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Sample records for plasmid curing

  1. Plasmid interference for curing antibiotic resistance plasmids in vivo.

    Science.gov (United States)

    Kamruzzaman, Muhammad; Shoma, Shereen; Thomas, Christopher M; Partridge, Sally R; Iredell, Jonathan R

    2017-01-01

    Antibiotic resistance increases the likelihood of death from infection by common pathogens such as Escherichia coli and Klebsiella pneumoniae in developed and developing countries alike. Most important modern antibiotic resistance genes spread between such species on self-transmissible (conjugative) plasmids. These plasmids are traditionally grouped on the basis of replicon incompatibility (Inc), which prevents coexistence of related plasmids in the same cell. These plasmids also use post-segregational killing ('addiction') systems, which poison any bacterial cells that lose the addictive plasmid, to guarantee their own survival. This study demonstrates that plasmid incompatibilities and addiction systems can be exploited to achieve the safe and complete eradication of antibiotic resistance from bacteria in vitro and in the mouse gut. Conjugative 'interference plasmids' were constructed by specifically deleting toxin and antibiotic resistance genes from target plasmids. These interference plasmids efficiently cured the corresponding antibiotic resistant target plasmid from different Enterobacteriaceae in vitro and restored antibiotic susceptibility in vivo to all bacterial populations into which plasmid-mediated resistance had spread. This approach might allow eradication of emergent or established populations of resistance plasmids in individuals at risk of severe sepsis, enabling subsequent use of less toxic and/or more effective antibiotics than would otherwise be possible, if sepsis develops. The generalisability of this approach and its potential applications in bioremediation of animal and environmental microbiomes should now be systematically explored.

  2. Plasmid interference for curing antibiotic resistance plasmids in vivo

    Science.gov (United States)

    Kamruzzaman, Muhammad; Shoma, Shereen; Thomas, Christopher M.; Partridge, Sally R.

    2017-01-01

    Antibiotic resistance increases the likelihood of death from infection by common pathogens such as Escherichia coli and Klebsiella pneumoniae in developed and developing countries alike. Most important modern antibiotic resistance genes spread between such species on self-transmissible (conjugative) plasmids. These plasmids are traditionally grouped on the basis of replicon incompatibility (Inc), which prevents coexistence of related plasmids in the same cell. These plasmids also use post-segregational killing (‘addiction’) systems, which poison any bacterial cells that lose the addictive plasmid, to guarantee their own survival. This study demonstrates that plasmid incompatibilities and addiction systems can be exploited to achieve the safe and complete eradication of antibiotic resistance from bacteria in vitro and in the mouse gut. Conjugative ‘interference plasmids’ were constructed by specifically deleting toxin and antibiotic resistance genes from target plasmids. These interference plasmids efficiently cured the corresponding antibiotic resistant target plasmid from different Enterobacteriaceae in vitro and restored antibiotic susceptibility in vivo to all bacterial populations into which plasmid-mediated resistance had spread. This approach might allow eradication of emergent or established populations of resistance plasmids in individuals at risk of severe sepsis, enabling subsequent use of less toxic and/or more effective antibiotics than would otherwise be possible, if sepsis develops. The generalisability of this approach and its potential applications in bioremediation of animal and environmental microbiomes should now be systematically explored. PMID:28245276

  3. Curing of plasmid pXO1 from Bacillus anthracis using plasmid incompatibility.

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    Xiankai Liu

    Full Text Available The large plasmid pXO1 encoding the anthrax toxin is important for the virulence of Bacillus anthracis. It is essential to cure pXO1 from B. anthracis to evaluate its role in the pathogenesis of anthrax infection. Because conventional methods for curing plasmids (e.g., curing agents or growth at elevated temperatures can induce mutations in the host chromosomal DNA, we developed a specific and reliable method to eliminate pXO1 from B. anthracis using plasmid incompatibility. Three putative replication origins of pXO1 were inserted into a temperature-sensitive plasmid to generate three incompatible plasmids. One of the three plasmids successfully eliminated the large plasmid pXO1 from B. anthracis vaccine strain A16R and wild type strain A16. These findings provided additional information about the replication/partitioning of pXO1 and demonstrated that introducing a small incompatible plasmid can generate plasmid-cured strains of B. anthracis without inducing spontaneous mutations in the host chromosome.

  4. An insight of traditional plasmid curing in Vibrio species

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    Vengadesh eLetchumanan

    2015-07-01

    Full Text Available As the causative agent of foodborne related illness, Vibrio species causes a huge impact on the public health and management. Vibrio species is often associated with seafood as the latter plays a role as a vehicle to transmit bacterial infections. Hence, antibiotics are used not to promote growth but rather to prevent and treat bacterial infections. The extensive use of antibiotics in the aquaculture industry and environment has led to the emerging of antibiotic resistant strains. This phenomenon has triggered an alarming public health concern due to the increase number of pathogenic Vibrio strains that are resistant to clinically used antibiotics and is found in the environment. Antibiotic resistance and the genes location in the strains can be detected through plasmid curing assay. The results derived from plasmid curing assay is fast, cost effective, sufficient in providing insights and influence the antibiotic management policies in the aquaculture industry. This presentation aims in discussing and providing insights on various curing agents in Vibrio species. To our best of knowledge, this is a first review written discussing on plasmid curing in Vibrio species.

  5. Plasmid Profiling and Curing of Lactobacillus Strains Isolated from Fermented Milk for Probiotic Applications

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    B. Lavanya

    2011-04-01

    Full Text Available In this study, the antimicrobial susceptibilities and presence of plasmids in 7 probiotics strains which had been isolated from the fermented milk were determined. Resistance to 8 commonly used antibiotics $- lactans (penicillin, ampicilin, gram positive spectrum (vanomycin, broad spectrum (rifampin, trimethoprim and aminogycosides (kanamycin, streptomycin, and bacitracin was assessed by disk diffusion. Among these strains 20, 20, 60, 70, 90 and 100% were found to be exhibit a significant degree of resistance to kanamycin, trimetroprim, rifampicin, kanamycin, amphicilin and penicillin respectively. Further, plasmid profile and curing of plasmid were performed for the seven isolates. Analysis of the plasmid profiles of the 7 cured derivatives revealed loss of plasmids except 2 strains where curing was partially effective. All the strains lost penicillin resistance after curing indicating that plasmids encodes for resistance character. However, vanomycin resistance is not lost upon curing which indicates that such resistance is usually intrinsic (chromosomally encoded and not transmissible. Finally, the antimicrobial susceptibility after curing was done to check the safety aspect of the isolates for their application as probiotics and among the 7 strains, 5 were proved to be potent probiotics.

  6. Two-step method for curing Escherichia coli of ColE1-derived plasmids

    DEFF Research Database (Denmark)

    Hove-Jensen, Bjarne

    2008-01-01

    To cure Escherichia coli for plasmids derived from the ColE1 replicon advantage is taken of the fact that maintenance of this replicon requires a wild-type allele of polA, encoding DNA polymerase I. Curing is achieved by cotransduction of a mutant polA allele with metE::Tn10, fadAB::Tn10 or other...

  7. The Native Plasmid pML21 Plays a Role in Stress Tolerance in Enterococcus faecalis ML21, as Analyzed by Plasmid Curing Using Plasmid Incompatibility.

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    Zuo, Fang-Lei; Chen, Li-Li; Zeng, Zhu; Feng, Xiu-Juan; Yu, Rui; Lu, Xiao-Ming; Ma, Hui-Qin; Chen, Shang-Wu

    2016-02-01

    To investigate the role of the native plasmid pML21 in Enterococcus faecalis ML21's response to abiotic stresses, the plasmid pML21 was cured based on the principle of plasmid incompatibility and segregational instability, generating E. faecalis mutant strain ML0. The mutant and the wild strains were exposed to abiotic stresses: bile salts, low pH, H2O2, ethanol, heat, and NaCl, and their survival rate was measured. We found that curing of pML21 lead to reduced tolerance to stress in E. faecalis ML0, especially oxidative and osmotic stress. Complementation analysis suggested that the genes from pML21 played different role in stress tolerance. The result indicated that pML21 plays a role in E. faecalis ML21's response to abiotic stresses.

  8. Curing of plasmid pBMB28 from Bacillus thuringiensis YBT-020 using an unstable replication region.

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    Wang, Pengxia; Zhu, Qian; Shang, Hui; Zhu, Yiguang; Sun, Ming

    2016-02-01

    Bacillus thuringiensis serovar finitimus strain YBT-020 is the well-studied spore-crystal association (SCA) phenotypic strain, whose parasporal crystals adhere to spore after lysis of the mother cell. Its endogenous plasmids pBMB26 and pBMB28 were proved essential for this SCA phenotype. In our previous study, using conventional methods, pBMB26 cured derivative and both pBMB26 and pBMB28 cured derivative of YBT-020 were obtained. However, YBT-020 solely cured of pBMB28 could not be obtained. In this study, an unstable replication region of pBMB28 was identified and was used to construct an incompatible plasmid pRep28B. This incompatible plasmid was successfully used to cure plasmid pBMB28 and was easily eliminated through segregational instability under the optimum growth temperature of YBT-020. Therefore, an endogenous plasmid was cured from the B. thuringiensis strain utilizing plasmid incompatibility. Moreover, using an unstable replication region instead of a temperature sensitive (Ts) replication region is better to cure the incompatible plasmid because it can avoid culturing at higher temperature. This method provides an efficient method for plasmid curing in B. thuringiensis and other bacteria.

  9. Development of Lactobacillus plantarum LL441 and its plasmid-cured derivatives in cheese.

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    Delgado, Susana; Mayo, Baltasar

    2003-04-01

    A wild Lactobacillus plantarum strain and two of its plasmid-cured derivatives were separately used as adjunct cultures in the manufacture of a Gouda-like traditional Spanish cheese. The wild strain, LL441, harbours seven plasmids and produces a lantibiotic-like bacteriocin. The LL441-B2 derivative has lost plasmids of 40 and 80 kb and the bacteriocin-producing capability. The LL441-B11 derivative has lost in addition a 70 kb plasmid encoding active alpha- and beta-galactosidases. All three strains could be used as adjunct cultures as none of the technological and biochemical parameters of the cheeses was affected. Both the wild-type and the two derivatives were recovered from experimental cheeses up to 30 days after manufacture at similar rates of nearly 20%. Thus, the phenotypic traits under examination were not essential for L. plantarum to grow into the cheese matrix.

  10. Plasmid-cured Chlamydia caviae activates TLR2-dependent signaling and retains virulence in the guinea pig model of genital tract infection.

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    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.

  11. A versatile one-step CRISPR-Cas9 based approach to plasmid-curing

    DEFF Research Database (Denmark)

    Lauritsen, Ida; Porse, Andreas; Sommer, Morten Otto Alexander

    2017-01-01

    tool enabling rapid removal of plasmids from bacterial cells is lacking. Results Based on replicon abundance and sequence conservation analysis, we show that the vast majority of bacterial cloning and expression vectors share sequence similarities that allow for broad CRISPR-Cas9 targeting. We have...... widely used for expression and engineering purposes. By virtue of the CRISPR-Cas9 targeting, our platform is highly expandable and can be applied in a broad host context. We exemplify the wide applicability of our system in Gram-negative bacteria by demonstrating the successful application in both...

  12. "Curing" of plasmid DNA in acetogen using microwave or applying an electric pulse improves cell growth and metabolite production as compared to the plasmid-harboring strain.

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    Berzin, Vel; Kiriukhin, Michael; Tyurin, Michael

    2013-03-01

    Plasmid-free acetogen Clostridium sp. MT962 electrotransformed with a small cryptic plasmid pMT351 was used to develop time- and cost-effective methods for plasmid elimination. Elimination of pMT351 restored production of acetate and ethanol to the levels of the plasmid-free strain with no dry cell weight changes. Destabilizing cell membrane via microwave at 2.45 GHz, or exposure to a single 12 ms square electric pulse at 35 kV cm⁻¹, eliminated pMT351 in 42-47 % of cells. Plasmid elimination with a single square electric pulse required 10 versus 0.1 J needed to introduce the same 3,202-bp plasmid into the cells as calculated per cell sample of Clostridium sp. MT962. Microwave caused visible changes in repPCR pattern and increased ethanol production at the expense of acetate. This is the first report on microwave of microwave ovens, wireless routers, and mobile devices causing chromosomal DNA aberrations in microbes along with carbon flux change.

  13. Development of a gene transfer system for curing of plasmids in the marine fish pathogen Vibrio salmonicida.

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    Valla, S; Frydenlund, K; Coucheron, D H; Haugan, K; Johansen, B; Jørgensen, T; Knudsen, G; Strøm, A

    1992-01-01

    All reported natural isolates of the marine fish pathogen Vibrio salmonicida contain plasmids, and in another marine fish pathogen, Vibrio anguillarum, it has been shown that a plasmid is important for expression of virulence by the organism. To study the function of the plasmids in V. salmonicida, we developed a gene transfer system based on the plasmid RSF1010 replicon. The gene transfer system was used to construct a plasmid-free strain, and this strain was found to behave similarly to the wild type in a fish pathogenicity test based on intraperitoneal injection of the bacteria. We were unable to detect any other phenotypic differences between the two strains. It could therefore be concluded that at least in the V. salmonicida strain tested, extrachromosomal DNA is not required for expression of virulence. Images PMID:1622274

  14. Plasmid curing and the loss of grip--the 65-kb replicon of Phaeobacter inhibens DSM 17395 is required for biofilm formation, motility and the colonization of marine algae.

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    Frank, Oliver; Michael, Victoria; Päuker, Orsola; Boedeker, Christian; Jogler, Christian; Rohde, Manfred; Petersen, Jörn

    2015-03-01

    Surface colonization is characteristic for a broad range of marine roseobacters and many strains have been isolated from biofilms, microbial mats and dinoflagellates. Phaeobacter inhibens DSM 17395, one of the best-studied representatives of the Roseobacter group, is an effective colonizer of marine surfaces, but the genetic basis of this trait is unknown. Based on the composition of its 65-kb RepA-I type plasmid that contains more than 20 genes for polysaccharide metabolism, including a rhamnose operon, which is required for O-antigen formation in Escherichia coli, it was hypothesized that this replicon was essential for surface attachment. Accordingly, a holistic approach was taken and the functional role of this extrachromosomal element in P. inhibens was investigated. Plasmid curing was performed with the homologous RepA-I replication system of Dinoroseobacter shibae DSM 16493(T). The Δ65-kb mutant completely lost its stickiness and could neither attach to artificial (glass, polystyrene) nor to natural surfaces (algae) and, consequently, its ability to form biofilms was impaired. Surprisingly, the mutant also lost the capacity for flagellar swimming motility required for surface colonization and the dispersal of biofilms. The data clearly showed that the 65-kb replicon of P. inhibens DSM 17395 was a genuine biofilm plasmid-mediating surface attachment. Homologous replicons are widely distributed among Rhodobacterales thus indicating the general importance of extrachromosomal elements for biofilm formation.

  15. Plasmid Biopharmaceuticals.

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    Prazeres, Duarte Miguel F; Monteiro, Gabriel A

    2014-12-01

    Plasmids are currently an indispensable molecular tool in life science research and a central asset for the modern biotechnology industry, supporting its mission to produce pharmaceutical proteins, antibodies, vaccines, industrial enzymes, and molecular diagnostics, to name a few key products. Furthermore, plasmids have gradually stepped up in the past 20 years as useful biopharmaceuticals in the context of gene therapy and DNA vaccination interventions. This review provides a concise coverage of the scientific progress that has been made since the emergence of what are called today plasmid biopharmaceuticals. The most relevant topics are discussed to provide researchers with an updated overview of the field. A brief outline of the initial breakthroughs and innovations is followed by a discussion of the motivation behind the medical uses of plasmids in the context of therapeutic and prophylactic interventions. The molecular characteristics and rationale underlying the design of plasmid vectors as gene transfer agents are described and a description of the most important methods used to deliver plasmid biopharmaceuticals in vivo (gene gun, electroporation, cationic lipids and polymers, and micro- and nanoparticles) is provided. The major safety issues (integration and autoimmunity) surrounding the use of plasmid biopharmaceuticals is discussed next. Aspects related to the large-scale manufacturing are also covered, and reference is made to the plasmid products that have received marketing authorization as of today.

  16. Vascular Cures

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    ... is Possible EVERY DOLLAR SAVES LIVES. Donate Now Vascular Cures innovates patient-centered research, catalyzes breakthrough collaborations and empowers people in their vascular health journey. what is vascular disease PATIENTS see ...

  17. Role of Plasmid in Production of Acetobacter Xylinum Biofilms

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    Abbas Rezaee

    2005-01-01

    Full Text Available Acetobacter xylinum has the ability to produce cellulotic biofilms. Bacterial cellulose is expected to be used in many industrial or biomedical materials for its unique characteristics. A. xylinum contains a complex system of plasmid DNA molecules. A 44 kilobases (kb plasmid was isolated in wild type of A. xylinum. To improve the cellulose producing ability of A. xylinum, role of the plasmid in production of cellulose was studied. The comparisons between wild type and cured cells of A. xylinum showed that there is considerably difference in cellulose production. In order to study the relationship between plasmid and the rate of cellulose production, bacteria were screened for plasmid profile by a modified method for preparation of plasmid. This method yields high levels of pure plasmid DNA that can be used for common molecular techniques, such as digestion and transformation, with high efficiency.

  18. Transformation of Haemophilus influenzae by plasmid RSF0885

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    Notani, N.K.; Setlow, J.K.; McCarthy, D.; Clayton, N.L.

    1981-12-01

    Plasmid RSF0885, which conferred ampicillin resistance, transformed competent Haemophilus influenzae cells with low efficiency (maximun, less than 0.01%). As judged by competition experiments and uptake of radioactivity, plasmid RSF0885 deoxyribonucleic acid was taken up into competent H. influenzae cells several orders of magnitude less efficiently than H. influenzae chromosomal deoxyribonucleic acid. Plasmid RSF0885 transformed cells with even lower efficiency than could be accounted for by the low uptake. Transformation was not affected by rec-1 and rec-2 mutations in the recipient, and strains cured of the plasmid did not show increased transformation. Plasmid molecules cut once with a restriction enzyme that made blunt ends did not transform. Transformation was favored by the closed circular form of the plasmid.

  19. Chemotherapy of Bacterial Plasmids

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    1979-01-29

    render them non-susceptible to K: z plasmid-encoded enzymes. (3) Development of drugs which are selective inhibitor! 1 4, of plasmid DNA replication. (4... Development of drugs which inhibit phenotypic as expression of plasmid genes, and (5) Development of drugs which are inhibitors o, drug-inactivating...Barnes [2] them non-susceptible to plasmid-encoded enzymes, tabulated data on the incidence of Gram-negative 3) development of drugs which are

  20. Plasmid segregation mechanisms

    DEFF Research Database (Denmark)

    Ebersbach, Gitte; Gerdes, Kenn; Charbon, Gitte Ebersbach

    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 ...

  1. Plasmid-associated sensitivity of Bacillus thuringiensis to UV light

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    Benoit, T.G.; Wilson, G.R.; Bull, D.L.; Aronson, A.I. (Department of Agriculture, College Station, TX (USA))

    1990-08-01

    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.

  2. Characterization of the Lactobacillus plantarum plasmid pCD033 and generation of the plasmid free strain L. plantarum 3NSH.

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    Heiss, Silvia; Grabherr, Reingard; Heinl, Stefan

    2015-09-01

    Lactobacillus plantarum CD033, a strain isolated from grass silage in Austria, harbors a 7.9 kb plasmid designated pCD033. Sequence analysis identified 14 open reading frames and 8 of these were supposed to be putative coding sequences. Gene annotation revealed no putative essential genes being plasmid encoded, but a plasmid addiction system based on a PemI/PemK-like toxin-antitoxin system, able to stabilize plasmid maintenance. Absence of a replication initiation protein, a double strand origin as well as a single strand origin on plasmid pCD033 suggests replication via a new type of theta mechanism, whereby plasmid replication is potentially initiated and regulated by non-coding RNA. Detailed examination of segregational stability of plasmid vectors consisting of pCD033-fragments, combined with a selection marker, resulted in definition of a stably maintained minimal replicon. A gene encoding a RepB/OrfX-like protein was found to be not essential for plasmid replication. Alignment of the amino acid sequence of this protein with related proteins unveiled a highly conserved amino acid motif (LLDQQQ). L. plantarum CD033 was cured of pCD033 resulting in the novel plasmid free strain L. plantarum 3NSH. Plasmid curing demonstrated that no essential features are provided by pCD033 under laboratory conditions.

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

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    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

  4. Optimal Composite Curing System

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    Handel, Paul; Guerin, Daniel

    The Optimal Composite Curing System (OCCS) is an intelligent control system which incorporates heat transfer and resin kinetic models coupled with expert knowledge. It controls the curing of epoxy impregnated composites, preventing part overheating while maintaining maximum cure heatup rate. This results in a significant reduction in total cure time over standard methods. The system uses a cure process model, operating in real-time, to determine optimal cure profiles for tool/part configurations of varying thermal characteristics. These profiles indicate the heating and cooling necessary to insure a complete cure of each part in the autoclave in the minimum amount of time. The system coordinates these profiles to determine an optimal cure profile for a batch of thermally variant parts. Using process specified rules for proper autoclave operation, OCCS automatically controls the cure process, implementing the prescribed cure while monitoring the operation of the autoclave equipment.

  5. Chlamydial plasmids and bacteriophages.

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    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.

  6. Plasmid mediated antibiotic resistance in isolated bacteria from burned patients.

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    Beige, Fahimeh; Baseri Salehi, Majid; Bahador, Nima; Mobasherzadeh, Sina

    2015-01-01

    Nowadays, the treatment of burned patients is difficult because of the high frequency of infection with antibiotic resistance bacteria. This study was conducted to evaluate the level of antibiotic resistance in Gram-negative bacteria and its relation with the existence of plasmid. The samples were collected from two hundred twenty hospitalized burned patients in Isfahan burn hospital during a three-month period (March 2012 to June 2012). The samples were isolated and the Gram-negative bacteria were identified using phenotypic method and API 20E System. Antibiotic susceptibility and plasmid profile were determined by standard Agar disc diffusion and plasmid spin column extraction methods. Totally 117 Gram-negative bacteria were isolated, the most common were Pseudomonas aerugionsa (37.6%), P. fluorescens (25.6%), Acinetobacter baumanii (20/5%) and Klebsiella pneumoniae (7.6%), respectively. The isolates showed high frequency of antibiotic resistance against ceftazidime and co-amoxiclave (100%) and low frequency of antibiotic resistance against amikacin with (70%).The results indicated that 60% of the isolates harboured plasmid. On the other hand, the patients infected with A. baumanii and P. aeruginosa were cured (with 60% frequency) whereas, those infected with P. fluorescens were not cured. Hence, probably antibiotic resistance markers of A. baumanii and P. aeruginosa are plasmid mediated; however, P. fluorescens is chromosomally mediated. Based on our findings, P. aerugionsa is a major causative agent of wound infections and amikacin could be considered as a more effective antibiotic for treatment of the burned patients.

  7. IncHI2 Plasmids Are Predominant in Antibiotic-Resistant Salmonella Isolates

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    Chen, Wenyao; Fang, Tingzi; Zhou, Xiujuan; Zhang, Daofeng; Shi, Xianming; Shi, Chunlei

    2016-01-01

    The wide usage of antibiotics contributes to the increase in the prevalence of antibiotic-resistant Salmonella. Plasmids play a critical role in horizontal transfer of antibiotic resistance markers in Salmonella. This study aimed to screen and characterize plasmid profiles responsible for antibiotic resistance in Salmonella and ultimately to clarify the molecular mechanism of transferable plasmid-mediated antibiotic resistance. A total of 226 Salmonella isolates were examined for antimicrobial susceptibility by a disk diffusion method. Thirty-two isolates (14.2%) were resistant to at least one antibiotic. The presence of plasmid-mediated quinolone resistance (PMQR) genes and β-lactamase genes were established by PCR amplification. PCR-based replicon typing revealed that these 32 isolates represented seven plasmid incompatibility groups (IncP, HI2, A/C, FIIs, FIA, FIB, and I1), and the IncHI2 (59.4%) was predominant. Antibiotic resistance markers located on plasmids were identified through plasmid curing. Fifteen phenotypic variants were obtained with the curing efficiency of 46.9% (15/32). The cured plasmids mainly belong to the HI2 incompatibility group. The elimination of IncHI2 plasmids correlated with the loss of β-lactamase genes (blaOXA-1 and blaTEM-1) and PMQR genes (qnrA and aac(6′)-Ib-cr). Both IncHI2 and IncI1 plasmids in a S. enterica serovar Indiana isolate SJTUF 10584 were lost by curing. The blaCMY -2-carrying plasmid pS10584 from SJTUF 10584 was fully sequenced. Sequence analysis revealed that it possessed a plasmid scaffold typical for IncI1 plasmids with the unique genetic arrangement of IS1294-ΔISEcp1-blaCMY -2-blc-sugE-ΔecnR inserted into the colicin gene cia. These data suggested that IncHI2 was the major plasmid lineage contributing to the dissemination of antibiotic resistance in Salmonella and the activity of multiple mobile genetic elements may contribute to antibiotic resistance evolution and dissemination between different plasmid

  8. Curing Behavior of Epoxy Asphalt

    Institute of Scientific and Technical Information of China (English)

    YU Jianying; CONG Peiliang; WU Shaopeng; CHENG Songbo

    2009-01-01

    The curing process of epoxy asphalt was analyzed by Fourier transform infrared (FT-IR)spectroscopy.Effect of curing temperature on viscosity of epoxy asphalt,and changes of mechanical properties with curing time were investigated.The evolution of concentration of epoxy band was followed as a function of the applied curing process.The experimental results indicate that the curing reaction rate of epoxy asphalt is invariable before 70 min at 120℃,and it decreases when curing time exceeds 70 min.The viscosity of epoxy asphalt increases slowly with curing time at initial curing stage.But it increases quickly after initial curing stage and the initial curing time decreases as the curing temperature increases.The tensile strength increases slowly at incipient curing stage and increases rapidly when curing time is form 20 min to 70 min.The elongation at break shows a decrease with curing time,but it exceeds 200%after cured.

  9. Mechanisms of plasmid segregation: have multicopy plasmids been overlooked?

    Science.gov (United States)

    Million-Weaver, Samuel; Camps, Manel

    2014-09-01

    Plasmids are self-replicating pieces of DNA typically bearing non-essential genes. Given that plasmids represent a metabolic burden to the host, mechanisms ensuring plasmid transmission to daughter cells are critical for their stable maintenance in the population. Here we review these mechanisms, focusing on two active partition strategies common to low-copy plasmids: par systems type I and type II. Both involve three components: an adaptor protein, a motor protein, and a centromere, which is a sequence area in the plasmid that is recognized by the adaptor protein. The centromere-bound adaptor nucleates polymerization of the motor, leading to filament formation, which can pull plasmids apart (par I) or push them towards opposite poles of the cell (par II). No such active partition mechanisms are known to occur in high copy number plasmids. In this case, vertical transmission is generally considered stochastic, due to the random distribution of plasmids in the cytoplasm. We discuss conceptual and experimental lines of evidence questioning the random distribution model and posit the existence of a mechanism for segregation in high copy number plasmids that moves plasmids to cell poles to facilitate transmission to daughter cells. This mechanism would involve chromosomally-encoded proteins and the plasmid origin of replication. Modulation of this proposed mechanism of segregation could provide new ways to enhance plasmid stability in the context of recombinant gene expression, which is limiting for large-scale protein production and for bioremediation.

  10. Plasmid-to-plasmid recombination in Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

    Balganesh, M.; Setlow, J.K.

    1986-01-01

    No recombination between plasmids was observed after conjugal transfer of a plasmid into a cell carrying another plasmid. Two types of such recombination took place after transformation, one type being Rec/sup +/ dependent and suggesting a preferred site of recombination. The other much rarer type was at least partially Rec/sup +/ independent.

  11. Radiation curing of epoxies

    Science.gov (United States)

    Dickson, Lawrence W.; Singh, Ajit

    The literature on radiation polymerization of epoxy compounds has been reviewed to assess the potential use of radiation for curing these industrially important monomers. Chemical curing of epoxies may proceed by either cationic or anionic mechanisms depending on the nature of the curing agent, but most epoxies polymerize by cationic mechanisms under the influence of high-energy radiation. Radiation-induced cationic polymerization of epoxy compounds is inhibited by trace quantities of water because of proton transfer from the chain-propagating epoxy cation to water. Several different methods with potential for obtaining high molecular weight polymers by curing epoxies with high-energy radiation have been studied. Polymeric products with epoxy-like properties have been produced by radiation curing of epoxy oligomers with terminal acrylate groups and mixtures of epoxies with vinyl monomers. Both of these types of resin have good potential for industrial-scale curing by radiation treatment.

  12. 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.

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

    Science.gov (United States)

    Kirchner, Miranda; Abuoun, Manal; Mafura, Muriel; Bagnall, Mary; Hunt, Theresa; Thomas, Christopher; Weile, Jan; Anjum, Muna F

    2013-01-01

    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.

  14. The Curing Meter

    DEFF Research Database (Denmark)

    Jensen, Ole Mejlhede

    2006-01-01

    Curing technology concerns adjustment and control of temperature and moisture conditions in hardening structures and elements of concrete. The curing technology comprises e.g. measurement, adjustment and control of the moisture conditions during the early hardening phase of the concrete to achiev...... from concrete surfaces in the early hardening phase. The Concrete Curing Meter is designed for simple and reliable monitoring of water loss from a wet surface of fresh concrete. This new measuring technique has been tested with several prototypes of the Curing Meter....

  15. Addiction protein Phd of plasmid prophage P1 is a substrate of the ClpXP serine protease of Escherichia coli.

    OpenAIRE

    Lehnherr, H; Yarmolinsky, M B

    1995-01-01

    Plasmid-encoded addiction genes augment the apparent stability of various low copy number bacterial plasmids by selectively killing plasmid-free (cured) segregants or their progeny. The addiction module of plasmid prophage P1 consists of a pair of genes called phd and doc. Phd serves to prevent host death when the prophage is retained and, should retention mechanisms fail, Doc causes death on curing. Doc acts as a cell toxin to which Phd is an antidote. In this study we show that host mutants...

  16. The Curing Meter

    DEFF Research Database (Denmark)

    Jensen, Ole Mejlhede

    2006-01-01

    Curing technology concerns adjustment and control of temperature and moisture conditions in hardening structures and elements of concrete. The curing technology comprises e.g. measurement, adjustment and control of the moisture conditions during the early hardening phase of the concrete to achiev...... from concrete surfaces in the early hardening phase. The Concrete Curing Meter is designed for simple and reliable monitoring of water loss from a wet surface of fresh concrete. This new measuring technique has been tested with several prototypes of the Curing Meter.......Curing technology concerns adjustment and control of temperature and moisture conditions in hardening structures and elements of concrete. The curing technology comprises e.g. measurement, adjustment and control of the moisture conditions during the early hardening phase of the concrete to achieve...... an optimal development of properties in the hardening concrete – so-called "moisture curing". Proper moisture curing is vital to concrete but laborious to accomplish and difficult to control at the construction site with present methods. This paper concerns a new method for site measurement of evaporation...

  17. Evidence for plasmid-encoded virulence factors in the phytopathogenic bacterium Clavibacter michiganensis subsp. michiganensis NCPPB382.

    Science.gov (United States)

    Meletzus, D; Bermphol, A; Dreier, J; Eichenlaub, R

    1993-01-01

    The tomato pathogen Clavibacter michiganensis subsp. michiganensis NCPPB382, which causes bacterial wilt, harbors two plasmids pCM1 (27.5 kb) and pCM2 (72 kb). After curing of the plasmids, bacterial derivatives were still proficient in the ability to colonize the host plant and in the production of exopolysaccharides but exhibited a reduced virulence. When one of the two plasmids is lost, there is a significant delay in the development of wilting symptoms after infection and a plasmid-free derivative is not able to induce disease symptoms. By cloning of restriction fragments of both plasmids in the plasmid-free strain CMM100, two DNA fragments which restored the virulent phenotype were identified. Further analysis suggested that a fragment of plasmid pCM1 encodes an endocellulase which is involved in the expression of the pathogenic phenotype. Images PMID:8458855

  18. C-CURE

    Data.gov (United States)

    US Agency for International Development — C-CURE system manages certain aspects of the access control system, including collecting employee and contractor names and photographs. The Office of Security uses...

  19. Curing of Concrete

    African Journals Online (AJOL)

    and pipes illustrate defects frequently caused by improper curing. Proper ... because in such pastes the layers of hydrated ... *3) When cement hydrates the solid products occupy a volume equal to the sum of volumes of anhydrous cement and.

  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 successful propagation and long-term continued existence of these extra-chromosomal elements is extensive. Apart from the accessory genetic elements that may provide plasmid-harboring cells a selective advantage, special focus is placed on the mechanisms conjugative plasmids employ to ensure their stable...... maintenance in the host cell. These importantly include the ability to self-mobilize in a process termed conjugative transfer, which may occur across species barriers. Other plasmid stabilizing mechanisms include the multimer resolution system, active partitioning, and post-segregational-killing of plasmid...

  1. Plasmid-Borne Antimicrobial Resistance of Staphylococcus aureus Isolated in a Hospital in Lisbon, Portugal.

    Science.gov (United States)

    Costa, Sofia Santos; Palma, Cláudia; Kadlec, Kristina; Fessler, Andrea T; Viveiros, Miguel; Melo-Cristino, José; Schwarz, Stefan; Couto, Isabel

    2016-12-01

    Plasmids play a key role in the genetic plasticity and survival of Staphylococcus aureus in challenging environments. Although many S. aureus plasmids have been described, still few studies portray the plasmid content of a given S. aureus population. The aim of this work was to characterize the plasmids carried by a collection of 53 S. aureus isolates collected in a large hospital in Lisbon, Portugal, and investigate their role in conferring resistance to several antimicrobial agents. Plasmids were present in 44 out of the 53 isolates and were grouped into eleven AccI restriction profiles. Plasmid curing of representative strains and comparison of antimicrobial susceptibility profiles between pairs of isogenic strains proved to be a valuable guidance tool in the identification of plasmid-located resistance genes. The plasmids harbored several resistance genes, namely blaZ (resistance to β-lactams), erm(C) (resistance to macrolides, lincosamides, and streptogramin B), cadA (resistance to cadmium and zinc), cadD (resistance to cadmium), and qacA and smr (resistance to biocides and dyes). This study demonstrates the impact of plasmids on the resistance properties of S. aureus, highlighting their role in the dissemination of antibiotic, heavy metal, and biocide resistance genes, and survival of this major pathogen in the hospital environment.

  2. Toxin Plasmids of Clostridium perfringens

    Science.gov (United States)

    Li, Jihong; Adams, Vicki; Bannam, Trudi L.; Miyamoto, Kazuaki; Garcia, Jorge P.; Uzal, Francisco A.; Rood, Julian I.

    2013-01-01

    SUMMARY In both humans and animals, Clostridium perfringens is an important cause of histotoxic infections and diseases originating in the intestines, such as enteritis and enterotoxemia. The virulence of this Gram-positive, anaerobic bacterium is heavily dependent upon its prolific toxin-producing ability. Many of the ∼16 toxins produced by C. perfringens are encoded by large plasmids that range in size from ∼45 kb to ∼140 kb. These plasmid-encoded toxins are often closely associated with mobile elements. A C. perfringens strain can carry up to three different toxin plasmids, with a single plasmid carrying up to three distinct toxin genes. Molecular Koch's postulate analyses have established the importance of several plasmid-encoded toxins when C. perfringens disease strains cause enteritis or enterotoxemias. Many toxin plasmids are closely related, suggesting a common evolutionary origin. In particular, most toxin plasmids and some antibiotic resistance plasmids of C. perfringens share an ∼35-kb region containing a Tn916-related conjugation locus named tcp (transfer of clostridial plasmids). This tcp locus can mediate highly efficient conjugative transfer of these toxin or resistance plasmids. For example, conjugative transfer of a toxin plasmid from an infecting strain to C. perfringens normal intestinal flora strains may help to amplify and prolong an infection. Therefore, the presence of toxin genes on conjugative plasmids, particularly in association with insertion sequences that may mobilize these toxin genes, likely provides C. perfringens with considerable virulence plasticity and adaptability when it causes diseases originating in the gastrointestinal tract. PMID:23699255

  3. A novel plasmid pEA68 of Erwinia amylovora and the description of a new family of plasmids.

    Science.gov (United States)

    Ismail, Emadeldeen; Blom, Jochen; Bultreys, Alain; Ivanović, Milan; Obradović, Aleksa; van Doorn, Joop; Bergsma-Vlami, Maria; Maes, Martine; Willems, Anne; Duffy, Brion; Stockwell, Virginia O; Smits, Theo H M; Puławska, Joanna

    2014-12-01

    Recent genome analysis of Erwinia amylovora, the causal agent of fire blight disease on Rosaceae, has shown that the chromosome is highly conserved among strains and that plasmids are the principal source of genomic diversity. A new circular plasmid, pEA68, was found in E. amylovora strain 692 (LMG 28361), isolated in Poland from Sorbus (mountain ash) with fire blight symptoms. Annotation of the 68,763-bp IncFIIa-type plasmid revealed that it contains 79 predicted CDS, among which two operons (tra, pil) are associated with mobility. The plasmid is maintained stably in E. amylovora and does not possess genes associated with antibiotic resistance or known virulence genes. Curing E. amylovora strain 692 of pEA68 did not influence its virulence in apple shoots nor amylovoran synthesis. Of 488 strains of E. amylovora from seventeen countries, pEA68 was only found in two additional strains from Belgium. Although the spread of pEA68 is currently limited to Europe, pEA68 comprises, together with pEA72 and pEA78 both found in North America, a new plasmid family that spans two continents.

