microbial...... community that can receive self-transmissible conjugal plasmids, has been studied, the intrinsic ability of a community to mobilize plasmids that lack conjugation systems is unexplored. Here, we present a novel framework and experimental method to estimate the mobilization potential of mixed communities. We...... of the donors receiving the conjugal plasmid in the first step. Further work is needed to establish how plasmid mobilization potential varies within and across microbial communities....

  5. Identification of pOENI-1 and Related Plasmids in Oenococcus oeni Strains Performing the Malolactic Fermentation in Wine

    Science.gov (United States)

    Favier, Marion; Bilhère, Eric; Lonvaud-Funel, Aline; Moine, Virginie; Lucas, Patrick M.

    2012-01-01

    Plasmids in lactic acid bacteria occasionally confer adaptive advantages improving the growth and behaviour of their host cells. They are often associated to starter cultures used in the food industry and could be a signature of their superiority. Oenococcus oeni is the main lactic acid bacteria species encountered in wine. It performs the malolactic fermentation that occurs in most wines after alcoholic fermentation and contributes to their quality and stability. Industrial O. oeni starters may be used to better control malolactic fermentation. Starters are selected empirically by virtue of their fermentation kinetics and capacity to survive in wine. This study was initiated with the aim to determine whether O. oeni contains plasmids of technological interest. Screening of 11 starters and 33 laboratory strains revealed two closely related plasmids, named pOENI-1 (18.3-kb) and pOENI-1v2 (21.9-kb). Sequence analyses indicate that they use the theta mode of replication, carry genes of maintenance and replication and two genes possibly involved in wine adaptation encoding a predicted sulphite exporter (tauE) and a NADH:flavin oxidoreductase of the old yellow enzyme family (oye). Interestingly, pOENI-1 and pOENI-1v2 were detected only in four strains, but this included three industrial starters. PCR screenings also revealed that tauE is present in six of the 11 starters, being probably inserted in the chromosome of some strains. Microvinification assays performed using strains with and without plasmids did not disclose significant differences of survival in wine or fermentation kinetics. However, analyses of 95 wines at different phases of winemaking showed that strains carrying the plasmids or the genes tauE and oye were predominant during spontaneous malolactic fermentation. Taken together, the results revealed a family of related plasmids associated with industrial starters and indigenous strains performing spontaneous malolactic fermentation that possibly

  6. Dissemination of imipenem-resistant Acinetobacter baumannii with new plasmid-borne bla(OXA-72) in Taiwan.

    Science.gov (United States)

    Kuo, Shu-Chen; Yang, Su-Pen; Lee, Yi-Tzu; Chuang, Han-Chuan; Chen, Chien-Pei; Chang, Chi-Ling; Chen, Te-Li; Lu, Po-Liang; Hsueh, Po-Ren; Fung, Chang-Phone

    2013-07-13

    The systemic surveillance of imipenem-resistant Acinetobacter baumannii (IRAB) from multicenters in Taiwan revealed the emergence of isolates with bla(OXA-72). This study described their genetic makeup, mechanism of spread, and contribution to carbapenem resistance. Two hundred and ninety-one non-repetitive isolates of A. baumannii were collected from 10 teaching hospitals from different geographical regions in Taiwan from June 2007 to September 2007. Minimal inhibitory concentrations (MICs) were determined by agar dilution. Clonality was determined by pulsed-field gel electrophoresis. Plasmid was extracted and digested by restriction enzymes, and subsequently analyzed by electrophoresis and Southern blot for bla(OXA-72). The flanking regions of bla(OXA-72) were determined by inverse PCR. The contribution of bla(OXA-72) to imipenem MIC was determined by transforming plasmids carrying bla(OXA-72) into imipenem-susceptible A. baumannii. Among 142 IRAB in Taiwan, 27 harbored bla(OXA-72); 22 originated from Southern Taiwan, 5 from Central Taiwan, and none from Northern Taiwan. There were two major clones. The bla(OXA-72) was identified in the plasmids of all isolates. Two genetic structures flanking plasmid-borne bla(OXA-72) were identified and shared identical sequences in certain regions; the one described in previous literature was present in only one isolate, and the new one was present in the remaining isolates. Introduction of bla(OXA-72) resulted in an increase of imipenem MIC in the transformants. The overexpression of bla(OXA-72) mRNA in response to imipenem further supported the contribution of bla(OXA-72). In conclusion, isolates with new plasmid-borne blaOXA-72 were found to be disseminated successfully in Southern Taiwan. The spread of the resistance gene depended on clonal spread and dissemination of a new plasmid. Bla(OXA-72) in these isolates directly led to their imipenem-resistance.

  7. Biofilm plasmids with a rhamnose operon are widely distributed determinants of the 'swim-or-stick' lifestyle in roseobacters.

    Science.gov (United States)

    Michael, Victoria; Frank, Oliver; Bartling, Pascal; Scheuner, Carmen; Göker, Markus; Brinkmann, Henner; Petersen, Jörn

    2016-10-01

    Alphaproteobacteria of the metabolically versatile Roseobacter group (Rhodobacteraceae) are abundant in marine ecosystems and represent dominant primary colonizers of submerged surfaces. Motility and attachment are the prerequisite for the characteristic 'swim-or-stick' lifestyle of many representatives such as Phaeobacter inhibens DSM 17395. It has recently been shown that plasmid curing of its 65-kb RepA-I-type replicon with >20 genes for exopolysaccharide biosynthesis including a rhamnose operon results in nearly complete loss of motility and biofilm formation. The current study is based on the assumption that homologous biofilm plasmids are widely distributed. We analyzed 33 roseobacters that represent the phylogenetic diversity of this lineage and documented attachment as well as swimming motility for 60% of the strains. All strong biofilm formers were also motile, which is in agreement with the proposed mechanism of surface attachment. We established transposon mutants for the four genes of the rhamnose operon from P. inhibens and proved its crucial role in biofilm formation. In the Roseobacter group, two-thirds of the predicted biofilm plasmids represent the RepA-I type and their physiological role was experimentally validated via plasmid curing for four additional strains. Horizontal transfer of these replicons was documented by a comparison of the RepA-I phylogeny with the species tree. A gene content analysis of 35 RepA-I plasmids revealed a core set of genes, including the rhamnose operon and a specific ABC transporter for polysaccharide export. Taken together, our data show that RepA-I-type biofilm plasmids are essential for the sessile mode of life in the majority of cultivated roseobacters.

  8. Characterization of IncN plasmids carrying blaCTX-M-1 and qnr genes in Escherichia coli and Salmonella from animals, the environment and humans

    DEFF Research Database (Denmark)

    Dolejska, Monika; Villa, Laura; Hasman, Henrik

    2013-01-01

    were compared using restriction fragment length polymorphism (RFLP), plasmid multilocus sequence typing (pMLST) and hybridization with repN, qnrS1, qnrB19 or blaCTX-M-1 probes. Plasmids pKT58A and pHHA45 were sequenced using the 454-Genome Sequencer FLX platform on a library constructed from plasmid...... DNA purified from the respective E. coli transformants.Results Three types of IncN plasmids carrying blaCTX-M-1, qnrS1 and qnrB19 genes were identified in strains isolated from the Czech Republic, Poland, Slovakia, Denmark, Italy and the Netherlands, corresponding to pMLST sequence type (ST) 1, ST3...