  4. Radiation curing in the eighties

    Energy Technology Data Exchange (ETDEWEB)

    Vrancken, A. (UCB, Drogenbos (Belgium). Chemical Specialities Div.)

    1984-05-01

    The subject is discussed under the headings: introduction; what is radiation curing; history; radiation curable resins (with properties of products); ultraviolet and electron beam curing; photoinitiation and the ultraviolet light curing process; electron beam curing (initiation; electron beam accelerators); end uses (graphic arts; wood finishing; paper upgrading; adhesives; metal finishing; electronic chemical; floor coatings).

  5. Conjugative plasmids of Neisseria gonorrhoeae.

    Directory of Open Access Journals (Sweden)

    Emilia Pachulec

    Full Text Available Many clinical isolates of the human pathogen Neisseria gonorrhoeae contain conjugative plasmids. The host range of these plasmids is limited to Neisseria species, but presence of a tetracycline (tetM determinant inserted in several of these plasmids is an important cause of the rapid spread of tetracycline resistance. Previously plasmids with different backbones (Dutch and American type backbones and with and without different tetM determinants (Dutch and American type tetM determinants have been identified. Within the isolates tested, all plasmids with American or Dutch type tetM determinants contained a Dutch type plasmid backbone. This demonstrated that tetM determinants should not be used to differentiate between conjugal plasmid backbones. The nucleotide sequences of conjugative plasmids with Dutch type plasmid backbones either not containing the tetM determinant (pEP5233 or containing Dutch (pEP5289 or American (pEP5050 type tetM determinants were determined. Analysis of the backbone sequences showed that they belong to a novel IncP1 subfamily divergent from the IncP1alpha, beta, gamma, delta and epsilon subfamilies. The tetM determinants were inserted in a genetic load region found in all these plasmids. Insertion was accompanied by the insertion of a gene with an unknown function, and rearrangement of a toxin/antitoxin gene cluster. The genetic load region contains two toxin/antitoxins of the Zeta/Epsilon toxin/antitoxin family previously only found in Gram positive organisms and the virulence associated protein D of the VapD/VapX toxin/antitoxin family. Remarkably, presence of VapX of pJD1, a small cryptic neisserial plasmid, in the acceptor strain strongly increased the conjugation efficiency, suggesting that it functions as an antitoxin for the conjugative plasmid. The presence of the toxin and antitoxin on different plasmids might explain why the host range of this IncP1 plasmid is limited to Neisseria species. The isolated plasmids

  6. Molecular analysis of the bacteriocin-encoding plasmid pDGL1 from Enterococcus durans and genetic characterization of the durancin locus

    Science.gov (United States)

    Enterococci constitute a significant component of lactic acid bacteria normally present in the intestinal microflora and include strains that produce bacteriocins. The genetic determinants for durancin GL in Enterococcus durans 41D were identified on the 8,347 bp plasmid pDGL1 by plasmid curing exp...

  7. Role of Plasmids in Lactobacillus brevis BSO 464 Hop Tolerance and Beer Spoilage

    Science.gov (United States)

    Bergsveinson, Jordyn; Baecker, Nina; Pittet, Vanessa

    2014-01-01

    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. PMID:25501474

  8. 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.

  9. Phenotypic plasticity in bacterial plasmids.

    Science.gov (United States)

    Turner, Paul E

    2004-01-01

    Plasmid pB15 was previously shown to evolve increased horizontal (infectious) transfer at the expense of reduced vertical (intergenerational) transfer and vice versa, a key trade-off assumed in theories of parasite virulence. Whereas the models predict that susceptible host abundance should determine which mode of transfer is selectively favored, host density failed to mediate the trade-off in pB15. One possibility is that the plasmid's transfer deviates from the assumption that horizontal spread (conjugation) occurs in direct proportion to cell density. I tested this hypothesis using Escherichia coli/pB15 associations in laboratory serial culture. Contrary to most models of plasmid transfer kinetics, my data show that pB15 invades static (nonshaking) bacterial cultures only at intermediate densities. The results can be explained by phenotypic plasticity in traits governing plasmid transfer. As cells become more numerous, the plasmid's conjugative transfer unexpectedly declines, while the trade-off between transmission routes causes vertical transfer to increase. Thus, at intermediate densities the plasmid's horizontal transfer can offset selection against plasmid-bearing cells, but at high densities pB15 conjugates so poorly that it cannot invade. I discuss adaptive vs. nonadaptive causes for the phenotypic plasticity, as well as potential mechanisms that may lead to complex transfer dynamics of plasmids in liquid environments. PMID:15166133

  10. Plasmid Rolling-Circle Replication.

    Science.gov (United States)

    Ruiz-Masó, J A; MachóN, C; Bordanaba-Ruiseco, L; Espinosa, M; Coll, M; Del Solar, G

    2015-02-01

    Plasmids are DNA entities that undergo controlled replication independent of the chromosomal DNA, a crucial step that guarantees the prevalence of the plasmid in its host. DNA replication has to cope with the incapacity of the DNA polymerases to start de novo DNA synthesis, and different replication mechanisms offer diverse solutions to this problem. Rolling-circle replication (RCR) is a mechanism adopted by certain plasmids, among other genetic elements, that represents one of the simplest initiation strategies, that is, the nicking by a replication initiator protein on one parental strand to generate the primer for leading-strand initiation and a single priming site for lagging-strand synthesis. All RCR plasmid genomes consist of a number of basic elements: leading strand initiation and control, lagging strand origin, phenotypic determinants, and mobilization, generally in that order of frequency. RCR has been mainly characterized in Gram-positive bacterial plasmids, although it has also been described in Gram-negative bacterial or archaeal plasmids. Here we aim to provide an overview of the RCR plasmids' lifestyle, with emphasis on their characteristic traits, promiscuity, stability, utility as vectors, etc. While RCR is one of the best-characterized plasmid replication mechanisms, there are still many questions left unanswered, which will be pointed out along the way in this review.

  11. Conjugation efficiency depends on intra and intercellular interactions between distinct plasmids: Plasmids promote the immigration of other plasmids but repress co-colonizing plasmids.

    Science.gov (United States)

    Gama, João Alves; Zilhão, Rita; Dionisio, Francisco

    2017-08-24

    Conjugative plasmids encode the genes responsible for the synthesis of conjugative pili and plasmid transfer. Expression of the conjugative machinery (including conjugative pili) may be costly to bacteria, not only due to the energetic/metabolic cost associated with their expression but also because they serve as receptors for certain viruses. Consequently, the presence of two plasmids in the same cell may be disadvantageous to each plasmid, because they may impose a higher fitness cost on the host. Therefore, plasmids may encode mechanisms to cope with co-resident plasmids. Moreover, it is possible that the transfer rate of a plasmid is affected by the presence of a distinct plasmid in the recipient cell. In this work, we measured transfer rates of twelve natural plasmids belonging to seven incompatibility groups in three situations, namely when: (i) donor cells contain a plasmid and recipient cells are plasmid-free; (ii) donor cells contain two unrelated plasmids and recipient cells are plasmid-free; and (iii) half of the cells contain a given plasmid and the other half contain another, unrelated, plasmid. In the third situation, recipient cells of a plasmid are the donor cells of the other plasmid. We show that there are more negative interactions (reduction of a plasmid's conjugative efficiency) between plasmids if they reside in the same cell than if they reside in different cells. However, if plasmids interacted intercellularly, the transfer rate of one of the plasmids was often higher (when the unrelated conjugative plasmid was present in the recipient cell) than if the recipient cell was plasmid-free - a positive effect. Experimental data retrieved from the study of mutant plasmids not expressing conjugative pili on the cell surface suggest that positive effects result from a higher efficiency of mating pair formation. Overall, our results suggest that negative interactions are significantly more frequent when plasmids occupy the same cell. Such

  12. Plasmid recombination in Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

    McCarthy, D.

    1982-01-01

    DNA recombination in exponential phase and competent Haemophilus influenzae was measured by an electron microscopic assay that relies on the conversion of plasmid RSF0885 monomers into multimeric forms. Dimer circles were present at a frequency of 2% in plasmid preparations from competent Rd (wild-type) cells; multimers were present at a frequency of 0.2% in preparations from exponential phase cells. Thus, plasmid recombination was stimulated in competent cells. Multimer formation occurred efficiently in cells of the transformation defective mutant rec2, implying that the rec2 gene product is not required for plasmid recombination. However, the absence of multimer plasmids in preparations from competent cells of the transformation defective mutant rec1 suggests that the rec1 gene product is required. Digestion of purified plasmids with restriction endonuclease PvuII, which makes a single cut in the monomer, revealed the presence of recombination intermediates composed of two linear plasmids joined to form two pairs of arms resembling the Greek letter chi. Length measurements of these arms taken from a population of recombination intermediates gave evidence that the plasmids were joined at sites of homology. The distributions of individual DNA strands, at the intersections of the four arms, could be resolved in some recombination intermediates and were of two types. The first type of junction appeared as a single-stranded arm appended to each corner. The second type of junction consisted of a single strand of DNA linking the two linear plasmids at a site of homology. The single-stranded linker was frequently situated at the edge of a short gap on one of the plasmids in the pair. The fine structures of the recombinational joints have been interpreted in terms of previously proposed models of recombination.

  13. Colonization of the respiratory tract by a virulent strain of avian Escherichia coli requires carriage of a conjugative plasmid.

    Science.gov (United States)

    Ginns, C A; Benham, M L; Adams, L M; Whithear, K G; Bettelheim, K A; Crabb, B S; Browning, G F

    2000-03-01

    The E3 strain of E. coli was isolated in an outbreak of respiratory disease in broiler chickens, and experimental aerosol exposure of chickens to this strain induced disease similar to that seen in the field. In order to establish whether the virulent phenotype of this strain was associated with carriage of particular plasmids, four plasmid-cured derivatives, each lacking two or more of the plasmids carried by the wild-type strain, were assessed for virulence. Virulence was found to be associated with one large plasmid, pVM01. Plasmid pVM01 was marked by introduction of the transposon TnphoA, carrying kanamycin resistance, and was then cloned by transformation of E. coli strain DH5alpha. The cloned plasmid was then reintroduced by conjugation into an avirulent plasmid-cured derivative of strain E3 which lacked pVM01. The conjugant was shown to be as virulent as the wild-type strain E3, establishing that this plasmid is required for virulence following aerosol exposure. This virulence plasmid conferred expression of a hydroxamate siderophore, but not colicins, on both strain E3 and strain DH5alpha. Carriage of this plasmid was required for strain E3 to colonize the respiratory tracts of chickens but was not necessary for colonization of the gastrointestinal tract. However, the virulence plasmid did not confer virulence, or the capacity to colonize the respiratory tract, on strain DH5alpha. Thus, these studies have established that infection of chickens with E. coli strain E3 by the respiratory route is dependent on carriage of a conjugative virulence plasmid, which confers the capacity to colonize specifically the respiratory tract and which also carries genes for expression of a hydroxymate siderophore. These findings will facilitate identification of the specific genes required for virulence in these pathogens.

  14. Dual cure photocatalyst systems

    Energy Technology Data Exchange (ETDEWEB)

    DeVoe, R.J.; Brown-Wensley, K.A.; Holmes, G.L.; Mathis, M.D.; McCormick, F.B.; Palazzotto, M.C.; Spurgeon, K.M. (Minnesota Mining and Mfg. Co., St. Paul, MN (USA). Corporate Research Labs.)

    1990-01-01

    A family of dual cure photocatalyst systems is being developed to be used in the solventless processing of organic coatings. The photocatalyst systems consist of organometallic compounds often in combination with other agents. Upon photolysis, the photocatalyst system generates a Lewis acid and a free radical. The Lewis acid can initiate the polymerization of epoxies or the addition of isocyanates and polyols to form polyurethanes while the free radical can initiate the polymerization of acrylates. The performance of the various photocatalyst systems will be compared on the basis of the physical properties of the cured compositions they produce. 17 figs.

  15. [Labelling of nif-plasmid pEA9 from Enterobacter agglomerans 339].

    Science.gov (United States)

    Liu, Cheng-jun; Klingmüller, Walter

    2002-07-01

    The authors describe the in vivo labelling of the plasmid pEA9 in Enterobacter agglomerans 339 with a kanamycin resistance gene. For labelling purposes the donor plasmid pST5 was constructed. This plasmid contains the nif ENX region from pEA9,in which a kanamycin resistance gene is cloned.pST5 was transformed into E.a.339 and subsequently cured from the host. Curing was achieved with AP medium. Fourty strains that had lost pST5,but retained the kanamycin resistance, could be isolated. It showed that none of these clones contained co-integrates of pST5 and pEA9. This is evident that in all clones the kanamycin resistance gene was integrated into pEA9 by homologous recombination.

  16. Plasmid control of 6-aminohexanoic acid cyclic dimer degradation enzymes of Flavobacterium sp. KI72.

    Science.gov (United States)

    Negoro, S; Shinagawa, H; Nakata, A; Kinoshita, S; Hatozaki, T; Okada, H

    1980-07-01

    Flavobacterium sp. K172, which is able to grow on 6-aminohexanoic acid cyclic dimer as the sole source of carbon and nitrogen, and plasmid control of the responsible enzymes, 6-aminohexanoic acid cyclic dimer hydrolase and 6-aminohexanoic acid linear oligomer hydrolase, were studied. The wild strain of K172 harbors three kinds of plasmid, pOAD1 (26.2 megadaltons), pOAD2 (28.8 megadaltons), and pOAD3 (37.2 megadaltons). The wild strain K172 was readily cured of its ability to grow on the cyclic dimer by mitomycin C, and the cyclic dimer hydrolase could not be detected either as catalytic activity or by antibody precipitation. No reversion of the cured strains was detected. pOAD2 was not detected in every cured strain tested but was restored in a transformant. The transformant recovered both of the enzyme activities, and the cyclic dimer hydrolase of the transformant was immunologically identical with that of the wild strain. All of the strains tested, including the wild, cured, and transformant ones, possessed identical pOAD3 irrespective of the metabolizing activity. Some of the cured strains possessed pOAD1 identical with the wild strain, but the others harbored plasmids with partially altered structures which were likely to be derived from pOAD1 by genetic rearrangements such as deletion, insertion, or substitution. These results suggested that the genes of the enzymes were borne on pOAD2.

  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. Curing the queue

    NARCIS (Netherlands)

    Zonderland, Maartje Elisabeth

    2012-01-01

    In this dissertation we study several problems related to the management of healthcare and the cure of disease. In each chapter a hospital capacity distribution problem is analyzed using techniques from operations research, also known as mathematical decision theory. The problems considered are insp

  19. 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...

  20. Hiccups: causes and cures.

    Science.gov (United States)

    Lewis, J H

    1985-12-01

    Hiccups result from a wide variety of conditions that act on the supraspinal hiccup center or that stimulate or disinhibit the limbs of its reflex arc. While scores of hiccup remedies have been reported over the centuries, no single "cure" stands out as being the most effective. Measures that stimulate the uvula or pharynx or disrupt diaphragmatic (respiratory) rhythm are simple to use and often help to speed the end of a bout of otherwise benign, self-limited hiccups. Such manueuvers may also terminate persistent hiccups. Drug therapy usually becomes necessary for more intractable hiccups; chlorpromazine and metoclopramide being two of the most widely employed agents for this purpose. Physical disruption of the phrenic nerve, hypnosis, and acupuncture are other modes of therapy that have been used in severe cases. Because so many reports of hiccup "cures" are based on anecdotal experience rather than controlled clinical studies, I review the available treatments to provide a rational approach for the management of hiccups.

  1. TEM-1-encoding small plasmids impose dissimilar fitness costs on Haemophilus influenzae and Haemophilus parainfluenzae.

    Science.gov (United States)

    Søndergaard, Annette; Lund, Marianne; Nørskov-Lauritsen, Niels

    2015-12-01

    Only two beta-lactamases, TEM-1 and ROB-1, have been observed in Haemophilus influenzae, while four different TEM but no ROB enzymes have been found in Haemophilus parainfluenzae. In order to investigate the mechanisms behind the dissemination of small beta-lactamase-encoding plasmids in H. influenzae and H. parainfluenzae, we assessed the fitness cost of three TEM-1- (pPN223, pA1209, pA1606), one TEM-15- (pSF3) and one ROB-1-bearing (pB1000) plasmid when expressed in either bacterial species. All plasmids were stable in H. influenzae and H. parainfluenzae except pB1000, which showed on average (sample mean) 76% curing in H. parainfluenzae after 5  days of subculture. Competition assays between isogenic strains with and without plasmid showed no competitive disadvantage of pPN223 and pA1606 in H. influenzae, or of pA1209 in H. parainfluenzae. In contrast, pSF3 and pB1000 were associated with significant competitive disadvantages in both species. Some of the competitive disadvantages may be related to differences in plasmid copy number and mRNA expression of the beta-lactamase genes, as revealed by quantitative PCR analysis. In conclusion, plasmids encoding TEM beta-lactamases isolated from H. influenzae and H. parainfluenzae can be stably transferred between species. The fast curing of pB1000 in H. parainfluenzae observed in this study correlates to the fact that ROB-1 has never been reported for this species. TEM-1-encoding plasmids are associated with the lowest level of fitness cost, but different TEM-1 plasmids confer different levels of fitness cost on the two hosts.

  2. Plasmid-Mediated Dimethoate Degradation by Bacillus licheniformis Isolated From a Fresh Water Fish Labeo rohita

    Directory of Open Access Journals (Sweden)

    Manisha Deb Mandal

    2005-01-01

    Full Text Available The Bacillus licheniformis strain isolated from the intestine of Labeo rohita by an enrichment technique showed capability of utilizing dimethoate as the sole source of carbon. The bacterium rapidly utilized dimethoate beyond 0.6 mg/mL and showed prolific growth in a mineral salts medium containing 0.45 mg/mL dimethoate. The isolated B licheniformis exhibited high level of tolerance of dimethoate (3.5 mg/mL in nutrient broth, while its cured mutant did not tolerate dimethoate beyond 0.45 mg/mL and it was unable to utilize dimethoate. The wild B licheniformis strain transferred dimethoate degradation property to E coli C600 (Nar, F− strain. The transconjugant harbored a plasmid of the same molecular size (approximately 54 kb as that of the donor plasmid; the cured strain was plasmid less. Thus a single plasmid of approximately 54 kb was involved in dimethoate degradation. Genes encoding resistance to antibiotic and heavy metal were also located on the plasmid.

  3. Plasmid-Mediated Dimethoate Degradation by Bacillus licheniformis Isolated From a Fresh Water Fish Labeo rohita

    Science.gov (United States)

    2005-01-01

    The Bacillus licheniformis strain isolated from the intestine of Labeo rohita by an enrichment technique showed capability of utilizing dimethoate as the sole source of carbon. The bacterium rapidly utilized dimethoate beyond 0.6 mg/mL and showed prolific growth in a mineral salts medium containing 0.45 mg/mL dimethoate. The isolated B licheniformis exhibited high level of tolerance of dimethoate (3.5 mg/mL) in nutrient broth, while its cured mutant did not tolerate dimethoate beyond 0.45 mg/mL and it was unable to utilize dimethoate. The wild B licheniformis strain transferred dimethoate degradation property to E coli C600 (Nar, F−) strain. The transconjugant harbored a plasmid of the same molecular size (approximately 54 kb) as that of the donor plasmid; the cured strain was plasmid less. Thus a single plasmid of approximately 54 kb was involved in dimethoate degradation. Genes encoding resistance to antibiotic and heavy metal were also located on the plasmid. PMID:16192686

  4. Modeling HIV Cure

    Science.gov (United States)

    Perelson, Alan; Conway, Jessica; Cao, Youfang

    A large effort is being made to find a means to cure HIV infection. I will present a dynamical model of post-treatment control (PTC) or ``functional cure'' of HIV-infection. Some patients treated with suppressive antiviral therapy have been taken off of therapy and then spontaneously control HIV infection such that the amount of virus in the circulation is maintained undetectable by clinical assays for years. The model explains PTC occurring in some patients by having a parameter regime in which the model exhibits bistability, with both a low and high steady state viral load being stable. The model makes a number of predictions about how to attain the low PTC steady state. Bistability in this model depends upon the immune response becoming exhausted when over stimulated. I will also present a generalization of the model in which immunotherapy can be used to reverse immune exhaustion and compare model predictions with experiments in SIV infected macaques given immunotherapy and then taken off of antiretroviral therapy. Lastly, if time permits, I will discuss one of the hurdles to true HIV eradication, latently infected cells, and present clinical trial data and a new model addressing pharmacological means of flushing out the latent reservoir. Supported by NIH Grants AI028433 and OD011095.

  5. Co-resident plasmids travel together.

    Science.gov (United States)

    Gama, João Alves; Zilhão, Rita; Dionisio, Francisco

    2017-08-24

    Conjugative plasmids encode genes that enable them to transfer, by conjugation, from a given host cell to another cell. Conjugative transfer, despite being an important feature of conjugative plasmids, is not constitutive for most plasmids, the reason being that genes involved in horizontal transfer are mostly repressed. Only upon their transient de-repression are plasmids able to transfer horizontally. If host cells harbour multiple plasmids, their simultaneous transfer depends on simultaneous transient de-repression of all plasmids. If de-repression of different plasmids was random and independent events, simultaneous de-repression should be a rare event because the probability of simultaneous de-repression would be the product of the probabilities of de-repression of each plasmid. Some previous observations support this hypothesis, while others show that co-transfer of plasmids is more frequent than this reasoning indicates. Here, we show that co-transfer of multiple plasmids mainly results from non-independent events: the probability that all plasmids within a cell become de-repressed is much higher than if de-repression of plasmids genes were independent. We found a simple model for the probability of co-transfer: the plasmid having the lowest conjugation rates is the one who limits co-transfer. In this sense, cells receiving the plasmid with the lower transfer rate also receive the other plasmid. If de-repression happens simultaneously on co-resident plasmids, common cues may stimulate de-repression of distinct plasmids. Copyright © 2017 Elsevier Inc. All rights reserved.

  6. The Chlamydophila felis plasmid is highly conserved.

    Science.gov (United States)

    Harley, Ross; Day, Sarinder; Di Rocco, Camillo; Helps, Chris

    2010-11-20

    The presence of a plasmid in the Chlamydiaceae is both species and strain specific. Knowledge of the prevalence of the plasmid in different Chlamydia species is important for future studies aiming to investigate the role of the plasmid in chlamydial biology and disease. Although strains of Chlamydophila felis with or without the plasmid have been identified, only a small number of laboratory-adapted strains have been analysed and the prevalence of the plasmid in field isolates has not been determined. This study aimed to determine the prevalence of the plasmid in C. felis-positive conjunctival and oropharyngeal clinical samples submitted for routine diagnosis of C. felis by real-time (Q)PCR. DNA extracts from four laboratory-adapted strains were also analysed. QPCR assays targeting regions of C. felis plasmid genes pCF01, pCF02 and pCF03 were developed for the detection of plasmid DNA. QPCR analysis of DNA extracts from C. felis-positive clinical samples found evidence of plasmid DNA in 591 of 595 samples representing 561 of 564 (99.5%) clinical cases. Plasmid DNA was also detected by QPCR in laboratory-adapted strains 1497V, K2487 and K2490, but not strain 905. We conclude that the plasmid is highly conserved in C. felis, and plasmid-deficient strains represent a rare but important population for future studies of chlamydial plasmid function.

  7. PLASMIDS FROM ANAEROCELLUM THERMOPHILUM AND USES THEREOF

    DEFF Research Database (Denmark)

    2003-01-01

    The present invention concerns the isolation of plasmids from extremely thermophilic anaerobic microorganisms and their use in genetic transformation of thermophilic and mesophilic microorganisms. More particular the invention concerns the use of thermostable plasmid vectors as tools for creating...

  8. Erwinia amylovora novel plasmid pEI70: complete sequence, biogeography, and role in aggressiveness in the fire blight phytopathogen.

    Directory of Open Access Journals (Sweden)

    Pablo Llop

    Full Text Available Comparative genomics of several strains of Erwinia amylovora, a plant pathogenic bacterium causal agent of fire blight disease, revealed that its diversity is primarily attributable to the flexible genome comprised of plasmids. We recently identified and sequenced in full a novel 65.8 kb plasmid, called pEI70. Annotation revealed a lack of known virulence-related genes, but found evidence for a unique integrative conjugative element related to that of other plant and human pathogens. Comparative analyses using BLASTN showed that pEI70 is almost entirely included in plasmid pEB102 from E. billingiae, an epiphytic Erwinia of pome fruits, with sequence identities superior to 98%. A duplex PCR assay was developed to survey the prevalence of plasmid pEI70 and also that of pEA29, which had previously been described in several E. amylovora strains. Plasmid pEI70 was found widely dispersed across Europe with frequencies of 5-92%, but it was absent in E. amylovora analyzed populations from outside of Europe. Restriction analysis and hybridization demonstrated that this plasmid was identical in at least 13 strains. Curing E. amylovora strains of pEI70 reduced their aggressiveness on pear, and introducing pEI70 into low-aggressiveness strains lacking this plasmid increased symptoms development in this host. Discovery of this novel plasmid offers new insights into the biogeography, evolution and virulence determinants in E. amylovora.

  9. Erwinia amylovora novel plasmid pEI70: complete sequence, biogeography, and role in aggressiveness in the fire blight phytopathogen.

    Science.gov (United States)

    Llop, Pablo; Cabrefiga, Jordi; Smits, Theo H M; Dreo, Tanja; Barbé, Silvia; Pulawska, Joanna; Bultreys, Alain; Blom, Jochen; Duffy, Brion; Montesinos, Emilio; López, María M

    2011-01-01

    Comparative genomics of several strains of Erwinia amylovora, a plant pathogenic bacterium causal agent of fire blight disease, revealed that its diversity is primarily attributable to the flexible genome comprised of plasmids. We recently identified and sequenced in full a novel 65.8 kb plasmid, called pEI70. Annotation revealed a lack of known virulence-related genes, but found evidence for a unique integrative conjugative element related to that of other plant and human pathogens. Comparative analyses using BLASTN showed that pEI70 is almost entirely included in plasmid pEB102 from E. billingiae, an epiphytic Erwinia of pome fruits, with sequence identities superior to 98%. A duplex PCR assay was developed to survey the prevalence of plasmid pEI70 and also that of pEA29, which had previously been described in several E. amylovora strains. Plasmid pEI70 was found widely dispersed across Europe with frequencies of 5-92%, but it was absent in E. amylovora analyzed populations from outside of Europe. Restriction analysis and hybridization demonstrated that this plasmid was identical in at least 13 strains. Curing E. amylovora strains of pEI70 reduced their aggressiveness on pear, and introducing pEI70 into low-aggressiveness strains lacking this plasmid increased symptoms development in this host. Discovery of this novel plasmid offers new insights into the biogeography, evolution and virulence determinants in E. amylovora.

  10. Plasmid required for virulence of Agrobacterium tumefaciens

    Energy Technology Data Exchange (ETDEWEB)

    Watson, B.; Currier, T.C.; Gordon, M.P.; Chilton, M.D.; Nester, E.W.

    1975-07-01

    The irreversible loss of crown gall-inducing ability of Agrobacterium tumefaciens strain C-58 during growth at 37/sup 0/C is shown to be due to loss of a large plasmid (1.2 x 10/sup 8/ daltons). The gene responsible for this high rate of plasmid loss at elevated temperatures seems to be located on the plasmid. In addition, another spontaneous avirulent variant, A. tumefaciens strain IIBNV6, is shown to lack the virulence plasmid which its virulent sibling strain, IIBV7, possesses. Deoxyribonucleic acid reassociation measurements prove that the plasmid is eliminated, not integrated into the chromosome, in both of the avirulent derivatives. Transfer of virulence from donor strain C-58 to avirulent recipient strain A136 results from the transfer of a plasmid, which appears identical to the donor plasmid by deoxyribonucleic acid reassociation measurements. The transfer of virulence in another cross, K27 x A136, was also shown to result from the transfer of a large plasmid. These findings establish unequivocally that the large plasmid determines virulence. Two additional genetic determinants have been located on the virulence plasmid of A. tumefaciens strain C-58: the ability to utilize nopaline and sensitivity to a bacteriocin produced by strain 84. The latter trait can be exploited for selection of avirulent plasmid-free derivatives of strain C-58. The trait of nopaline utilization appears to be on the virulence plasmid also in strains IIBV7 and K27.

  11. Controversies in HIV cure research

    Directory of Open Access Journals (Sweden)

    Johnston Rowena

    2012-03-01

    Full Text Available Abstract Background Antiretroviral therapy significantly reduces HIV viral burden and prolongs life, but does not cure HIV infection. The major scientific barrier to a cure is thought to be the persistence of the virus in cellular and/or anatomical reservoirs. Discussion Most efforts to date, including pharmaco, immuno or gene therapy, have failed to cure patients, with the notable exception of a stem cell transplant recipient commonly known as the Berlin patient. This case has revived interest in the potential to cure HIV infection and has highlighted the need to resolve critical questions in the basic, pre-clinical and clinical research spheres as they pertain specifically to efforts to eradicate HIV from the body of an infected person (a sterilizing cure or at least render the need for lifelong antiretroviral therapy obsolete (functional cure. This paper describes ongoing debates in each of these research spheres as they were presented and discussed at a satellite session that took place at the 6th International AIDS Society Conference on HIV Pathogenesis, Treatment and Prevention in Rome in July 2011. Summary The resolution of these debates may have important implications for the search for a cure, the most efficient ways to identify and test promising interventions, and ultimately the availability of such a cure to diverse groups of HIV patients around the world.

  12. Mixture proportioning for internal curing

    DEFF Research Database (Denmark)

    Bentz, Dale P.; Pietro, Lura; Roberts, John W.

    2005-01-01

    hydration and strength development, reduced autogenous shrinkage and cracking, reduced permeability, and increased durability. The impact of internal curing begins immediately with the initial hydration of the cement, with benefits that are observed at ages as early as two days. Internal curing...

  13. Influence of native plasmids to fitness of Pantoea vagans strain C9-1

    Science.gov (United States)

    Pantoea vagans strain C9-1 is a biological control agent for fire blight caused by Erwinia amylovora. We cured C9-1 of two of its three plasmids: pPag2, pPag3, and both pPag2 and pPag3, tested phenotypes of the derivatives, and evaluated blossom colonization in the field. pPag2 (166 kb) encodes for ...

  14. CURING EFFICIENCY OF DUAL-CURE RESIN CEMENT UNDER ZIRCONIA WITH TWO DIFFERENT LIGHT CURING UNITS

    Directory of Open Access Journals (Sweden)

    Pınar GÜLTEKİN

    2015-04-01

    Full Text Available Purpose: Adequate polymerization is a crucial factor in obtaining optimal physical properties and a satisfying clinical performance from composite resin materials. The aim of this study was to evaluate the polymerization efficiency of dual-cure resin cement cured with two different light curing units under zirconia structures having differing thicknesses. Materials and Methods: 4 zirconia discs framework in 4 mm diameter and in 0.5 mm, 1 mm and 1.5 mm thickness were prepared using computer-aided design system. One of the 0.5 mm-thick substructures was left as mono-layered whereas others were layered with feldspathic porcelain of same thickness and ceramic samples with 4 different thicknesses (0.5, 1, 1.5 and 2.0 mm were prepared. For each group (n=12 resin cement was light cured in polytetrafluoroethylene molds using Light Emitting Diode (LED or Quartz-Tungsten Halogen (QHT light curing units under each of 4 zirconia based discs (n=96. The values of depth of cure (in mm and the Vickers Hardness Number values (VHN were evaluated for each specimen. Results: The use of LED curing unit produced a greater depth of cure compared to QTH under ceramic discs with 0.5 and 1 mm thickness (p<0.05.At 100μm and 300 μm depth, the LED unit produced significantly greater VHN values compared to the QTH unit (p<0.05. At 500 μm depth, the difference between the VHN values of LED and QTH groups were not statistically significant. Conclusion: Light curing may not result in adequate resin cement polymerization under thick zirconia structures. LED light sources should be preferred over QTH for curing dual-cure resin cements, especially for those under thicker zirconia restorations.

  15. Origin and Evolution of Rickettsial Plasmids.

    Science.gov (United States)

    El Karkouri, Khalid; Pontarotti, Pierre; Raoult, Didier; Fournier, Pierre-Edouard

    2016-01-01

    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 transfer as well as

  16. The effect of light curing units, curing time, and veneering materials on resin cement microhardness

    Directory of Open Access Journals (Sweden)

    Nurcan Ozakar Ilday

    2013-06-01

    Conclusion: Light-curing units, curing time, and veneering materials are important factors for achieving adequate dual cure resin composite microhardness. High-intensity light and longer curing times resulted in the highest microhardness values.

  17. Chlamydophila felis: plasmid detection in Italian isolates.

    Science.gov (United States)

    Di Francesco, Antonietta; Donati, Manuela; Salvatore, Daniela; Cevenini, Roberto; Di Paolo, Maria; Baldelli, Raffaella

    2010-04-01

    Plasmids have been detected in the majority of strains in the genus Chlamydia and in many Chlamydophila species. Previous studies showed that FP Pring and FP Cello Chlamydophila felis strains have an extrachromosomial plasmid, whereas the FP Baker strain does not. Azuma et al. recently sequenced the entire genomic DNA sequence of the Japanese Cp. felis strain Fe/C-56 and described a 7,552 base pair circular plasmid. In the present study a highly conserved plasmid gene was detected in 11 Italian Cp. felis isolates, showing 100% nucleotide identity with the plasmid gene of Fe/C-56 Cp. felis strain.

  18. Plasmid-mediated biodegradation of the anionic surfactant sodium dodecyl sulphate, by Pseudomonas aeruginosa S7.

    Science.gov (United States)

    Yeldho, Deepthi; Rebello, Sharrel; Jisha, M S

    2011-01-01

    Sodium dodecyl sulphate (SDS), an anionic surfactant, has been used extensively due to its low cost and excellent foaming properties. Fifteen different bacterial isolates capable of degrading SDS were isolated from detergent contaminated soil by enrichment culture technique and the degradation efficiency was assessed by Methylene Blue Active Substances (MBAS) assay. The most efficient SDS degrading isolate was selected and identified as Pseudomonas aeruginosa S7. The selected isolate was found to harbor a single 6-kb plasmid. Acridine orange, ethidium bromide, SDS and elevated temperatures of incubation failed to cure the plasmid. The cured derivatives of SDS degrading Pseudomonas aeruginosa were obtained only when ethidium bromide and elevated temperature (40 °C) were used together. Transformation of E. coli DH5α with plasmid isolated from S7 resulted in subsequent growth of the transformants on minimal salt media with SDS (0.1%) as the sole source of carbon. The SDS degradation ability of S7 and the transformant was found to be similar as assessed by Methylene Blue Active Substance Assay. The antibiotic resistance profiles of S7, competent DH5α and transformant were analyzed and it was noted that the transfer of antibiotic resistance correlated with the transfer of plasmid as well as SDS degrading property.

  19. Towards an HIV cure

    Directory of Open Access Journals (Sweden)

    Steven Deeks

    2014-11-01

    Full Text Available Given the challenge of delivering complex, expensive and potentially harmful antiretroviral therapy (ART on a global level, there is intense interest in the development of short-term, well-tolerated regimens that allow individuals to interrupt therapy indefinitely without experiencing a rebound in viremia. This so-called “cure” or “remission” might be due to complete eradication of all replication-competent HIV during ART or durable host-mediated control of persistent virus in absence of ART. Recent heroic interventions such as hematopoietic stem cell transplant and very early initiation of antiretroviral therapy suggest that dramatic reductions in the reservoir size can be achieved, but that complete eradication will be difficult if not impossible to achieve. Most attempts to stimulate effective host-mediated control of HIV have failed. It is likely that for a true cure to be achieved, both approaches – reductions in the reservoir size and durable immune surveillance – will be needed, a state that is similar to that observed in “elite” controllers and post-treatment controllers. The implications for recent advances and setbacks in achieving HIV remission for future research priorities will be discussed.

  20. Towards a 'cure' for IBD.

    Science.gov (United States)

    Fiocchi, Claudio

    2012-01-01

    To discuss a cure for IBD, one should first define the concept 'a cure'. If it is intended as the general restoration of health, this is already possible, as many current therapies do a good job in inducing long periods of remission in Crohn's disease, and colectomy can technically cure ulcerative colitis. If it is more strictly defined as the complete and permanent elimination of the cause, predisposing and permissive factors, reinstatement of normal microbial ecology and restoration of mucosal immune homeostasis, then a cure for IBD is out of reach, at least for now. Regardless of the definition, major strides have been made in attempting to cure IBD by addressing the key components of its pathogenesis: the environment (exposome), the genetic makeup (genome), the gut microbiota (microbiome) and the immune system (immunome). However, the isolated modulation of each component is insufficient to provide a cure, and different requirements may be needed depending on the stage of the disease and each patient subset. To achieve a cure, one key approach is currently missing: the integration of knowledge from all the pathogenic components. We continue to learn more and more about each component using traditional 'canonical' systems, which allow the accumulation of data without taking into consideration the other components. We are still not studying the 'omes' of IBD, we should be using 'omics' technologies that can generate a more global vision of IBD pathogenesis on which to base novel, multiple pathway-integrated therapies. Copyright © 2012 S. Karger AG, Basel.

  1. Curing efficacy of light emitting diodes of dental curing units

    Directory of Open Access Journals (Sweden)

    Seyd Mostafa Mousavinasab

    2009-03-01

    Full Text Available Background and aims. The aim of the present study was to compare the efficacy of quartz tungsten halogen (QTH and light emitting diode (LED curing lights on polymerization of resin composite. Materials and Methods. A hybrid resin composite was used to prepare samples which were cured using two QTH and ten LED light curing sources. Twelve groups, each containing ten samples, were prepared using each light source. The cured depth of the resin was determined using ISO 4049 method and Vickers hardness values were determined at 1.0 mm intervals. Data was analyzed by ANOVA and Tukey test. Results. Data analysis demonstrated a significant difference between light sources for depth of cure. At 1.0 mm below the surface all the tested light sources and at 2.0-mm intervals all light sources except two (Optilux 501 and LEDemetron I and at 3.0-mm intervals only two light sources (PenCure and LEDemetron II could produce hardness values higher than 80% of superficial layer values. Conclusion. This study showed that a variety of LED light sources used in the present study are as effective as the high-intensity QTH lights in polymerization of resin composite.

  2. Novel techniques for concrete curing

    DEFF Research Database (Denmark)

    Kovler, Konstantin; Jensen, Ole Mejlhede

    2005-01-01

    It is known that some high-strength/high-performance concretes (HSC/HPC) are prone to cracking at an early age unless special precautions are taken. The paper deals with the methods of curing as one of the main strategies to ensure good performance of concrete. Curing by both external (conventional......) and internal methods is reviewed and analyzed, among other methods of mitigating shrinkage and cracking of concrete. The focus is on the mitigation of autogenous shrinkage of low water to binder ratio (w/b) concrete by means of internal curing. The concepts of internal curing are based on using lightweight...... aggregate, superabsorbent polymers or water-soluble chemicals, which reduce water evaporation (so called "internal sealing"). These concepts have been intensively researched in the 90s, but still are not widespread among contractors and concrete suppliers. The differences between conventional methods...

  3. Correlation of cure monitoring techniques

    Science.gov (United States)

    Chang, S. S.; Mopsik, F. I.; Hunston, D. L.

    Six different composite matrix or neat resin cure-monitoring methods are presently used to follow the cure process in a model epoxy system, and the results obtained are compared. Differential scanning calorimetry, viscosity monitoring, the ultrasonic shear wave propagation technique, dielectric spectrometry, and two different fluorescence intensity techniques are compared with a view to common traits and differences. Dielectric fluorescence and ultrasonic measurement techniques are noted to be applicable to on-line process monitoring.

  4. Optimization of a plasmid electroporation protocol for Aeromonas salmonicida subsp. salmonicida.

    Science.gov (United States)

    Dallaire-Dufresne, Stéphanie; Emond-Rheault, Jean-Guillaume; Attéré, Sabrina A; Tanaka, Katherine H; Trudel, Mélanie V; Frenette, Michel; Charette, Steve J

    2014-03-01

    Aeromonas salmonicida subsp. salmonicida is a major fish pathogen. Molecular tools are required to study the virulence and genomic stability of this bacterium. An efficient electroporation-mediated transformation protocol for A. salmonicida subsp. salmonicida would make genetic studies faster and easier. In the present study, we designed the 4.1-kb pSDD1 plasmid as a tool for optimizing an electroporation protocol for A. salmonicida subsp. salmonicida. We systematically tested the electroporation conditions to develop a protocol that generates the maximum number of transformants. Under these optimal conditions (25 kV/cm, 200 Ω, 25 μF), we achieved an electroporation efficiency of up to 1×10(5) CFU/μg DNA. The electroporation protocol was also tested using another plasmid of 10.6-kb and three different strains of A. salmonicida subsp. salmonicida. The strains displayed significant differences in their electro-transformation competencies. Strain 01-B526 was the easiest to electroporate, especially with the pSDD1 plasmid. This plasmid was stably maintained in the 01-B526 transformants, as were the native plasmids, but could be easily cured by removing the selection conditions. This is the first efficient electroporation protocol reported for A. salmonicida subsp. salmonicida, and offers new possibilities for studying this bacterium.

  5. Temperature Depended Role of Shigella flexneri Invasion Plasmid on the Interaction with Acanthamoeba castellanii

    Directory of Open Access Journals (Sweden)

    Amir Saeed

    2012-01-01

    Full Text Available Shigella flexneri is a Gram-negative bacterium causing the diarrhoeal disease shigellosis in humans. The virulence genes required for invasion are clustered on a large 220 kb plasmid encoding type three secretion system (TTSS apparatus and virulence factors such as adhesions and invasion plasmid antigens (Ipa. The bacterium is transmitted by contaminated food, water, or from person to person. Acanthamoebae are free-living amoebae (FLA which are found in diverse environments and isolated from various water sources. Different bacteria interact differently with FLA since Francisella tularensis, Vibrio cholerae, Shigella sonnei, and S. dysenteriae are able to grow inside A. castellanii. In contrast, Pseudomonas aeruginosa induces both necrosis and apoptosis to kill A. castellanii. The aim of this study is to examine the role of invasion plasmid of S. flexneri on the interaction with A. castellanii at two different temperatures. A. castellanii in the absence or presence of wild type, IpaB mutant, or plasmid-cured strain S. flexneri was cultured at 30∘C and 37∘C and the interaction was analysed by viable count of both bacteria and amoebae, electron microscopy, flow cytometry, and statistical analysis. The outcome of the interaction was depended on the temperature since the growth of A. castellanii was inhibited at 30∘C, and A. castellanii was killed by invasion plasmid mediated necrosis at 37∘C.

  6. Curing units' ability to cure restorative composites and dual-cured composite cements under composite overlay.

    Science.gov (United States)

    Park, Sung-Ho; Kim, Su-Sun; Cho, Yong-Sik; Lee, Chang-Kyu; Noh, Byng-Duk

    2004-01-01

    This study compared the efficacy of using conventional low-power density QTH (LQTH) units, high-power density QTH (HQTH) units, argon (Ar) laser and Plasma arc curing (PAC) units for curing dual-cured resin cements and restorative resin composites under a pre-cured resin composite overlay. The microhardness of the two types of restorative resins (Z100 and Tetric Ceram) and a dual-cured resin cement (Variolink II) were measured after they were light cured for 60 seconds in a 2 mm Teflon mold. The recorded microhardness was determined to be the optimum microhard-ness (OM). Either one of the two types of restorative resins (Z100, Tetric Ceram) or the dual cured resin cement (Variolink II) were placed under a 1.5-mm thick and 8 mm diameter pre-cured Targis (Vivadent/Ivoclar AG, Schaan, Liechtenstein) overlay. The specimens that were prepared for each material were divided into four groups depending upon the curing units used (HQTH, PAC, Laser or LQTH) and were further subdi-vided into subgroups according to light curing time. The curing times used were 30, 60, 90 and 120 seconds for HQTH; 12, 24, 36 and 48 seconds for the PAC unit; 15, 30, 45 and 60 for the Laser and 60, 120 or 180 seconds for the LQTH unit. Fifteen specimens were assigned to each sub- group. The microhardness of the upper and and lower composite surfaces under the Targis overlay were measured using an Optidur Vickers hardness-measuring instrument (Göttfert Feinwerktechnik GmbH, Buchen, Germany). In each material, for each group, a three-way ANOVA with Tukey was used at the 0.05 level of significance to compare the microhardnesses of the upper and lower composite surfaces and the previously measured OM of the material. From the OM of each material, 80% OM was calculated and the time required for the microhardness of the upper and lower surface of the specimen to reach 100% and 80% of OM was determined. In Z100 and Tetric Ceram, when the composites were light cured for 120 seconds using the HQTH lamp

  7. Cold-Curing Structural Epoxy Resins: Analysis of the Curing Reaction as a Function of Curing Time and Thickness

    Directory of Open Access Journals (Sweden)

    Carola Esposito Corcione

    2014-09-01

    Full Text Available The curing reaction of a commercial cold-curing structural epoxy resin, specifically formulated for civil engineering applications, was analyzed by thermal analysis as a function of the curing time and the sample thickness. Original and remarkable results regarding the effects of curing time on the glass transition temperature and on the residual heat of reaction of the cold-cured epoxy were obtained. The influence of the sample thickness on the curing reaction of the cold-cured resin was also deeply investigated. A highly exothermal reaction, based on a self-activated frontal polymerization reaction, was supposed and verified trough a suitable temperature signal acquisition system, specifically realized for this measurement. This is one of the first studies carried out on the curing behavior of these peculiar cold-cured epoxy resins as a function of curing time and thickness.

  8. Cold-Curing Structural Epoxy Resins: Analysis of the Curing Reaction as a Function of Curing Time and Thickness.

    Science.gov (United States)

    Corcione, Carola Esposito; Freuli, Fabrizio; Frigione, Mariaenrica

    2014-09-22

    The curing reaction of a commercial cold-curing structural epoxy resin, specifically formulated for civil engineering applications, was analyzed by thermal analysis as a function of the curing time and the sample thickness. Original and remarkable results regarding the effects of curing time on the glass transition temperature and on the residual heat of reaction of the cold-cured epoxy were obtained. The influence of the sample thickness on the curing reaction of the cold-cured resin was also deeply investigated. A highly exothermal reaction, based on a self-activated frontal polymerization reaction, was supposed and verified trough a suitable temperature signal acquisition system, specifically realized for this measurement. This is one of the first studies carried out on the curing behavior of these peculiar cold-cured epoxy resins as a function of curing time and thickness.

  9. Persistence of Antibiotic Resistance Plasmids in Biofilms

    Science.gov (United States)

    2014-10-01

    plasmids* in*populations*of* Gram > negative *bacteria*grown*in*biofilms*and*well>mixed*liquid*cultures.** * Task2:*Characterize*the*evolution*of*plasmid...R.! Edwards.! 2005.! Overview! of! nosocomial! infections! caused! by! gramP negative ! bacilli .!Clin.!Infect.!Dis.!41:848P854.! LoftiePEaton,!W.,!A... negative ! interaction!between!one!of! its!chromosomal!segments!and!the!plasmid! by!simply!deleting!the!appropriate!chromosomal!segment.!! 7. None

  10. The function of three indigenous plasmids in Mesorhizobium huakuii 2020 and its symbiotic inter-action with Sym pJB5JI of Rhizobium leguminosarum

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    A Mesorhizobium huakuii strain 2020, isolated from a rice-growing field in southern China, contains three indigenous plasmids named p2020a, p2020b and p2020c, respectively. The plasmids were deleted via Tn5-sacB insertion, and two cured derivatives were obtained. Interestingly, the mutant 2020D29 curing of p2020c could significantly enhance the capacity of symbiotic nitrogen fixation. But the mutant 2020D8 curing of p2020b lost the ability to nodulate Astragalus sinicus. Furthermore, the third plasmid p2020a could be hardly eliminated, suggesting that some house-keeping genes necessary for strain growth located on this plasmid. Then the Sym plasmid pJB5JI of R. leguminosarum bv. viciae was transferred into 2020 and its cured derivatives. The pot plant test showed that the ability of competition and symbiotic nitrogen fixation of transconjugant 2020-137 (pJB5JI) was increased evidently in con-trast to 2020. pJB5JI could not restore the ability of 2020D8 to nodulate Astragalus sinicus. 2020D8-8 (pJB5JI) could form ineffective nodules on peas, which implied that the symbiotic plasmid pJB5JI could express its function at the chromosomal background of Mesorhizobium huakuii 2020. The plas-mid stability was checked in transconjugants under free-living and during symbiosis. The results indi-cated that pJB5JI failed to be detected in some nodule isolates. That Km resistance gene could be am-plified from all transconjugants and nodule isolates suggested that pJB5JI was fully or partially inte-grated into the chromosome of recipients.

  11. Plasmid profiles of Moraxella bovis isolates.

    Science.gov (United States)

    McDonald, T J; Pugh, G W

    1986-04-01

    Two-hundred isolates of Moraxella bovis were selected at random and examined for the presence of plasmid DNA by a rapid alkaline-detergent lysis method. All isolates contained from 1 to 6 plasmids, with varying agarose-gel electrophoretic migration patterns. Most (80%) isolates carried 2 to 4 plasmids, which ranged in molecular weight from 2.6 to 80 megadaltons. Seemingly, plasmid profiles can be used as a simple, reliable epizootiologic tool to establish a strain identification scheme for M bovis.

  12. Plasmid transfer systems in the rhizobia.

    Science.gov (United States)

    Ding, Hao; Hynes, Michael F

    2009-08-01

    Rhizobia are agriculturally important bacteria that can form nitrogen-fixing nodules on the roots of leguminous plants. Agricultural application of rhizobial inoculants can play an important role in increasing leguminous crop yields. In temperate rhizobia, genes involved in nodulation and nitrogen fixation are usually located on one or more large plasmids (pSyms) or on symbiotic islands. In addition, other large plasmids of rhizobia carry genes that are beneficial for survival and competition of rhizobia in the rhizosphere. Conjugative transfer of these large plasmids thus plays an important role in the evolution of rhizobia. Therefore, understanding the mechanism of conjugative transfer of large rhizobial plasmids provides foundations for maintaining, monitoring, and predicting the behaviour of these plasmids during field release events. In this minireview, we summarize two types of known rhizobial conjugative plasmids, including quorum sensing regulated plasmids and RctA-repressed plasmids. We provide evidence for the existence of a third type of conjugative plasmid, including pRleVF39c in Rhizobium leguminosarum bv. viciae strain VF39SM, and we provide a comparison of the different types of conjugation genes found in members of the rhizobia that have had their genomes sequenced so far.

  13. Effect of chromosome homology an plasmid transformation and plasmid conjugal transfer in Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

    Balganesh, M.; Setlow, J.K.

    1984-05-14

    The pairing between plasmid and the homologous part of the chromosome associated with plasmid establishment may differ from the pairing which results from integration of a homologous region of the plasmid into the chromosome. Thus the rate of novobiocin transformation decreases with duplication of the chromosomal portion in pMB2, but the rate of establishment of the plasmid increases with this duplication. A model to explain these data is given. 17 references, 5 figures, 4 tables.

  14. Plasmid-Mediated OqxAB Is an Important Mechanism for Nitrofurantoin Resistance in Escherichia coli.

    Science.gov (United States)

    Ho, Pak-Leung; Ng, Ka-Ying; Lo, Wai-U; Law, Pierra Y; Lai, Eileen Ling-Yi; Wang, Ya; Chow, Kin-Hung

    2015-11-09

    Increasing consumption of nitrofurantoin (NIT) for treatment of acute uncomplicated urinary tract infections (UTI) highlights the need to monitor emerging NIT resistance mechanisms. This study investigated the molecular epidemiology of the multidrug-resistant efflux gene oqxAB and its contribution to nitrofurantoin resistance by using Escherichia coli isolates originating from patients with UTI (n = 205; collected in 2004 to 2013) and food-producing animals (n = 136; collected in 2012 to 2013) in Hong Kong. The oqxAB gene was highly prevalent among NIT-intermediate (11.5% to 45.5%) and -resistant (39.2% to 65.5%) isolates but rare (0% to 1.7%) among NIT-susceptible (NIT-S) isolates. In our isolates, the oqxAB gene was associated with IS26 and was carried by plasmids of diverse replicon types. Multilocus sequence typing revealed that the clones of oqxAB-positive E. coli were diverse. The combination of oqxAB and nfsA mutations was found to be sufficient for high-level NIT resistance. Curing of oqxAB-carrying plasmids from 20 NIT-intermediate/resistant UTI isolates markedly reduced the geometric mean MIC of NIT from 168.9 μg/ml to 34.3 μg/ml. In the plasmid-cured variants, 20% (1/5) of isolates with nfsA mutations were NIT-S, while 80% (12/15) of isolates without nfsA mutations were NIT-S (P = 0.015). The presence of plasmid-based oqxAB increased the mutation prevention concentration of NIT from 128 μg/ml to 256 μg/ml and facilitated the development of clinically important levels of nitrofurantoin resistance. In conclusion, plasmid-mediated oqxAB is an important nitrofurantoin resistance mechanism. There is a great need to monitor the dissemination of this transferable multidrug-resistant efflux pump.

  15. The expression of a plasmid-specified exported protein causes structural plasmid instability in Bacillus subtilis

    NARCIS (Netherlands)

    Cordes, C.; Meima, R; Twiest, B; Kazemier, B; Venema, G; vanDijl, JM; Bron, S

    The rolling-circle plasmid pGP1 was used to study the effects of the expression of a plasmid-specified exported protein on structural plasmid stability in Bacillus subtilis. pGP1 contains a fusion between the Bacillus licheniformis penP gene, encoding a C-terminally truncated penicillinase, and the

  16. Curing efficiency of various types of light-curing units.

    Science.gov (United States)

    Rahiotis, Chris; Kakaboura, Afrodite; Loukidis, Michalis; Vougiouklakis, George

    2004-02-01

    This study compared monomer conversion (DC), the per cent linear polymerization shrinkage (%LS), the wall-to-wall contraction pattern (per cent of peripheral opening, %DM, and maximal marginal gap, MG) and depth of cure (DOC), of a hybrid resin composite (Spectrum TPH) exposed to different types of light-curing units and exposure modes (Virtuoso-PAC, Elipar TriLight-QTH, and FreeLight-LED). The QTH and LED units were used in two curing modes: the exponential ramp and the continuous output modes. Monomer conversion was investigated by micro Multiple Internal Reflection (MIR)-Fourier-transform infrared (FTIR) spectroscopy and %LS was measured by the deflective disc method. The wall-to-wall contraction method used a cylindrical cavity model in extracted human teeth. The per cent debonded margins relative to the cavity periphery (%DM) and the width of maximum gap (MG) was evaluated. The DOC was determined using Vickers microhardness measurements (200 g load, 20 s) at the top surface (H0), at 2 mm (H2) and at 4 mm (H4) depths, and the results expressed as H2/H0 and H4/H0 ratios. Significantly lower %DC and %LS values were provided by PAC and LED units. No differences were found in %DM among the curing units and PAC exhibited the highest MG. No significant differences were noted among light-curing groups in terms of H2/H0 microhardness values. The QTH, operated in exponential mode, resulted in the highest H4/H0 value. The exponential mode of the QTH demonstrated superior performance for the total of the characteristics evaluated.

  17. Role of the virulence plasmid pR99 and the metalloprotease Vvp in resistance of Vibrio vulnificus serovar E to eel innate immunity.

    Science.gov (United States)

    Valiente, E; Lee, C T; Lamas, J; Hor, L; Amaro, C

    2008-01-01

    Vibrio vulnificus biotype 2 serovar E (VSE) is a bacterial pathogen that produces a haemorrhagic septicaemia called vibriosis in eels. Its ability to grow in blood is conferred by a recently described virulence plasmid [Lee CT, Amaro C, Wu KM, Valiente E, Chang YF, Tsai SF, et al. A common virulence plasmid in biotype 2 Vibrio vulnificus and its dissemination aided by a conjugal plasmid. Journal of Bacteriology, submitted for publication.]. In this study, we analyzed the role of this plasmid together with the role played by the metalloprotease (Vvp) in the interaction between bacteria and eel innate immunity. To this end, we compared and statistically analyzed the differences in resistance to serum and mucus factors (complement, selected antimicrobial peptides, transferrin and lysozyme) and also to phagocytosis/opsonophagocytosis between one VSE strain and its derivatives: a plasmid-cured strain and a vvp-deficient mutant. The wild-type and the metalloprotease-deficient strains were resistant to both the bactericidal action of fresh serum and the phagocytosis and opsonophagocytosis by eel phagocytes, confirming that Vvp is not involved in resistance to eel innate immunity. In contrast, the cured strain was sensitive to both the bactericidal action of eel serum activated by the alternative pathway and phagocytosis/opsonophagocytosis. Since no plasmid-encoded ORF, with homology to known genes, is related to the resistance to innate immunity [Lee CT, Amaro C, Wu KM, Valiente E, Chang YF, Tsai SF, et al. A common virulence plasmid in biotype 2 Vibrio vulnificus and its dissemination aided by a conjugal plasmid. Journal of Bacteriology, submitted for publication.], this function could be codified by one or more new genes. Further studies are underway to characterize the plasmid-encoded system responsible for V. vulnificus resistance to the innate immune system of eels.

  18. Breather cloth for vacuum curing

    Science.gov (United States)

    Reed, M. W.

    1979-01-01

    Finely-woven nylon cloth that has been treated with Teflon improves vacuum adhesive bonding of coatings to substrates. Cloth is placed over coating; entire assembly, including substrate, coating, and cloth, is placed in plastic vacuum bag for curing. Cloth allows coating to "breathe" when bag is evacuated. Applications include bonding film coatings to solar concentrators and collectors.

  19. Key Impact of an Uncommon Plasmid on Bacillus amyloliquefaciens subsp. plantarum S499 Developmental Traits and Lipopeptide Production

    Science.gov (United States)

    Molinatto, Giulia; Franzil, Laurent; Steels, Sébastien; Puopolo, Gerardo; Pertot, Ilaria; Ongena, Marc

    2017-01-01

    The rhizobacterium Bacillus amyloliquefaciens subsp. plantarum S499 (S499) is particularly efficient in terms of the production of cyclic lipopeptides, which are responsible for the high level of plant disease protection provided by this strain. Sequencing of the S499 genome has highlighted genetic differences and similarities with the closely related rhizobacterium B. amyloliquefaciens subsp. plantarum FZB42 (FZB42). More specifically, a rare 8008 bp plasmid (pS499) harboring a rap-phr cassette constitutes a major distinctive element between S499 and FZB42. By curing this plasmid, we demonstrated that its presence is crucial for preserving the typical physiology of S499 cells. Indeed, the growth rate and extracellular proteolytic activity were significantly affected in the cured strain (S499 P−). Furthermore, pS499 made a significant contribution to the regulation of cyclic lipopeptide production. Surfactins and fengycins were produced in higher quantities by S499 P−, whereas lower amounts of iturins were detected. In line with the increase in surfactin release, bacterial motility improved after curing, whereas the ability to form biofilm was reduced in vitro. The antagonistic effect against phytopathogenic fungi was also limited for S499 P−, most probably due to the reduction of iturin production. With the exception of this last aspect, S499 P− behavior fell between that of S499 and FZB42, suggesting a role for the plasmid in shaping some of the phenotypic differences observed in the two strains. PMID:28154555

  20. Susan G. Komen for the Cure

    Science.gov (United States)

    ... an Event Komen Race for the Cure Susan G. Komen 3 Day Find Local Events Fundraise Fundraise ... 2016 Search Events Friday, September 22, 2017 Susan G. Komen Italia Race for the Cure® - Bologna Bologna, ...

  1. Accelerated Cure Project for Multiple Sclerosis

    Science.gov (United States)

    ... main content Accelerating research toward a cure for multiple sclerosis Home Contact Us Search form Search Connect Volunteer ... is to accelerate efforts toward a cure for multiple sclerosis by rapidly advancing research that determines its causes ...

  2. Rapid Development of the Radiation Curing Sector

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    @@ Radiation curing is an advanced material surface treatment technology using ultraviolet (UV) radiation and electronic beams (EB). With the greater attention paid to environ mentel protection in recent years ,radiation curing has developed rapidly.

  3. Plasmid typing of Shigella sonnei epidemic strains and molecular relationship of their R-plasmids.

    Science.gov (United States)

    Mendoza, M C; Gonzalez, A J; Mendez, F J; Hardisson, C

    1988-06-01

    We conducted a surveillance program on epidemic and/or endemic Shigella strains in Asturias (Spain), their frequency and dispersion in our community, and their R-plasmids. We analyzed initial isolates of Shigella sonnei from two epidemic outbreaks using antibiotic resistance patterns and plasmid profile analysis as epidemiological markers. We found that the 2 outbreaks were caused by different S. sonnei strains, which respectively carried one and two R-plasmids together with other plasmids. The molecular relationship among these and three other R-plasmids from two S. sonnei strains isolated during a previous outbreak, were studied by restriction enzyme analysis and DNA-DNA hybridizations. We were able to establish different levels of relationship among the six R-plasmids.

  4. Techniques for internal water curing of concrete

    DEFF Research Database (Denmark)

    Jensen, Ole Mejlhede; Pietro, Lura

    2003-01-01

    This paper gives an overview of different techniques for incorporation of internal curing water in concrete. Internal curing can be used to mitigate self-desiccation and self-desiccation shrinkage. Some concretes may need 50 kg/m3 of internal curing water for this purpose. The price of the internal...

  5. Prevalence and molecular characterization of plasmid- mediated ...

    African Journals Online (AJOL)

    lactamase genes among nosocomial Staphylococcus aureus drug resistance isolates in Taiwan. .... Table 2: Plasmid profiles of the clinical antibiotic-resistant pathogens. Strain. Profile .... Madec J. Characterization of clinical canine methicillin-.

  6. antimicrobial susceptibility and plasmids from escherichia coli ...

    African Journals Online (AJOL)

    2001-10-10

    Oct 10, 2001 ... transmission to humans of E. coli containing antibiotic resistance plasmids ... resistant micro-organisms, which may in turn transfer resistance to .... cells were washed with sterile normal saline to remove leached. Я-lactamase ...

  7. Freud's psychoanalysis: a moral cure.

    Science.gov (United States)

    Eriksson, Johan

    2014-08-01

    That psychoanalytical treatment in its classical Freudian sense is primarily a moral or ethical cure is not a very controversial claim. However, it is far from obvious how we are to understand precisely the moral character of psychoanalysis. It has frequently been proposed that this designation is valid because psychoanalysis strives neither to cure psychological symptoms pharmaceutically, nor to superficially modify the behaviour of the analysand, but to lead the analysand through an interpretive process during which he gradually gains knowledge of the unconscious motives that determine his behaviour, a process that might ideally liberate him to obtain, in relation to his inner desires, the status of a moral agent. There resides something appealing in these claims. But it is the author's belief that there is an even deeper moral dimension applying to psychoanalytical theory and praxis. Freudian psychoanalysis is a moral cure due to its way of thematizing psychological suffering as moral suffering. And this means that the moral subject - the being that can experience moral suffering - is not primarily something that the psychoanalytical treatment strives to realize, but rather the presupposition for the way in which psychoanalysis theorizes psychological problems as such. Copyright © 2014 Institute of Psychoanalysis.

  8. From HCV To HBV Cure.

    Science.gov (United States)

    Schinazi, Raymond F; Asselah, Tarik

    2017-01-01

    Approximately 170 million people are chronically infected with HCV and 350 million are chronically infected with HBV worldwide. It is estimated that more than one million patients die from complications related to chronic viral hepatitis, mainly HCC which is one of the most frequent cancers in many countries, especially Africa, the Middle East and Asia. HCV drug development has been impressive, and this revolution led to several direct-acting antiviral agents achieving an HCV cure after only 6-12 weeks. This progress could theorically lead to HCV global elimination making HCV and its consequences a rarity. HBV research and development programs can learn from the HCV experience, to achieve an HBV functional or sterilizing cure. This review will summarize key steps which have been realized for an HCV cure, and discuss the next steps to achieve for an HCV elimination. And also, how this HCV revolution has inspired scientists and clinicians to achieve the same for HBV.

  9. Protein diversity confers specificity in plasmid segregation.

    Science.gov (United States)

    Fothergill, Timothy J G; Barillà, Daniela; Hayes, Finbarr

    2005-04-01

    The ParG segregation protein (8.6 kDa) of multidrug resistance plasmid TP228 is a homodimeric DNA-binding factor. The ParG dimer consists of intertwined C-terminal domains that adopt a ribbon-helix-helix architecture and a pair of flexible, unstructured N-terminal tails. A variety of plasmids possess partition loci with similar organizations to that of TP228, but instead of ParG homologs, these plasmids specify a diversity of unrelated, but similarly sized, partition proteins. These include the proteobacterial pTAR, pVT745, and pB171 plasmids. The ParG analogs of these plasmids were characterized in parallel with the ParG homolog encoded by the pseudomonal plasmid pVS1. Like ParG, the four proteins are dimeric. No heterodimerization was detectable in vivo among the proteins nor with the prototypical ParG protein, suggesting that monomer-monomer interactions are specific among the five proteins. Nevertheless, as with ParG, the ParG analogs all possess significant amounts of unordered amino acid residues, potentially highlighting a common structural link among the proteins. Furthermore, the ParG analogs bind specifically to the DNA regions located upstream of their homologous parF-like genes. These nucleoprotein interactions are largely restricted to cognate protein-DNA pairs. The results reveal that the partition complexes of these and related plasmids have recruited disparate DNA-binding factors that provide a layer of specificity to the macromolecular interactions that mediate plasmid segregation.

  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......-like apparatus in prokaryotes. The identification of chromosomal homologues of the well-characterized plasmid partitioning genes indicates that there could be a general mechanism of bacterial DNA partitioning. Udgivelsesdato: July 1...

  11. Multiple plasmid interference - Pledging allegiance to my enemy's enemy.

    Science.gov (United States)

    Gama, João Alves; Zilhão, Rita; Dionisio, Francisco

    2017-08-24

    As shown in the previous article, two distinct conjugative plasmids sometimes interact within bacterial cells, implicating changes of transfer rates. In most cases of interactions within bacteria, the transfer of one of the plasmids decreases. Less frequently, the transfer rate of one of the plasmids increases. Here we analyse what happens if three distinct conjugative plasmids colonize the same bacterial cell. Our aim is to understand how interactions between two plasmids affect the transfer rate of the third plasmid. After showing that plasmids interact in 59 out of 84 possible interactions we show that, with some exceptions, if the transfer rate of a plasmid decreases in the presence of a second plasmid, a decrease is also observed in the presence of a third plasmid. Moreover, if the conjugation rate of a plasmid increases in the presence of another, an increase is also observed if there is a third plasmid in the cell. Both types of interactions are mostly independent of the third plasmid's identity, even if sometimes the third plasmid quantitatively distorts the interaction of the other two plasmids. There is a bias towards negative intensifying interactions, which provide good news concerning the spread conjugative plasmids encoding antibiotic-resistance genes and virulence factors. Copyright © 2017 Elsevier Inc. All rights reserved.

  12. Clostridium perfringens type A–E toxin plasmids

    Science.gov (United States)

    Freedman, John C.; Theoret, James R.; Wisniewski, Jessica A.; Uzal, Francisco A.; Rood, Julian I.; McClane, Bruce A.

    2014-01-01

    Clostridium perfringens relies upon plasmid-encoded toxin genes to cause intestinal infections. These toxin genes are associated with insertion sequences that may facilitate their mobilization and transfer, giving rise to new toxin plasmids with common backbones. Most toxin plasmids carry a transfer of clostridial plasmids locus mediating conjugation, which likely explains the presence of similar toxin plasmids in otherwise unrelated C. perfringens strains. The association of many toxin genes with insertion sequences and conjugative plasmids provides virulence flexibility when causing intestinal infections. However, incompatibility issues apparently limit the number of toxin plasmids maintained by a single cell. PMID:25283728

  13. Composite cure monitoring with Bragg grating sensors

    Science.gov (United States)

    Slattery, Kerry T.; Corona-Bittick, Kelli; Dorr, Donald J.

    1998-03-01

    Residual stress is induced in fiber composite materials during the cure process because the thermal expansion coefficient of the fiber is generally much lower than that of the polymer matrix. The two materials are 'locked' together at the cure temperature. Then, as they cool, the matrix attempts to contract more than the fiber leading to tension in the matrix and compression in the fiber. This can lead to the formation of microcracks parallel to the fibers in thick composite piles or yarns. The magnitude of residual stress can be reduced by modifying the cure cycle; however, optimizing the cure cycle requires a complete understanding of the state of cure throughout the composite. This is a complex problem -- especially in thick composites. Pilot studies have been performed placing Bragg gratin sensors in glass fabric preforms and monitoring the response of the grating during resin infusion and cure. The typical response shows the initial thermal expansion of the Bragg grating, a rapid contraction of the grating as the resin gels, slower contraction during cure, and thermal contraction at the composite thermal expansion coefficient during cool down. This data is then sued with micromechanical models of the fiber/matrix interaction during cure to establish material parameters for cure simulation. Once verified, these cure simulation methods will be used to optimize tooling design and cure cycles in composite components.

  14. Historical Events That Spawned the Field of Plasmid Biology.

    Science.gov (United States)

    Kado, Clarence I

    2014-10-01

    This chapter revisits the historical development and outcome of studies focused on the transmissible, extrachromosomal genetic elements called plasmids. Early work on plasmids involved structural and genetic mapping of these molecules, followed by the development of an understanding of how plasmids replicate and segregate during cell division. The intriguing property of plasmid transmission between bacteria and between bacteria and higher cells has received considerable attention. The utilitarian aspects of plasmids are described, including examples of various plasmid vector systems. This chapter also discusses the functional attributes of plasmids needed for their persistence and survival in nature and in man-made environments. The term plasmid biology was first conceived at the Fallen Leaf Lake Conference on Promiscuous Plasmids, 1990, Lake Tahoe, California. The International Society for Plasmid Biology was established in 2004 (www.ISPB.org).

  15. Characterization of the replication, transfer, and plasmid/lytic phage cycle of the Streptomyces plasmid-phage pZL12.

    Science.gov (United States)

    Zhong, Li; Cheng, Qiuxiang; Tian, Xinli; Zhao, Liqian; Qin, Zhongjun

    2010-07-01

    We report here the isolation and recombinational cloning of a large plasmid, pZL12, from endophytic Streptomyces sp. 9R-2. pZL12 comprises 90,435 bp, encoding 112 genes, 30 of which are organized in a large operon resembling bacteriophage genes. A replication locus (repA) and a conjugal transfer locus (traA-traC) were identified in pZL12. Surprisingly, the supernatant of a 9R-2 liquid culture containing partially purified phage particles infected 9R-2 cured of pZL12 (9R-2X) to form plaques, and a phage particle (phiZL12) was observed by transmission electron microscopy. Major structural proteins (capsid, portal, and tail) of phiZL12 virions were encoded by pZL12 genes. Like bacteriophage P1, linear phiZL12 DNA contained ends from a largely random pZL12 sequence. There was also a hot end sequence in linear phiZL12. phiZL12 virions efficiently infected only one host, 9R-2X, but failed to infect and form plaques in 18 other Streptomyces strains. Some 9R-2X spores rescued from lysis by infection of phiZL12 virions contained a circular pZL12 plasmid, completing a cycle comprising autonomous plasmid pZL12 and lytic phage phiZL12. These results confirm pZL12 as the first example of a plasmid-phage in Streptomyces.

  16. Cure-A-Phobia感

    Institute of Scientific and Technical Information of China (English)

    黄乔

    2015-01-01

    爵士乐源于二十世纪的非洲黑人音乐,被公认为欧洲现代流行音乐的开端,随着二十一世纪多元化文化的不断呈现,爵士乐在我国也愈来愈受到关注.特别是在各艺术类院校,相关爵士乐教学及演出活动也越来越多.本文在此将对瑞典爵士乐队Cure-A-Phobia的一场音乐会略抒己见.

  17. Bacteriophage selection against a plasmid-encoded sex apparatus leads to the loss of antibiotic-resistance plasmids

    OpenAIRE

    Jalasvuori, Matti; Friman, Ville-Petri; Nieminen, Anne; Jaana K.H. Bamford; Buckling, Angus

    2011-01-01

    Antibiotic-resistance genes are often carried by conjugative plasmids, which spread within and between bacterial species. It has long been recognized that some viruses of bacteria (bacteriophage; phage) have evolved to infect and kill plasmid-harbouring cells. This raises a question: can phages cause the loss of plasmid-associated antibiotic resistance by selecting for plasmid-free bacteria, or can bacteria or plasmids evolve resistance to phages in other ways? Here, we show that multiple ant...

  18. An Invertron-Like Linear Plasmid Mediates Intracellular Survival and Virulence in Bovine Isolates of Rhodococcus equi.