  9. Plasmid-mediated quinolone resistance in Salmonella serotypes isolated from chicken carcasses in Turkey

    Directory of Open Access Journals (Sweden)

    Zafer Ata

    2014-01-01

    Full Text Available Quinolones have been extensively used for treatment of a variety of invasive and systemic infections of salmonellosis. Widespread use of these agents has been associated with the emergence and dissemination of quinolone-resistant pathogens. The quinolone resistance and plasmid-mediated quinolone resistance determinants (qnrA, qnrB, qnrS and aac(6’-Ib-cr of 85 Salmonella isolates from chicken carcasses were investigated in this study. Isolates were serotyped according to the Kauffman-White-Le Minor scheme, and broth microdilution method was used to determine quinolone resistance. Plasmid-mediated quinolone resistance genes were investigated by real-time PCR and positive results were confirmed by sequencing. Among the Salmonella isolates, 30/85 (35% and 18/85 (21% were found to be resistant to enrofloxacin (MIC ≥ 2 mg/ml, and danofloxacin (MIC ≥ 2 mg/ml, respectively. All the isolates were negative for qnrA, qnrB and aac(6’-Ib-cr genes, nevertheless 2% (S. Brandenburg and S. Dabou were positive for qnrS (qnrS1 determinant. This study is the first and unique investigating the plasmid- mediated quinolone resistance determinants of Salmonella isolated from chicken carcasses in Turkey.

  10. Solid lipid nanoparticles mediate non-viral delivery of plasmid DNA to dendritic cells

    Science.gov (United States)

    Penumarthi, Alekhya; Parashar, Deepti; Abraham, Amanda N.; Dekiwadia, Chaitali; Macreadie, Ian; Shukla, Ravi; Smooker, Peter M.

    2017-06-01

    There is an increasing demand for novel DNA vaccine delivery systems, mainly for the non-viral type as they are considered relatively safe. Therefore, solid lipid nanoparticles (SLNs) were investigated for their suitability as a non-viral DNA vaccine delivery system. SLNs were synthesised by a modified solvent-emulsification method in order to study their potential to conjugate with plasmid DNA and deliver them in vitro to dendritic cells using eGFP as the reporter plasmid. The DNA-SLN complexes were characterised by electron microscopy, gel retardation assays and dynamic light scattering. The cytotoxicity assay data supported their biocompatibility and was used to estimate safe threshold concentration resulting in high transfection rate. The transfection efficiency of these complexes in a dendritic cell line was shown to increase significantly compared to plasmid alone, and was comparable to that mediated by lipofectamine. Transmission electron microscopy studies delineated the pathway of cellular uptake. Endosomal escape was observed supporting the mechanism of transfection.

  11. Protection of vanillin derivative VND3207 on plasmid DNA damage induced by different LET ionizing radiation

    International Nuclear Information System (INIS)

    Xu Huihui; Wang Li; Sui Li; Guan Hua; Wang Yu; Liu Xiaodan; Zhang Shimeng; Xu Qinzhi; Wang Xiao; Zhou Pingkun

    2011-01-01

    Objective: To evaluate the radioprotective effect of vanillin derivative VND3207 on DNA damage induced by different LET ionizing radiation. Methods: The plasmid DNA in liquid was irradiated by 60 Co γ-rays, proton or 7 Li heavy ion with or without VND3207. The conformation changes of plasmid DNA were assessed by agarose gel electrophoresis and the quantification was done using gel imaging system. Results: The DNA damage induced by proton and 7 Li heavy ion was much more serious as compared with that by 60 Co γ-rays, and the vanillin derivative VND3207 could efficiently decrease the DNA damage induced by all three types of irradiation sources, which was expressed as a significantly reduced ratio of open circular form (OC) of plasmid DNA. The radioprotective effect of VND3207 increased with the increasing of drug concentration. The protective efficiencies of 200 μmol/L VND3207 were 85.3% (t =3.70, P=0.033), 73.3% (t=10.58, P=0.017) and 80.4% (t=8.57, P=0.008) on DNA damage induction by 50 Gy of γ-rays, proton and 7 Li heavy ion, respectively. It seemed that the radioprotection of VND3207 was more effective on DNA damage induced by high LET heavy ion than that by proton. Conclusions: VND3207 has a protective effect against the genotoxicity of different LET ionizing radiation, especially for γ-rays and 7 Li heavy ion. (authors)

  12. Strand breaks and lethal damage in plasmid DNA subjected to 60CO-γirradiation

    International Nuclear Information System (INIS)

    Klimczak, U.

    1992-01-01

    Experiments with calf thymus DNA subjected to extracellular irradiation yield information on the role of direct and indirect effects in single-strand breakage, if this is evaluated with reference to the scavenger activity in respect of OH radicals. The role of the two processes in the occurrence of double-stand breaks and further damage leading to cell decay has so far remained largely obscure. It was the aim of the study described here to contribute to research in this field by performing in vitro experiments on biologically active DNA. For this purpose, DNA from pBR322 plasmids was irradiated in the presence of OH-radical scavengers. The number of single-strand and double-strand breaks was determined on the basis of the system's ability to eliminate OH radicals. In order to asses the influence of irradiation processes on the biological activity of DNA, investigations were carried out in E. coli for transformations caused by irradiated plasmid DNA. The results were interpreted in the light of theories about inhomogenous reaction kinetics put forward by Mark et al. (1989). It was finally discussed, which of the gamma-irradiation injuries occurring in DNA was to be held responsible for the inactivation of plasmid DNA and which enzymatic processes were additionally at work here. (orig./MG) [de

  13. DNA sequence analysis of plasmids from multidrug resistant Salmonella enterica serotype Heidelberg isolates.

    Directory of Open Access Journals (Sweden)

    Jing Han

    Full Text Available Salmonella enterica serovar Heidelberg is among the most detected serovars in swine and poultry, ranks among the top five serotypes associated with human salmonellosis and is disproportionately associated with invasive infections and mortality in humans. Salmonella are known to carry plasmids associated with antimicrobial resistance and virulence. To identify plasmid-associated genes in multidrug resistant S. enterica serovar Heidelberg, antimicrobial resistance plasmids from five isolates were sequenced using the 454 LifeSciences pyrosequencing technology. Four of the isolates contained incompatibility group (Inc A/C multidrug resistance plasmids harboring at least eight antimicrobial resistance genes. Each of these strains also carried a second resistance plasmid including two IncFIB, an IncHI2 and a plasmid lacking an identified Inc group. The fifth isolate contained an IncI1 plasmid, encoding resistance to gentamicin, streptomycin and sulfonamides. Some of the IncA/C plasmids lacked the full concert of transfer genes and yet were able to be conjugally transferred, likely due to the transfer genes carried on the companion plasmids in the strains. Several non-IncA/C resistance plasmids also carried putative virulence genes. When the sequences were compared to previously sequenced plasmids, it was found that while all plasmids demonstrated some similarity to other plasmids, they were unique, often due to differences in mobile genetic elements in the plasmids. Our study suggests that Salmonella Heidelberg isolates harbor plasmids that co-select for antimicrobial resistance and virulence, along with genes that can mediate the transfer of plasmids within and among other bacterial isolates. Prevalence of such plasmids can complicate efforts to control the spread of S. enterica serovar Heidelberg in food animal and human populations.