    Science.gov (United States)

    Valero-Rello, Ana; Hapeshi, Alexia; Anastasi, Elisa; Alvarez, Sonsiray; Scortti, Mariela; Meijer, Wim G; MacArthur, Iain; Vázquez-Boland, José A

    2015-07-01

    We report a novel host-associated virulence plasmid in Rhodococcus equi, pVAPN, carried by bovine isolates of this facultative intracellular pathogenic actinomycete. Surprisingly, pVAPN is a 120-kb invertron-like linear replicon unrelated to the circular virulence plasmids associated with equine (pVAPA) and porcine (pVAPB variant) R. equi isolates. pVAPN is similar to the linear plasmid pNSL1 from Rhodococcus sp. NS1 and harbors six new vap multigene family members (vapN to vapS) in a vap pathogenicity locus presumably acquired via en bloc mobilization from a direct predecessor of equine pVAPA. Loss of pVAPN rendered R. equi avirulent in macrophages and mice. Mating experiments using an in vivo transconjugant selection strategy demonstrated that pVAPN transfer is sufficient to confer virulence to a plasmid-cured R. equi recipient. Phylogenetic analyses assigned the vap multigene family complement from pVAPN, pVAPA, and pVAPB to seven monophyletic clades, each containing plasmid type-specific allelic variants of a precursor vap gene carried by the nearest vap island ancestor. Deletion of vapN, the predicted "bovine-type" allelic counterpart of vapA, essential for virulence in pVAPA, abrogated pVAPN-mediated intramacrophage proliferation and virulence in mice. Our findings support a model in which R. equi virulence is conferred by host-adapted plasmids. Their central role is mediating intracellular proliferation in macrophages, promoted by a key vap determinant present in the common ancestor of the plasmid-specific vap islands, with host tropism as a secondary trait selected during coevolution with specific animal species. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  19. R-plasmid transfer in a wastewater treatment plant.

    Science.gov (United States)

    Mach, P A; Grimes, D J

    1982-12-01

    Enteric bacteria have been examined for their ability to transfer antibiotic resistance in a wastewater treatment plant. Resistant Salmonella enteritidis, Proteus mirabilis, and Escherichia coli were isolated from clinical specimens and primary sewage effluent. Resistance to ampicillin, chloramphenicol, streptomycin, sulfadiazine, and tetracycline was demonstrated by spread plate and tube dilution techniques. Plasmid mediation of resistance was shown by ethidium bromide curing, agarose gel electrophoresis, and direct cell transfer. Each donor was mated with susceptible E. coli and Shigella sonnei. Mating pairs (and recipient controls) were suspended in unchlorinated primary effluent that had been filtered and autoclaved. Suspensions were added to membrane diffusion chambers which were then placed in the primary and secondary setting tanks of the wastewater treatment plant. Resistant recombinants were detected by replica plating nutrient agar master plates onto xylose lysine desoxycholate agar plates that contained per milliliter of medium 10 micrograms of ampicillin, 30 micrograms of chloramphenicol, 10 micrograms of streptomycin, 100 micrograms of sulfadiazine, or 30 micrograms of tetracycline. Mean transfer frequencies for laboratory matings were 2.1 X 10(-3). In situ matings for primary and secondary settling resulted in frequencies of 4.9 X 10(-5) and 7.5 X 10(-5), respectively. These values suggest that a significant level of resistance transfer occurs in wastewater treatment plants in the absence of antibiotics as selective agents.

  20. ASRM test report: Autoclave cure process development

    Science.gov (United States)

    Nachbar, D. L.; Mitchell, Suzanne

    1992-01-01

    ASRM insulated segments will be autoclave cured following insulation pre-form installation and strip wind operations. Following competitive bidding, Aerojet ASRM Division (AAD) Purchase Order 100142 was awarded to American Fuel Cell and Coated Fabrics Company, Inc. (Amfuel), Magnolia, AR, for subcontracted insulation autoclave cure process development. Autoclave cure process development test requirements were included in Task 3 of TM05514, Manufacturing Process Development Specification for Integrated Insulation Characterization and Stripwind Process Development. The test objective was to establish autoclave cure process parameters for ASRM insulated segments. Six tasks were completed to: (1) evaluate cure parameters that control acceptable vulcanization of ASRM Kevlar-filled EPDM insulation material; (2) identify first and second order impact parameters on the autoclave cure process; and (3) evaluate insulation material flow-out characteristics to support pre-form configuration design.

  1. Distribution of small native plasmids in Streptococcus pyogenes in India.

    Science.gov (United States)

    Bergmann, René; Nerlich, Andreas; Chhatwal, Gursharan S; Nitsche-Schmitz, D Patric

    2014-05-01

    Complete characterization of a Streptococcus pyogenes population from a defined geographic region comprises information on the plasmids that circulate in these bacteria. Therefore, we determined the distribution of small plasmids (pyogenes isolates from India, where diversity of strains and incidence rates of S. pyogenes infections are high. The collection comprised 77 emm-types. For plasmid detection and discrimination, we developed PCRs for different plasmid replication initiation protein genes, the putative repressor gene copG and bacteriocin genes dysA and scnM57. Plasmid distribution was limited to 13 emm-types. Co-detection analysis using aforementioned PCRs revealed four distinct plasmid sub-types, two of which were previously unknown. Representative plasmids pA852 and pA996 of the two uncharacterized plasmid sub-types were sequenced. These two plasmids could be assigned to the pMV158 and the pC194/pUB110 family of rolling-circle plasmids, respectively. The majority of small plasmids found in India belonged to the two newly characterized sub-types, with pA852- and pA996-like plasmids amounting to 42% and 22% of all detected plasmids, respectively. None of the detected plasmids coded for a known antibiotic resistance gene. Instead, all of the four plasmid sub-types carried known or potential bacteriocin genes. These genes may have influence on the evolutionary success of certain S. pyogenes genotypes. Notably, pA852-like plasmids were found in all isolates of the most prevalent emm-type 11.0. Together, a priori fitness of this genotype and increased fitness due to the acquired plasmids may have rendered type emm11.0 successful and caused the prevalence of pA852-like plasmids in India.

  2. Interfacial Properties of Electron Beam Cured Composites

    Energy Technology Data Exchange (ETDEWEB)

    Eberle, C.C.

    1999-12-30

    The objectives of the CRADA are to: Confirm that fiber-resin adhesion is responsible for the observed poor shear properties; Determine the mechanism(s) responsible for poor adhesion between carbon fibers and epoxy resins after e-beam curing; Develop and evaluate resin systems and fiber treatments to improve the properties of e-beam cured, carbon-fiber-reinforced composites; and Develop refined methods for processing e-beam cured, carbon-fiber-reinforced composites.

  3. Microwave Cure of Phenol-Formaldehyde Adhesive

    OpenAIRE

    高谷, 政広; 田平, 英敏; 岡本, 忠

    2006-01-01

    [Synopsis] Phenol-formaldehyde resin has been used as a versatile material for adhesives and coatings of a wide range of adherends because of its excellent performance in water- resistance, strength against abrasion, and so on. However, it has a drawback of slow rate of cure and relevant emission of formaldehyde gas after bonding. We studied the curing performance under irradiation of microwave for the purpose of looking for a way of accelerating the cure rate of phenol formaldehyde resin. Th...

  4. Electron Beam Curing of Advanced Composites

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    The fundamental concept of electron beam method and the application in cure of composites are elaborated in this paper. The components of electron beam curing system are introduced. The mechanisms of interaction between electron beam and polymer matrix composites are presented. Recent studies reported including work of authors themselves on electron beam curing of composites are also discussed. Moreover, the authors believe that it is necessary to do the basic research about understanding how electron beam affects cured network and the mechanical/physical properties of the composites, for establishing a quantitative or semi-quantitative formulation.

  5. Genes from plasmid pKM101 in Haemophilus influenzae: separation of functions of mucA and mucB

    Energy Technology Data Exchange (ETDEWEB)

    Balganesh, M.; Setlow, J.K.

    1985-11-01

    Haemophilus influenzae, normally not mutable by UV, became UV mutable with a recombinant plasmid insertion. A 7.8-kilobase-pair (kbp) fragment of the plasmid pKM101 containing the mucA and mucB genes was ligated to the shuttle vector pDM2, and a Rec- strain of H. influenzae was transformed with the ligated mixture. All of the transformants, unlike the parent Rec- strain, were resistant to UV, could carry out postreplication repair and Weigle reactivation, showed greatly increased spontaneous mutation, and contained a plasmid carrying an insert of only 1.2 rather than 7.8 kbp. This plasmid in a umuC mutant strain of Escherichia coli complemented a pKM101 derivative lacking mucA function but with an intact mucB gene, although there was no complementation with a mucA+ mucB- plasmid, suggesting that the newly constructed plasmid coded for the mucA protein; this is in accord with the restriction analysis and hybridization between the plasmid and a probe containing all of the mucA gene but only a small fraction of mucB. When one of the H. influenzae Rec- transformants lost the plasmid, the resistance to UV was retained but the high spontaneous mutation and UV mutability were not. The fact that there was hybridization between the chromosome of the cured strain and a probe containing both muc genes but none when almost no mucB was present suggested that at least part of the mucB gene had been integrated into the Rec- chromosome. Five different postreplication repair-proficient strains became UV mutable and had high spontaneous mutation rates caused by the putative mucA plasmid, indicating that these strains already possessed a chromosomal equivalent of the mucB gene.

  6. Plasmid profile of bacteria isolated from tears of HIV/AIDS patients

    Directory of Open Access Journals (Sweden)

    O B Ajayi

    2009-01-01

    Full Text Available Objective: The purpose of this study is to determine the presence and transfer of plasmids in bacteria isolated from tears of HIV/AIDS patients, their sensitivity and resistance to commercially available antibiotics. Design: This was a cross sectional experimental study. Materials and methods:One hundred tears samples from HIV/ AIDS patients and fifty tears samples from HIV/AIDS negative patients were screened for resistance to 14 commercially available antibiotics using disc diffusion method. Result: Three multiple antibiotics resistant strains of staphylococcus aureus and four multiple antibiotics resistance strains of Pseudomonas aeruginosa were identified. staphylococcus aureus strains showed 100% resistance to Ampiclox and erythromycin, 66.6% to Perfloxacin, amoxicillin and septrin, 33.33% to ciprofloxacin. Pseudomonas aeruginosa strains showed 100% resistance to streptomycin, amoxicillin, septrin and chloramphenicol. Only I strain of staphylococcus aureus showed presence of plasmid which was not transferable to Escherichia coli because of presence of disulphide cross--linked cell wall. Other strains of both staphylococcus aureus and Pseudomonas aeruginosa remained resistant after curing . Conclusion: Further studies are needed in this area to show if antibiotic resistance in HIV/AIDS positive patients could be as a result of plasmid as well as other factors.

  7. Curing efficiency of three light emitting diode units at different curing profiles

    Directory of Open Access Journals (Sweden)

    Priyanka Verma

    2016-01-01

    Conclusions: Reduction of exposure time to 6 s with high-intensity curing light seemed to be clinically acceptable and should be recommended. Curing of metal brackets with single exposure from buccal side showed lower shear bond strength values.

  8. Prevention is better than cure

    CERN Multimedia

    Anaïs Schaeffer

    2012-01-01

    Throughout the year, members of the Safety Inspections Services section of HSE Unit devote themselves to ensuring the safety of all infrastructure and equipment that represent a specific hazard within the Organization. They regularly comb through all CERN's infrastructure to forestall any accidents and their potential impact, true to the adage that prevention is better than cure.   This site has a few (!) slight safety problems... Spot the mistakes! (Details of the game below.) Ensuring that an HV electrical installation is properly earthed, that a system under pressure has no weak points, that an item of lifting equipment can be used without risk, that safety valves operate at the right pressure threshold as well as checking that a heavy object that could inflict injury if it fell is not stored on top of a cupboard: such are the types of inspections performed by the Safety Inspection Service (DGS-SEE-SIS). "These checks reassure those in charge of equipment and infrastruct...

  9. Hepatitis C, stigma and cure.

    Science.gov (United States)

    Marinho, Rui Tato; Barreira, David Pires

    2013-10-28

    The infection with hepatitis C virus (HCV) is one of the most important global chronic viral infections worldwide. It is estimated to affect around 3% of the world population, about 170-200 million people. Great part of the infections are asymptomatic, the patient can be a chronic carrier for decades without knowing it. The most severe consequences of the chronic infection are liver cirrhosis and hepatocellular carcinoma, which appears in 20%-40% of the patients, leading to hepatic failure and death. The HCV was discovered 25 years ago in 1989, is a RNA virus and classified by the World Health Organization as an oncogenic one. Hepatocellular carcinoma is one of the most important cancers, the fifth worldwide in terms of mortality. It has been increasing in the Ocidental world, mainly due to chronic hepatitis C. Hepatitis C is not only a liver disease and a cause of cirrhosis, but also a mental, psychological, familiar, and social disease. The stigma that the infected person sometimes carries is tremendous having multiple consequences. The main cause is lack of adequate information, even in the health professionals setting. But, besides the "drama" of being infected, health professionals, family, society and the infected patients, must be aware of the chance of real cure and total and definitive elimination of the virus. The treatment for hepatitis C has begun in the last 80's with a percentage of cure of 6%. Step by step the efficacy of the therapy for hepatitis C is rapidly increasing and nowadays with the very new medications, the so called Direct Antiviral Agents-DAAs of new generation, is around 80%-90%.

  10. Stress responses and replication of plasmids in bacterial cells

    Directory of Open Access Journals (Sweden)

    Wegrzyn Alicja

    2002-05-01

    Full Text Available Abstract Plasmids, DNA (or rarely RNA molecules which replicate in cells autonomously (independently of chromosomes as non-essential genetic elements, play important roles for microbes grown under specific environmental conditions as well as in scientific laboratories and in biotechnology. For example, bacterial plasmids are excellent models in studies on regulation of DNA replication, and their derivatives are the most commonly used vectors in genetic engineering. Detailed mechanisms of replication initiation, which is the crucial process for efficient maintenance of plasmids in cells, have been elucidated for several plasmids. However, to understand plasmid biology, it is necessary to understand regulation of plasmid DNA replication in response to different environmental conditions in which host cells exist. Knowledge of such regulatory processes is also very important for those who use plasmids as expression vectors to produce large amounts of recombinant proteins. Variable conditions in large-scale fermentations must influence replication of plasmid DNA in cells, thus affecting the efficiency of recombinant gene expression significantly. Contrary to extensively investigated biochemistry of plasmid replication, molecular mechanisms of regulation of plasmid DNA replication in response to various environmental stress conditions are relatively poorly understood. There are, however, recently published studies that add significant data to our knowledge on relations between cellular stress responses and control of plasmid DNA replication. In this review we focus on plasmids derived from bacteriophage λ that are among the best investigated replicons. Nevertheless, recent results of studies on other plasmids are also discussed shortly.

  11. Curing Mechanism of Condensed Polynuclear Aromatic Resin and Thermal Stability of Cured Resin

    Institute of Scientific and Technical Information of China (English)

    Li Shibin; Sun Qiqian; Wang Yuwei; Wu Mingbo; Zhang Zailong

    2015-01-01

    In order to improve the thermal stability of condensed polynuclear aromatic (COPNA) resin synthesized from vacuum residue, 1,4-benzenedimethanol was added to cure COPNA resin. The curing mechanism was investigated by pro-ton nuclear magnetic resonance spectrometry, solid carbon-13 nuclear magnetic resonance spectrometry and Fourier trans-form infrared spectroscopy. Microstructures of the uncured and the cured COPNA resins were studied by scanning electron microscopy and X-ray diffractometry. The thermal stability of COPNA resins before and after curing was tested by thermo-gravimetric analysis. The element composition of the cured COPNA resin heated at different temperatures was analyzed by an element analyzer. The results showed that the uncured COPNA resin reacted with the cross-linking agent during the cur-ing process, and the curing mechanism was conifrmed to be the electrophilic substitution reaction. Compared with the un-cured COPNA resin, the cured COPNA resin had a smooth surface, well-ordered and streamlined sheet structure with more crystalline solids, better molecular arrangement and orientation. The weight loss process of the uncured and cured COPNA resins was divided into three stages. Carbon residue of the cured COPNA resin was 41.65%at 600℃, which was much higher than 25.02%of the uncured COPNA resin, which indicated that the cured COPNA resin had higher thermal stability.

  12. 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

  13. 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 veter

  14. 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 veter

  15. Plasmids spread very fast in heterogeneous bacterial communities.

    Science.gov (United States)

    Dionisio, Francisco; Matic, Ivan; Radman, Miroslav; Rodrigues, Olivia R; Taddei, François

    2002-01-01

    Conjugative plasmids can mediate gene transfer between bacterial taxa in diverse environments. The ability to donate the F-type conjugative plasmid R1 greatly varies among enteric bacteria due to the interaction of the system that represses sex-pili formations (products of finOP) of plasmids already harbored by a bacterial strain with those of the R1 plasmid. The presence of efficient donors in heterogeneous bacterial populations can accelerate plasmid transfer and can spread by several orders of magnitude. Such donors allow millions of other bacteria to acquire the plasmid in a matter of days whereas, in the absence of such strains, plasmid dissemination would take years. This "amplification effect" could have an impact on the evolution of bacterial pathogens that exist in heterogeneous bacterial communities because conjugative plasmids can carry virulence or antibiotic-resistance genes. PMID:12524329

  16. Dental resin cure monitoring by inherent fluorescence

    Science.gov (United States)

    Li, Qun; Zhou, Jack X.; Li, Qingxiong; Wang, Sean X.

    2008-02-01

    It is demonstrated that the inherent fluorescence of a dental composite resin can be utilized to monitor the curing status, i.e. degree of conversion of the resin. The method does not require any sample preparation and is potentially very fast for real time cure monitoring. The method is verified by Raman spectroscopy analysis.

  17. 7 CFR 29.6010 - Cured.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Cured. 29.6010 Section 29.6010 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections, Marketing... INSPECTION Standards Definitions § 29.6010 Cured. Tobacco dried of its sap by either natural or...

  18. Endogenous mutagenesis in recombinant sulfolobus plasmids.

    Science.gov (United States)

    Sakofsky, Cynthia J; Grogan, Dennis W

    2013-06-01

    Low rates of replication errors in chromosomal genes of Sulfolobus spp. demonstrate that these extreme thermoacidophiles can maintain genome integrity in environments with high temperature and low pH. In contrast to this genetic stability, we observed unusually frequent mutation of the β-D-glycosidase gene (lacS) of a shuttle plasmid (pJlacS) propagated in Sulfolobus acidocaldarius. The resulting Lac(-) mutants also grew faster than the Lac(+) parent, thereby amplifying the impact of the frequent lacS mutations on the population. We developed a mutant accumulation assay and corrections for the effects of copy number and differential growth for this system; the resulting measurements and calculations yielded a corrected rate of 5.1 × 10(-4) mutational events at the lacS gene per plasmid replication. Analysis of independent lacS mutants revealed three types of mutations: (i) G · C-to-A · T transitions, (ii) slipped-strand events, and (iii) deletions. These mutations were frequent in plasmid-borne lacS expressed at a high level but not in single-copy lacS in the chromosome or at lower levels of expression in a plasmid. Substitution mutations arose at only two of 12 potential priming sites of the DNA primase of the pRN1 replicon, but nearly all these mutations created nonsense (chain termination) codons. The spontaneous mutation rate of plasmid-borne lacS was 175-fold higher under high-expression than under low-expression conditions. The results suggest that important DNA repair or replication fidelity functions are impaired or overwhelmed in pJlacS, with results analogous to those of the "transcription-associated mutagenesis" seen in bacteria and eukaryotes.

  19. Plasmid-mediated tetracycline resistance in Haemophilus ducreyi.

    OpenAIRE

    Albritton, W L; Maclean, I W; Slaney, L A; Ronald, A. R.; Deneer, H G

    1984-01-01

    Clinical isolates of Haemophilus ducreyi were shown to be resistant to tetracycline. Resistance was associated in some strains with a 30-megadalton plasmid capable of transferring resistance in conjugative matings with other strains of H. ducreyi and other species of Haemophilus. Restriction endonuclease digestion patterns suggest a relationship between H. ducreyi plasmids and other tetracycline resistance plasmids in Haemophilus. The presence of plasmid-mediated resistance to the tetracyclin...

  20. Plasmid DNA entry into postmitotic nuclei of primary rat myotubes.

    OpenAIRE

    Dowty, M E; Williams, P.; G. Zhang; Hagstrom, J E; Wolff, J A

    1995-01-01

    These studies were initiated to elucidate the mechanism of DNA nuclear transport in mammalian cells. Biotin- or gold-labeled plasmid and plasmid DNA expression vectors for Escherichia coli beta-galactosidase or firefly luciferase were microinjected into the cytoplasm of primary rat myotubes in culture. Plasmid DNA was expressed in up to 70% of the injected myotubes, which indicates that it entered intact, postmitotic nuclei. The nuclear transport of plasmid DNA occurred through the nuclear po...

  1. Replication of plasmids in gram-negative bacteria.

    OpenAIRE

    1989-01-01

    Replication of plasmid deoxyribonucleic acid (DNA) is dependent on three stages: initiation, elongation, and termination. The first stage, initiation, depends on plasmid-encoded properties such as the replication origin and, in most cases, the replication initiation protein (Rep protein). In recent years the understanding of initiation and regulation of plasmid replication in Escherichia coli has increased considerably, but it is only for the ColE1-type plasmids that significant biochemical d...

  2. 9 CFR 319.103 - Cured beef tongue.

    Science.gov (United States)

    2010-01-01

    ... 9 Animals and Animal Products 2 2010-01-01 2010-01-01 false Cured beef tongue. 319.103 Section 319... Cured beef tongue. In preparing “Cured Beef Tongue,” the application of curing solution to the fresh beef tongue shall not result in an increase in the weight of the cured beef tongue of more than...

  3. Curing agent for polyepoxides and epoxy resins and composites cured therewith. [preventing carbon fiber release

    Science.gov (United States)

    Serafini, T. T.; Delvigs, P.; Vannucci, R. D. (Inventor)

    1981-01-01

    A curing for a polyepoxide is described which contains a divalent aryl radical such as phenylene a tetravalent aryl radical such as a tetravalent benzene radical. An epoxide is cured by admixture with the curing agent. The cured epoxy product retains the usual properties of cured epoxides and, in addition, has a higher char residue after burning, on the order of 45% by weight. The higher char residue is of value in preventing release to the atmosphere of carbon fibers from carbon fiber-epoxy resin composites in the event of burning of the composite.

  4. Plasmid Segregation: Spatial Awareness at the Molecular Level

    DEFF Research Database (Denmark)

    Møller-Jensen, Jakob; Gerdes, Kenn

    2007-01-01

    In bacteria, low-copy number plasmids ensure their stable inheritance by partition loci (par), which actively distribute plasmid replicates to each side of the cell division plane. Using time-lapse fluorescence microscopic tracking of segregating plasmid molecules, a new study provides novel insi...

  5. Cloning of Two Bacteriocin Genes from a Lactococcal Bacteriocin Plasmid

    NARCIS (Netherlands)

    Belkum, Marco J. van; Hayema, Bert Jan; Geis, Arnold; Kok, Jan; Venema, Gerard

    1989-01-01

    Lactococcus lactis subsp. cremoris 9B4 plasmid p9B4-6 (60 kilobases [kb]), which specifies bacteriocin production and immunity, was analyzed with restriction endonucleases, and fragments of this plasmid were cloned into shuttle vectors based on the broad-host-range plasmid pWVO1. Two regions on p9B4

  6. Multilocus sequence typing of IncN plasmids

    DEFF Research Database (Denmark)

    García-Fernández, Aurora; Villa, Laura; Moodley, Arshnee

    2011-01-01

    categorization of IncN plasmids. METHODS: Twelve fully sequenced IncN plasmids available at GenBank were analysed in silico for selecting the loci for the IncN-specific pMLST. A total of 58 plasmids originating from different reservoirs (human, pig, poultry, cattle and horses) and geographic regions (Italy...

  7. Fasciation induction by the phytopathogen Rhodococcus fascians depends upon a linear plasmid encoding a cytokinin synthase gene.

    Science.gov (United States)

    Crespi, M; Messens, E; Caplan, A B; van Montagu, M; Desomer, J

    1992-01-01

    Rhodococcus fascians is a nocardiform bacteria that induces leafy galls (fasciation) on dicotyledonous and several monocotyledonous plants. The wild-type strain D188 contained a conjugative, 200 kb linear extrachromosomal element, pFiD188. Linear plasmid-cured strains were avirulent and reintroduction of this linear element restored virulence. Pulsed field electrophoresis indicated that the chromosome might also be a linear molecule of 4 megabases. Three loci involved in phytopathogenicity have been identified by insertion mutagenesis of this Fi plasmid. Inactivation of the fas locus resulted in avirulent strains, whereas insertions in the two other loci affected the degree of virulence, yielding attenuated (att) and hypervirulent (hyp) bacteria. One of the genes within the fas locus encoded an isopentenyltranferase (IPT) with low homology to analogous proteins from Gram-negative phytopathogenic bacteria. IPT activity was detected after expression of this protein in Escherichia coli cells. In R.fascians, ipt expression could only be detected in bacteria induced with extracts from fasciated tissue. R.fascians strains without the linear plasmid but containing this fas locus alone could not provoke any phenotype on plants, indicating additional genes from the linear plasmid were also essential for virulence. These studies, the first genetic analysis of the interaction of a Gram-positive bacterium with plants, suggest that a novel mechanism for plant tumour induction has evolved in R.fascians independently from the other branches of the eubacteria. Images PMID:1547783

  8. A Transmissible Plasmid-Borne Pathogenicity Island Confers Piscibactin Biosynthesis in the Fish Pathogen Photobacterium damselae subsp. piscicida

    Science.gov (United States)

    Rivas, Amable J.; Balado, Miguel; Fuentes-Monteverde, Juan Carlos; Rodríguez, Jaime; Jiménez, Carlos; Lemos, Manuel L.; Waldor, Matthew K.

    2015-01-01

    The fish pathogen Photobacterium damselae subsp. piscicida produces the siderophore piscibactin. A gene cluster that resembles the Yersinia high-pathogenicity island (HPI) encodes piscibactin biosynthesis. Here, we report that this HPI-like cluster is part of a hitherto-uncharacterized 68-kb plasmid dubbed pPHDP70. This plasmid lacks homologs of genes that mediate conjugation, but we found that it could be transferred at low frequencies from P. damselae subsp. piscicida to a mollusk pathogenic Vibrio alginolyticus strain and to other Gram-negative bacteria, likely dependent on the conjugative functions of the coresident plasmid pPHDP60. Following its conjugative transfer, pPHDP70 restored the capacity of a vibrioferrin mutant of V. alginolyticus to grow under low-iron conditions, and piscibactin became detectable in its supernatant. Thus, pPHDP70 appears to harbor all the genes required for piscibactin biosynthesis and transport. P. damselae subsp. piscicida strains cured of pPHDP70 no longer produced piscibactin, had impaired growth under iron-limited conditions, and exhibited markedly decreased virulence in fish. Collectively, our findings highlight the importance of pPHDP70, with its capacity for piscibactin-mediated iron acquisition, in the virulence of P. damselae subsp. piscicida. Horizontal transmission of this plasmid-borne piscibactin synthesis gene cluster in the marine environment may facilitate the emergence of new pathogens. PMID:26092457

  9. Transfection of bone marrow mesenchymal stem cells using green fluorescence protein labeled hVEGF165 recombinant plasmid mediated by liposome

    Institute of Scientific and Technical Information of China (English)

    Tao Wang; Tian-An Liao; Shao-Bo Zhong

    2013-01-01

    Objective:To study the role of bone marrow mesenchymal stem cells (BMSCs) in construction of vascularized engineered tissue. Methods: hVEGF165 was amplified via RT-PCR before recombinant with pShuttle-green fluorescence protein;green fluorescent protein (GFP)-CMV. Then the recombinant shuttle plasmid was transfected into BMSCs with LipofectamineTM 2000 for packaging and amplifying. hVEGF165 mRNA expression in BMSCs cells was tested. Results:The sequence of hVEGF165 in pShuttle-GFP-hVEGF165 plasmid was confirmed by double-enzyme cleavage method and sequencing. hVEGF165 was highly expressed in BMSCs. Conclusions:The GFP/hVEGF165 recombinant plasmid vector was constructed successfully and expressed effectively in host cells, which may be helpful for discussing the possibility of the application of VEGF165-BMSCs in tissue engineering and ischemic disease cure.

  10. Bacteriophages limit the existence conditions for conjugative plasmids.

    Science.gov (United States)

    Harrison, Ellie; Wood, A Jamie; Dytham, Calvin; Pitchford, Jonathan W; Truman, Julie; Spiers, Andrew; Paterson, Steve; Brockhurst, Michael A

    2015-06-02

    Bacteriophages are a major cause of bacterial mortality and impose strong selection on natural bacterial populations, yet their effects on the dynamics of conjugative plasmids have rarely been tested. We combined experimental evolution, mathematical modeling, and individual-based simulations to explain how the ecological and population genetics effects of bacteriophages upon bacteria interact to determine the dynamics of conjugative plasmids and their persistence. The ecological effects of bacteriophages on bacteria are predicted to limit the existence conditions for conjugative plasmids, preventing persistence under weak selection for plasmid accessory traits. Experiments showed that phages drove faster extinction of plasmids in environments where the plasmid conferred no benefit, but they also revealed more complex effects of phages on plasmid dynamics under these conditions, specifically, the temporary maintenance of plasmids at fixation followed by rapid loss. We hypothesized that the population genetic effects of bacteriophages, specifically, selection for phage resistance mutations, may have caused this. Further mathematical modeling and individual-based simulations supported our hypothesis, showing that conjugative plasmids may hitchhike with phage resistance mutations in the bacterial chromosome. Conjugative plasmids are infectious loops of DNA capable of transmitting DNA between bacterial cells and between species. Because plasmids often carry extra genes that allow bacteria to live in otherwise-inhospitable environments, their dynamics are central to understanding bacterial adaptive evolution. The plasmid-bacterium interaction has typically been studied in isolation, but in natural bacterial communities, bacteriophages, viruses that infect bacteria, are ubiquitous. Using experiments, mathematical models, and computer simulations we show that bacteriophages drive plasmid dynamics through their ecological and evolutionary effects on bacteria and ultimately

  11. Bacteriophage selection against a plasmid-encoded sex apparatus leads to the loss of antibiotic-resistance plasmids.

    Science.gov (United States)

    Jalasvuori, Matti; Friman, Ville-Petri; Nieminen, Anne; Bamford, Jaana K H; Buckling, Angus

    2011-12-23

    Antibiotic-resistance genes are often carried by conjugative plasmids, which spread within and between bacterial species. It has long been recognized that some viruses of bacteria (bacteriophage; phage) have evolved to infect and kill plasmid-harbouring cells. This raises a question: can phages cause the loss of plasmid-associated antibiotic resistance by selecting for plasmid-free bacteria, or can bacteria or plasmids evolve resistance to phages in other ways? Here, we show that multiple antibiotic-resistance genes containing plasmids are stably maintained in both Escherichia coli and Salmonella enterica in the absence of phages, while plasmid-dependent phage PRD1 causes a dramatic reduction in the frequency of antibiotic-resistant bacteria. The loss of antibiotic resistance in cells initially harbouring RP4 plasmid was shown to result from evolution of phage resistance where bacterial cells expelled their plasmid (and hence the suitable receptor for phages). Phages also selected for a low frequency of plasmid-containing, phage-resistant bacteria, presumably as a result of modification of the plasmid-encoded receptor. However, these double-resistant mutants had a growth cost compared with phage-resistant but antibiotic-susceptible mutants and were unable to conjugate. These results suggest that bacteriophages could play a significant role in restricting the spread of plasmid-encoded antibiotic resistance.

  12. The Plasmid of Escherichia coli Strain S88 (O45:K1:H7) That Causes Neonatal Meningitis Is Closely Related to Avian Pathogenic E. coli Plasmids and Is Associated with High-Level Bacteremia in a Neonatal Rat Meningitis Model▿

    Science.gov (United States)

    Peigne, Chantal; Bidet, Philippe; Mahjoub-Messai, Farah; Plainvert, Céline; Barbe, Valérie; Médigue, Claudine; Frapy, Eric; Nassif, Xavier; Denamur, Erick; Bingen, Edouard; Bonacorsi, Stéphane

    2009-01-01

    A new Escherichia coli virulent clonal group, O45:K1, belonging to the highly virulent subgroup B21 was recently identified in France, where it accounts for one-third of E. coli neonatal meningitis cases. Here we describe the sequence, epidemiology and function of the large plasmid harbored by strain S88, which is representative of the O45:K1 clonal group. Plasmid pS88 is 133,853 bp long and contains 144 protein-coding genes. It harbors three different iron uptake systems (aerobactin, salmochelin, and the sitABCD genes) and other putative virulence genes (iss, etsABC, ompTP, and hlyF). The pS88 sequence is composed of several gene blocks homologous to avian pathogenic E. coli plasmids pAPEC-O2-ColV and pAPEC-O1-ColBM. PCR amplification of 11 open reading frames scattered throughout the plasmid was used to investigate the distribution of pS88 and showed that a pS88-like plasmid is present in other meningitis clonal groups such as O18:K1, O1:K1, and O83:K1. A pS88-like plasmid was also found in avian pathogenic strains and human urosepsis strains belonging to subgroup B21. A variant of S88 cured of its plasmid displayed a marked loss of virulence relative to the wild-type strain in a neonatal rat model, with bacteremia more than 2 log CFU/ml lower. The salmochelin siderophore, a known meningovirulence factor, could not alone explain the plasmid's contribution to virulence, as a salmochelin mutant displayed only a minor fall in bacteremia (0.9 log CFU/ml). Thus, pS88 is a major virulence determinant related to avian pathogenic plasmids that has spread not only through meningitis clonal groups but also human urosepsis and avian pathogenic strains. PMID:19307211

  13. Curing efficiency of modern LED units.

    Science.gov (United States)

    Rencz, Adam; Hickel, Reinhard; Ilie, Nicoleta

    2012-02-01

    Recent reports claim that modern light-emitting diode (LED) curing units improve curing efficiency by increasing the units' irradiance. In this context also, short polymerisation times up to 5 s are proposed. The aim of this study was to examine whether there are differences in the curing efficiency of modern LED curing units by assessing their effect on two different composite materials and by varying the irradiation time. A nano- and a micro-hybrid resin-based composite (RBC) were polymerised for 5, 10 and 20 s with three commercial and a Prototype LED unit (Elipar™ S10). Cylindrical specimens (6 mm in depth, 4 mm in diameter) were prepared in three increments, each 2-mm thick, and were consecutively cured. Degree of cure was measured for 20 min in real time at the bottom of the samples, starting with the photoinitiation. The micro-mechanical properties (modulus of elasticity, E and Vickers hardness, HV) were measured as a function of depth, in 100-μm steps, on the above described samples stored in distilled water for 24 h at 37°C. Data were analysed with multivariate ANOVA followed by Tukey's test, t test and partial eta-squared statistics. In descending order of the strength of their effect, the type of RBC, depth, polymerisation time and curing unit were significant factors affecting the micro-mechanical parameters (p surface, a minimum of 20 s of irradiation is necessary for an adequate polymerisation 2 mm beyond the surface.

  14. 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.

  15. 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.

  16. One-step microwave foaming and curing

    Science.gov (United States)

    Gagliani, J.; Lee, R.; Sorathia, U. A. K.; Wilcoxson, A. L.

    1981-01-01

    Process that combines microwave foaming and curing of polyimide precursors in single step produces fire-resistant foam slabs of much larger volume than has previously been possible. By adding selected conductive fillers to powder precursors and by using high-power microwave oven, foam slabs with dimensions in excess of 61 by 61 by 7.6 cm are made. Typical foaming and curing and curing time is 35 minutes in microwave oven with additional 1 to 2 hour postcure in conventional oven.

  17. Acceleration of curing of resin composite at the bottom surface using slow-start curing methods.

    Science.gov (United States)

    Yoshikawa, Takako; Morigami, Makoto; Sadr, Alireza; Tagami, Junji

    2013-01-01

    The aim of this study was to evaluate the effect of two slow-start curing methods on acceleration of the curing of resin composite specimens at the bottom surface. The light-cured resin composite was polymerized using one of three curing techniques: (1) 600 mW/cm(2) for 60 s, (2) 270 mW/cm(2) for 10 s+0-s interval+600 mW/cm(2) for 50 s, and (3) 270 mW/cm(2) for 10 s+5-s interval+600 mW/cm(2) for 50 s. After light curing, Knoop hardness number was measured at the top and bottom surfaces of the resin specimens. The slow-start curing method with the 5-s interval caused greater acceleration of curing of the resin composite at the bottom surface of the specimens than the slow-start curing method with the 0-s interval. The light-cured resin composite, which had increased contrast ratios during polymerization, showed acceleration of curing at the bottom surface.

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

    DEFF Research Database (Denmark)

    Klümper, Uli

    at high frequencies from diverse donors, I showed plasmid or donor dependence of plasmid transfer to other species. Additionally, environmental factors like stress also impact the permissiveness of phylogenetic groups towards plasmids. The developed method and results increase our ability to predict......Horizontal transfer of mobile genetic elements facilitates adaptive and evolutionary processes in bacteria. Among the known mobile genetic elements, plasmids can confer their hosts with accessory adaptive traits, such as antibiotic or heavy metal resistances, or additional metabolic pathways...... and the extent of bacterial phyla permissive towards plasmid receipt are largely unknown. Historically, methods exploring the underlying genetic and environmental factors of plasmid transfer have been heavily reliant on cultivation and expression of plasmid encoded phenotypes. This has provided an incomplete...

  19. Modeling sRNA-Regulated Plasmid Maintenance

    Science.gov (United States)

    Klumpp, Stefan

    2017-01-01

    We study a theoretical model for the toxin-antitoxin (hok/sok) mechanism for plasmid maintenance in bacteria. Toxin-antitoxin systems enforce the maintenance of a plasmid through post-segregational killing of cells that have lost the plasmid. Key to their function is the tight regulation of expression of a protein toxin by an sRNA antitoxin. Here, we focus on the nonlinear nature of the regulatory circuit dynamics of the toxin-antitoxin mechanism. The mechanism relies on a transient increase in protein concentration rather than on the steady state of the genetic circuit. Through a systematic analysis of the parameter dependence of this transient increase, we confirm some known design features of this system and identify new ones: for an efficient toxin-antitoxin mechanism, the synthesis rate of the toxin’s mRNA template should be lower that of the sRNA antitoxin, the mRNA template should be more stable than the sRNA antitoxin, and the mRNA-sRNA complex should be more stable than the sRNA antitoxin. Moreover, a short half-life of the protein toxin is also beneficial to the function of the toxin-antitoxin system. In addition, we study a therapeutic scenario in which a competitor mRNA is introduced to sequester the sRNA antitoxin, causing the toxic protein to be expressed. PMID:28085919

  20. Modeling sRNA-regulated Plasmid Maintenance

    CERN Document Server

    Gong, Chen Chris

    2016-01-01

    We study a theoretical model for the toxin-antitoxin (hok/sok) mechanism for plasmid maintenance in bacteria. Toxin-antitoxin systems enforce the maintenance of a plasmid through post-segregational killing of cells that have lost the plasmid. Key to their function is the tight regulation of expression of a protein toxin by an sRNA antitoxin. Here, we focus on the nonlinear nature of the regulatory circuit dynamics of the toxin-antitoxin mechanism. The mechanism relies on a transient increase in protein concentration rather than on the steady state of the genetic circuit. Through a systematic analysis of the parameter dependence of this transient increase, we confirm some known design features of this system and identify new ones: for an efficient toxin-antitoxin mechanism, the synthesis rate of the toxin's mRNA template should be lower that of the sRNA antitoxin, the mRNA template should be more stable than the sRNA antitoxin, and the mRNA-sRNA complex should be more stable than the sRNA antitoxin. Moreover, ...

  1. [Mineral water as a cure].

    Science.gov (United States)

    Nocco, Priska Binz

    2008-01-01

    The treatment of diseases with mineral spring water belongs to the oldest medical therapies. The "remedy" mineral water is therefore of importance also within the pharmacy. The present pharmacy historical work examines the impact of the use of mineral waters, as well as of their dried components, as therapeutic agents in the 19th and early 20th centuries, i.e. from approx. 1810 to 1930, as well as the contributions given by pharmacists in the development and analysis of mineral water springs. Beside these aspects, the aim here is also to describe the role played by pharmacists in the production of artificial mineral water as well as in the sale and wholesale of natural and artificial mineral water. In the first part of this work the situation in Switzerland and its surrounding countries, such as Germany, France, Italy and Austria, is discussed. The second part contains a case-study of the particular situation in the Canton Tessin. It is known from the scientific literature published at that time that information on mineral water was frequently reported. Starting from the beginning of the 19th century the number of such publications increased tremendously. The major part of them were publications in scientific journals or contributions to medical and pharmaceutical manuals and reference books. In particular the spa-related literature, such as spa-guides, was of growing interest to a broad public. The inclusion of monographs into the Swiss, the Cantonal as well the foreign pharmacopoeias granted a legal frame for the mineral waters and their dried components. These works are of major importance from a pharmacy historical standpoint and represent a unique proof of historical evidence of the old medicinal drug heritage. The most frequently used therapies based on mineral waters were drinking and bath cures. Several diseases, particularly those of a chronic character, were treated with mineral waters. The positive influence of these cures on the recovery of the patients

  2. Light-cured resin for post patterns.

    Science.gov (United States)

    Waldmeier, M D; Grasso, J E

    1992-09-01

    A method of using light-cured acrylic resin as an alternative to the use of chemically-cured acrylic resins with elastomeric impressions for direct post patterns is presented. The GC Unifast LC acrylic resin is a powder/liquid type resin cured by exposure to visible light. The polymerization process has four stages before final curing: slurry, stringy, dough-like (plastic), and rubber-like (elastic). Advantages over current direct and indirect procedures include ease of manipulation of the material and no change in laboratory handling procedures. While in the dough-like state, the material can be contoured. In the rubber-like state, it is flexible to disengage from minor undercut areas.

  3. Fast Curing of Composite Wood Products

    Energy Technology Data Exchange (ETDEWEB)

    Dr. Arthur J. Ragauskas

    2006-04-26

    The overall objective of this program is to develop low temperature curing technologies for UF and PF resins. This will be accomplished by: • Identifying the rate limiting UF and PF curing reactions for current market resins; • Developing new catalysts to accelerate curing reactions at reduced press temperatures and times. In summary, these new curing technologies will improve the strength properties of the composite wood products and minimize the detrimental effects of wood extractives on the final product while significantly reducing energy costs for wood composites. This study is related to the accelerated curing of resins for wood composites such as medium density fiberboard (MDF), particle board (PB) and oriented strandboard (OSB). The latter is frequently manufactured with a phenol-formaldehyde resin whereas ureaformaldehyde (UF) resins are usually used in for the former two grades of composite wood products. One of the reasons that hinder wider use of these resins in the manufacturing of wood composites is the slow curing speed as well as inferior bondability of UF resin. The fast curing of UP and PF resins has been identified as an attractive process development that would allow wood to be bonded at higher moisture contents and at lower press temperatures that currently employed. Several differing additives have been developed to enhance cure rates of PF resins including the use of organic esters, lactones and organic carbonates. A model compound study by Conner, Lorenz and Hirth (2002) employed 2- and 4-hydroxymethylphenol with organic esters to examine the chemical basis for the reported enhanced reactivity. Their studies suggested that the enhance curing in the presence of esters could be due to enhanced quinone methide formation or enhanced intermolecular SN2 reactions. In either case the esters do not function as true catalysts as they are consumed in the reaction and were not found to be incorporated in the polymerized resin product. An

  4. Effect of cure cycle on curing process and hardness for epoxy resin

    Directory of Open Access Journals (Sweden)

    2009-09-01

    Full Text Available A 3-dimensional finite element model is developed to simulate and analyze the temperature and degree of cure field of epoxy casting part during cure process. The present model based on general finite element software ABAQUS is verified by literature example and experimental data. The numerical results show good agreement with literature example and measured data, and are even more accurate than the simulation of literature. After modeling successfully, the influence of temperature cure cycle ramps have on the temperature and degree of cure gradient is investigated. Moreover, the effect of non-uniform temperature and degree of cure field within epoxy casting part on hardness is demonstrated. The present model provides an accurate and novel method that allows further insight into the process of cure for epoxy resin.

  5. EVALUATION OF DIELECTRIC CURING MONITORING INVESTIGATING LIGHT-CURING DENTAL FILLING COMPOSITES

    Directory of Open Access Journals (Sweden)

    Johannes Steinhaus

    2011-05-01

    Full Text Available The aim of this study is the evaluation of a dielectric analysis (DEA method monitoring the curing behaviour of a light curing dental filling material in real-time. The evaluation is to extract the influence of light intensity on the photo-curing process of dental composite filling materials. The intensity change is obtained by measuring the curing process at different sample depth. It could be shown that increasing sample thickness, and therefore exponentially decreasing light intensity, causes a proportional decrease in the initial curing rate. Nevertheless, the results give rise to the assumption that lower illumination intensities over a long period cause higher overall conversion, and thus better mechanical properties. This would allow for predictions of the impact of different curing-rates on the final mechanical properties.

  6. High Power UV LED Industrial Curing Systems

    Energy Technology Data Exchange (ETDEWEB)

    Karlicek, Robert, F., Jr; Sargent, Robert

    2012-05-14

    UV curing is a green technology that is largely underutilized because UV radiation sources like Hg Lamps are unreliable and difficult to use. High Power UV LEDs are now efficient enough to replace Hg Lamps, and offer significantly improved performance relative to Hg Lamps. In this study, a modular, scalable high power UV LED curing system was designed and tested, performing well in industrial coating evaluations. In order to achieve mechanical form factors similar to commercial Hg Lamp systems, a new patent pending design was employed enabling high irradiance at long working distances. While high power UV LEDs are currently only available at longer UVA wavelengths, rapid progress on UVC LEDs and the development of new formulations designed specifically for use with UV LED sources will converge to drive more rapid adoption of UV curing technology. An assessment of the environmental impact of replacing Hg Lamp systems with UV LED systems was performed. Since UV curing is used in only a small portion of the industrial printing, painting and coating markets, the ease of use of UV LED systems should increase the use of UV curing technology. Even a small penetration of the significant number of industrial applications still using oven curing and drying will lead to significant reductions in energy consumption and reductions in the emission of green house gases and solvent emissions.

  7. 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.

  8. Comparative study on compressive strength of Self cured SCC and Normally cured SCC

    Directory of Open Access Journals (Sweden)

    Ms. Akanksha A. Patil

    2014-11-01

    Full Text Available Curing is the process of maintaining proper moisture content particularly within 28 days to promote optimum cement hydration immediately after placement. Self-compacting concrete is made up of admixture i.e. superplasticizer. In recent years, self-compacting concrete (SCC has gained wide use for placement in congested reinforced concrete structures with difficult casting conditions. Also various curing methods are adopted in the construction industry especially for vertical structures, inaccessible areas s.a. high rise buildings, water scarce areas etc. In such structures conventional curing is not practically possible in most of the cases. But we need efficient curing which improves the strength and durability of concrete. In the present work, comparison of compressive strength of normally cured SCC and SCC cured with self curing material i.e. wax based, white pigmented, membrane forming concrete curing compound has been done. This study is investigating that weather the use of self curing compound is economical or not in remote areas of water without compromising with the compressive strength of concrete.

  9. Cure fraction estimation from the mixture cure models for grouped survival data.

    Science.gov (United States)

    Yu, Binbing; Tiwari, Ram C; Cronin, Kathleen A; Feuer, Eric J

    2004-06-15

    Mixture cure models are usually used to model failure time data with long-term survivors. These models have been applied to grouped survival data. The models provide simultaneous estimates of the proportion of the patients cured from disease and the distribution of the survival times for uncured patients (latency distribution). However, a crucial issue with mixture cure models is the identifiability of the cure fraction and parameters of kernel distribution. Cure fraction estimates can be quite sensitive to the choice of latency distributions and length of follow-up time. In this paper, sensitivity of parameter estimates under semi-parametric model and several most commonly used parametric models, namely lognormal, loglogistic, Weibull and generalized Gamma distributions, is explored. The cure fraction estimates from the model with generalized Gamma distribution is found to be quite robust. A simulation study was carried out to examine the effect of follow-up time and latency distribution specification on cure fraction estimation. The cure models with generalized Gamma latency distribution are applied to the population-based survival data for several cancer sites from the Surveillance, Epidemiology and End Results (SEER) Program. Several cautions on the general use of cure model are advised.

  10. Plasmid transfer between bacteria in soil microcosms and the field

    Directory of Open Access Journals (Sweden)

    Eric Smit

    1997-01-01

    Full Text Available In ibis review factors influencing conjugal plasmid transfer between bacteria and the possible role of naturally occurring selftransmissible plasmide for the dissemination of recombinant DNA in soil will be discussed. In microcosm studies, plasmid transfer between various species of introduced bacteria has been detected. Moreover, plamid transfer to indigenous soil micoorganisms was observed. Soil is an oligotrophic environment and plasmid transfer occurred mainly under conditions which were nutritionally favourable for bacteria, such as in the plant rhizosphere and in the presence of clay minerais or added nutrients. Mobilizable plasmids, lacking the ability to transfer themselves, have been reported to be transferred in the presence of selftransmissible plasmids. A study comparing conjugal transfer in microcosme with those in the field revealed that the transfer rates found in microcosme and in the field were similar. Transfer of chromosomal DNA by plasmid RP4 could only be shown on filters and was not observed in soil. Transfer of plasmids carrying biodegradative genes appeared to be favoured in the presence of the compound that can be degraded. Evidence was found for the presence of naturally-occurring selftransmissible plasmids in bacteria in the rhizosphere which could mobilize recombinant plasmids.

  11. Isolation and screening of plasmids from the epilithon which mobilize recombinant plasmid pD10.

    OpenAIRE

    Hill, K E; A. J. Weightman; Fry, J C

    1992-01-01

    This study examined the potential of bacteria from river epilithon to mobilize a recombinant catabolic plasmid, pD10, encoding 3-chlorobenzoate degradation and kanamycin resistance. Fifty-four mobilizing plasmids were exogenously isolated by triparental matings between strains of Pseudomonas putida and epilithic bacteria from the River Taff (South Wales, United Kingdom). Frequencies for mobilization ranged from 1.7 x 10(-8) to 4.5 x 10(-3) per recipient at 20 degrees C. The sizes of the mobil...

  12. Effect of Promoters and Plasmid Copy Number on Cyt1A Synthesis and Crystal Assembly in Bacillus thuringiensis.

    Science.gov (United States)

    Park, Hyun-Woo; Hice, Robert H; Federici, Brian A

    2016-01-01

    Cyt1Aa is a major mosquitocidal protein synthesized during sporulation of Bacillus thuringiensis subsp. israelensis, composing more than 50% of its parasporal body. This high level of synthesis is due to several factors including three strong sporulation-dependent promoters, a strong transcription termination sequence, and an associated 20-kDa helper protein. Cyt1Aa's toxicity is low compared to the Cry proteins of this species, namely, Cry4Aa, Cry4Ba, and Cry11Aa, but it nevertheless plays an important role in the biology of B. thuringiensis subsp. israelensis in that it synergizes their mosquitocidal toxicity and suppresses the evolution of resistance. In the present study, the effects of using different cyt1Aa promoter combinations and plasmid copy number on synthesis of Cyt1Aa were evaluated. Using the 4Q7 (plasmid-cured) strain of B. thuringiensis subsp. israelensis as an experimental host, a plasmid copy number of two or three yielded no Cyt1Aa, whereas a copy number of four yielded only small crystals, even when expression was driven by one of the wild-type promoters. However, using all three wild-type promoters and a plasmid copy number of 20 yielded Cyt1A crystals tenfold larger than those produced by one promoter and a plasmid copy number of four. High levels of Cyt1Aa synthesis resulted in significantly fewer spores per unit medium and imperfectly formed crystals. Similar results were obtained when Cyt1Aa synthesis was evaluated using the same expression constructs in a mutant strain of B. thuringiensis subsp. israelensis that lacks the cyt1Aa gene.

  13. Isolation of clinical strains of Pseudomonas aeruginosa harboring different plasmids.

    Science.gov (United States)

    Ranjbar, R; Owlia, P; Saderi, H; Bameri, Z; Izadi, M; Jonaidi, N; Morovvati, S

    2007-09-01

    Aim of this study was to investigate the presence of plasmids among the strains of P. aeruginosa isolated from clinically diagnosed cases in Tehran in 2006. A total of 38 strains of P. aeruginosa were isolated. With the exception of one isolate, all P. aeruginosa strains harbored at least one plasmid band. The electrophoretic analysis of plasmid DNAs showed different number of plasmid bands among the strains tested. The DNA band of 1.4 kbp was evident in 84.2% of the strains. Approximately 71 and 21% of the isolates harbored concomitantly two and three plasmids, respectively. Isolation of strains with diverse types of plasmids suggests the different cluster of P. aeruginosa might be disseminated during the current study period.

  14. Conjugative botulinum neurotoxin-encoding plasmids in Clostridium botulinum.

    Directory of Open Access Journals (Sweden)

    Kristin M Marshall

    Full Text Available BACKGROUND: Clostridium botulinum produces seven distinct serotypes of botulinum neurotoxins (BoNTs. The genes encoding different subtype neurotoxins of serotypes A, B, F and several dual neurotoxin-producing strains have been shown to reside on plasmids, suggesting that intra- and interspecies transfer of BoNT-encoding plasmids may occur. The objective of the present study was to determine whether these C. botulinum BoNT-encoding plasmids are conjugative. METHODOLOGY/PRINCIPAL FINDINGS: C. botulinum BoNT-encoding plasmids pBotCDC-A3 (strain CDC-A3, pCLJ (strain 657Ba and pCLL (strain Eklund 17B were tagged with the erythromycin resistance marker (Erm using the ClosTron mutagenesis system by inserting a group II intron into the neurotoxin genes carried on these plasmids. Transfer of the tagged plasmids from the donor strains CDC-A3, 657Ba and Eklund 17B to tetracycline-resistant recipient C. botulinum strains was evaluated in mating experiments. Erythromycin and tetracycline resistant transconjugants were isolated from donor:recipient mating pairs tested. Transfer of the plasmids to the transconjugants was confirmed by pulsed-field gel electrophoresis (PFGE and Southern hybridizations. Transfer required cell-to-cell contact and was DNase resistant. This indicates that transfer of these plasmids occurs via a conjugation mechanism. CONCLUSIONS/SIGNIFICANCE: This is the first evidence supporting conjugal transfer of native botulinum neurotoxin-encoding plasmids in C. botulinum, and provides a probable mechanism for the lateral distribution of BoNT-encoding plasmids to other C. botulinum strains. The potential transfer of C. botulinum BoNT-encoding plasmids to other bacterial hosts in the environment or within the human intestine is of great concern for human pathogenicity and necessitates further characterization of these plasmids.

  15. Photonic plasmid stability of transformed Salmonella Typhimurium: A comparison of three unique plasmids

    Directory of Open Access Journals (Sweden)

    Lay Donald

    2009-07-01

    Full Text Available Abstract Background Acquiring a highly stable photonic plasmid in transformed Salmonella Typhimurium for use in biophotonic studies of bacterial tracking in vivo is critical to experimental paradigm development. The objective of this study was to determine stability of transformed Salmonella Typhimurium (S. typh-lux using three different plasmids and characterize their respective photonic properties. Results In presence of ampicillin (AMP, S. typh-lux with pCGLS-1, pAK1-lux and pXEN-1 plasmids exhibited 100% photon-emitting colonies over a 10-d study period. Photon emitters of S. typh-lux with pCGLS-1, pAK1-lux and pXEN-1 without AMP selection decreased over time (P 7 to 1 × 109 CFU, P 0.05; although photonic emissions across a range of bacterial concentrations were not different (1 × 104 to 1 × 106 CFU, P > 0.05. For very low density bacterial concentrations imaged in 96 well plates photonic emissions were positively correlated with bacterial concentration (P 3 to 1 × 105 CFU low to high were different in the 96-well plate format (P Conclusion These data characterize photon stability properties for S. typh-lux transformed with three different photon generating plasmids that may facilitate real-time Salmonella tracking using in vivo or in situ biophotonic paradigms.

  16. HEAT-RESISTANT COMPOSITES CURED BY ELECTRON BEAM

    Institute of Scientific and Technical Information of China (English)

    Jian-wen Bao; Yang Li; Xiang-bao Chen; Feng-mei Li

    2001-01-01

    Electron beam (EB) curing of composites has many advantages. Heat-resistant EB-curing composites could substitute polyimide composites used in aeronautical engines. In this paper, the effects of catalyst and dose on the cured resin were investigated. The heat-resistance of the resin cured by EB was evaluated by dynamic mechanical analysis (DMA). The experimental results show that the mechanical properties of the composites cured by EB could meet the specifications of aeronautical engines at 250°C.

  17. Bacteriophages Limit the Existence Conditions for Conjugative Plasmids

    Science.gov (United States)

    Wood, A. Jamie; Dytham, Calvin; Pitchford, Jonathan W.; Truman, Julie; Spiers, Andrew; Paterson, Steve; Brockhurst, Michael A.

    2015-01-01

    ABSTRACT Bacteriophages are a major cause of bacterial mortality and impose strong selection on natural bacterial populations, yet their effects on the dynamics of conjugative plasmids have rarely been tested. We combined experimental evolution, mathematical modeling, and individual-based simulations to explain how the ecological and population genetics effects of bacteriophages upon bacteria interact to determine the dynamics of conjugative plasmids and their persistence. The ecological effects of bacteriophages on bacteria are predicted to limit the existence conditions for conjugative plasmids, preventing persistence under weak selection for plasmid accessory traits. Experiments showed that phages drove faster extinction of plasmids in environments where the plasmid conferred no benefit, but they also revealed more complex effects of phages on plasmid dynamics under these conditions, specifically, the temporary maintenance of plasmids at fixation followed by rapid loss. We hypothesized that the population genetic effects of bacteriophages, specifically, selection for phage resistance mutations, may have caused this. Further mathematical modeling and individual-based simulations supported our hypothesis, showing that conjugative plasmids may hitchhike with phage resistance mutations in the bacterial chromosome. PMID:26037122

  18. Plasmid genes required for microcin B17 production.

    Science.gov (United States)

    San Millán, J L; Kolter, R; Moreno, F

    1985-09-01

    The production of the antibiotic substance microcin B17 (Mcc) is determined by a 3.5-kilobase DNA fragment from plasmid pMccB17. Several Mcc- mutations on plasmid pMccB17 were obtained by both transposon insertion and nitrosoguanidine mutagenesis. Plasmids carrying these mutations were tested for their ability to complement Mcc- insertion or deletion mutations on pMM102 (pMM102 is a pBR322 derivative carrying the region encoding microcin B17). Results from these experiments indicate that at least four plasmid genes are required for microcin production.

  19. 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...

  20. [Isolation of the R'his plasmids of Vibrio cholerae].

    Science.gov (United States)

    Rusina, O Iu; Tiganova, I G; Aleshkin, G I; Andreeva, I V; Skavronskaia, A G

    1987-06-01

    V. cholerae strain VT5104 capable of donor activity in conjugation has been constructed by the genetic technique based on plasmid RP4::Mucts62 integration into V. cholerae chromosome due to plasmid homology with Mucts62 inserted into the chromosome. The gene for histidine synthesis has been mobilized and transferred into the recipient cells from VT5104 donor. The conjugants obtained are able to efficiently transfer his+ gene included into the plasmid structure in conjugation with eltor recipient. Thus, the constructed strain VT5104 generates R' plasmids carrying V. cholerae chromosomal genes.

  1. Assessment of Curing Efficiency and Effect of Moist Curing on Performance of Fly Ash Concrete

    Institute of Scientific and Technical Information of China (English)

    LI Meili; QIAN Jueshi; WANG Lixia; XU Shanshan; JIA Xingwen; FAN Yunyan

    2011-01-01

    This study was conducted to evaluate the sensitivity of compressive strength,water permeability and electrical resistance of near-surface layer concrete with different fly ash contents to curing conditions. It is shown that the sensitivity to curing condition and fly ash content descends in the following order: difference between internal and surface resistivity (ρ)at 28 days, water permeability and compressive strength; both of longer duration of moist curing and use of fly ash in concrete enhanced the water penetration resistance. It is indicated that the resistivity difference p at 28 days can reflect accurately the curing history of fly ash concrete regardless of mix proportions; and use of fly ash in concrete requires longer moist curing duration.

  2. Degradative Plasmid and Heavy Metal Resistance Plasmid Naturally Coexist in Phenol and Cyanide Assimilating Bacteria

    Directory of Open Access Journals (Sweden)

    Bahig E.  Deeb

    2009-01-01

    Full Text Available Problem statement: Heavy metals are known to be powerful inhibitors of xenobiotics biodegradation activities. Alleviation the inhibitory effect of these metals on the phenol biodegradation activities in presence of heavy metals resistant plasmid was investigated. Approach: Combination of genetic systems of degradation of xenobiotic compound and heavy metal resistance was one of the approaches to the creation of polyfunctional strains for bioremediation of soil after co-contamination with organic pollutants and heavy metals. Results: A bacterial strain Pseudomonas putida PhCN (pPhCN1, pPhCN2 had been obtained. This bacterium contained two plasmids, a 120 Kb catabolic plasmid that encode for breakdown of phenol (pPhCN1 and pPhCN2 plasmid (100 Kb that code for cadmium and copper resistant. Cyanide assimilation by this bacterium was encoded by chromosomal genes. The inhibitory effect of cadmium (Cd2+ or copper (Cu2+ on the degradation of phenol and cyanide by P. putida strains PhCN and PhCN1 (contained pPhCN1 were investigated. The resistant strain PhCN showed high ability to degrade phenol and cyanide in presence of Cd2+ or Cu2+ comparing with the sensitive strain PhCN1. In addition, Cd2+ or Cu2+ was also found to exert a strong inhibitory effect on the C23O dioxygenase enzyme activity in the presence of cyanide as a nitrogen source. Conclusion: The presence of heavy metal resistance plasmid alleviated the inhibitory effect of metals on the phenol and cyanide assimilation by resistant strain.

  3. [A novel Salmonella Typhimurium plasmid, pAnkS: an example for plasmid evolution in antibiotic resistance].

    Science.gov (United States)

    Sahin, Fikret; Karasartova, Djursun; Gerçeker, Devran; Aysev, A Derya; Erdem, Birsel

    2008-07-01

    In this study, a plasmid, carrying ampicillin resistance (ampR) gene, isolated from a clinical isolate of Salmonella enterica serotype Typhimurium presenting ACSSuT (ampicilin, chloramphenicol, streptomycin, sulphonamide, tetracycline) resistance phenotype, was defined. The length of complete sequence of this plasmid was 8271 base pairs (bp), and it was named as pAnkS owing to its isolation place (plasmid-Ankara- Salmonella). The plasmid was analyzed for potential reading frames and structural features indicative of transposons and transposon relics. The Xmnl enzyme restriction fragments of pAnkS were cloned into E. coli plasmid vectors (pBSK), sequenced and analyzed with the BLAST programs. Plasmid pAnkS has contained a previously defined enterohemorrhagic E. coli (EHEC) plasmid p4821 as a core region and also contained a complete Tn3-like transposon of 4950 bp consisting of the left terminal repeat, Tn3-related tnpR and tnpA genes for transposition functions, ampicillin resistance gene bla(TEM), and the right terminal repeats, pAnkS showed strong homology with another Salmonella plasmid, pNTP16, for sequences that belong to p4821 and partial Tn3 segments. It was found that pNTP16 also carries kanamycin resistance gene (kanR) in addition to ampR gene. Plasmid pAnkS is one of the few completely sequenced plasmids from Salmonella Typhimurium and is in the middle of the pathway of evolution of plasmid from p4821 to pNTP16. The identification of pAnkS might help better understanding of plasmid evolution.

  4. Plasmid mediated antibiotic resistance ofVibrio cholerae O1 biotype El Tor serotype Ogawa associated with an outbreak in Kolkata, India

    Institute of Scientific and Technical Information of China (English)

    Shyamapada Mandal; Manisha DebMandal; Nishith Kumar Pal

    2010-01-01

    Objective:To determine the antibiotic resistance ofVibrio cholerae (V. cholerae)O1 biotype El Tor serotype Ogawa isolates involved in an outbreak of watery diarrhea in Kolkata, and to explore the role of plasmid in mediating antibiotic resistance.Methods: Antibiotic susceptibility and minimum inhibitory concentration(MIC) values of antibiotics for the isolated V. choleraeO1 Ogawa (n=12) were determined by disk diffusion and agar dilution methods, respectively, using ampicillin (Am), chloramphenicol (C), trimethoprim (Tm), tetracycline (T), erythromycine (Er), nalidixic acid (Nx), ciprofloxacin (Cp), amikacin (Ak) and cefotaxime (Cf). Plasmid curing of multidrug resistant(MDR)V. choleraeO1 Ogawa strains was done following ethidium bromide treatment. Following electrophoresis, the plasmidDNAs, extracted from the isolatedMDRV. choleraeO1 Ogawa strains and their cured derivatives, were visualized and documented in‘gel doc’ system.Results: The outbreak causingV. choleraeO1 Ogawa isolates wereMDR as determined by disk diffusion susceptibility test, andMIC determination. The isolates showed three different drug resistance patterns: AmTmTErNx (for6 isolates), TmTErCp (for 5 isolates), and AmTmNx (for one isolate), and showed uniform sensitivity to C, Ak and Cf. The loss of plasmids with the concomitant loss of resistance to Am, Tm, T and Er of the isolates occurred following ethidium bromide treatment.Conclusions: The current findings suggest that theV. choleraeO1Ogawa associated with the cholera outbreak wereMDR, and resistance to Am, Tm, T and Er among the isolates were plasmid mediated.

  5. Effect of plasmid R391 and other IncJ plasmids on the survival of Escherichia coli after UV irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Pembroke, J.T.; Stevens, E. (University Coll., Galway (Ireland))

    1984-07-01

    The presence of the IncJ plasmids R391, R997, R705, R706, R748, and R749 was shown to sensitize Escherichia coli AB1157 and both its uvr A and lexA derivatives to UV irradiation. No alteration in post-irradiation survival was observed in a recA mutant containing these plasmids, compared with the non-plasmid-containing recA strain. Analysis of recombination frequency in Hfr crosses to recA/sup +/ cells containing plasmid R391 indicated a reduction in recombination frequency compared with that obtained in similar crosses to a non-plasmid-containing strain. This effect was not due to plasmid-encoded restriction or entry exclusion systems and therefore must be considered as a real block in recombination. When cells containing plasmid R391 were irradiated and allowed to photoreactivate, an increase in survival was observed which was comparable to that observed in the non-plasmid-containing derivative. This indicated that post-irradiation processing of UV-induced damage, or lack of such processing, by mechanisms other than photoreactivation was responsible for the UV sensitivity associated with plasmid R391.

  6. Curing mechanism of flexible aqueous polymeric coatings.

    Science.gov (United States)

    Irfan, Muhammad; Ahmed, Abid Riaz; Dashevskiy, Andriy; Kolter, Karl; Bodmeier, Roland

    2017-02-25

    The objective of this study was to explain curing phenomena for pellets coated with a flexible polymeric coating based on poly(vinyl acetate) (Kollicoat(®) SR 30D) with regard to the effect of starter cores, thickness of drug layer, adhesion of coating to drug-layered-cores as well as coating properties. In addition, appropriate approaches to eliminate the curing effect were identified. Sugar or MCC cores were layered with the model drugs carbamazepine, theophylline, propranolol HCl, tramadol HCl and metoprolol HCl using HPMC (5 or 25% w/w, based on drug) as a binder. Drug-layered pellets were coated with Kollicoat(®) SR 30D in a fluidized bed coater using TEC (10% w/w) as plasticizer and talc (35-100% w/w) as anti-tacking agent. Drug release, pellet properties (morphology, water uptake-weight loss and osmolality) and adhesion of the coating to the drug layer were investigated as a function of curing at 60 °C or 60 °C/75% RH for 24 h. The film formation of the aqueous dispersion of Kollicoat(®) SR 30D was complete, and therefore, a strong curing effect (decrease in drug release) at elevated temperature and humidity (60 °C/75% RH) could not be explained by the well-known hydroplasticization and the further gradual coalescence of the colloidal polymer particles. According to the provided mechanistic explanation, the observed curing effect was associated with 1) high flexibility of coating, 2) adhesion between coating and drug layer, 3) water retaining properties of the drug layer, and 4) osmotically active cores. Unwanted curing effects could be minimized/eliminated by the addition of talc or/and pore-forming water soluble polymers in the coating, increasing binder amount or applying an intermediate coating, by increasing the thickness of drug layer or using non-osmotic cores. A new insight into curing phenomena mainly associated with the adhesion between drug layer and coating was provided. Appropriate approaches to avoid unwanted curing effect were identified.

  7. Prevent and cure disuse bone loss

    Science.gov (United States)

    Jee, Webster S. S.

    1994-01-01

    Anabolic agents like parathyroid hormone and postagladin E-like substances were studied in dogs and rats to determine their effectiveness in the prevention and cure of bone loss due to immobilization. It was determined that postagladin E2 administration prevented immobilization while at the same time it added extra bone in a dose responsive manner. Although bone mass returns, poor trabecular architecture remains after normal ambulation recovery from immobilization. Disuse related bone loss and poor trabecular architecture were cured by post-immobilization postagladin E2 treatment.

  8. Curing kinetics of visible light curing dental resin composites investigated by dielectric analysis (DEA).

    Science.gov (United States)

    Steinhaus, Johannes; Hausnerova, Berenika; Haenel, Thomas; Großgarten, Mandy; Möginger, Bernhard

    2014-03-01

    During the curing process of light curing dental composites the mobility of molecules and molecule segments is reduced leading to a significant increase of the viscosity as well as the ion viscosity. Thus, the kinetics of the curing behavior of 6 different composites was derived from dielectric analysis (DEA) using especially redesigned flat sensors with interdigit comb electrodes allowing for irradiation at the top side and measuring the ion viscosity at the bottom side. As the ion viscosities of dental composites change 1-3 orders of magnitude during the curing process, DEA provides a sensitive approach to evaluate their curing behavior, especially in the phase of undisturbed chain growth. In order to determine quantitative kinetic parameters a kinetic model is presented and examined for the evaluation of the ion viscosity curves. From the obtained results it is seen that DEA might be employed in the investigation of the primary curing process, the quality assurance of ingredients as well as the control of processing stability of the light curing dental composites. Copyright © 2014 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

  9. Strength of Geopolymer Cement Curing at Ambient Temperature by Non-Oven Curing Approaches: An Overview

    Science.gov (United States)

    Wattanachai, Pitiwat; Suwan, Teewara

    2017-06-01

    At the present day, a concept of environmentally friendly construction materials has been intensively studying to reduce the amount of releasing greenhouse gases. Geopolymer is one of the cementitious binders which can be produced by utilising pozzolanic wastes (e.g. fly ash or furnace slag) and also receiving much more attention as a low-CO2 emission material. However, to achieve excellent mechanical properties, heat curing process is needed to apply to geopolymer cement in a range of temperature around 40 to 90°C. To consume less oven-curing energy and be more convenience in practical work, the study on geopolymer curing at ambient temperature (around 20 to 25°C) is therefore widely investigated. In this paper, a core review of factors and approaches for non-oven curing geopolymer has been summarised. The performance, in term of strength, of each non-oven curing method, is also presented and analysed. The main aim of this review paper is to gather the latest study of ambient temperature curing geopolymer and to enlarge a feasibility of non-oven curing geopolymer development. Also, to extend the directions of research work, some approaches or techniques can be combined or applied to the specific properties for in-field applications and embankment stabilization by using soil-cement column.

  10. Light-Cured Self-Etch Adhesives Undergo Hydroxyapatite-Triggered Self-Cure.

    Science.gov (United States)

    Liu, Y; Bai, X; Liu, Y W; Wang, Y

    2016-03-01

    Light cure is a popular mode of curing for dental adhesives. However, it suffers from inadequate light delivery when the restoration site is less accessible, in which case a self-cure mechanism is desirable to salvage any compromised polymerization. We previously reported a novel self-cure system mediated by ethyl 4-(dimethylamino)-benzoate (4E) and hydroxyapatite (HAp). The present work aims to investigate if such self-cure phenomenon takes place in adhesives that underwent prior inadequate light cure and to elucidate if HAp released from the dental etching process is sufficient to trigger it. Model self-etch adhesives were formulated with various components, including bis[2-methacryloyloxy)ethyl]-phosphate (2MP) as acidic monomer and trimethylbenzoyl-diphenylphosphine oxide (TPO) as photoinitiator. In vitro evolution of degree of conversion (DC) of HAp-incorporated adhesives was monitored by infrared spectroscopy during light irradiation and dark storage. Selected adhesives were allowed to etch and extract HAp from enamel, light-cured in situ, and stored in the dark, after which Raman line mapping was used to obtain spatially resolved DC across the enamel-resin interface. Results showed that TPO+4E adhesives reached DC similar to TPO-only counterparts upon completion of light irradiation but underwent another round of initiation that boosted DC to ~100% regardless of HAp level or prior light exposure. When applied to enamel, TPO-only adhesives had ~80% DC in resin, which gradually descended to ~50% in enamel, whereas TPO+4E adhesives consistently scored ~80% DC across the enamel-resin interface. These observations suggest that polymerization of adhesives that underwent insufficient light cure is salvaged by the novel self-cure mechanism, and such salvaging effect can be triggered by HAp released from dental substrate during the etching process.

  11. Chromosomal targeting of replicating plasmids in the yeast Hansenula polymorpha

    NARCIS (Netherlands)

    Faber, Klaas Nico; Swaving, Gert Jan; Faber, Folkert; Ab, Geert; Harder, Willem; Veenhuis, Marten; Haima, Pieter

    1992-01-01

    Using an optimized transformation protocol we have studied the possible interactions between transforming plasmid DNA and the Hansenula polymorpha genome. Plasmids consisting only of a pBR322 replicon, an antibiotic resistance marker for Escherichia coli and the Saccharomyces cerevisiae LEU2 gene we

  12. Genomic comparison of archaeal conjugative plasmids from Sulfolobus

    DEFF Research Database (Denmark)

    Greve, Bo Bjørn

    2004-01-01

    All of the known self-transmissable plasmids of the Archaea have been found in the genus Sulfolobus. To gain more insight into archaeal conjugative processes, four newly isolated self-transmissable plasmids, pKEF9, pHVE14, pARN3 and pARN4, were sequenced and subjected to a comparative sequence...