  14. Characterization of pLAC1, a cryptic plasmid isolated from Lactobacillus acidipiscis and comparative analysis with its related plasmids.

    Science.gov (United States)

    Asteri, Ioanna-Areti; Papadimitriou, Konstantinos; Boutou, Effrossyni; Anastasiou, Rania; Pot, Bruno; Vorgias, Constantinos E; Tsakalidou, Effie

    2010-07-15

    The pLAC1 plasmid of Lactobacillus acidipiscis ACA-DC 1533, a strain isolated from traditional Kopanisti cheese, was characterised. Nucleotide sequence analysis revealed a circular molecule of 3478bp with a G+C content of 37.2%. Ab initio annotation indicated four putative open reading frames (orfs). orf1 and orf4 were found to encode a replication initiation protein (Rep) and a mobilization protein (Mob), respectively. The deduced products of orf2 and orf3 revealed no significant homology to other known proteins. However, in silico examination of the plasmid sequence supported the existence of a novel operon that includes rep, orf2 and orf3 in pLAC1 and that this operon is highly conserved also in plasmids pLB925A02, pSMA23, pLC88 and pC7. RT-PCR experiments allowed us to verify that these three genes are co-transcribed as a single polycistronic mRNA species. Furthermore, phylogenetic analysis of pLAC1 Rep and Mob proteins demonstrated that they may have derived from different plasmid origins, suggesting that pLAC1 is a product of a modular evolution process. Comparative analysis of full length nucleotide sequences of pLAC1 and related Lactobacillus plasmids showed that pLAC1 shares a very similar replication backbone with pLB925A02, pSMA23 and pLC88. In contrast, mob of pLAC1 was almost identical with the respective gene of plasmids pLAB1000, pLB4 and pPB1. These findings lead to the conclusion that pLAC1 acquired mob probably via an ancestral recombination event. Our overall work highlights the importance of characterizing plasmids deriving from non-starter 'wild' isolates in order to better appreciate plasmid divergence and evolution of lactic acid bacteria. 2010 Elsevier B.V. All rights reserved.

  15. Effective removal of a range of Ti/Ri plasmids using a pBBR1-type vector having a repABC operon and a lux reporter system.

    Science.gov (United States)

    Yamamoto, Shinji; Sakai, Ayako; Agustina, Vita; Moriguchi, Kazuki; Suzuki, Katsunori

    2018-02-01

    Ti and Ri plasmids of pathogenic Agrobacterium strains are stably maintained by the function of a repABC operon and have been classified into four incompatibility groups, namely, incRh1, incRh2, incRh3, and incRh4. Removal of these plasmids from their bacterial cells is an important step in determining strain-specific virulence characteristics and to construct strains useful for transformation. Here, we developed two powerful tools to improve this process. We first established a reporter system to detect the presence and absence of Ti/Ri plasmids in cells by using an acetosyringone (AS)-inducible promoter of the Ti2 small RNA and luxAB from Vibrio harveyi. This system distinguished a Ti/Ri plasmid-free cell colony among plasmid-harboring cell colonies by causing the latter colonies to emit light in response to AS. We then constructed new "Ti/Ri eviction plasmids," each of which carries a repABC from one of four Ti/Ri plasmids that belonged to incRh1, incRh2, incRh3, and incRh4 groups in the suicidal plasmid pK18mobsacB and in a broad-host-range pBBR1 vector. Introduction of the new eviction plasmids into Agrobacterium cells harboring the corresponding Ti/Ri plasmids led to Ti/Ri plasmid-free cells in every incRh group. The Ti/Ri eviction was more effective by plasmids with the pBBR1 backbone than by those with the pK18mobsacB backbone. Furthermore, the highly stable cryptic plasmid pAtC58 in A. tumefaciens C58 was effectively evicted by the introduction of a pBBR1 vector containing the repABC of pAtC58. These results indicate that the set of pBBR1-repABC plasmids is a powerful tool for the removal of stable rhizobial plasmids.

  16. Growth of a plasmid-bearing (pYV) Yersinia pestis KIM5 in retail raw ground pork

    Science.gov (United States)

    Yersinia pestis can cause oro-pharyngeal plague as a result of consumption or handling of meat from infected animals. Thus, food naturally or intentionally contaminated can have a role in the dissemination of human plague. The growth of a conditionally virulent plasmid (pYV)-bearing rifampicin-res...

  17. Comparative genomics of an IncA/C multidrug resistance plasmid from Escherichia coli and Klebsiella isolates from intensive care unit patients and the utility of whole-genome sequencing in health care settings.

    Science.gov (United States)

    Hazen, Tracy H; Zhao, LiCheng; Boutin, Mallory A; Stancil, Angela; Robinson, Gwen; Harris, Anthony D; Rasko, David A; Johnson, J Kristie

    2014-08-01

    The IncA/C plasmids have been implicated for their role in the dissemination of β-lactamases, including gene variants that confer resistance to expanded-spectrum cephalosporins, which are often the treatment of last resort against multidrug-resistant, hospital-associated pathogens. A bla(FOX-5) gene was detected in 14 Escherichia coli and 16 Klebsiella isolates that were cultured from perianal swabs of patients admitted to an intensive care unit (ICU) of the University of Maryland Medical Center (UMMC) in Baltimore, MD, over a span of 3 years. Four of the FOX-encoding isolates were obtained from subsequent samples of patients that were initially negative for an AmpC β-lactamase upon admission to the ICU, suggesting that the AmpC β-lactamase-encoding plasmid was acquired while the patient was in the ICU. The genomes of five E. coli isolates and six Klebsiella isolates containing bla(FOX-5) were selected for sequencing based on their plasmid profiles. An ∼ 167-kb IncA/C plasmid encoding the FOX-5 β-lactamase, a CARB-2 β-lactamase, additional antimicrobial resistance genes, and heavy metal resistance genes was identified. Another FOX-5-encoding IncA/C plasmid that was nearly identical except for a variable region associated with the resistance genes was also identified. To our knowledge, these plasmids represent the first FOX-5-encoding plasmids sequenced. We used comparative genomics to describe the genetic diversity of a plasmid encoding a FOX-5 β-lactamase relative to the whole-genome diversity of 11 E. coli and Klebsiella isolates that carry this plasmid. Our findings demonstrate the utility of whole-genome sequencing for tracking of plasmid and antibiotic resistance gene distribution in health care settings. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  18. Plasmid containing a DNA ligase gene from Haemophilus influenzae

    International Nuclear Information System (INIS)

    McCarthy, D.; Griffin, K.; Setlow, J.K.

    1984-01-01

    A ligase gene from Haemophilus influenzae was cloned into the shuttle vector pDM2. Although the plasmid did not affect X-ray sensitivity, it caused an increase in UV sensitivity of the wild-type but not excision-defective H. influenzae and a decrease in UV sensitivity of the rec-1 mutant. 14 references, 2 figures

  19. Antibiogram and plasmid profiling of carbapenemase and extended ...

    African Journals Online (AJOL)

    Background: The increased reports of ESBL dissemination from various centres in south western, Nigeria and the recent emergence of carbapenem resistant bacteria prompted the conception of this study. Objectives: To demonstrate the relationship between high molecular weight plasmids and the expression of antibiotic ...