  13. Linear plasmid in the genome of Clavibacter michiganensis subsp. sepedonicus.

    Science.gov (United States)

    Brown, Susan E; Knudson, Dennis L; Ishimaru, Carol A

    2002-05-01

    Contour-clamped homogeneous electric field gel analysis of genomic DNA of the plant pathogen Clavibacter michiganensis subsp. sepedonicus revealed the presence of a previously unreported extrachromosomal element. This new element was demonstrated to be a linear plasmid. Of 11 strains evaluated, all contained either a 90-kb (pCSL1) or a 140-kb (pCSL2) linear plasmid.

  14. Homology of plasmids in strains of unicellular cyanobacteria

    NARCIS (Netherlands)

    Hondel, C.A.M.J.J. van den; Keegstra, W.; Borrias, W.E.; Arkel, G.A. van

    1979-01-01

    Six strains of unicellular cyanobacteria were examined for the presence of plasmids. Analysis of lysates of these strains by CsCl-ethidium bromide density centrifugation yielded a major chromosomal DNA band and a minor band containing covalently closed circular plasmid DNA, as shown by electron micr

  15. Examination of uropathogenic Escherichia coli strains conferring large plasmids

    Directory of Open Access Journals (Sweden)

    SUHARTONO

    2010-04-01

    Full Text Available Suhartono (2010 Examination of uropathogenic Escherichia coli strains conferring large plasmids. Biodiversitas 11: 59-64. Of major uropathogens, Escherichia coli has been widely known as a main pathogen of UTIs globally and has considerable medical and financial consequences. A strain of UPEC, namely E. coli ST131, confers a large plasmid encoding cephalosporinases (class C β-lactamase or AmpC that may be disseminated through horizontal transfer among bacterial populations. Therefore, it is worth examining such large plasmids by isolating, purifying, and digesting the plasmid with restriction enzymes. The examination of the large plasmids was conducted by isolating plasmid DNA visualized by agarose gel electrophoresis as well as by PFGE. The relationship of plasmids among isolates was carried out by HpaI restriction enzyme digestion. Of 36 isolates of E. coli ST 131, eight isolates possessed large plasmids, namely isolates 3, 9, 10, 12, 17, 18, 26 and 30 with the largest molecular size confirmed by agarose gel electrophoresis and PFGE was ~42kb and ~118kb respectively. Restriction enzyme analysis revealed that isolates 9, 10, 12, 17 and 18 have the common restriction patterns and those isolates might be closely related.

  16. 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 papil

  17. 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 papil

  18. Plasmid cloning vehicle for Haemophilus influenzae and Escherichia coli

    Energy Technology Data Exchange (ETDEWEB)

    McCarthy, D.; Clayton, N.L.; Setlow, J.K.

    1982-09-01

    A new plasmid cloning vehicle (pDM2) was used to introduce a library of Haemophilus influenzae chromosomal fragments into H. influenzae. Transformants of the higly recombination-defective rec-1 mutant were more likely to contain exclusively recombinant plasmids after exposure to ligated DNA mixtures than was the wild type. pDM2 could replicate in Escherichia coli K-12.

  19. 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. Th

  20. 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 blaCMY and blaNDM Current typing methods for IncA/C plasmids offer limited resolution. In this study, we present the complete sequence of a blaNDM-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 blaNDM-positive plasmids examined (15/17; 88%) fall into ST1, suggesting acquisition and subsequent expansion of this blaNDM-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.

  1. 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...... diversity of recipient bacterial phyla for the plasmid was observed, especially in WWTP outlets. We also identified permissive bacteria potentially able to cross WWTPs and engage in conjugation before and after water treatment. Bacterial activity and lifestyle seem to influence conjugation extent...

  2. 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...

  3. [Influence of spv plasmid genes group in Salmonella Enteritidis virulence for chickens. I. Occurrence of spv plasmid genes group in Salmonella Enteritidis large virulence plasmid].

    Science.gov (United States)

    Madajczak, Grzegorz; Binek, Marian

    2005-01-01

    Many Salmonella Enteritidis virulence factors are encoded by genes localized on plasmids, especially large virulence plasmid, in highly conserved fragment, they create spv plasmid gene group. The aims of realized researches were spv genes occurrence evaluation and composition analysis among Salmonella Enteritidis strains caused infection in chickens. Researches were realized on 107 isolates, where in every cases large virulence plasmid 59 kbp size were detected. Specific nucleotides sequences of spv genes (spvRABCD) were detected in 47.7% of isolates. In the rest of examined bacteria spv genes occurred variably. Most often extreme genes of spv group, like spvR and spvD were absent, what could indicate that factors encoded by them are not most important for Salmonella Enteritidis live and their expressed virulence.

  4. Mechanical characterization and modeling of curing thermosets

    NARCIS (Netherlands)

    Van 't Hof, C.

    2006-01-01

    Chemical shrinkage and simultaneous build-up of mechanical properties in curing thermosets leads to the build-up of residual stresses and strains. Depending on the constraints these may cause interface failure, dimensional inaccuracy or failure in the thermoset or its surrounding structure. The pre

  5. Creep measurements on curing epoxy systems

    DEFF Research Database (Denmark)

    Kammer, Charlotte; Szabo, Peter

    1998-01-01

    The chemical curing of a stoichiometric mixture of the diglycidyl ether of bisphenol A and a 1,3-bis-(aminomethyl)-cyclohexane is studied.Creep experiments are combined with measurements in a Differential Scanning Calorimeter (DSC) to determine the change in bulk viscosity due to network formation....

  6. Internal water curing with Liapor aggregates

    DEFF Research Database (Denmark)

    Lura, Pietro

    2005-01-01

    Internal water curing is a very efficient way to counteract self-desiccation and autogenous shrinkage in high performance concrete, thereby reducing the likelihood of early-age cracking. This paper deals with early-age volume changes and moisture transport in lightweight aggregate concrete realiz...

  7. Creep measurements on curing epoxy systems

    DEFF Research Database (Denmark)

    Kammer, Charlotte; Szabo, Peter

    1998-01-01

    The chemical curing of a stoichiometric mixture of the diglycidyl ether of bisphenol A and a 1,3-bis-(aminomethyl)-cyclohexane is studied.Creep experiments are combined with measurements in a Differential Scanning Calorimeter (DSC) to determine the change in bulk viscosity due to network formation....

  8. Sample displacement chromatography of plasmid DNA isoforms.

    Science.gov (United States)

    Černigoj, Urh; Martinuč, Urška; Cardoso, Sara; Sekirnik, Rok; Krajnc, Nika Lendero; Štrancar, Aleš

    2015-10-02

    Sample displacement chromatography (SDC) is a chromatographic technique that utilises different relative binding affinities of components in a sample mixture and has been widely studied in the context of peptide and protein purification. Here, we report a use of SDC to separate plasmid DNA (pDNA) isoforms under overloading conditions, where supercoiled (sc) isoform acts as a displacer of open circular (oc) or linear isoform. Since displacement is more efficient when mass transfer between stationary and mobile chromatographic phases is not limited by diffusion, we investigated convective interaction media (CIM) monoliths as stationary phases for pDNA isoform separation. CIM monoliths with different hydrophobicities and thus different binding affinities for pDNA (CIM C4 HLD, CIM-histamine and CIM-pyridine) were tested under hydrophobic interaction chromatography (HIC) conditions. SD efficiency for pDNA isoform separation was shown to be dependent on column selectivity for individual isoform, column efficiency and on ammonium sulfate (AS) concentration in loading buffer (binding strength). SD and negative mode elution often operate in parallel, therefore negative mode elution additionally influences the efficiency of the overall purification process. Optimisation of chromatographic conditions achieved 98% sc pDNA homogeneity and a dynamic binding capacity of over 1mg/mL at a relatively low concentration of AS. SDC was successfully implemented for the enrichment of sc pDNA for plasmid vectors of different sizes, and for separation of linear and and sc isoforms, independently of oc:sc isoform ratio, and flow-rate used. This study therefore identifies SDC as a promising new approach to large-scale pDNA purification, which is compatible with continuous, multicolumn chromatography systems, and could therefore be used to increase productivity of pDNA production in the future. Copyright © 2015 Elsevier B.V. All rights reserved.

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

    DEFF Research Database (Denmark)

    Klümper, Uli

    larger than previously assumed. I was able to show abundant plasmid transfer from the Gram negative donor strains to a wide diversity of Gram positive soil bacteria, formerly thought to constitute distinct clusters of gene transfer. Moreover, among the observed transconjugants, I identified a core super...... environmental factors that modulate plasmid transfer in soil microbial communities. In order to attain these goals, I developed a high-throughput method that enabled me to evaluate the permissiveness of bacterial communities towards introduced plasmids. This new approach is based on the introduction...... fraction of soil the bacteria (up to 1 in 10,000) were able to take up any of these broad host range conjugal plasmids. The transconjugal pools comprised 11 bacterial phyla. This finding indicates that the realized transfer range of broad host range plasmids in environmental microbial communities is much...

  10. Complex nature of enterococcal pheromone-responsive plasmids.

    Science.gov (United States)

    Wardal, Ewa; Sadowy, Ewa; Hryniewicz, Waleria

    2010-01-01

    Pheromone-responsive plasmids constitute a unique group of approximately 20 plasmids identified, as yet, only among enterococcal species. Several of their representatives, e.g. pAD1, pCF10, pPD1 and pAM373 have been extensively studied. These plasmids possess a sophisticated conjugation mechanism based on response to sex pheromones--small peptides produced by plasmid-free recipient cells. Detailed analysis of regulation and function of the pheromone response process revealed its great complexity and dual role--in plasmid conjugation and modulation of enterococcal virulence. Among other functional modules identified in pheromone plasmids, the stabilization/partition systems play a crucial role in stable maintenance of the plasmid molecule in host bacteria. Among them, the par locus of pAD1 is one of the exceptional RNA addiction systems. Pheromone-responsive plasmids contribute also to enterococcal phenotype being an important vehicle of antibiotic resistance in this genus. Both types of acquired vancomycin resistance determinants, vanA and vanB, as well many other resistant phenotypes, were found to be located on these plasmids. They also encode two basic agents of enterococcal virulence, i.e. aggregation substance (AS) and cytolysin. AS participates in mating-pair formation during conjugation but can also facilitate the adherence ofenterococci to human tissues during infection. The second protein, cytolysin, displays hemolytic activity and helps to invade eukaryotic cells. There are still many aspects of the nature of pheromone plasmids that remain unclear and more detailed studies are needed to understand their uniqueness and complexity.

  11. Talking Cure Models: A Framework of Analysis

    Science.gov (United States)

    Marx, Christopher; Benecke, Cord; Gumz, Antje

    2017-01-01

    Psychotherapy is commonly described as a “talking cure,” a treatment method that operates through linguistic action and interaction. The operative specifics of therapeutic language use, however, are insufficiently understood, mainly due to a multitude of disparate approaches that advance different notions of what “talking” means and what “cure” implies in the respective context. Accordingly, a clarification of the basic theoretical structure of “talking cure models,” i.e., models that describe therapeutic processes with a focus on language use, is a desideratum of language-oriented psychotherapy research. Against this background the present paper suggests a theoretical framework of analysis which distinguishes four basic components of “talking cure models”: (1) a foundational theory (which suggests how linguistic activity can affect and transform human experience), (2) an experiential problem state (which defines the problem or pathology of the patient), (3) a curative linguistic activity (which defines linguistic activities that are supposed to effectuate a curative transformation of the experiential problem state), and (4) a change mechanism (which defines the processes and effects involved in such transformations). The purpose of the framework is to establish a terminological foundation that allows for systematically reconstructing basic properties and operative mechanisms of “talking cure models.” To demonstrate the applicability and utility of the framework, five distinct “talking cure models” which spell out the details of curative “talking” processes in terms of (1) catharsis, (2) symbolization, (3) narrative, (4) metaphor, and (5) neurocognitive inhibition are introduced and discussed in terms of the framework components. In summary, we hope that our framework will prove useful for the objective of clarifying the theoretical underpinnings of language-oriented psychotherapy research and help to establish a more comprehensive

  12. Antibiotic resistance of vibrio cholerae: special considerations of R-plasmids.

    Science.gov (United States)

    Kuwahara, S

    1978-09-01

    Studies on the transmission of R plasmid by conjugation between enterobacteria and vibrio or related bacteria were reviewed. The majority of the reports confirmed successful transmission from enterobacteria to Vibrio cholerae and related species, although the transmission frequencies were extremely low and the transmitted R plasmid was very unstable except for thermosensitive kanamycin plasmid and usual R plasmid coexisting with P plasmid. Strains of V. cholerae and Aeromonas liquefaciens as well as A. salmonicida bearing R plasmid were detected in nature. R plasmid was relatively unstable in V. cholerae strains with which transmission of R plasmid to enterobacteria was confirmed. At present, only 3 R plasmids have been obtained from naturally occurring strains of V. cholerae. Although the 2 European plasmids belong to the C incompatibility group with 98 megadalton closed covalent circular DNA molecule, one plasmid belongs to the J group with more than 25 megadalton molecular weight, and no CCC of satelite DNA was detected in bacteria harboring this plasmid.

  13. Bacterial Mitosis: ParM of Plasmid R1 Moves Plasmid DNA by an Actin-like Insertional Polymerization Mechanism

    DEFF Research Database (Denmark)

    Møller-Jensen, Jakob; Borch, Jonas; Dam, Mette

    2003-01-01

    Bacterial DNA segregation takes place in an active and ordered fashion. In the case of Escherichia coli plasmid R1, the partitioning system (par) separates paired plasmid copies and moves them to opposite cell poles. Here we address the mechanism by which the three components of the R1 par system...

  14. COMPARISON OF THE E44 EPOXY RESINS IN DIFFERENT CONTENTS OF CURING AGENT BY MICROWAVE AND THERMAL CURING METHODS

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    This paper discusses the fundamental principle of microwave heating, and based on the advantages of microwave heating, use maleic anhydride as curing agent.The technology of microwave curing E44 epoxy resins is investigated,the mechanical properties of cured epoxy resin samples in different contents of curing agent by microwave and thermal curing methods are measured respectively, and then some experimental results for which are obtained.At last, this paper analyses why microwave curing can improve mechanical property of epoxy resin.

  15. Plasmid DNA studies in Lactobacillus plantarum strains isolated from olive fermentations: production of and immunity to plantaricin OL15 is associated to a 9.6 Kb plasmid (pOL15

    Directory of Open Access Journals (Sweden)

    Mourad, Kacem

    2007-06-01

    Full Text Available Previously 12 Lactobacillus plantarum strains were isolated from fermented olives. Among these, only L. plantarum OL15 produced bacteriocin (plantaricin OL15. In this study, the 12 strains were examined for plasmid DNA content. Of these, 9 strains have shown one to three plasmid bands ranging in size from 5.4 to 12.2 kb. L. plantarum OL15 exhibited one plasmid (9.6 kb which was named pOL15. After curing with novobiocin and ethidium bromide, the plasmid profile analysis of non producing derivatives, showed that the 9.6 kb plasmid pOL15 harbored by the parental strain had been lost in all cases and none of them regained the ability to produce plantaricin OL15 suggesting that the production of plantaricin OL15 is plasmid linked. Plantaricin OL15 was not inactived by amylase and lipase suggesting that plantaricin OL15 activity was not dependent on the presence of either a carbohydrate or lipid moiety. Plantaricin OL15 showed activity against lactic acid bacteria of different species and also against olive spoilage and phytopathogenic bacteria, including Pseudomonas and Erwinia.En un estudio previo, se aislaron 12 cepas de Lactobacillus plantarum a partir de aceitunas fermentadas. Entre ellas, solo L. plantarum OL15 produjo bacteriocinas (plantaricin OL15. En este estudio, se examinó el contenido de AND plásmido en las 12 cepas citadas. Entre ellas, 9 cepas han mostrado de una a tres bandas de plásmido con tamaños en el rango de 5.4 a 12.2 kb. L. plantarum OL15 exhibió un plásmido (9.6 kb que se denominó pOL15. Después del curado con novobiocina y bromuro de etidio, la pérdida del plásmido pOL15 asociada a la pérdida de su facultad para producir plantaricin OL15, sugiere que la producción de plantaricina OL15 está ligada al plásmido. La plantaricin OL15 no se inactivó por amilasa ni por lipasa sugiriendo que su actividad no es dependiente de la presencia de carbohidratos o lípidos. La plantaricina OL15 mostró actividad frente a

  16. Bond strength of a light-cured and two auto-cured glass ionomer liners.

    Science.gov (United States)

    Holtan, J R; Nystrom, G P; Olin, P S; Rudney, J; Douglas, W H

    1990-10-01

    Ninety-nine extracted human molar teeth were used in this study comparing the shear bond strengths on dentine of one light-cured and two auto-cured polyalkenoate (glass ionomer) cements. Bond strength can be influenced by differences in tooth structure. A balanced-incomplete block design (Hull and Nie, 1981) was used to reduce variation attributable to such differences. Cements were applied to paired dentine surfaces in combinations such that 66 tooth sides were treated with each material. A light-cured dentinal adhesive and composite resin restorative material were then placed and shear bond strength testing was conducted exactly 24 h after the completion of each specimen. Mean forces (MPa) for the three materials were compared using an appropriate analysis of variance model (balanced-incomplete-blocks) The shear bond strengths (MPa) of the light-cured liner (Espe, Seefeld/Oberbay, FRG) was 4.71 +/- 1.16. Vitrabond showed the greatest variance of all three materials tested, however this material's average bond strength was greater than the maximum achieved for the other materials. Student-Newman-Keuls comparison of means showed that all cements differed significantly from each other (alpha = 0.05). It is concluded that the light-cured glass ionomer liner exhibited significantly better shear bond strength performance than the two auto-cured glass ionomers tested.

  17. Thermal and Cure Kinetics of Epoxy Molding Compounds Cured with Thermal Latency Accelerators

    Directory of Open Access Journals (Sweden)

    Chean-Cheng Su

    2013-01-01

    Full Text Available The cure kinetics and mechanisms of a biphenyl type epoxy molding compounds (EMCs with thermal latency organophosphine accelerators were studied using differential scanning calorimetry (DSC. Although the use of triphenylphosphine-1,4-benzoquinone (TPP-BQ and triphenylphosphine (TPP catalysts in biphenyl type EMCs exhibited autocatalytic mechanisms, thermal latency was higher in the TPP-BQ catalyst in EMCs than in the TPP catalyst in EMCs. Analyses of thermal characteristics indicated that TPP-BQ is inactive at low temperatures. At high temperatures, however, TPP-BQ increases the curing rate of EMC in dynamic and isothermal curing experiments. The reaction of EMCs with the TPP-BQ latent catalyst also had a higher temperature sensitivity compared to the reaction of EMCs with TPP catalyst. In resin transfer molding, EMCs containing the TPP-BQ thermal latency accelerator are least active at a low temperature. Consequently, EMCs have a low melt viscosity before gelation, and the resins and filler are evenly mixed in the kneading process. Additionally, flowability is increased before the EMCs form a network structure in the molding process. The proposed kinetic model adequately describes curing behavior in EMCs cured with two different organophosphine catalysts up to the rubber state in the progress of curing.

  18. Carriage of extended-spectrum beta-lactamase-plasmids does not reduce fitness but enhances virulence in some strains of pandemic E. coli lineages

    Directory of Open Access Journals (Sweden)

    Katharina eSchaufler

    2016-03-01

    Full Text Available Pathogenic ESBL-producing E. coli lineages occur frequently worldwide, not only in a human health context but in animals and the environment, also in settings with low antimicrobial pressures. This study investigated the fitness costs of ESBL-plasmids and their influence on chromosomally encoded features associated with virulence, such as those involved in the planktonic and sessile behaviors of ST131 and ST648 E. coli. ESBL-plasmid-carrying wild-type E. coli strains, their corresponding ESBL-plasmid-cured variants (PCV, and complementary ESBL-carrying transformants were comparatively analyzed using growth curves, Omnilog® phenotype microarray (PM assays, macrocolony and biofilm formation, swimming motility, and RNA sequence analysis. Growth curves and PM results pointed towards similar growth and metabolic behaviors among the strains. Phenotypic differences in some strains were detected, including enhanced curli fimbriae and/or cellulose production as well as a reduced swimming capacity of some ESBL-carrying strains, as compared to their respective PCVs. RNA sequencing mostly confirmed the phenotypic results, suggesting that the chromosomally encoded csgD pathway is a key factor involved. These results contradict the hypothesis that ESBL-plasmid-carriage leads to a fitness loss in ESBL-carrying strains. Instead, the results indicate an influence of some ESBL-plasmids on chromosomally encoded features associated with virulence in some E. coli strains. In conclusion, apart from antibiotic resistance selective advantages, ESBL-plasmid-carriage may also lead to enhanced virulence or adaption to specific habitats in some strains of pandemic ESBL-producing E. coli lineages.

  19. Effect of curing on clinical retention. A 7-year evaluation

    DEFF Research Database (Denmark)

    van Dijken, Jan WV; Pallesen, Ulla

    Objective: To evaluate in a prospective evaluation the retention of Class V compomer and hybrid resin composite restorations placed with a 1-step self etch system and cured with continuous, soft-start and pulse-delay curing. Methods: 139 Class V restorations were placed and cured at random in 60...... subjects with a compomer (Dyract AP;68) or resin composite (Tetric Ceram:67) and cured for 40s with continuous, soft-start and pulse-delay modes. The restorations were evaluated with slightly modified USPHS criteria yearly during 7 years. Results: Of 135 restorations evaluated at 7 years, 29 were lost, 17...... compomer (25%) and 12 Tetric Ceram (18%). The cumulative loss rates for the compomer and resin composite restorations cured with the three curing modes were. continuous cure: 26.1%/18.2%, soft-start: 22.7%/19.0%, pulse delay: 26.1%/16.7%. No statistical difference were seen between the curing modes...

  20. INTRODUCTION Curing is holding fruits at temperatures and ...

    African Journals Online (AJOL)

    One of the. EFFECT OF CURING ON THE SHELF LIFE OF AMBERSWEET ORANGES ... Alternaria citri. Incidence was lowest in fruits cured in the shade to 7% weight loss. .... Period of Storage (Weeks)/ Incidence of Decay (%). (% Wt. loss). 1.

  1. Drinking Peroxide as 'Natural' Cure Leads to Dangerous Blood Clots

    Science.gov (United States)

    ... https://medlineplus.gov/news/fullstory_163513.html Drinking Peroxide as 'Natural' Cure Leads to Dangerous Blood Clots ... 9, 2017 (HealthDay News) -- Ingesting high-concentration hydrogen peroxide as a "natural cure" or cleansing agent may ...

  2. [Dimensional accuracy of microwave-cured denture base resin].

    Science.gov (United States)

    Uchida, K; Okamoto, F; Ogata, K; Sato, T

    1989-02-01

    Recently, microwave-cured denture base resin was developed, and the resin solved the problem of internal porosity which had been generated by curing the conventional denture base resins with microwave irradiation. In this study, the dimensional accuracy of microwave-cured denture base resin was compared with that of other denture base resins, such as pour-type resin, heat-cured resin and heat-shock resin. From the experiment, the following results were obtained. 1. Dimensional accuracy of microwave-cured denture base resin was better than that of heat-cured resin and heat-shock resin, and was similar to that of pour-type resin. 2. Dimensional accuracy of microwave-cured denture base resin by slow cooling method and rapid cooling method was almost the same. Those findings suggest that microwave-cured denture base resin is valuable in clinic.

  3. Determinant of flexible Parametric Estimation of Mixture Cure ...

    African Journals Online (AJOL)

    PROF. OLIVER OSUAGWA

    2015-12-01

    Dec 1, 2015 ... Gastric Cancer data from 76 patients received adjuvant CRT and 125 receiving ... Keywords: Cure Fraction Model, Gastric Cancer, flexible Model, AIC, Mean time to cure. 1.0 Introduction ..... finish cancer registry. Int J Cancer ...

  4. Plasmid DNA Manufacturing for Indirect and Direct Clinical Applications.

    Science.gov (United States)

    Schmeer, Marco; Buchholz, Tatjana; Schleef, Martin

    2017-10-01

    Plasmid DNA is currently gaining increasing importance for clinical research applications in gene therapy and genetic vaccination. For direct gene transfer into humans, good manufacturing practice (GMP)-grade plasmid DNA is mandatory. The same holds true if the drug substance contains a genetically modified cell, for example chimeric antigen receptor (CAR) T cells, where these cells as well as the contained plasmids are used. According to the responsible regulatory agencies, they have to be produced under full GMP. On the other hand, for GMP production of, for example, mRNA or viral vectors (lentiviral vectors, adeno-associated virus vectors, etc.), in many cases, High Quality Grade plasmid DNA is accepted as a starting material. The manufacturing process passes through different production steps. To ensure the right conditions are used for the plasmid, a pilot run must be conducted at the beginning. In this step, a followed upscaling with respect to reproducibility and influences on product quality is performed. Subsequently, a cell bank of the transformed productions strain is established and characterized. This cell bank is used for the cultivation process. After cell harvesting and lysis, several chromatography steps are conducted to receive a pure plasmid product. Depending on the respective required quality grade, the plasmid product is subject to several quality controls. The last step consists of formulation and filling of the product.

  5. In vitro replication of cyanobacterial plasmids from Synechocystis PCC 6803.

    Science.gov (United States)

    Yang, X; Daniell, H; McFadden, B

    1994-09-01

    Little knowledge of DNA replication in cyanobacteria is available. In this study, we report the development and characterization of an in vitro system for studies of replication of the endogenous plasmids from the unicellular cyanobacterium Synechocystis 6803. This system (fraction III) was isolated at high salt concentrations and partially purified on a heparin-agarose column. DNA polymerases in Synechocystis 6803 appeared to be associated with membranes and could be released by the addition of ammonium sulfate to 20% saturation. DNA synthesis in fraction III was dependent on the addition of cyanobacterial plasmids isolated from the same strain. The in vitro replication products consist mostly of the supercoiled form of the plasmids. Unlike replication of many Escherichia coli plasmids, replication of cyanobacterial plasmids did not require added ATP, was not inhibited by omission of the ribonucleotides, and was insensitive to the RNA polymerase inhibitor rifampicin and the gyrase inhibitor novobiocin, but was inhibited by ethidium bromide. These data suggest that RNA may not be involved in the initiation of replication of cyanobacterial plasmids from Synechocystis 6803. In addition, intermediates of replication have been detected by two-dimensional gel electrophoresis. Density labeling experiments also indicate that cyanobacterial plasmid synthesis in vitro occurs by a semiconservative replication.

  6. Electron Beam Curing of Polymer Matrix Composites - CRADA Final Report

    Energy Technology Data Exchange (ETDEWEB)

    Janke, C. J. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Howell, Dave [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Norris, Robert E. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)

    1997-05-01

    The major cost driver in manufacturing polymer matrix composite (PMC) parts and structures, and one of the elements having the greatest effect on their quality and performance, is the standard thermal cure process. Thermal curing of PMCs requires long cure times and high energy consumption, creates residual thermal stresses in the part, produces volatile toxic by-products, and requires expensive tooling that is tolerant of the high cure temperatures.

  7. Techniques and materials for internal water curing of concrete

    DEFF Research Database (Denmark)

    Jensen, Ole Mejlhede; Lura, Pietro

    2006-01-01

    This paper gives an overview of different techniques for incorporation of internal curing water in concrete. Internal water curing can be used to mitigate self-desiccation and selfdesiccation shrinkage. Some concretes may need 50 kg/m3 of internal curing water for this purpose. The price of the i...

  8. 7 CFR 29.6002 - Air-cured.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Air-cured. 29.6002 Section 29.6002 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections, Marketing... INSPECTION Standards Definitions § 29.6002 Air-cured. Tobacco cured under natural atmospheric...

  9. 7 CFR 30.12 - Fire-cure.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Fire-cure. 30.12 Section 30.12 Agriculture Regulations... AND STANDARDS Classification of Leaf Tobacco Covering Classes, Types and Groups of Grades § 30.12 Fire-cure. To cure tobacco under artificial atmospheric conditions by the use of open fires, the smoke and...

  10. Curing Reaction Model of Epoxy Asphalt Binder

    Institute of Scientific and Technical Information of China (English)

    QIAN Zhendong; CHEN Leilei; WANG Yaqi; SHEN Jialin

    2012-01-01

    In order to understand the strength developing law of the epoxy asphalt mixture,a curing reaction model of the epoxy asphalt binder was proposed based upon the thermokinetic analysis.Given some assumptions,the model was developed by applying the Kissinger law as well as Arrhenius equation,and the differential scanning calorimetry was performed for estimating the model parameters.To monitor the strength development of the epoxy asphalt mixture,a strength test program was employed and then results were compared to those produced from the proposed model.The comparative evaluation shows that a good consistency exists between the outputs from test program and the proposed model,indicating that the proposed model can be used effectively for simulating the curing reaction process for the epoxy asphalt binder and predicting the strength development for the epoxy asphalt mixture.

  11. Contactless optoelectronic technique for monitoring epoxy cure.

    Science.gov (United States)

    Cusano, A; Buonocore, V; Breglio, G; Calabrò, A; Giordano, M; Cutolo, A; Nicolais, L

    2000-03-01

    We describe a novel noninvasive optical technique to monitor the refractive-index variation in an epoxy-based resin that is due to the polymerization process. This kind of resin is widely used in polymer matrix composites. It is well known that the process of fabricating a thermoset-based composite involves mass and heat transfer coupled with irreversible chemical reactions that induce physical changes. To improve the quality and the reliability of these materials, monitoring the cure and optimization of the manufacturing process are of key importance. We discuss the basic operating principles of an optical system based on angle deflection measurements and present typical cure-monitoring results obtained from optical characterization. The method provides a flexible, high-sensitivity, material-independent, low-cost, noninvasive tool for monitoring real-time refractive-index variation.

  12. Cure of HCV related liver disease.

    Science.gov (United States)

    Shiffman, Mitchell L; Benhamou, Yves

    2015-01-01

    Chronic hepatitis C virus (HCV) causes chronic liver injury and can lead to cirrhosis and hepatocellular carcinoma (HCC). HCV can also interact with the immune system to cause several HCV related disorders including essential mixed cryoglobulinemia, vasculitis, dermatitis, glomerulonephritis and lymphoma. A strong association between HCV and diabetes mellitus also exists. These extrahepatic features may lead to increased fatigue and a reduced quality of life. It is now possible to cure most patients with chronic HCV using oral antiviral therapy. Many of these HCV-related disorders and symptoms can be cured when HCV is eradicated. However, some patients may have irreversible injury to extrahepatic sites, cirrhosis that cannot resolve, an increased risk for HCC, persistent fatigue and a reduced quality of life, despite achieving sustained virological response.

  13. Alternative cancer cures: "unproven" or "disproven"?

    Science.gov (United States)

    Vickers, Andrew

    2004-01-01

    Oncology has always coexisted with therapies offered outside of conventional cancer treatment centers and based on theories not found in biomedicine. These alternative cancer cures have often been described as "unproven," suggesting that appropriate clinical trials have not been conducted and that the therapeutic value of the treatment is unknown. Contrary to much popular and scientific writing, many alternative cancer treatments have been investigated in good quality clinical trials, and they have been shown to be ineffective. In this article, clinical trial data on a number of alternative cancer cures including Livingston-Wheeler, Di Bella Multitherapy, antineoplastons, vitamin C, hydrazine sulfate, Laetrile, and psychotherapy are reviewed. The label "unproven" is inappropriate for such therapies; it is time to assert that many alternative cancer therapies have been "disproven."

  14. Aloe vera as cure for lichen planus.

    Science.gov (United States)

    Patil, Bharati A; Bhaskar, Hebbar Pragati; Pol, Jyoti S; Sodhi, Amandeep; Madhu, Asha V

    2013-01-01

    Oral lichen planus is a difficult condition to treat because of its chronic nature. Various treatment modalities have resulted in partial regression of symptoms but not a complete cure. Aloe vera, a product with minimal adverse effects, can be tried to treat this disorder. A 38-year-old male patient diagnosed with lichen planus of the skin and the oral mucosa was suffering from severe pain and a burning sensation intraorally and pruritus of the skin lesions. Considering the extensive involvement, an herbal alternative was considered. The patient was prescribed aloe vera juice and gel application for two months. At the nine-month follow-up, the patient was symptom-free and totally cured of the intraoral and skin lesions.

  15. Light-Curing Adhesive Repair Tapes

    Science.gov (United States)

    Allred, Ronald; Haight, Andrea Hoyt

    2009-01-01

    Adhesive tapes, the adhesive resins of which can be cured (and thereby rigidized) by exposure to ultraviolet and/or visible light, are being developed as repair patch materials. The tapes, including their resin components, consist entirely of solid, low-outgassing, nonhazardous or minimally hazardous materials. They can be used in air or in vacuum and can be cured rapidly, even at temperatures as low as -20 C. Although these tapes were originally intended for use in repairing structures in outer space, they can also be used on Earth for quickly repairing a wide variety of structures. They can be expected to be especially useful in situations in which it is necessary to rigidize tapes after wrapping them around or pressing them onto the parts to be repaired.

  16. Plasmid-determined resistance to fosfomycin in Serratia marcescens.

    Science.gov (United States)

    Mendoza, C; Garcia, J M; Llaneza, J; Mendez, F J; Hardisson, C; Ortiz, J M

    1980-08-01

    Multiple-antibiotic-resistant strains of Serratia marcescens isolated from hospitalized patients were examined for their ability to transfer antibiotic resistance to Escherichia coli by conjugation. Two different patterns of linked transferable resistance were found among the transconjugants. The first comprised resistance to carbenicillin, streptomycin, and fosfomycin; the second, and more common, pattern included resistance to carbenicillin, streptomycin, kanamycin, gentamicin, tetracycline, chloramphenicol, sulfonamide, and fosfomycin. The two types of transconjugant strains carried a single plasmid of either 57 or 97 megadaltons in size. Both of these plasmids are present in parental S. marcescens strains resistant to fosfomycin. The 57-megadalton plasmid was transformed into E. coli.

  17. [Epidemiologic study of 2 S. typhimurium outbreaks using plasmid fingerprints].

    Science.gov (United States)

    Baumgartner, A; Breer, C; Schopfer, K

    1989-04-05

    An outbreak of salmonellosis in an old people's home is reported. The infectious agent, S. typhi-murium, was isolated not only from several inmates but also from sick cows of the farm belonging to the home, in animal feed, from employees of the local butcher's shop, and finally in sludge from the local sewage plant. Plasmid analysis provided evidence of a common origin for the isolated S. typhi-murium strains. The incriminated strains harboured, together with two low-molecular-weight plasmids, a plasmid of approximately 50 Mdal, which was also demonstrated in some other S. typhi-murium strains isolated from clinical cases in the area around St. Gallen.

  18. Effect of Plasmid Incompatibility on DNA Transfer to Streptococcus cremoris

    OpenAIRE

    Van Der Lelie, Daniel; Vossen, Jos M.B.M. van der; Venema, Gerard

    1988-01-01

    Several Streptococcus cremoris strains were used in protoplast transformation and interspecific protoplast fusion experiments with Streptococcus lactis and Bacillus subtilis, with pGKV110, pGKV21, and ΔpAMβ1 as the marker plasmids. ΔpAMβ1 is a 15.9-kilobase nonconjugative, deletion derivative of pAMβ1, which is considerably larger than the pGKV plasmids (approximately 4.5 kilobases). In general, ΔpAMβ1 was transferred more efficiently than the pGKV plasmids. Using electroporation, we were abl...

  19. Separation of plasmid DNA topoisomers by multimodal chromatography.

    Science.gov (United States)

    Silva-Santos, A Rita; Alves, Cláudia P A; Prazeres, Duarte Miguel F; Azevedo, Ana M

    2016-06-15

    The ability to analyze the distribution of topoisomers in a plasmid DNA sample is important when evaluating the quality of preparations intended for gene therapy and DNA vaccination or when performing biochemical studies on the action of topoisomerases and gyrases. Here, we describe the separation of supercoiled (sc) and open circular (oc) topoisomers by multimodal chromatography. A medium modified with the ligand N-benzyl-N-methyl ethanolamine and an elution scheme with increasing NaCl concentration are used to accomplish the baseline separation of sc and oc plasmid. The utility of the method is demonstrated by quantitating topoisomers in a purified plasmid sample. Copyright © 2016 Elsevier Inc. All rights reserved.

  20. Fast curing ethylene vinyl acetate films with dual curing agent towards application as encapsulation materials for photovoltaic modules

    Directory of Open Access Journals (Sweden)

    W-H. Ruan

    2014-02-01

    Full Text Available To speed up curing of ethylene vinyl acetate (EVA films as encapsulation materials for photovoltaic modules, a dual curing agent of benzoyl peroxide (BPO and butylperoxy 2-ethylhexyl carbonate (TBEC was introduced in this work. The experimental results indicated that for the weight ratio of BPO/TBEC of 0.6/2.4, over 80% gel content of EVA was yielded after curing at 130°C for 12 min. Compared with the case of single curing agent, the present one obviously operated at much lower temperature with faster rate. By carefully studying the influence of curing agent proportion and curing conditions on gel content of EVA films, as well as rheology and curing kinetics, the mechanism involved was analyzed and verified. The results are believed to be useful for developing new curing system of EVA encapsulation films with improved processability.

  1. Coating and curing apparatus and methods

    Energy Technology Data Exchange (ETDEWEB)

    Brophy, Brenor L.; Gonsalves, Peter R.; Maghsoodi, Sina; Colson, Thomas E.; Yang, Yu S.; Abrams, Ze' ev R.

    2016-04-19

    Disclosed is a coating apparatus including flow coating and roll-coating that may be used for uniform sol-gel coating of substrates such as glass, solar panels, windows or part of an electronic display. Also disclosed are methods for substrate preparation, flow coating and roll coating. Lastly, systems and methods for curing sol-gel coatings deposited onto the surface of glass substrates using high temperature air-knives, infrared emitters and direct heat applicators are disclosed.

  2. Photothermal Monitoring Of Curing Of Polymers

    Science.gov (United States)

    Rooney, Michael

    1993-01-01

    Time-resolved infrared radiometry (TRIR) adapted to monitoring curing of some polymers in production. Proposal part of continuing effort to perfect production of hydroxy-terminated polybutadiene for use in liners of solid-fuel rocket motors. Applicable to monitoring changing states of many other materials in process. TRIR, non-contact technique implemented with remotely situated equipment and better suited to use in production.

  3. Using Sex to Cure the Genome.

    Directory of Open Access Journals (Sweden)

    Eduardo P C Rocha

    2016-03-01

    Full Text Available The diversification of prokaryotes is accelerated by their ability to acquire DNA from other genomes. However, the underlying processes also facilitate genome infection by costly mobile genetic elements. The discovery that cells can uptake DNA by natural transformation was instrumental to the birth of molecular biology nearly a century ago. Surprisingly, a new study shows that this mechanism could efficiently cure the genome of mobile elements acquired through previous sexual exchanges.

  4. Sage Gene Expression Profiles Characterizing Cure

    Science.gov (United States)

    2006-10-01

    therapies to improve breast cancer cure frequency. Down Regulation of VEGF in MTCL Cells. RNA interference has been a powerful genetic tool to study...Science 296, 550 (2002). 100. D. A. Rubinson et al., Nat Genet 33, 401 (2003). 101. E. Devroe, P. A. Silver, BMC Biotechnol 2 (2002). 102. J. P...double- plotted along the estrous cycle. Double plotting of rhythmic patterns is a standard chronobiological technique that allows visualization of

  5. Cure Behavior and Thermal Properties of Diepoxidized Cardanol Resin Cured by Electron Beam Process

    Energy Technology Data Exchange (ETDEWEB)

    Cho, Donghwan; Cheon, Jinsil [Kumoh National Institute of Technology, Gumi (Korea, Republic of)

    2013-07-01

    Thermal curing of epoxy resin requires high temperature, time-consuming process and the volatilization of hardener. It has known that electron beam curing of epoxy resin is a fast process and occurs at low or room temperature that help reduce residual mechanical stresses in thermosetting polymers. Diepoxidized cardanol (DEC) can be synthesized by an enzymatic method from cashew nut shell liquid (CNSL), that constitutes nearly one-third of the total nut weight. A large amount of CNSL can be formed as a byproduct of the mechanical processes used to render the cashew kerneledible and its total production approaches one million tons annually, which can be bio-degradable and replace the industrial thermosetting plastics. It is expected that DEC may be cured as in an epoxy resin, which was constituted on two epoxide group and long alkyl chain, and two-types of onium salts (cationic initiator) were used as a photo-initiator. The experimental variables of this study are type and concentration of photo-initiators and electron beam dosage. In this study, the effects of initiator type and concentration on the cure behavior and the thermal properties of DEC resin processed by using electron beam technology were studied using FT-IR, TGA, TMA, DSC, and DMA. Figure 1 is the FT-IR results, showing the change of chemical structure of pure DEC and electron beam cured DEC. The characteristic absorption peak of epoxide group appeared at 850cm{sup -1}. The shape and the height were reduced when the sample was irradiated with electron beam. From this result, the epoxide groups is DEC were opened by electron beam and cured. After then, electron beam cured DEC was investigated the effect of forming 3-dimensional network.

  6. An investigation on the effect of light cure and self cure composite resins on bonding strength of light cure glass ionomer to dentin (In-vitro

    Directory of Open Access Journals (Sweden)

    Pahlavan A

    2004-02-01

    Full Text Available Composite reins have recently become popular for posterior teeth restorations. Gap"nformation and subsequent microleakage are of the complications resulting from such restorations. One of the"ntechniques to overcome polymerization shrinkage of composite resins is sandwich technique (application of"nglass ionomer as a base beneath the composite resin. Since polymerization patterns in two types of composite"nresins (light cure and self cure differ from each other, various effects on the bond strength between glass"nionomer and dentin are expected."nPurpose: The aim of this in vitro study was to evaluate the effects of self- cure and light- cure composite"nresins in sandwich technique on the bond strength of light cure glass ionomer and dentin."nMaterials and Methods: 40 extracted human premolars were selected and divided into four groups:"nGroup 1: Light cure glass ionomer of 1mm thickness was placed on dentin."nGroup 2: 1mm thickness of light cure glass ionomer plus a mass of self cure composite resin of 2mm"nthickness were placed."nGroup 3: 1mm thickness of light cure glass ionomer plus light cure composite resin as two separate 1mm layer"nwere placed."nGroup 4: 1mm thickness of light cure glass ionomer with 37% phosphoric acid etching followed by two"nseparate layers of light cure composite resin of 1 mm thickness were placed."nSEM was used to determine gap size ai Gl- dentin and Gi- composite interlaces. The findings were analyzed"nby ANOVA and t-student tests."nResults: Groups 1 and 2 showed no gap at Gl-dentm interface and also cracks were not observed in all these"nspecimens. In group 3, there was gap between light cure GI and light cure composite resin and cracks were"nseen in GI, too. Group 4 showed gap at both interfaces and more cracks were seen in GI. Groups I and 2"nshowed the least gap formation and group 4 showed the most. Statistically significant difference was found"nbetween groups 3, 4 and group 1 (control, 2."nConclusion: Base

  7. Bacterial mitosis: ParM of plasmid R1 moves plasmid DNA by an actin-like insertional polymerization mechanism.

    Science.gov (United States)

    Møller-Jensen, Jakob; Borch, Jonas; Dam, Mette; Jensen, Rasmus B; Roepstorff, Peter; Gerdes, Kenn

    2003-12-01

    Bacterial DNA segregation takes place in an active and ordered fashion. In the case of Escherichia coli plasmid R1, the partitioning system (par) separates paired plasmid copies and moves them to opposite cell poles. Here we address the mechanism by which the three components of the R1 par system act together to generate the force required for plasmid movement during segregation. ParR protein binds cooperatively to the centromeric parC DNA region, thereby forming a complex that interacts with the filament-forming actin-like ParM protein in an ATP-dependent manner, suggesting that plasmid movement is powered by insertional polymerization of ParM. Consistently, we find that segregating plasmids are positioned at the ends of extending ParM filaments. Thus, the process of R1 plasmid segregation in E. coli appears to be mechanistically analogous to the actin-based motility operating in eukaryotic cells. In addition, we find evidence suggesting that plasmid pairing is required for ParM polymerization.

  8. Influenza Plasmid DNA Vaccines: Progress and Prospects.

    Science.gov (United States)

    Bicho, Diana; Queiroz, João António; Tomaz, Cândida Teixeira

    2015-01-01

    Current influenza vaccines have long been used to fight flu infectious; however, recent advances highlight the importance of produce new alternatives. Even though traditional influenza vaccines are safe and usually effective, they need to be uploaded every year to anticipate circulating flu viruses. This limitation together with the use of embryonated chicken eggs as the substrate for vaccine production, is time-consuming and could involve potential biohazards in growth of new virus strains. Plasmid DNA produced by prokaryote microorganisms and encoding foreign proteins had emerged as a promising therapeutic tool. This technology allows the expression of a gene of interest by eukaryotic cells in order to induce protective immune responses against the pathogen of interest. In this review, we discuss the strategies to choose the best DNA vaccine to be applied in the treatment and prevention of influenza. Specifically, we give an update of influenza DNA vaccines developments, all involved techniques, their main characteristics, applicability and technical features to obtain the best option against influenza infections.

  9. Plasmid-Mediated Sulfamethoxazole Resistance Encoded by the sul2 Gene in the Multidrug-Resistant Shigella flexneri 2a Isolated from Patients with Acute Diarrhea in Dhaka, Bangladesh

    Science.gov (United States)

    Iqbal, Mohd S.; Rahman, Mostafizur; Islam, Rafiad; Banik, Atanu; Amin, M. Badrul; Akter, Fatema; Talukder, Kaisar Ali

    2014-01-01

    In this study, mechanisms of plasmid-mediated sulfamethoxazole resistances in the clinical strains of multi-drug resistant (MDR) Shigella flexneri 2a were elucidated for the first time in Bangladesh. From 2006 to 2011, a total of 200 S. flexneri 2a strains were randomly selected from the stock of the Enteric and Food Microbiology Laboratory of icddr,b. Antimicrobial susceptibility of the strains showed 73%, 98%, 93%, 58%, 98%, 64% and 4% resistance to trimethoprim-sulfamethoxazole, nalidixic acid, ampicillin, erythromycin, tetracycline, ciprofloxacin and ceftriaxone respectively. Plasmid profiling revealed heterogeneous patterns and interestingly, all the trimethoprim-sulfamethoxazole resistant (SXTR) strains yielded a distinct 4.3 MDa plasmid compared to that of the trimethoprim-sulfamethoxazole susceptible (SXTS) strains. Curing of this 4.3 MDa plasmid resulted in the susceptibility to sulfamethoxazole alone suggesting the involvement of this plasmid in the resistance of sulfamethoxazole. Moreover, PCR analysis showed the presence of sul2 gene in SXTR strains which is absent in SXTS strains as well as in the 4.3 MDa plasmid-cured derivatives, confirming the involvement of sul2 in the resistance of sulfamethoxazole. Furthermore, pulsed-field gel electrophoresis (PFGE) analysis revealed that both the SXTR and SXTS strains were clonal. This study will significantly contributes to the knowledge on acquired drug resistance of the mostly prevalent S. flexneri 2a and further warrants continuous monitoring of the prevalence and correlation of this resistance determinants amongst the clinical isolates of Shigella and other enteric pathogens around the world to provide effective clinical management of the disease. PMID:24416393

  10. Plasmid-mediated sulfamethoxazole resistance encoded by the sul2 gene in the multidrug-resistant Shigella flexneri 2a isolated from patients with acute diarrhea in Dhaka, Bangladesh.

    Directory of Open Access Journals (Sweden)

    Mohd S Iqbal

    Full Text Available In this study, mechanisms of plasmid-mediated sulfamethoxazole resistances in the clinical strains of multi-drug resistant (MDR Shigella flexneri 2a were elucidated for the first time in Bangladesh. From 2006 to 2011, a total of 200 S. flexneri 2a strains were randomly selected from the stock of the Enteric and Food Microbiology Laboratory of icddr,b. Antimicrobial susceptibility of the strains showed 73%, 98%, 93%, 58%, 98%, 64% and 4% resistance to trimethoprim-sulfamethoxazole, nalidixic acid, ampicillin, erythromycin, tetracycline, ciprofloxacin and ceftriaxone respectively. Plasmid profiling revealed heterogeneous patterns and interestingly, all the trimethoprim-sulfamethoxazole resistant (SXT(R strains yielded a distinct 4.3 MDa plasmid compared to that of the trimethoprim-sulfamethoxazole susceptible (SXT(S strains. Curing of this 4.3 MDa plasmid resulted in the susceptibility to sulfamethoxazole alone suggesting the involvement of this plasmid in the resistance of sulfamethoxazole. Moreover, PCR analysis showed the presence of sul2 gene in SXT(R strains which is absent in SXT(S strains as well as in the 4.3 MDa plasmid-cured derivatives, confirming the involvement of sul2 in the resistance of sulfamethoxazole. Furthermore, pulsed-field gel electrophoresis (PFGE analysis revealed that both the SXT(R and SXT(S strains were clonal. This study will significantly contributes to the knowledge on acquired drug resistance of the mostly prevalent S. flexneri 2a and further warrants continuous monitoring of the prevalence and correlation of this resistance determinants amongst the clinical isolates of Shigella and other enteric pathogens around the world to provide effective clinical management of the disease.

  11. Accurate Cure Modeling for Isothermal Processing of Fast Curing Epoxy Resins

    Directory of Open Access Journals (Sweden)

    Alexander Bernath

    2016-11-01

    Full Text Available In this work a holistic approach for the characterization and mathematical modeling of the reaction kinetics of a fast epoxy resin is shown. Major composite manufacturing processes like resin transfer molding involve isothermal curing at temperatures far below the ultimate glass transition temperature. Hence, premature vitrification occurs during curing and consequently has to be taken into account by the kinetic model. In order to show the benefit of using a complex kinetic model, the Kamal-Malkin kinetic model is compared to the Grindling kinetic model in terms of prediction quality for isothermal processing. From the selected models, only the Grindling kinetic is capable of taking into account vitrification. Non-isothermal, isothermal and combined differential scanning calorimetry (DSC measurements are conducted and processed for subsequent use for model parametrization. In order to demonstrate which DSC measurements are vital for proper cure modeling, both models are fitted to varying sets of measurements. Special attention is given to the evaluation of isothermal DSC measurements which are subject to deviations arising from unrecorded cross-linking prior to the beginning of the measurement as well as from physical aging effects. It is found that isothermal measurements are vital for accurate modeling of isothermal cure and cannot be neglected. Accurate cure predictions are achieved using the Grindling kinetic model.

  12. Post-cure depth of cure of bulk fill dental resin-composites.

    Science.gov (United States)

    Alrahlah, A; Silikas, N; Watts, D C

    2014-02-01

    To determine the post-cure depth of cure of bulk fill resin composites through using Vickers hardness profiles (VHN). Five bulk fill composite materials were examined: Tetric EvoCeram(®) Bulk Fill, X-tra base, Venus(®) Bulk Fill, Filtek™ Bulk Fill, SonicFill™. Three specimens of each material type were prepared in stainless steel molds which contained a slot of dimensions (15 mm × 4 mm × 2 mm), and a top plate. The molds were irradiated from one end. All specimens were stored at 37°C for 24h, before measurement. The Vickers hardness was measured as a function of depth of material, at 0.3mm intervals. Data were analysed by one-way ANOVA using Tukey post hoc tests (α=0.05). The maximum VHN ranged from 37.8 to 77.4, whilst the VHN at 80% of max.VHN ranged from 30.4 to 61.9. The depth corresponding to 80% of max.VHN, ranged from 4.14 to 5.03 mm. One-way ANOVA showed statistically significant differences between materials for all parameters tested. SonicFill exhibited the highest VHN (presin composites can be cured to an acceptable post-cure depth, according to the manufacturers' claims. SonicFill and Tetric EvoCeram Bulk Fill had the greatest depth of cure among the composites examined. Copyright © 2013 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

  13. 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.

  14. Plasmid Conjugation in E. coli and Drug Resistance

    African Journals Online (AJOL)

    Prof. Ogunji

    respiratory infections etc) or prescribing the 'newest' antibiotics in the market when older “brands” may ..... influence an increase in mortality rate; high economic burden and longer hospital ... Conjugating plasmids into bacteria; Tri Parental.

  15. Construction and Identification of Plasmid pTA-TUB2

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    An about 1.40 Kb target gene fragment was yielded by PCR amplification with the plasmid pRB 129,which was identified by restriction enzyme digestion that the PCR product was TU B2 gene.The gene was digested by the restriction enzyme and was linked with pTA plasmid to construct pTA-TU B2 plasmid.The plasmid was transformed into Chaetomium spp.by PEG method and the transformation rate was 27/(2×105) and it is nine times higher than that of pRB 129.The transformants can grow on the PDA containing 1 000 μg*mL-1 carbendazim,which is 1 000 times higher than the original Chaetomium spp.The resistance was stable after 10 times transfer on non-selective medium.

  16. Cure mechanisms in materials for use in esthetic dentistry.

    Science.gov (United States)

    Kwon, Tae-Yub; Bagheri, Rafat; Kim, Young K; Kim, Kyo-Han; Burrow, Michael F

    2012-02-01

    The current paper reviews the curing mechanisms found in resin-based materials used in dentistry. Historical aspects of dental products and the associated curing mechanisms are reviewed. In comparison with common industrial procedures, curing methods employed for dental materials are relatively limited because of the need to polymerize quickly in the oral cavity at an ambient temperature. Heat-cure and self-cure dental resins utilize benzoyl peroxide initiator alone with a tertiary amine co-initiator. At present, most dental restorative composites use a camphorquinone-amine complex initiation, visible light-cure, one-component systems, although alternative photoinitiators have been researched and developed. A multiple curing mode in a dual-cure material is a complex combination of various initiation systems. The use of aryl sulfinic acid sodium salt to overcome adverse chemical interactions between simplified adhesives and self- or dual-cure composites is based on another self-cure polymerization mechanism, sulfinic acid-initiated polymerization, proposed by Hagger in 1948. The sodium salt of aryl sulfinic acid reacts with an acidic monomer in simplified adhesives, and is believed to produce radicals. Clinically, it is important to try to optimize the degree of conversion of resin-based materials using proper manipulation and adequate light-curing techniques to ensure the best outcome for materials used to restore teeth.

  17. Transfer of conjugative plasmids among bacteria under environmentally relevant conditions

    DEFF Research Database (Denmark)

    Musovic, Sanin

    at spredningskapacitet af en konjugerbare plasmid, der koder for kviksølv resistens via merA genet, finder sted under substrat begrænsede forhold til syntetisk bakterielt samfund. Plasmid overførsel var meget forhøjet ved kontinuert udsættelse af mikrokosms for en høj koncentration af kviksølv. De forskellige vækstrater...

  18. Rubber curing chemistry governing the orientation of layered silicate

    Directory of Open Access Journals (Sweden)

    2007-11-01

    Full Text Available The effect of curing systems on the orientation and the dispersion of the layered silicates in acrylonitrile butadiene rubber nanocomposite is reported. Significant differences in X-ray diffraction pattern between peroxide curing and sulfur curing was observed. Intense X-ray scattering values in the XRD experiments from peroxide cured vulcanizates indicate an orientation of the layers in a preferred direction as evinced by transmission electron micrographs. However, sulfur cured vulcanizates show no preferential orientation of the silicate particles. Nevertheless, a closer inspection of transmission electron microscopy (TEM images of peroxide and sulfur cured samples shows exfoliated silicate layers in the acrylonitrile butadiene rubber (NBR matrix. It was revealed in the prevailing study that the use of an excess amount of stearic acid in the formulation of the sulfur curing package leads to almost exfoliated type X-ray scattering pattern.

  19. A New Shuttle Plasmid That Stably Replicates in Clostridium acetobutylicum.

    Science.gov (United States)

    Lee, Sang-Hyun; Kwon, Min-A; Choi, Sunwha; Kim, Sooah; Kim, Jungyeon; Shin, Yong-An; Kim, Kyoung Heon

    2015-10-01

    We have developed a new shuttle plasmid, designated as pLK1-MCS that can replicate in both Clostridium acetobutylicum and Escherichia coli, by combining the pUB110 and pUC19 plasmids. Plasmid pLK1-MCS replicated more stably than previously reported plasmids containing either the pIM13 or the pAMβ1 replicon in the absence of antibiotic selective pressure. The transfer frequency of pLK1-MCS into C. acetobutylicum was similar to the transfer frequency of other shuttle plasmids. We complemented C. acetobutylicum ML1 (that does not produce solvents such as acetone, butanol, and ethanol owing to loss of the megaplasmid pSOL1 harboring the adhE1-ctfAB-adc operon) by introducing pLK1-MCS carrying the adhE1-ctfAB-adc operon into C. acetobutylicum ML1. The transformed cells were able to resume anaerobic solvent production, indicating that the new shuttle plasmid has the potential for practical use in microbial biotechnology.

  20. Plasmid copy number noise in monoclonal populations of bacteria

    Science.gov (United States)

    Wong Ng, Jérôme; Chatenay, Didier; Robert, Jérôme; Poirier, Michael Guy

    2010-01-01

    Plasmids are extra chromosomal DNA that can confer to their hosts’ supplementary characteristics such as antibiotic resistance. Plasmids code for their copy number through their own replication frequency. Even though the biochemical networks underlying the plasmid copy number (PCN) regulation processes have been studied and modeled, no measurement of the heterogeneity in PCN within a whole population has been done. We have developed a fluorescent-based measurement system, which enables determination of the mean and noise in PCN within a monoclonal population of bacteria. Two different fluorescent protein reporters were inserted: one on the chromosome and the other on the plasmid. The fluorescence of these bacteria was measured with a microfluidic flow cytometry device. We show that our measurements are consistent with known plasmid characteristics. We find that the partitioning system lowers the PCN mean and standard deviation. Finally, bacterial populations were allowed to grow without selective pressure. In this case, we were able to determine the plasmid loss rate and growth inhibition effect.

  1. CURING OF POLYMERIC COMPOSITES USING MICROWAVE RESIN TRANSFER MOULDING (RTM

    Directory of Open Access Journals (Sweden)

    R. YUSOFF

    2007-08-01

    Full Text Available The main objective of this work is to compare the difference between microwave heating and conventional thermal heating in fabricating carbon/epoxy composites. Two types of epoxy resin systems were used as matrices, LY5052-HY5052 and DGEBA-HY917-DY073. All composite samples were fabricated using resin transfer moulding (RTM technique. The curing of the LY5052-HY5052-carbon and the DGEBA-HY917-DY073-carbon composite systems, were carried out at 100 °C and 120 °C, respectively. Microwave heating showed better temperature control than conventional heating, however, the heating rate of the microwave cured samples were slower than the conventionally cured samples. This was attributed to the lower power (250 W used when heating with microwaves compared to 2000 W used in conventional heating. Study of thermal characteristics as curing progressed showed that the polymerisation reaction occurred at a faster rate during microwave curing than in conventional curing for both the DGEBA and the LY/HY5052 carbon composite systems. The actual cure cycle was reduced from 60 minutes to 40 minutes when using microwaves for curing DGEBA-carbon composites. As for LY/HY5052-carbon composites, the actual cure cycle was reduced from 3 hours to 40 minutes. Both conventional and microwave heating yielded similar glass transition temperatures (120 °C for DGEBA systems and 130 °C for LY/HY5052 systems. Microwave cured composites had higher void contents than conventionally cured composites (2.2-2.8% and 1.8-2.4% for DGEBA and LY/HY5052 microwave cured composites, respectively, compared to 0.2-0.4% for both DGEBA and LY/HY5052 thermally cured composites. C-scan traces showed that all composites, regardless of methods of curing, had minimal defects.

  2. Cure shrinkage effects in epoxy and polycyanate matrix composites

    Energy Technology Data Exchange (ETDEWEB)

    Spellman, G.P.

    1995-12-22

    A relatively new advanced composite matrix, polycyanate ester, was evaluated for cure shrinkage. The chemical cure shrinkage of composites is difficult to model but a number of clever experimental techniques are available to the investigator. In this work the method of curing a prepreg layup on top of a previously cured laminate of identical ply composition is utilized. The polymeric matrices used in advanced composites have been primarily epoxies and therefore a common system of this type, Fiberite 3501-6, was used as a base case material. Three polycyanate matrix systems were selected for the study. These are: Fiberite 954-2A, YLA RS-3, and Bryte Technology BTCy-1. The first three of these systems were unidirectional prepreg with carbon fiber reinforcement. The Bryte Technology material was reinforced with E-glass fabric. The technique used to evaluate cure shrinkage results in distortion of the flatness of an otherwise symmetric laminate. The first laminate is cured in a conventional fashion. An identical layup is cured on this first laminate. During the second cure all constituents are exposed to the same thermal cycles. However, only the new portion of the laminate will experience volumetric changes associate with matrix cure. The additional strain of cure shrinkage results in an unsymmetric distribution of residual stresses and an associated warpage of the laminate. The baseline material, Fiberite 3501-6, exhibited cure shrinkage that was in accordance with expectations. Cure strains were {minus}4.5E-04. The YLA RS-3 material had cure strains somewhat lower at {minus}3.2E-04. The Fiberite 954-2A cure strain was {minus}1.5E-04 that is 70% lower than the baseline material. The glass fabric material with the Bryte BTCy-1 matrix did not result in meaningful results because the processing methods were not fully compatible with the material.

  3. CURING KINETICS AND PROPERTIES OF ACRYLIC RESIN CURED WITH AZIRIDINE CROSSLINKER

    Institute of Scientific and Technical Information of China (English)

    Fei Xie; Zong-hui Liu; De-qing Wei

    2002-01-01

    A kind of aziridine crosslinkers was synthesized and used to crosslink acrylate copolymers. The crosslinking properties and curing kinetics of the resin were studied. It was found that with the increase of the content of crosslinker in the emulsion, the mechanical properties and solvent resistance of the resin will be apparently improved, but its glass transition temperature (Tg) is very low. The lowest amount of crosslinker used in the acrylic resin emulsion is 0.25%. Curing kinetics studied by DSC show that this curing reaction occurs readily because the apparent activation energy of the reaction is low(65.1 KJ/mol). These results demonstrate that the aziridine crosslinker is indeed a low temperature crosslinking agent and can be used at room temperature.

  4. Experimental Study on the Curing Effect of Dredged Sediments with Three Types of Curing Agents

    Directory of Open Access Journals (Sweden)

    Yan Lei-Ming

    2016-01-01

    Full Text Available Sediment solidification technology is widely used to dispose dredged sediment, three types of curing agents were used in this study to solidified the dredged sediment from shallows in Nantong with three types of curing agents: JY, ZL and FJ. The results showed that the optimal additive amounts of these three curing agents were 140g JY, 16g ZL, 2.0g FJ per 1000g of the dredged sediment respectively, their 28d USC were up to 2.48 MPa, 2.96 MPa and 3.00 MPa. JY has obvious early strength effect, which of FJ is not that obvious, but the later-stage strength of sediment solidified by FJ are relatively higher.

  5. Curing Study of Epoxy/clay Nanocomposite by a New Aromatic Diamine Functional Curing Agent

    Directory of Open Access Journals (Sweden)

    Seyed Mahdi Saadati

    2013-12-01

    Full Text Available The curing behavior of an epoxy/clay nanocomposite system composed of diglycidyl ether of bisphenol A (DGEBA epoxy resin with a new aromatic diamine curing agent and organically modified clay was investigated. Epoxy/clay nanocomposite samples were prepared by mixing DGEBA, different percentages of nanoclay (1, 3, 5, 7, and 9% and 4-(4-(2,6-diphenylpyridin-4-ylphenoxybenzene-1,3-diamine (DAP. The optimum ratio of the epoxy/clay nanocomposite system (3% was investigated by total enthalpy of the curing reaction from differential scanning calorimetry (DSC thermograms. The kinetic parameters such as activation energy, pre-exponential factor, and rate constant were obtained from DSC data.

  6. Construction and Use of Flow Cytometry Optimized Plasmid-Sensor Strains

    DEFF Research Database (Denmark)

    Bahl, Martin Iain; Oregaard, Gunnar; Sørensen, Søren Johannes;

    2009-01-01

    stability of the plasmid is high. The method presented here relies on a phenotypic (green fluorescence protein) marker, which is switched on if the host bacteria loses the residing plasmid. The incorporation of flow cytometry for single-cell detection and discrimination between plasmid-free and plasmid...

  7. Allelopathy of plasmid-bearing and plasmid-free organisms competing for two complementary resources in a chemostat.

    Science.gov (United States)

    Bhattacharyya, Joydeb; Smith, Hal L; Pal, Samares

    2012-01-01

    We consider a model of competition between plasmid-bearing and plasmid-free organisms for two complementary nutrients in a chemostat. We assume that the plasmid-bearing organism produces an allelopathic agent at the cost of its reproductive abilities which is lethal to plasmid-free organism. Our analysis leads to different thresholds in terms of the model parameters acting as conditions under which the organisms associated with the system cannot thrive even in the absence of competition. Local stability of the system is obtained in the absence of one or both the organisms. Also, global stability of the system is obtained in the presence of both the organisms. Computer simulations have been carried out to illustrate various analytical results.

  8. Improving a self-curing dental resin by eliminating oxygen, hydroquinone and water from its curing process.

    Science.gov (United States)

    Keh, En-Sheng; Hayakawa, Iwao; Takahashi, Hidekazu; Watanabe, Akihiko; Iwasaki, Yasuhiko; Akiyoshi, Kazunari; Nakabayashi, Nobuo

    2002-12-01

    Self-curing dental resins are always manipulated in the presence of curing inhibitory factors such as oxygen, hydroquinone, water and another contaminants such as saliva and blood. The purpose of this study was to elucidate the effects of eliminating these curing inhibitory factors on resin properties. Several clinically relevant characteristics of an experimental resin cured in the absence of inhibitory factors (purified group) were determined and compared with the resin cured conventionally (control group). The purified group showed a significantly shorter induction period and higher reaction exotherm. It also showed significantly higher tensile strength and lower strain to failure. With regard to the amount of residual monomer, fracture toughness and fatigue strength the improvements were not significant. These findings suggest that the polymerization outcomes and mechanical properties of the purified group resin can be improved in general. Dental clinicians should pay attention to the storage and curing environments while manipulating the self-curing resin.

  9. 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

    of these plasmids within pathogenic hosts are poorly understood. Here we study plasmid-host adaptations following transfer of a 73 kb conjugative multidrug resistance plasmid to naïve clinical isolates of Klebsiella pneumoniae and Escherichia coli We use experimental evolution, mathematical modelling and population...... 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...... of plasmid adaptation. While insertion sequences are well known to supply plasmids with adaptive traits, our findings suggest that they also play an important role in plasmid evolution by maintaining the plasticity necessary to alleviate plasmid-host constrains. Further, the observed evolutionary strategy...

  10. Multiple Pathways of Plasmid DNA Transfer in Helicobacter pylori

    Science.gov (United States)

    Rohrer, Stefanie; Holsten, Lea; Weiss, Evelyn; Benghezal, Mohammed; Fischer, Wolfgang; Haas, Rainer

    2012-01-01

    Many Helicobacter pylori (Hp) strains carry cryptic plasmids of different size and gene content, the function of which is not well understood. A subgroup of these plasmids (e.g. pHel4, pHel12), contain a mobilisation region, but no cognate type IV secretion system (T4SS) for conjugative transfer. Instead, certain H. pylori strains (e.g. strain P12 carrying plasmid pHel12) can harbour up to four T4SSs in their genome (cag-T4SS, comB, tfs3, tfs4). Here, we show that such indigenous plasmids can be efficiently transferred between H. pylori strains, even in the presence of extracellular DNaseI eliminating natural transformation. Knockout of a plasmid-encoded mobA relaxase gene significantly reduced plasmid DNA transfer in the presence of DNaseI, suggesting a DNA conjugation or mobilisation process. To identify the T4SS involved in this conjugative DNA transfer, each individual T4SS was consecutively deleted from the bacterial chromosome. Using a marker-free counterselectable gene deletion procedure (rpsL counterselection method), a P12 mutant strain was finally obtained with no single T4SS (P12ΔT4SS). Mating experiments using these mutants identified the comB T4SS in the recipient strain as the major mediator of plasmid DNA transfer between H. pylori strains, both in a DNaseI-sensitive (natural transformation) as well as a DNaseI-resistant manner (conjugative transfer). However, transfer of a pHel12::cat plasmid from a P12ΔT4SS donor strain into a P12ΔT4SS recipient strain provided evidence for the existence of a third, T4SS-independent mechanism of DNA transfer. This novel type of plasmid DNA transfer, designated as alternate DNaseI-Resistant (ADR) mechanism, is observed at a rather low frequency under in vitro conditions. Taken together, our study describes for the first time the existence of three distinct pathways of plasmid DNA transfer between H. pylori underscoring the importance of horizontal gene transfer for this species. PMID:23029142

  11. The cure for employee malaise--motivation.

    Science.gov (United States)

    Dawson, K M; Dawson, S N

    1991-01-01

    Although working conditions, hours, pay, and advancement opportunities are better now than in the 1950s--the "golden age" of American business--today's workers are significantly less satisfied. Why? The authors believe the cause of this malaise is lack of motivation. This article examines several techniques to cure employee malaise and discusses the long-term benefits of these techniques, which include empowerment, recognition, career development, the Pygmalion effect, incentives, and rewards. By making a commitment to these motivational techniques, managers will boost the morale and enthusiasm of their employees and their organization. This motivational process is not quick and easy; developing your employees is an ongoing process.

  12. Cure behavior of epoxy polymers used in microelectronics

    Science.gov (United States)

    Taweeplengsangsuke, Jantrawan

    2000-10-01

    Underfill resins are used to reduce solder fatigue in flip-chip assemblies. Both underfilling and curing processes are critical to achieve reliable finished products. This study included two main themes; the development of processing diagrams for commercial underfill resins and the fundamental understanding of cure kinetics and evolution of cure stresses in model epoxies. A variety of techniques have been used to characterize the fundamental phenomena involved in the processing of underfill resins. The flow behavior, cure kinetics, cure stresses, outgassing phenomena and void formation of underfill resins were studied. Processing diagrams for underfill resins were developed by mapping fundamental phenomena involved to provide a guideline for underfilling and curing steps. With these processing diagrams one should be able to design his underfilling and curing process with scientific approach. Two commercial underfill resins, which were evaluated as part of a SEMATECH program, where shown to possess significantly different processing characteristics. However, the appropriate explanation could not be made without knowing the proprietary chemistry of the systems. Therefore, model epoxies were formulated. Three different systems were investigated; amine, imidazole, and anhydride curing agent systems. With the known chemistry, the reaction kinetics and developed cure stress can be explained more clearly. Vitrification, which severely retards the rate of reaction, was found in amine (N-aminoethylpiperazine) and anhydride (Hexahydro-4-methylphthalic anhydride) cured systems when the cure temperatures were lower than the glass transition temperature (T g) of fully cured samples; however, imidazole(2-ethyl-4-methyl-imidazole) did not show vitrification when it was cured at 20 degrees below the T g of fully cured sample due to the large exotherms. Interestingly for imidazole system, the higher cure temperature exhibited a lower Tg. This could possibly be explained by the

  13. Altered Murine Tissue Colonization by Borrelia burgdorferi following Targeted Deletion of Linear Plasmid 17-Carried Genes

    OpenAIRE

    Casselli, Timothy; Tourand, Yvonne; Bankhead, Troy

    2012-01-01

    The causative agent of Lyme disease, Borrelia burgdorferi, possesses a segmented genome comprised of a single linear chromosome and upwards of 23 linear and circular plasmids. Much of what is known about plasmid-borne genes comes from studying laboratory clones that have spontaneously lost one or more plasmids during in vitro passage. Some plasmids, including the linear plasmid lp17, are never or rarely reported to be lost during routine culture; therefore, little is known about the requireme...

  14. Mechanical modeling of stress generation during cure of encapsulating resins

    Energy Technology Data Exchange (ETDEWEB)

    Lagasse, R.R.; Chambers, R.S.; Guess, T.R. (Sandia National Labs., Albuquerque, NM (USA)); Plazek, D.J.; Bero, C. (Pittsburgh Univ., PA (USA). Dept. of Materials Science and Engineering)

    1991-01-01

    We have developed a numerical model for calculating stresses generated during cure of shrinking encapsulating resins. Mechanical modeling of polymer encapsulated electronic devices usually focuses on stress generated during cooling after cure. The stress developed during cure, due to shrinkage of the encapsulant, is normally neglected. That assumption is valid if both the shear and bulk moduli of the encapsulant at the cure temperature are negligible with respect to the moduli at lower temperatures. Our measurements on a model epoxy encapsulant show that the shear modulus during cure, varying from 0 to 6 MPa, is at least 100 times smaller than that at ambient temperature. In contrast, the bulk modulus at the cure temperature is only 2.5 times smaller. Since the bulk modulus during cure cannot be neglected, significant stress can be produced if volume shrinkage is constrained by a stiff mold or embedded elements. In fact, mechanical failure of encapsulating materials during cure has been evident in some of our experiments. Using measurements of shear and bulk moduli plus volume shrinkage as inputs to a finite element model, we have successfully predicted the shrinkage strains and stresses developed during cure of a model epoxy resin inside a cylindrical tube. Consideration of cure shrinkage stress has led to a process modification that appears to reduce mechanical failures in a real encapsulated device. 6 refs., 6 figs.

  15. Critical parameters for electron beam curing of cationic epoxies and property comparison of electron beam cured cationic epoxies versus thermal cured resins and composites

    Energy Technology Data Exchange (ETDEWEB)

    Janke, C.J.; Norris, R.E.; Yarborough, K. [Oak Ridge National Lab., TN (United States). Oak Ridge Centers for Manufacturing Technology; Havens, S.J. [Oak Ridge Inst. for Science and Education, TN (United States); Lopata, V.J. [Atomic Energy of Canada Ltd., Pinawa, Manitoba (Canada). Whiteshell Labs.