  20. Quinolones Resistance And R-Plasmids Of Clinical Isolates Of ...

    African Journals Online (AJOL)

    Background: There has been reported incidence in the emergence of. Quinolones resistance in clinical isolates in Nigeria and the level in resistance has been on the increase. Objective: To determine the antimicrobial resistance patterns and plasmids profiles of 67 clinical Pseudomonas species from a teaching hospital ...

  1. Chromosomal context and replication properties of ARS plasmids in ...

    Indian Academy of Sciences (India)

    2015-11-28

    Nov 28, 2015 ... plasmid but only a subset of them functions as replication origins in their ... except that they are rich in A + T content (As on one strand and Ts .... different unique, terminal, PCR-generated restriction sites used for cloning each fragment are ..... Hall TA 1999 BioEdit: a user-friendly biological sequence align-.

  2. a positive control plasmid for reporter gene assay

    African Journals Online (AJOL)

    STORAGESEVER

    2008-07-04

    Jul 4, 2008 ... qualification as a positive control for luciferase reporter gene assays. Key words: Reporter gene plasmid, luciferase assay, cytomegalovirus promoter/enhancer, human melanoma cell line. INTRODUCTION. Reporter genes, often called reporters, have become a precious tool in studies of gene expression ...

  3. Pharmaceutical development of the plasmid DNA vaccine pDERMATT

    NARCIS (Netherlands)

    Quaak, S.G.L.

    2009-01-01

    The discovery of tumor specific antigens and self tolerance mechanisms against these antigens led to the assumption that antigens circulating at sufficient concentration levels could break this self tolerance mechanism and evoke immunological antitumor effects. pDERMATT (plasmid DNA encoding

  4. plasmid mediated resistance in multidrug resistant bacteria isolated

    African Journals Online (AJOL)

    User

    PLASMID MEDIATED RESISTANCE IN MULTIDRUG RESISTANT BACTERIA. ISOLATED FROM CHILDREN WITH SUSPECTED SEPTICAEMIA IN ZARIA,. NIGERIA. AbdulAziz, Z. A.,1* Ehinmidu, J. O.,1 Adeshina, G. O.,1 Pala, Y. Y2., Yusuf, S. S2. and. Bugaje, M. A.3. 1Department of Pharmaceutics and Pharmaceutical ...

  5. Resistant plasmid profile analysis of multidrug resistant Escherichia ...

    African Journals Online (AJOL)

    Background: Multi-drug resistant Escherichia coli has become a major threat and cause of many urinary tract infections (UTIs) in Abeokuta, Nigeria. Objectives: This study was carried out to determine the resistant plasmids of multidrug resistant Escherichia coli isolated from (Urinary tract infections)UTIs in Abeokuta.

  6. Effect of Surfactants on Plasmid DNA Stability and Release from ...

    African Journals Online (AJOL)

    Purpose: To evaluate the effect of surfactants on plasmid DNA during preparation and release from polylactic glycolide (PLGA) microspheres. Methods: Various surfactants, both ionic and non-ionic (Span, Tween, Triton X100, cetyltrimethylammonium bromide and sodium dodecyl sulphate), were added during the ...

  7. Screening of degradative plasmids from Arthrobacter sp. HW08 and ...

    African Journals Online (AJOL)

    Administrator

    2011-06-08

    Jun 8, 2011 ... Media were solidified, if necessary, by the addition of 15 g agar ... genome extraction reagent kit, plasmid DNA fast extraction kit and. DNA segments ... spectrophotometer (Spectronic Instruments, Rochester, NY) and. SW content .... cultivation on LB slant for 100 times at 30 °C for 2 days, it was found that ...

  8. Antibiogram and plasmid profiling of carbapenemase and extended ...

    African Journals Online (AJOL)

    EB

    susceptibility was recorded against the Quinolone class of antibiotics; Meropenem remained the most active antibiotic against ESBL isolates ... Conclusion: Due to the relationship between high molecular weight plasmids and multi-drug resistance, we hereby recommend ..... Agents. Chemotherapy 2005; 49: 2137-. 2139. 7.

  9. Plasmid-encoded diacetyl (acetoin) reductase in Leuconostoc pseudomesenteroides

    DEFF Research Database (Denmark)

    Rattray, Fergal P; Myling-Petersen, Dorte; Larsen, Dianna

    2003-01-01

    A plasmid-borne diacetyl (acetoin) reductase (butA) from Leuconostoc pseudomesenteroides CHCC2114 was sequenced and cloned. Nucleotide sequence analysis revealed an open reading frame encoding a protein of 257 amino acids which had high identity at the amino acid level to diacetyl (acetoin...

  10. Comparative assessment of plasmid DNA delivery by encapsulation ...

    African Journals Online (AJOL)

    Purpose: To compare the gene delivery effectiveness of plasmid DNA (pDNA) encapsulated within poly (D,L-lactide-co-glycolide) (PLGA) nanoparticles with that adsorbed on PLGA nanoparticles. Methods: PLGA nanoparticles were prepared using solvent-evaporation method. To encapsulate pDNA within the particles, ...

  11. Physical structure and genetic expression of the sulfonamide-resistance plasmid pLS80 and its derivatives in Streptococcus pneumoniae and Bacillus subtilis

    Energy Technology Data Exchange (ETDEWEB)

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

    1984-01-01

    The 10-kb chromosomal fragment of Streptococcus pneumoniae cloned in pLS80 contains the sul-d allele of the pneumococcal gene for dihydropteroate synthase. As a single copy in the chromosome this allele confers resistance to sulfanilamide at 0.2 mg/ml; in the multicopy plasmid it confers resistance to 2.0 mg/ml. The sul-d mutation was mapped by restriction analysis to a 0.4-kb region. A spontaneous deletion beginning approx. 1.5 kb to the right of the sul-d mutation prevented gene function, possibly by removing a promoter. This region could be restored by chromosomal facilitation and be demonstrated in the plasmid by selection for sulfonamide resistance. Under selection for a vector marker, tetracycline resistance, only the deleted plasmid was detectable, apparently as a result of plasmid segregation and the advantageous growth rates of cells with smaller plasmids. When such cells were selected for sulfonamide resistance, the deleted region returned to the plasmid, presumably by equilibration between the chromosome and the plasmid pool, to give a low frequency (approx. 10/sup -3/) of cells resistant to sulfanilamide at 2.0 mg/ml. Models for the mechanisms of chromosomal facilitation and equilibration are proposed. Several derivatives of pLS80 could be transferred to Bacillus subtilis, where they conferred resistance to sulfanilamide at 2 mg/ml, thereby demonstrating cross-species expression of the pneumococcal gene. Transfer of the plasmids to B. subtilis gave rise to large deletions to the left of the sul-d marker, but these deletions did not interfere with the sul-d gene function. Restriction maps of pLS80 and its variously deleted derivatives are presented.

  12. Elimination of indigenous linear plasmids in Streptomyces hygroscopicus var. jinggangensis and Streptomyces sp. FR008 to increase validamycin A and candicidin productivities.

    Science.gov (United States)

    Lu, Chenyang; Wu, Hang; Su, Xiurong; Bai, Linquan

    2017-05-01

    Giant linear plasmids, which replicate independently of the chromosomes, widely exist in actinobacteria. Previous studies mostly focused on the replication and evolution of the linear plasmids or the secondary metabolite gene clusters and the resistance gene clusters therein. However, the relationships of the linear plasmids to the productivities of secondary metabolites have not been studied. In this work, we developed a method to eliminate the indigenous linear plasmid pSHJG1 in Streptomyces hygroscopicus var. jinggangensis, and validamycin A titer increased by 12.5% (from 19.16 ± 1.93 to 21.56 ± 2.25 g/L) in the high-yielding strain TL01 and 43.7% (from 4.67 ± 0.05 to 6.71 ± 0.21 g/L) in the wild-type strain 5008, whereas the cellular growth of the plasmid-cured mutant was reduced. Subsequently, the plasmid-cured mutant was complemented with three structure genes involved in cellular growth in pSHJG1 under the control of a strong PvalA promoter. Among them, the complementation of genes pSHJG1.069 and pSHJG1.072, encoding a putative hydrolase and putative P-loop ATPase, respectively, resulted in the restoration of cellular growth and validamycin A titer. Furthermore, the elimination of indigenous linear plasmid pHZ228 in the candicidin producer Streptomyces sp. FR008 also led to enhanced candicidin production and reduced cellular growth. Because of the wide distribution of indigenous linear plasmids in actinobacteria, the engineering strategy described here could be implemented in a variety of strains for the overproduction of various natural products.

  13. The detection and sequencing of a broad-host-range conjugative IncP-1β plasmid in an epidemic strain of Mycobacterium abscessus subsp. bolletii.

    Directory of Open Access Journals (Sweden)

    Sylvia Cardoso Leão

    Full Text Available BACKGROUND: An extended outbreak of mycobacterial surgical infections occurred in Brazil during 2004-2008. Most infections were caused by a single strain of Mycobacterium abscessus subsp. bolletii, which was characterized by a specific rpoB sequevar and two highly similar pulsed-field gel electrophoresis (PFGE patterns differentiated by the presence of a ∼50 kb band. The nature of this band was investigated. METHODOLOGY/PRINCIPAL FINDINGS: Genomic sequencing of the prototype outbreak isolate INCQS 00594 using the SOLiD platform demonstrated the presence of a 56,267-bp [corrected] circular plasmid, designated pMAB01. Identity matrices, genetic distances and phylogeny analyses indicated that pMAB01 belongs to the broad-host-range plasmid subgroup IncP-1β and is highly related to BRA100, pJP4, pAKD33 and pB10. The presence of pMAB01-derived sequences in 41 M. abscessus subsp. bolletii isolates was evaluated using PCR, PFGE and Southern blot hybridization. Sixteen of the 41 isolates showed the presence of the plasmid. The plasmid was visualized as a ∼50-kb band using PFGE and Southern blot hybridization in 12 isolates. The remaining 25 isolates did not exhibit any evidence of this plasmid. The plasmid was successfully transferred to Escherichia coli by conjugation and transformation. Lateral transfer of pMAB01 to the high efficient plasmid transformation strain Mycobacterium smegmatis mc(2155 could not be demonstrated. CONCLUSIONS/SIGNIFICANCE: The occurrence of a broad-host-range IncP-1β plasmid in mycobacteria is reported for the first time. Thus, genetic exchange could result in the emergence of specific strains that might be better adapted to cause human disease.

  14. High Prevalence of Plasmid-Mediated Quinolone Resistance and IncQ Plasmids Carrying qnrS2 Gene in Bacteria from Rivers near Hospitals and Aquaculture in China

    OpenAIRE

    Wen, Yanping; Pu, Xiaoying; Zheng, Wei; Hu, Guang

    2016-01-01

    Effluents from hospital and aquaculture are considered important sources of quinolone resistance. However, little information is available on the impact of this effluent on nearby rivers. In this study, 188 ciprofloxacin-resistant bacterial isolates obtained from rivers near hospitals and aquaculture were screened for plasmid-mediated quinolone resistance (PMQR) genes. Species identification, antibiotic susceptibility testing, and PMQR gene transferability assessment were conducted for PMQR-p...

  15. Survival and evolution of a large multidrug resistance plasmid in new clinical bacterial hosts

    DEFF Research Database (Denmark)

    Porse, Andreas; Schønning, Kristian; Munck, Christian

    2016-01-01

    Large conjugative plasmids are important drivers of bacterial evolution and contribute significantly to the dissemination of antibiotic resistance. Although plasmid borne multidrug resistance is recognized as one of the main challenges in modern medicine, the adaptive forces shaping the evolution...

  16. Plasmid profiling of bacterial isolates from confined environments

    Science.gov (United States)

    van Houdt, Rob; Provoost, Ann; Coninx, Ilse; Leys, Natalie; Mergeay, Max

    Plasmid profiling of bacterial isolates from confined environments R. Van Houdt, I. Coninx, A. Provoost, N. Leys, and M. Mergeay Expertise group for Molecular and Cellular Biology, Institute for Environment, Health and Safety, Belgian Nuclear Research Centre (SCK•CEN), Boeretang 200, B-2400 Mol, Belgium. Human exploration of extreme and isolated hostile environments such as space requires special confined small volume habitats to protect and house the crew. However, human confinement in such small volume habitats has restrictions on waste disposal and personal hygiene and inevitably generates a particular community of microorganisms within the habitat. These microorganisms are mainly originating from the crew (skin, mucous membranes, upper respiratory tract, mouth, and gastrointestinal tract) but also include the residing environmental microorganisms. Earth-based confined habitats such as the Antarctic Research Station Concordia are used as test beds for long-duration spaceflights to study the physiologic and psychological adaptation to isolated environments. The dynamics of the environmental microbial population in such a test bed could render additional insights in assessing the potential health risks in long-duration space missions. Not only total bacterial contamination levels are important, but it is essential to identify also the predominant microbial taxa and their mobile genetic elements (MGE). These MGEs could be exchanged between bacteria by horizontal gene transfer and may alter the pathogenic potential since they often carry antibiotic resistance or more in general adaptation-enhancing traits. In this study several bacterial strains isolated in the Concordia research station were examined for their plasmid content. An optimized protocol for extraction of large plasmids showed the present of at least one plasmid in 50% of the strains. For all strains the minimal inhibitory concentration of a range of antibiotics was determined indicating resistance to

  17. Survival and evolution of a large multidrug resistance plasmid in new clinical bacterial hosts

    DEFF Research Database (Denmark)

    Porse, Andreas; Schønning, Kristian; Munck, Christian

    2016-01-01

    sequencing to show that the long-term persistence and molecular integrity of the plasmid is highly influenced by multiple factors within a 25 kb plasmid region constituting a host-dependent burden. In the E. coli hosts investigated here, improved plasmid stability readily evolves via IS26 mediated deletions...... consistently followed by all evolved E. coli lineages exposes a trade-off between horizontal and vertical transmission that may ultimately limit the dissemination potential of clinical multidrug resistance plasmids in these hosts....

  18. Presence of Glycopeptide-Encoding Plasmids in Enterococcal Isolates from Food and Humans in Denmark

    DEFF Research Database (Denmark)

    Migura, Lourdes Garcia; Valenzuela, Antonio Jesus Sanchez; Jensen, Lars Bogø

    2011-01-01

    developed techniques for classification of plasmids. Replicons associated with sex pheromone-inducible plasmids were detected in all GR E. faecalis, whereas GR Enterococcus faecium contained plasmids known to be widely distributed among enterococci. vanA resistance is common in E. faecium isolates from meat...... and animals in Europe and is rarely found in E. faecalis. This article describes the first characterization of MGE from vanA mediated E. faecalis, thus linking this resistance genotype to pheromone responding plasmids....

  19. Phytotracker, an information management system for easy recording and tracking of plants, seeds and plasmids

    Directory of Open Access Journals (Sweden)

    Nieuwland Jeroen

    2012-10-01

    Full Text Available Abstract Background A large number of different plant lines are produced and maintained in a typical plant research laboratory, both as seed stocks and in active growth. These collections need careful and consistent management to track and maintain them properly, and this is a particularly pressing issue in laboratories undertaking research involving genetic manipulation due to regulatory requirements. Researchers and PIs need to access these data and collections, and therefore an easy-to-use plant-oriented laboratory information management system that implements, maintains and displays the information in a simple and visual format would be of great help in both the daily work in the lab and in ensuring regulatory compliance. Results Here, we introduce ‘Phytotracker’, a laboratory management system designed specifically to organise and track plasmids, seeds and growing plants that can be used in mixed platform environments. Phytotracker is designed with simplicity of user operation and ease of installation and management as the major factor, whilst providing tracking tools that cover the full range of activities in molecular genetics labs. It utilises the cross-platform Filemaker relational database, which allows it to be run as a stand-alone or as a server-based networked solution available across all workstations in a lab that can be internet accessible if desired. It can also be readily modified or customised further. Phytotracker provides cataloguing and search functions for plasmids, seed batches, seed stocks and plants growing in pots or trays, and allows tracking of each plant from seed sowing, through harvest to the new seed batch and can print appropriate labels at each stage. The system enters seed information as it is transferred from the previous harvest data, and allows both selfing and hybridization (crossing to be defined and tracked. Transgenic lines can be linked to their plasmid DNA source. This ease of use and flexibility

  20. Plasmid vectors for Xylella fastidiosa utilizing a toxin-antitoxin system for plasmid stability in the absence of antibiotic selection

    Science.gov (United States)

    The phytopathogen Xylella fastidiosa causes disease in a variety of important crop and landscape plants. Functional genetic studies have led to a broader understanding of virulence mechanisms used by this pathogen in the grapevine host. Plasmid shuttle vectors are important tools in studies of bacte...

  1. Construction and expression of pEgr-sHemopexin recombinant plasmid induced by ionizing radiation in vitro

    International Nuclear Information System (INIS)

    Wang Guiquan; Jilin Univ., Changchun; Xu Chuanjie; Yang Wen; Piao Chunji; Dong Zhen

    2005-01-01

    Objective: To clone mouse secretable Hemopexin (sPEX) cDNA, construct pEgr-sPEX recombinant plasmid and detect the expression of recombinant plasmid in B16F10 cells. Methods: Hemopexin cDNA was amplified from the NIH3T3 cells by RT-PCR. After the cDNA identified by sequencing, the pEgr-sPEX recombinant plasmid was constructed and the plasmid was transfected into B16F10 cells with liposome and the expression of PEX induced by ionizing radiation in B16F10 cells was detected by Western blotting. Results: The sequencing results proved the cloned sPEX cDNA to be completely identical with that reported in the GenBank. The mouse sPEX cDNA was inserted correctly into expression vector and expressed successfully. Conclusion: The mouse sPEX cDNA is cloned successfully and it is confirmed that pEgr-sPEX possesses the radiation inducing expression characteristics in vitro. (authors)

  2. Structural and functional analysis of the kid toxin protein from E. coli Plasmid R1

    NARCIS (Netherlands)

    Hargreaves, D.; Santos-Sierra, S.; Giraldo, R.; Sabariegos-Jareño, R.; de la Cueva-Méndez, G.; Boelens, R.|info:eu-repo/dai/nl/070151407; Díaz-Orejas, R.; Rafferty, J.B.

    2002-01-01

    We have determined the structure of Kid toxin protein from E. coli plasmid R1 involved in stable plasmid inheritance by postsegregational killing of plasmid-less daughter cells. Kid forms a two-component system with its antagonist, Kis antitoxin. Our 1.4 Å crystal structure of Kid reveals a 2-fold

  3. Studies on the expression of plasmid-borne genes in the endosymbiotic state of Rhizobium leguminosarum

    NARCIS (Netherlands)

    Krol, A.J.M.

    1982-01-01

    The subject matter of the research reported in this thesis is the role of plasmid-borne genes of Rhizobium in symbiosis and nitrogen fixation. Plasmid DNA was isolated from Rhizobium leguminosarum strain PRE and the expression of plasmid DNA in nitrogen

  4. Generation of Induced Pluripotent Stem Cells from Frozen Buffy Coats using Non-integrating Episomal Plasmids.

    Science.gov (United States)

    Meraviglia, Viviana; Zanon, Alessandra; Lavdas, Alexandros A; Schwienbacher, Christine; Silipigni, Rosamaria; Di Segni, Marina; Chen, Huei-Sheng Vincent; Pramstaller, Peter P; Hicks, Andrew A; Rossini, Alessandra

    2015-06-05

    Somatic cells can be reprogrammed into induced pluripotent stem cells (iPSCs) by forcing the expression of four transcription factors (Oct-4, Sox-2, Klf-4, and c-Myc), typically expressed by human embryonic stem cells (hESCs). Due to their similarity with hESCs, iPSCs have become an important tool for potential patient-specific regenerative medicine, avoiding ethical issues associated with hESCs. In order to obtain cells suitable for clinical application, transgene-free iPSCs need to be generated to avoid transgene reactivation, altered gene expression and misguided differentiation. Moreover, a highly efficient and inexpensive reprogramming method is necessary to derive sufficient iPSCs for therapeutic purposes. Given this need, an efficient non-integrating episomal plasmid approach is the preferable choice for iPSC derivation. Currently the most common cell type used for reprogramming purposes are fibroblasts, the isolation of which requires tissue biopsy, an invasive surgical procedure for the patient. Therefore, human peripheral blood represents the most accessible and least invasive tissue for iPSC generation. In this study, a cost-effective and viral-free protocol using non-integrating episomal plasmids is reported for the generation of iPSCs from human peripheral blood mononuclear cells (PBMNCs) obtained from frozen buffy coats after whole blood centrifugation and without density gradient separation.

  5. results

    Directory of Open Access Journals (Sweden)

    Salabura Piotr

    2017-01-01

    Full Text Available HADES experiment at GSI is the only high precision experiment probing nuclear matter in the beam energy range of a few AGeV. Pion, proton and ion beams are used to study rare dielectron and strangeness probes to diagnose properties of strongly interacting matter in this energy regime. Selected results from p + A and A + A collisions are presented and discussed.

  6. Poly(hydroxyethyl methacrylate) based magnetic nanoparticles for plasmid DNA purification from Escherichia coli lysate

    Energy Technology Data Exchange (ETDEWEB)

    Percin, Is Latin-Small-Letter-Dotless-I k [Department of Biology, Hacettepe University, Ankara (Turkey); Karakoc, Veyis [Department of Chemistry, Biochemistry Division, Hacettepe University, Ankara (Turkey); Akgoel, Sinan [Department of Biochemistry, Ege University, Izmir (Turkey); Aksoez, Erol [Department of Biology, Hacettepe University, Ankara (Turkey); Denizli, Adil, E-mail: denizli@hacettepe.edu.tr [Department of Chemistry, Biochemistry Division, Hacettepe University, Ankara (Turkey)

    2012-07-01

    The aim of this study is to prepare poly(hydroxyethyl methacrylate-N-methacryloyl-(L)-histidine) [PHEMAH] magnetic nanoparticles for plasmid DNA (pDNA) purification from Escherichia coli (E. coli) cell lysate. Magnetic nanoparticles were produced by surfactant free emulsion polymerization. mPHEMAH nanoparticles were characterized by elemental analysis, Fourier transform infrared spectroscopy (FTIR), atomic force microscopy (AFM), vibrating sample magnetometer (VSM), electron spin resonance (ESR), thermogravimetric analyses (TGA) and transmission electron microscopy (TEM). Surface area, average particle size and size distribution were also performed. Specific surface area of the mPHEMAH nanoparticles was found to be 1180 m{sup 2}/g. Elemental analysis of MAH for nitrogen was estimated as 0.18 mmol/g polymer. The amount of pDNA adsorbed onto the mPHEMAH nanoparticles first increased and then reached a saturation value at around 1.0 mg/mL of pDNA concentration. Compared with the mPHEMA nanoparticles (50 {mu}g/g polymer), the pDNA adsorption capacity of the mPHEMAH nanoparticles (154 mg/g polymer) was improved significantly due to the MAH incorporation into the polymeric matrix. The maximum pDNA adsorption was achieved at 25 Degree-Sign C. The overall recovery of pDNA was calculated as 92%. The mPHEMAH nanoparticles could be used six times without decreasing the pDNA adsorption capacity significantly. The results indicate that the PHEMAH nanoparticles promise high selectivity for pDNA. - Highlights: Black-Right-Pointing-Pointer Magnetic nanoparticles have several advantages over conventional adsorbents. Black-Right-Pointing-Pointer MAH acted as the pseudospecific ligand, ligand immobilization step was eliminated. Black-Right-Pointing-Pointer pDNA adsorption amount was 154 mg/g. Black-Right-Pointing-Pointer Fifty-fold capacity increase was obtained when compared to conventional matrices.

  7. Poly(hydroxyethyl methacrylate) based magnetic nanoparticles for plasmid DNA purification from Escherichia coli lysate

    International Nuclear Information System (INIS)

    Perçin, Işık; Karakoç, Veyis; Akgöl, Sinan; Aksöz, Erol; Denizli, Adil

    2012-01-01

    The aim of this study is to prepare poly(hydroxyethyl methacrylate-N-methacryloyl-(L)-histidine) [PHEMAH] magnetic nanoparticles for plasmid DNA (pDNA) purification from Escherichia coli (E. coli) cell lysate. Magnetic nanoparticles were produced by surfactant free emulsion polymerization. mPHEMAH nanoparticles were characterized by elemental analysis, Fourier transform infrared spectroscopy (FTIR), atomic force microscopy (AFM), vibrating sample magnetometer (VSM), electron spin resonance (ESR), thermogravimetric analyses (TGA) and transmission electron microscopy (TEM). Surface area, average particle size and size distribution were also performed. Specific surface area of the mPHEMAH nanoparticles was found to be 1180 m 2 /g. Elemental analysis of MAH for nitrogen was estimated as 0.18 mmol/g polymer. The amount of pDNA adsorbed onto the mPHEMAH nanoparticles first increased and then reached a saturation value at around 1.0 mg/mL of pDNA concentration. Compared with the mPHEMA nanoparticles (50 μg/g polymer), the pDNA adsorption capacity of the mPHEMAH nanoparticles (154 mg/g polymer) was improved significantly due to the MAH incorporation into the polymeric matrix. The maximum pDNA adsorption was achieved at 25 °C. The overall recovery of pDNA was calculated as 92%. The mPHEMAH nanoparticles could be used six times without decreasing the pDNA adsorption capacity significantly. The results indicate that the PHEMAH nanoparticles promise high selectivity for pDNA. - Highlights: ► Magnetic nanoparticles have several advantages over conventional adsorbents. ► MAH acted as the pseudospecific ligand, ligand immobilization step was eliminated. ► pDNA adsorption amount was 154 mg/g. ► Fifty-fold capacity increase was obtained when compared to conventional matrices.

  8. Statistical data and results obtained on irradiated transistors 2N.2221 Sesco and 2N.2907 SGS; Donnees de fiabilite et resultats statistiques obtenus sur des transistors 2n.2221 Sesco et 2n.2907 SGS irradies

    Energy Technology Data Exchange (ETDEWEB)

    Blin, A; Le Ber, J

    1966-07-01

    This document provides results obtained on many samples of transistors irradiated in the laboratories of the Institut of Nuclear Physic of Lyon. The physical aspects of the irradiation,the statistical aspects of the study and the reliability under irradiation have been studied, but the accent is done on the statistical analysis. (A.L.B.)

  9. To report the obtained results in the operation multicycle study of the L V U-1 using design data with the FCSII (1D) and PRESTO (3D) codes of the FMS system

    International Nuclear Information System (INIS)

    Montes T, J.L.; Cortes C, C.C.

    1991-07-01

    This work is to carry out a multicycle study for the Laguna Verde U-1 reactor with the FCS-II (1 - 20 cycles) and PRESTO (1 - 6 cycles) codes and to compare the obtained results against those reported by General Electric. (Author)

  10. Do the results of respirable dust samples obtained from direct-on-filter X-ray diffraction, direct-on-filter infrared and indirect infrared (KBr pellet) methods correlate?

    CSIR Research Space (South Africa)

    Pretorius, C

    2010-11-01

    Full Text Available The objective of this study was to determine whether a correlation exists between the quartz results obtained from direct-on-filter X-ray Diffraction analysis, direct-on-filter Fourier-Transform Infrared analysis and indirect analysis (Potassium...

  11. Construction of recombinant plasmid pIRESEgr-IFN γ and its expression in Lewis lung carcinoma induced by irradiation

    International Nuclear Information System (INIS)

    Yang Wei; Li Xiuyi; Gong Shouliang; Sun Ting; Gong Pingsheng

    2007-01-01

    Objective: To construct the recombinant plasmid pIRESEgr-IFN γ and detect its expression in Lewis lung carcinoma induced by irradiation in vitro. Methods: The recombinant plasmid pIRESEgr-IFN γ containing Egr-1 promoter and IFN γ gene was constructed with gene recombinant technique. The plasmid was transferred into Lewis lung carcinoma by liposome in vitro. The correlations of dose- and time-effects in the expression of IFN γ gene induced by X-ray were detected by ELISA. Results: The identification with enzymes proved that Egr-1 promoter and IFN γ gene were inserted into vector pIRESlneo correctly. After X-ray irradiation with different doses, the expression of IFN γ in the supernatant of Lewis lung carcinoma transfected by pIRESEgr-IFN γ was significantly higher than that in 0 Gy group (P<0.001). After 5 Gy X-ray irradiation, the expression of IFN γ was the highest, being 4.39 times as much as that in 0 Gy group. The expression of IFN γ in the supernatant increased after 5 Gy X-ray irradiation, being 6.27 times as much as that in 0 h group 36 h after irradiation. Conclusion: The recombinant plasmid pIRESEgr-IFN γ is constructed successfully, and it has the property of enhancing the expression of IFN γ gene induced by irradiation. (authors)

  12. pEVL: A Linear Plasmid for Generating mRNA IVT Templates With Extended Encoded Poly(A Sequences

    Directory of Open Access Journals (Sweden)

    Alexandra E Grier

    2016-01-01

    Full Text Available Increasing demand for large-scale synthesis of in vitro transcribed (IVT mRNA is being driven by the increasing use of mRNA for transient gene expression in cell engineering and therapeutic applications. An important determinant of IVT mRNA potency is the 3′ polyadenosine (poly(A tail, the length of which correlates with translational efficiency. However, present methods for generation of IVT mRNA rely on templates derived from circular plasmids or PCR products, in which homopolymeric tracts are unstable, thus limiting encoded poly(A tail lengths to ≃120 base pairs (bp. Here, we have developed a novel method for generation of extended poly(A tracts using a previously described linear plasmid system, pJazz. We find that linear plasmids can successfully propagate poly(A tracts up to ≃500 bp in length for IVT mRNA production. We then modified pJazz by removing extraneous restriction sites, adding a T7 promoter sequence upstream from an extended multiple cloning site, and adding a unique type-IIS restriction site downstream from the encoded poly(A tract to facilitate generation of IVT mRNA with precisely defined encoded poly(A tracts and 3′ termini. The resulting plasmid, designated pEVL, can be used to generate IVT mRNA with consistent defined lengths and terminal residue(s.

  13. Association and Expression of Virulence from Plasmids of the Group B Strain in Pseudomonas syringae pv. eriobotryae

    Directory of Open Access Journals (Sweden)

    Tran Dang Khanh

    2018-04-01

    Full Text Available Pseudomonas syringae pv. eriobotryae causes serious stem canker in loquat (Eriobotrya japonica trees. This study was conducted to determine whether plasmids are involved with its virulence. The strain NAE89, which belonged to the B group, harbored two plasmids at approximately 6.2 and 50 Mdal that caused stem canker and halo leaf spots on loquat plants. Following digestion with BamHI and ligation into the BamHI cloning site of the broad range host cosmid pLAFR3, four DNA fragments at 3.8, 6.6, 12.3, and 22.8 kb were generated. Although the plasmid-encoded virulence gene psvA was undigested with the BamHI, the halo leaf spot gene may be adjacent to the psvA gene was digested. A pLAFR3 cosmid clone was introduced into the non-pathogenic PE0 and NAE89-1 strains by triparental matings and the pathogenicity was recovered. As a result, the pLAFR3 cosmid clone was introduced into the largest size DNA fragment of 22.8 kb and determined to be the causal agent of canker on the stem of the loquat. This study revealed that the psvA gene, previously found in the 50 Mdal plasmid, was also observed in the 22.8 kb DNA fragment.

  14. High-throughput screening of cellulase F mutants from multiplexed plasmid sets using an automated plate assay on a functional proteomic robotic workcell

    Directory of Open Access Journals (Sweden)

    Qureshi Nasib

    2006-05-01

    Full Text Available Abstract Background The field of plasmid-based functional proteomics requires the rapid assay of proteins expressed from plasmid libraries. Automation is essential since large sets of mutant open reading frames are being cloned for evaluation. To date no integrated automated platform is available to carry out the entire process including production of plasmid libraries, expression of cloned genes, and functional testing of expressed proteins. Results We used a functional proteomic assay in a multiplexed setting on an integrated plasmid-based robotic workcell for high-throughput screening of mutants of cellulase F, an endoglucanase from the anaerobic fungus Orpinomyces PC-2. This allowed us to identify plasmids containing optimized clones expressing mutants with improved activity at lower pH. A plasmid library of mutagenized clones of the celF gene with targeted variations in the last four codons was constructed by site-directed PCR mutagenesis and transformed into Escherichia coli. A robotic picker integrated into the workcell was used to inoculate medium in a 96-well deep well plate, combining the transformants into a multiplexed set in each well, and the plate was incubated on the workcell. Plasmids were prepared from the multiplexed culture on the liquid handler component of the workcell and used for in vitro transcription/translation. The multiplexed expressed recombinant proteins were screened for improved activity and stability in an azo-carboxymethylcellulose plate assay. The multiplexed wells containing mutants with improved activity were identified and linked back to the corresponding multiplexed cultures stored in glycerol. Spread plates were prepared from the glycerol stocks and the workcell was used to pick single colonies from the spread plates, prepare plasmid, produce recombinant protein, and assay for activity. The screening assay and subsequent deconvolution of the multiplexed wells resulted in identification of improved Cel

  15. One Novel Multiple-Target Plasmid Reference Molecule Targeting Eight Genetically Modified Canola Events for Genetically Modified Canola Detection.

    Science.gov (United States)

    Li, Zhuqing; Li, Xiang; Wang, Canhua; Song, Guiwen; Pi, Liqun; Zheng, Lan; Zhang, Dabing; Yang, Litao

    2017-09-27

    Multiple-target plasmid DNA reference materials have been generated and utilized as good substitutes of matrix-based reference materials in the analysis of genetically modified organisms (GMOs). Herein, we report the construction of one multiple-target plasmid reference molecule, pCAN, which harbors eight GM canola event-specific sequences (RF1, RF2, MS1, MS8, Topas 19/2, Oxy235, RT73, and T45) and a partial sequence of the canola endogenous reference gene PEP. The applicability of this plasmid reference material in qualitative and quantitative PCR assays of the eight GM canola events was evaluated, including the analysis of specificity, limit of detection (LOD), limit of quantification (LOQ), and performance of pCAN in the analysis of various canola samples, etc. The LODs are 15 copies for RF2, MS1, and RT73 assays using pCAN as the calibrator and 10 genome copies for the other events. The LOQ in each event-specific real-time PCR assay is 20 copies. In quantitative real-time PCR analysis, the PCR efficiencies of all event-specific and PEP assays are between 91% and 97%, and the squared regression coefficients (R 2 ) are all higher than 0.99. The quantification bias values varied from 0.47% to 20.68% with relative standard deviation (RSD) from 1.06% to 24.61% in the quantification of simulated samples. Furthermore, 10 practical canola samples sampled from imported shipments in the port of Shanghai, China, were analyzed employing pCAN as the calibrator, and the results were comparable with those assays using commercial certified materials as the calibrator. Concluding from these results, we believe that this newly developed pCAN plasmid is one good candidate for being a plasmid DNA reference material in the detection and quantification of the eight GM canola events in routine analysis.

  16. Investigation of diversity of plasmids carrying the blaTEM-52 gene

    DEFF Research Database (Denmark)

    Bielak, Eliza Maria; Bergenholtz, Rikke D.; Jørgensen, Mikael Skaanning

    2011-01-01

    OBJECTIVES: To investigate the diversity of plasmids that carry blaTEM-52 genes among Escherichia coli and Salmonella enterica originating from animals, meat products and humans. METHODS: A collection of 22 blaTEM-52-encoding plasmids was characterized by restriction fragment length polymorphism...... of self-transfer to a plasmid-free E. coli recipient. CONCLUSIONS: The blaTEM-52 gene found in humans c