    1997-01-16

    Electron beam curing of composites is a nonthermal, nonautoclave curing process offering the following advantages compared to conventional thermal curing: substantially reduced manufacturing costs and curing times; improvements in part quality and performance; reduced environmental and health concerns; and improvements in material handling. In 1994 a Cooperative Research and Development Agreement (CRADA), sponsored by the Department of Energy Defense Programs and 10 industrial partners, was established to advance electron beam curing of composites. The CRADA has successfully developed hundreds of new toughened and untoughened resins, offering unlimited formulation and processing flexibility. Several patent applications have been filed for this work. Composites made from these easily processable, low shrinkage material match the performance of thermal cured composites and exhibit: low void contents comparable to autoclave cured composites (less than 1%); superb low water absorption values in the same range as cyanate esters (less than 1%); glass transition temperatures rivaling those of polyimides (greater than 390 C); mechanical properties comparable to high performance, autoclave cured composites; and excellent property retention after cryogenic and thermal cycling. These materials have been used to manufacture many composite parts using various fabrication processes including hand lay-up, tow placement, filament winding, resin transfer molding and vacuum assisted resin transfer molding.

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

    Directory of Open Access Journals (Sweden)

    Joseph J Gillespie

    Full Text Available BACKGROUND: 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. METHODOLOGY/PRINCIPAL FINDINGS: 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. CONCLUSION/SIGNIFICANCE: 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

  17. Influence of curing protocol on selected properties of light-curing polymers

    DEFF Research Database (Denmark)

    Dewaele, Magali; Asmussen, Erik; Peutzfeldt, Anne;

    2009-01-01

    The purpose of this study was to investigate the effect of light-curing protocol on degree of conversion (DC), volume contraction (C), elastic modulus (E), and glass transition temperature (T(g)) as measured on a model polymer. It was a further aim to correlate the measured values with each other....

  18. Heavy metal content in flue cured and air cured tobaccos from main production areas in Serbia

    Directory of Open Access Journals (Sweden)

    Radojičić Vesna

    2004-01-01

    Full Text Available The aim of this work was to examine the influence of growing conditions on heavy metal content in virginia (flue cured and burley (air cured tobacco type. Moreover, some of these metals could appear in the tobacco cigarette smoke. This fact may cause negative consequences for cigarette consumers. Examinations were carried out in five various production areas, for each tobacco type, during two years, 1998 and 1999; those were extreme years for production.Considering the results, it can be concluded: The most important factor for heavy metal content in tobacco leaves are weather conditions especially the amount of rainfall, since there is a direct correlation between the rainfall and size of roots, a dominant vegetation organ. It is inevitable that the metals content in flue cured and air cured tobaccos grown at different experimental lots, is still below the legally prescribed values. Tobacco leaves from Šabac production area have a little higher metal content than tobacco grown in other production areas, because a larger amount of copper was found. The metal content in plants can increase because of its absorption from soil, atmosphere, or mineral fertilizers and pesticides, and, what is very important, the content of metals can be high due to the vicinity of large industrial centers.

  19. CURE: Clean use of reactor energy

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1990-05-01

    This paper presents the results of a joint Westinghouse Hanford Company (Westinghouse Hanford)-Pacific Northwest Laboratory (PNL) study that considered the feasibility of treating radioactive waste before disposal to reduce the inventory of long-lived radionuclides, making the waste more suitable for geologic disposal. The treatment considered here is one in which waste would be chemically separated so that long-lived radionuclides can be treated using specific processes appropriate for the nuclide. The technical feasibility of enhancing repository performance by this type of treatment is considered in this report. A joint Westinghouse Hanford-PNL study group developed a concept called the Clean Use of Reactor Energy (CURE), and evaluated the potential of current technology to reduce the long-lived radionuclide content in waste from the nuclear power industry. The CURE process consists of three components: chemical separation of elements that have significant quantities of long-lived radioisotopes in the waste, exposure in a neutron flux to transmute the radioisotopes to stable nuclides, and packaging of radionuclides that cannot be transmuted easily for storage or geologic disposal. 76 refs., 32 figs., 24 tabs.

  20. Radiation curing of carbon fibre composites

    Science.gov (United States)

    Spadaro, G.; Alessi, S.; Dispenza, C.; Sabatino, M. A.; Pitarresi, G.; Tumino, D.; Przbytniak, G.

    2014-01-01

    Epoxy/carbon fibre reinforced composites were produced by means of e-beam irradiation through a pulsed 10 MeV electron beam accelerator. The matrix consisted of a difunctional epoxy monomer (DGEBA) and an initiator of cationic polymerisation, while the reinforcement was a unidirectional high modulus carbon fibre fabric. Dynamic mechanical thermal analysis was carried out in order to determine the cross-linking degree. The analysis pointed out a nonuniformity in the cross-linking degree of the e-beam cured panels, with the formation of clusters at low Tg (glass transition temperature) and clusters at high Tg. An out-of-mould post irradiation thermal treatment on e-beam cured samples provides a higher uniformity in the network although some slight degradation effects. Mode I delamination fracture toughness and Interlaminar Shear Strength (ISS) were also investigated by means of Double Cantilever Beam (DCB) and Short Beam Shear tests, respectively. Results from this mechanical characterisation allowed to correlate fracture toughness of the bulk matrix resin, cross-linking density and fibre/matrix interaction to the delamination fracture behaviour of the fibre reinforced material.

  1. Out-of-Autoclave Cure Composites

    Science.gov (United States)

    Hayes, Brian S.

    2015-01-01

    As the size of aerospace composite parts exceeds that of even the largest autoclaves, the development of new out-of-autoclave processes and materials is necessary to ensure quality and performance. Many out-of-autoclave prepreg systems can produce high-quality composites initially; however, due to long layup times, the resin advancement commonly causes high void content and variations in fiber volume. Applied Poleramic, Inc. (API), developed an aerospace-grade benzoxazine matrix composite prepreg material that offers more than a year out-time at ambient conditions and provides exceptionally low void content when out-of-autoclave cured. When compared with aerospace epoxy prepreg systems, API's innovation offers significant improvements in terms of out-time at ambient temperature and the corresponding tack retention. The carbon fiber composites developed with the optimized matrix technology have significantly better mechanical performance in terms of hot-wet retention and compression when compared with aerospace epoxy matrices. These composites also offer an excellent overall balance of properties. This matrix system imparts very low cure shrinkage, low coefficient of thermal expansion, and low density when compared with most aerospace epoxy prepreg materials.

  2. 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

  3. In situ cure monitoring of advanced fiber reinforced composites

    Science.gov (United States)

    Powell, Graham R.; Crosby, Peter A.; Fernando, Gerard F.; France, Chris M.; Spooncer, Ronald C.; Waters, David N.

    1995-04-01

    This paper describes a comparative study of in-situ cure monitoring and cure modelling by three methods: (a) evanescent wave spectroscopy, (b) refractive index change, (c) near- infrared spectroscopy. Optical fibers were embedded into aerospace epoxy resins during the manufacturing process of the composite. The cure characteristics were then tracked in real- time during the processing of the material via evanescent wave interaction. This technique is based upon monitoring of characteristic infrared absorption bands of the resin system to find the concentration of the epoxy and amine hardener as a function of cure time. Hence this technique is suitable for on-line process monitoring and optimization. Results obtained from the optical fiber sensors were used to model the curing behavior of the resin system. The results were compared with near-infrared spectroscopy and differential scanning calorimetry experiments carried out under similar conditions. The feasibility of utilizing refractive index changes to monitor the extent of cure has also been demonstrated.

  4. Photoacoustic monitoring of inhomogeneous curing processes in polystyrene emulsions

    Energy Technology Data Exchange (ETDEWEB)

    Vargas-Luna, M.; Gutierrez-Juarez, G.; Rodriguez-Vizcaino, J.M.; Varela-Nsjera, J.B.; Rodriguez-Palencia, J.M.; Bernal-Alvarado, J.; Sosa, M. [Instituto de Fisica, Universidad de Guanajuato, Leon, Guanajuato (Mexico); Alvarado-Gil, J.J. [Centro de Investigacion y de Estudios Avanzados del IPN, Unidad Merida, Antigua Carretera a Progreso, Merida, Yucatan (Mexico)

    2002-07-07

    The time evolution of the inhomogeneous curing process of polystyrene emulsions is studied using a variant of the conventional photoacoustic (PA) technique. The thermal effusivity, as a function of time, is determined in order to monitor the sintering process of a styrene emulsion in different steps of the manufacturing procedure. PA measurements of thermal effusivity show a sigmoidal growth as a function of time during the curing process. The parameterization of these curves permits the determination of the characteristic curing time and velocity of the process. A decreasing of the curing time and an increasing curing velocity for the final steps of the manufacturing process are observed. The feasibility of our approach and its potentiality for the characterization of other curing process are discussed. (author)

  5. Transcription-replication collision increases recombination efficiency between plasmids.

    Science.gov (United States)

    Jialiang, Li; Feng, Chen; Zhen, Xu; Jibing, Chen; Xiang, Lv; Lingling, Zhang; Depei, Liu

    2013-11-01

    It has been proposed that the stalling of the replication forks can induce homologous recombination in several organisms, and that arrested replication forks may offer nuclease targets, thereby providing a substrate for proteins involved in double-strand repair. In this article, we constructed a plasmid with the potential for transcription-replication collision (TRC), in which DNA replication and RNA transcription occur on the same DNA template simultaneously. Theoretically, transcription will impede DNA replication and increase homologous recombination. To validate this hypothesis, another plasmid was constructed that contained a homologous sequence with the exception of some mutated sites. Co-transfection of these two plasmids into 293T cells resulted in increased recombination frequency. The ratio of these two plasmids also affected the recombination frequency. Moreover, we found high expression levels of RAD51, which indicated that the increase in the recombination rate was probably via the homologous recombination pathway. These results indicate that mutant genes in plasmids can be repaired by TRC-induced recombination.

  6. Functional amyloids as inhibitors of plasmid DNA replication

    Science.gov (United States)

    Molina-García, Laura; Gasset-Rosa, Fátima; Moreno-del Álamo, María; Fernández-Tresguerres, M. Elena; Moreno-Díaz de la Espina, Susana; Lurz, Rudi; Giraldo, Rafael

    2016-01-01

    DNA replication is tightly regulated to constrain the genetic material within strict spatiotemporal boundaries and copy numbers. Bacterial plasmids are autonomously replicating DNA molecules of much clinical, environmental and biotechnological interest. A mechanism used by plasmids to prevent over-replication is ‘handcuffing’, i.e. inactivating the replication origins in two DNA molecules by holding them together through a bridge built by a plasmid-encoded initiator protein (Rep). Besides being involved in handcuffing, the WH1 domain in the RepA protein assembles as amyloid fibres upon binding to DNA in vitro. The amyloid state in proteins is linked to specific human diseases, but determines selectable and epigenetically transmissible phenotypes in microorganisms. Here we have explored the connection between handcuffing and amyloidogenesis of full-length RepA. Using a monoclonal antibody specific for an amyloidogenic conformation of RepA-WH1, we have found that the handcuffed RepA assemblies, either reconstructed in vitro or in plasmids clustering at the bacterial nucleoid, are amyloidogenic. The replication-inhibitory RepA handcuff assembly is, to our knowledge, the first protein amyloid directly dealing with DNA. Built on an amyloid scaffold, bacterial plasmid handcuffs can bring a novel molecular solution to the universal problem of keeping control on DNA replication initiation. PMID:27147472

  7. Dcm methylation is detrimental to plasmid transformation in Clostridium thermocellum

    Energy Technology Data Exchange (ETDEWEB)

    Guss, Adam M [ORNL; Olson, Daniel G. [Thayer School of Engineering at Dartmouth; Caiazza, Nicky [Mascoma Corporation; Lynd, Lee R [Thayer School of Engineering at Dartmouth

    2012-01-01

    BACKGROUND: Industrial production of biofuels and other products by cellulolytic microorganisms is of interest but hindered by the nascent state of genetic tools. Although a genetic system for Clostridium thermocellum DSM1313 has recently been developed, available methods achieve relatively low efficiency and similar plasmids can transform C. thermocellum at dramatically different efficiencies. RESULTS: We report an increase in transformation efficiency of C. thermocellum for a variety of plasmids by using DNA that has been methylated by Escherichia coli Dam but not Dcm methylases. When isolated from a dam+ dcm+ E. coli strain, pAMG206 transforms C. thermocellum 100-fold better than the similar plasmid pAMG205, which contains an additional Dcm methylation site in the pyrF gene. Upon removal of Dcm methylation, transformation with pAMG206 showed a four- to seven-fold increase in efficiency; however, transformation efficiency of pAMG205 increased 500-fold. Removal of the Dcm methylation site from the pAM205 pyrF gene via silent mutation resulted in increased transformation efficiencies equivalent to that of pAMG206. Upon proper methylation, transformation efficiency of plasmids bearing the pMK3 and pB6A origins of replication increased ca. three orders of magnitude. CONCLUSION: E. coli Dcm methylation decreases transformation efficiency in C. thermocellum DSM1313. The use of properly methylated plasmid DNA should facilitate genetic manipulation of this industrially relevant bacterium.

  8. Dcm methylation is detrimental to plasmid transformation in Clostridium thermocellum

    Directory of Open Access Journals (Sweden)

    Guss Adam M

    2012-05-01

    Full Text Available Abstract Background Industrial production of biofuels and other products by cellulolytic microorganisms is of interest but hindered by the nascent state of genetic tools. Although a genetic system for Clostridium thermocellum DSM1313 has recently been developed, available methods achieve relatively low efficiency and similar plasmids can transform C. thermocellum at dramatically different efficiencies. Results We report an increase in transformation efficiency of C. thermocellum for a variety of plasmids by using DNA that has been methylated by Escherichia coli Dam but not Dcm methylases. When isolated from a dam+dcm+E. coli strain, pAMG206 transforms C. thermocellum 100-fold better than the similar plasmid pAMG205, which contains an additional Dcm methylation site in the pyrF gene. Upon removal of Dcm methylation, transformation with pAMG206 showed a four- to seven-fold increase in efficiency; however, transformation efficiency of pAMG205 increased 500-fold. Removal of the Dcm methylation site from the pAMG205 pyrF gene via silent mutation resulted in increased transformation efficiencies equivalent to that of pAMG206. Upon proper methylation, transformation efficiency of plasmids bearing the pMK3 and pB6A origins of replication increased ca. three orders of magnitude. Conclusions E. coli Dcm methylation decreases transformation efficiency in C. thermocellum DSM1313. The use of properly methylated plasmid DNA should facilitate genetic manipulation of this industrially relevant bacterium.

  9. Process Formulations And Curing Conditions That Affect Saltstone Properties

    Energy Technology Data Exchange (ETDEWEB)

    Reigel, M. M.; Pickenheim, B. R.; Daniel, W. E.

    2012-09-28

    The first objective of this study was to analyze saltstone fresh properties to determine the feasibility of reducing the formulation water to premix (w/p) ratio while varying the amount of extra water and admixtures used during processing at the Saltstone Production Facility (SPF). The second part of this study was to provide information for understanding the impact of curing conditions (cure temperature, relative humidity (RH)) and processing formulation on the performance properties of cured saltstone.

  10. The curing process of layered silicate/epoxy nanocomposites

    OpenAIRE

    Nieminen, Ilkka

    2009-01-01

    The industrial methods of processing polymeric thermoset matrix nanocomposites require a good knowledge of the matrix curing process. The dispersion of nanoparticles in the matrix effects the curing process due to the interaction between nanoparticles and the matrix at the atomic level, especially when the nanoparticles have been treated with a surfactant agent ( alquilammonium) that is in competition with the reactive groups of the hardener. This thesis investigates the curing process o...

  11. Curing Methods and Their Effects on The Strength of Concrete

    OpenAIRE

    Nuruddeen Usman; Muhammad Nura Isa

    2015-01-01

    There are a lot of arguments on which method of curing concrete gives good strength. These different opinions results into this study, which aim at investigating the effects of different curing methods on the strength of concrete. Laboratory test was employed for this study. Normal concretes were prepared using specified mix ratio of 1:2:4 and 1:3:6. The cubes tested for compressive strength at 3, 7, 21, and 28 days of curing respectively using four curing methods namely immersion...

  12. Optical fibre grating refractometers for resin cure monitoring

    Science.gov (United States)

    Buggy, S. J.; Chehura, E.; James, S. W.; Tatam, R. P.

    2007-06-01

    The use of fibre grating refractometers as a means of monitoring the cure of a UV-cured epoxy resin is presented. The wavelength shift of the attenuation bands of a long period grating and the spectral response of a tilted fibre Bragg grating sensor were measured simultaneously during the cure of the resin and compared with measurements made using a fibre optic Fresnel-based refractometer. The results showed a good correlation (6 × 10-3 rius) and illustrate the potential of the techniques for non-invasive composite material cure monitoring.

  13. The conjugative plasmid of a bean-nodulating Sinorhizobium fredii strain is assembled from sequences of two Rhizobium plasmids and the chromosome of a Sinorhizobium strain

    Directory of Open Access Journals (Sweden)

    Brom Susana

    2011-06-01

    Full Text Available Abstract Background Bean-nodulating Rhizobium etli originated in Mesoamerica, while soybean-nodulating Sinorhizobium fredii evolved in East Asia. S. fredii strains, such as GR64, have been isolated from bean nodules in Spain, suggesting the occurrence of conjugative transfer events between introduced and native strains. In R. etli CFN42, transfer of the symbiotic plasmid (pRet42d requires cointegration with the endogenous self-transmissible plasmid pRet42a. Aiming at further understanding the generation of diversity among bean nodulating strains, we analyzed the plasmids of S. fredii GR64: pSfr64a and pSfr64b (symbiotic plasmid. Results The conjugative transfer of the plasmids of strain GR64 was analyzed. Plasmid pSfr64a was self-transmissible, and required for transfer of the symbiotic plasmid. We sequenced pSfr64a, finding 166 ORFs. pSfr64a showed three large segments of different evolutionary origins; the first one presented 38 ORFs that were highly similar to genes located on the chromosome of Sinorhizobium strain NGR234; the second one harbored 51 ORFs with highest similarity to genes from pRet42d, including the replication, but not the symbiosis genes. Accordingly, pSfr64a was incompatible with the R. etli CFN42 symbiotic plasmid, but did not contribute to symbiosis. The third segment contained 36 ORFs with highest similarity to genes localized on pRet42a, 20 of them involved in conjugative transfer. Plasmid pRet42a was unable to substitute pSfr64a for induction of pSym transfer, and its own transfer was significantly diminished in GR64 background. The symbiotic plasmid pSfr64b was found to differ from typical R. etli symbiotic plasmids. Conclusions S. fredii GR64 contains a chimeric transmissible plasmid, with segments from two R. etli plasmids and a S. fredii chromosome, and a symbiotic plasmid different from the one usually found in R. etli bv phaseoli. We infer that these plasmids originated through the transfer of a symbiotic-conjugative-plasmid

  14. Resolution of Multimeric Forms of Circular Plasmids and Chromosomes.

    Science.gov (United States)

    Crozat, Estelle; Fournes, Florian; Cornet, François; Hallet, Bernard; Rousseau, Philippe

    2014-10-01

    One of the disadvantages of circular plasmids and chromosomes is their high sensitivity to rearrangements caused by homologous recombination. Odd numbers of crossing-over occurring during or after replication of a circular replicon result in the formation of a dimeric molecule in which the two copies of the replicon are fused. If they are not converted back to monomers, the dimers of replicons may fail to correctly segregate at the time of cell division. Resolution of multimeric forms of circular plasmids and chromosomes is mediated by site-specific recombination, and the enzymes that catalyze this type of reaction fall into two families of proteins: the serine and tyrosine recombinase families. Here we give an overview of the variety of site-specific resolution systems found on circular plasmids and chromosomes.

  15. Conjugation of plasmids of Neisseria gonorrhoeae to other Neisseria species: potential reservoirs for the beta-lactamase plasmid.

    Science.gov (United States)

    Genco, C A; Knapp, J S; Clark, V L

    1984-09-01

    The discovery that penicillinase production in Neisseria gonorrhoeae was plasmid mediated and the spread of the beta-lactamase encoding plasmids in gonococcal isolates since 1976, raise the possibility that a nonpathogenic indigenous bacterium could serve as a reservoir for these plasmids. We initiated studies to define the ability of commensal Neisseria species and Branhamella catarrhalis strains, as well as strains of the pathogen Neisseria meningitidis, to serve as recipients in conjugation with Neisseria gonorrhoeae. We found that with N. gonorrhoeae as the donor, 3 of 5 Neisseria cinerea, 2 of 5 Neisseria flava, 0 of 1 Neisseria flavescens, 1 of 3 Neisseria subflava, 0 of 6 B. catarrhalis, 0 of 7 Neisseria lactamica, 1 of 5 Neisseria mucosa, 1 of 7 Neisseria perflava/sicca, and 0 of 13 N. meningitidis strains gave detectable conjugation frequencies (greater than 10(-8). N. cinerea was the only species found to maintain the gonococcal conjugal plasmid (pLE2451). A N. cinerea transconjugant containing pLE2451 was observed to transfer both the beta-lactamase plasmid and pLE2451 to N. gonorrhoeae at high frequency.

  16. Brane polarization is no cure for tachyons

    CERN Document Server

    Bena, Iosif

    2015-01-01

    Anti-M2 and anti-D3 branes placed in regions with charges dissolved in fluxes have a tachyon in their near-horizon region, which causes these branes to repel each other. If the branes are on the Coulomb branch this tachyon gives rise to a runaway behavior, but when the branes are polarized into five-branes this tachyon only appears to lower the energy of the polarized branes, without affecting its stability. We analyze brane polarization in the presence of a brane-brane-repelling tachyon and show that when the branes are polarized along the direction of the tachyon the polarized shell is unstable. This implies that tachyons cannot be cured by brane polarization and indicates that, at least in a certain regime of parameters, anti-D3 branes polarized into NS5 branes at the bottom of the Klebanov-Strassler solution have an instability.

  17. Ambient Cured Alkali Activated Flyash Masonry Units

    Science.gov (United States)

    Venugopal, K.; Radhakrishna; Sasalatti, Vinod M.

    2016-09-01

    Geopolymers belong to a category of non-conventional and non-Portland cement based cementitious binders which are produced using industrial by products like fly ash and ground granulated blast furnace slag (GGBFS). This paper reports on the development of geopolymer mortars for production of masonry units. The geopolymer mortars were prepared by mixing various by products with manufactured sand and a liquid mixture of sodium silicate and sodium hydroxide solutions. After curing at ambient conditions, the masonry units were tested for strength properties such as water absorption, initial rate of absorption, compression, shear- bond, and stress-strain behaviour etc. It was observed that the flexural strength of the blocks is more than 2 MPa and shear bond strength is more than 0.4MPa. It was found that the properties of geopolymer blocks were superior to the traditional masonry units. Hence they can be recommended for structural masonry.

  18. Helicobacter pylori: Basic Mechanisms to Clinical Cure

    Directory of Open Access Journals (Sweden)

    ABR Thomson

    1995-01-01

    Full Text Available Since its rediscovery 10 years ago, Helicobacter pylori has reshaped our thinking about the course of peptic ulcer disease. Our approach to the patient with a duodenal ulcer has become one of attempting eradication therapy at the time of first diagnosis, in the hope of curing the ulcer disease. Gastric and duodenal ulceration are only two of the manifestations of this chronic antral infection; other complications of H pylori include gastritis, gastric cancer and possible maltomas. Therapy of H pylori infection is complicated and involves dual therapy with an antibiotic plus a protein pump inhibitor, such as omeprazole 20 mg bid plus amoxicillin 1 g bid for two weeks, triple or quadruple therapy with bismuth, two antibiotics and an H2-receptor antagonist. Vaccination against H pylori is on the far horizon.

  19. Anion exchange purification of plasmid DNA using expanded bed adsorption.

    Science.gov (United States)

    Ferreira, G N; Cabral, J M; Prazeres, D M

    2000-01-01

    Recent developments in gene therapy with non-viral vectors and DNA vaccination have increased the demand for large amounts of pharmaceutical-grade plasmid DNA. The high viscosity of process streams is of major concern in the purification of plasmids, since it can cause high back pressures in column operations, thus limiting the throughput. In order to avoid these high back pressures, expanded bed anion exchange chromatography was evaluated as an alternative to fixed bed chromatography. A Streamline 25 column filled with 100 ml of Streamline QXL media, was equilibrated with 0.5 M NaCl in TE (10 mM Tris, 1 mM EDTA, pH = 8.0) buffer at an upward flow of 300 cmh-1, E. coli lysates (obtained from up to 3 liters of fermentation broth) were injected in the column. After washing out the unbound material, the media was allowed to sediment and the plasmid was eluted with 1 M NaCl in TE buffer at a downward flow of 120 cmh-1. Purification factors of 36 +/- 1 fold, 26 +/- 0.4 plasmid purity, and close to 100% yields were obtained when less than one settled column volume of plasmid feed was injected. However, both recovery yield and purity abruptly decreased when larger amounts were processed-values of 35 +/- 2 and 5 +/- 0.7 were obtained for the recovery yield and purity, respectively, when 250 ml of feedstock were processed. In these cases, gel clogging and expansion collapse were observed. The processing of larger volumes, thus larger plasmid quantities, was only possible by performing an isopropanol precipitation step prior to the chromatographic step. This step led to an enhancement of the purification step.

  20. Curing of epoxy matrix composite in stratosphere

    Science.gov (United States)

    Kondyurin, Alexey; Kondyurina, Irina; Bilek, Marcela

    Large structures for habitats, greenhouses, space bases, space factories are needed for next stage of space exploitation. A new approach enabling large-size constructions in space relies on the use of the polymerization technology of fiber-filled composites with a curable polymer matrix applied in the free space environment. The polymerisation process is proposed for the material exposed to high vacuum, dramatic temperature changes, space plasma, sun irradiation and atomic oxygen (in low Earth orbit), micrometeorite fluence, electric charging and microgravitation. The stratospheric flight experiments are directed to an investigation of the curing polymer matrix under the stratospheric conditions on. The unique combination of low atmospheric pressure, high intensity UV radiation including short wavelength UV and diurnal temperature variations associated with solar irradiation strongly influences the chemical processes in polymeric materials. The first flight experiment with uncured composites was a part of the NASA scientific balloon flight program realised at the NASA stratospheric balloon station in Alice Springs, Australia. A flight cassette installed on payload was lifted with a “zero-pressure” stratospheric balloon filled with Helium. Columbia Scientific Balloon Facility (CSBF) provided the launch, flight telemetry and landing of the balloon and payload. A cassette of uncured composite materials with an epoxy resin matrix was exposed 3 days in the stratosphere (40 km altitude). The second flight experiment was realised in South Australia in 2012, when the cassette was exposed in 27 km altitude. An analysis of the chemical structure of the composites showed, that the space irradiations are responsible for crosslinking of the uncured polymers exposed in the stratosphere. The first prepreg in the world was cured successfully in stratosphere. The investigations were supported by Alexander von Humboldt Foundation, NASA and RFBR (12-08-00970) grants.

  1. Stratospheric experiments on curing of composite materials

    Science.gov (United States)

    Chudinov, Viacheslav; Kondyurin, Alexey; Svistkov, Alexander L.; Efremov, Denis; Demin, Anton; Terpugov, Viktor; Rusakov, Sergey

    2016-07-01

    Future space exploration requires a large light-weight structure for habitats, greenhouses, space bases, space factories and other constructions. A new approach enabling large-size constructions in space relies on the use of the technology of polymerization of fiber-filled composites with a curable polymer matrix applied in the free space environment on Erath orbit. In orbit, the material is exposed to high vacuum, dramatic temperature changes, plasma of free space due to cosmic rays, sun irradiation and atomic oxygen (in low Earth orbit), micrometeorite fluence, electric charging and microgravitation. The development of appropriate polymer matrix composites requires an understanding of the chemical processes of polymer matrix curing under the specific free space conditions to be encountered. The goal of the stratospheric flight experiment is an investigation of the effect of the stratospheric conditions on the uncured polymer matrix of the composite material. The unique combination of low residual pressure, high intensity UV radiation including short-wave UV component, cosmic rays and other aspects associated with solar irradiation strongly influences the chemical processes in polymeric materials. We have done the stratospheric flight experiments with uncured composites (prepreg). A balloon with payload equipped with heater, temperature/pressure/irradiation sensors, microprocessor, carrying the samples of uncured prepreg has been launched to stratosphere of 25-30 km altitude. After the flight, the samples have been tested with FTIR, gel-fraction, tensile test and DMA. The effect of cosmic radiation has been observed. The composite was successfully cured during the stratospheric flight. The study was supported by RFBR grants 12-08-00970 and 14-08-96011.

  2. blaCMY-2-positive IncA/C plasmids from Escherichia coli and Salmonella enterica are a distinct component of a larger lineage of plasmids.

    Science.gov (United States)

    Call, Douglas R; Singer, Randall S; Meng, Da; Broschat, Shira L; Orfe, Lisa H; Anderson, Janet M; Herndon, David R; Kappmeyer, Lowell S; Daniels, Joshua B; Besser, Thomas E

    2010-02-01

    Large multidrug resistance plasmids of the A/C incompatibility complex (IncA/C) have been found in a diverse group of Gram-negative commensal and pathogenic bacteria. We present three completed sequences from IncA/C plasmids that originated from Escherichia coli (cattle) and Salmonella enterica serovar Newport (human) and that carry the cephamycinase gene blaCMY-2. These large plasmids (148 to 166 kbp) share extensive sequence identity and synteny. The most divergent plasmid, peH4H, has lost several conjugation-related genes and has gained a kanamycin resistance region. Two of the plasmids (pAM04528 and peH4H) harbor two copies of blaCMY-2, while the third plasmid (pAR060302) harbors a single copy of the gene. The majority of single-nucleotide polymorphisms comprise nonsynonymous mutations in floR. A comparative analysis of these plasmids with five other published IncA/C plasmids showed that the blaCMY-2 plasmids from E. coli and S. enterica are genetically distinct from those originating from Yersinia pestis and Photobacterium damselae and distal to one originating from Yersinia ruckeri. While the overall similarity of these plasmids supports the likelihood of recent movements among E. coli and S. enterica hosts, their greater divergence from Y. pestis or Y. ruckeri suggests less recent plasmid transfer among these pathogen groups.

  3. Cure Kinetics of DGEBA with Hyperbranched Poly(3-hydroxyphenyl) Phosphate as Curing Agent Studied by Non-isothermal DSC

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    The cure kinetics of diglycidyl ether of bisphenol A(DGEBA) with hyperbranched poly(3-hydroxyphenyl) phosphate(HHPP) as the curing agent was investigated by means of non-isothermal differential scanning calorimetry(DSC) at various heating rates. The results were compared with the corresponding results by using 1,3-dihydroxybenzene(DHB) as a model compound. The results show that HHPP can enhance the cure reaction of DGEBA, resulting in the decrease of the peak temperature of the curing curve as well as the decrease of the activation energy because of the flexible -P-O- groups in the backbone of HHPP. However, both the activation energy of the cured polymer and the peak temperature of the curing curve are increased with DHB as a curing agent. The cure kinetics of the DGEBA/HHPP system was calculated by using the isoconversional method given by Málek. It was found that the two-parameter autocatalytic model(esták-Berggren equation) is the most adequate one to describe the cure kinetics of the studied system at various heating rates. The obtained non-isothermal DSC curves from the experimental data show the results being accordant with those theoretically calculated.

  4. The effect of curing time and curing method on microleakge of conservative adhesive resin restorations: an in vitro study

    Directory of Open Access Journals (Sweden)

    Alireza Heidari

    2012-01-01

    Full Text Available Background and Aims: Using the conservative adhesive resin restoration (CAR in uncooperative children lead to numerous problems because of being time consuming. The purpose of this study was to compare the microleakage of conservative adhesive resin restoration under separate curing and co-curing.Materials and Methods: In this experimental study, 120 intact premolar teeth were collected and 120 vertical grooves were prepared on them. Then the teeth were divided into four groups: group 1, separated curing of bonding agent, flowable composite and sealant; group 2, co-curing of all materials for 60 seconds; group 3,co-curing of all materials for 40 seconds and group 4, co-curing of all materials for 20 seconds. Then the specimens were thermocycled and immersed in basic fuchsin solution. The teeth were sectioned horizontally and dye penetration was evaluated with stereomicroscope. Date were analyzed using one-way ANOVA and Scheffe test.Results: Mean value of dye penetration in groups 1, 2, 3, and 4 was 1.53±0.6, 2.06±0.6, 2.5±0.7 and 3.53±0.6, respectively. There was a statistically significant difference between group 1 and the other groups (P=0.0001.Conclusion: Considering the problems caused by microleakage in conservative resin adhesive restorations, co-curing method should not be used. In the case of using co-curing method, 60 second curing time is suggested for sufficient polymerization.

  5. Influence of curing tip distance on resin composite Knoop hardness number, using three different light curing units.

    Science.gov (United States)

    Caldas, Danilo Biazzetto de Menezes; de Almeida, Janaina Bertoncelo; Correr-Sobrinho, Lourenço; Sinhoreti, Mário Alexandre Coelho; Consani, Simonides

    2003-01-01

    This in vitro study evaluated the influence of curing tip distance on the Knoop Hardness Number (KHN) of a resin composite when using three different light curing units: (1) a halogen light (XL 1500 curing unit-3M), (2) a "softstart-polymerization" (Elipar Trilight curing in an exponential mode-ESPE) and (3) a PAC (Apolo 95E curing unit-DMD). The resin composite, Filtek Z250 (3M), was cured by these curing units at three light-tip distances from the resin composite: 0 mm, 6 mm and 12 mm. The resin composite specimens were flattened to their middle portion and submitted to 18 KHN measurements perspecimen. The results showed that for the Elipar Trilight unit, the hardness of the resin composite decreased as the light tip distance increased. The XL 1500 unit presented a significant decrease in hardness as the depth of cure of the resin composite increased. Apolo 95E caused a decrease in the resin composite hardness values when the depth of cure and light tip distance increased.

  6. 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 t

  7. 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

  8. 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

  9. Ribonucleases, antisense RNAs and the control of bacterial plasmids.

    Science.gov (United States)

    Saramago, Margarida; Bárria, Cátia; Arraiano, Cecília M; Domingues, Susana

    2015-03-01

    In the last decade regulatory RNAs have emerged as powerful tools to regulate the expression of genes both in prokaryotes and in eukaryotes. RNases, by degrading these RNA molecules, control the right amount of regulatory RNAs, which is fundamental for an accurate regulation of gene expression in the cell. Remarkably the first antisense RNAs identified were plasmid-encoded and their detailed study was crucial for the understanding of prokaryotic antisense RNAs. In this review we highlight the role of RNases in the precise modulation of antisense RNAs that control plasmid replication, maintenance and transfer.

  10. Micromechanical properties of veneer luting resins after curing through ceramics.

    Science.gov (United States)

    Oztürk, Elif; Hickel, Reinhard; Bolay, Sükran; Ilie, Nicoleta

    2012-02-01

    The aim of this study was to assess the performance of light-cured luting resin after curing under the ceramic restoration in comparison to dual-cured luting resin, by evaluating the micromechanical properties. Two hundred seventy thin luting composite films of ca. 170 μm in thickness were prepared by using two light-cured luting resins (Variolink Veneer, Ivoclar Vivadent; RelyX Veneer, 3M ESPE) and a dual-cured luting resin (Variolink II, Ivoclar Vivadent). The composites were cured by using a LED-unit (Bluephase®, Ivoclar Vivadent) with three different curing times (10, 20, and 30 s) under two ceramics (IPS e.max Press, Ivoclar Vivadent; IPS Empress® CAD, Ivoclar Vivadent) of different thicknesses (0, 0.75, and 2 mm). Forty-five groups were included, each containing six thin films. The samples were stored after curing for 24 h at 37°C by maintaining moisture conditions with distilled water. Micromechanical properties of the composites were measured with an automatic microhardness indenter (Fisherscope H100C, Germany). For each sample, ten indentations were made, thus totalizing 60 measurements per group. Micromechanical properties of the luting resins were statistically analyzed (SPSS 17.0). Significant differences were observed between the micromechanical properties of the luting resins (p mechanical properties compared to the light-cured luting resins. The effect of luting resin type on the micromechanical properties of the luting resins was higher than the effect of curing time, ceramic type and ceramic thickness respectively (*The values of reference without ceramics for 30 s curing time).

  11. Hardening of a dual-cure resin cement using QTH and LED curing units

    OpenAIRE

    Santos,Maria Jacinta Moraes Coelho; Passos, Sheila Pestana; Encarnação, Monalisa Olga Lessa da; Santos Junior,Gildo Coelho; Bottino, Marco Antonio [UNESP

    2010-01-01

    p. 110-115 OBJECTIVE: This study evaluated the surface hardness of a resin cement (RelyX ARC) photoactivated through indirect composite resin (Cristobal) disks of different thicknesses using either a light-emitting diode (LED) or quartz tungsten halogen (QTH) light source. MATERIAL AND METHODS: Eighteen resin cement specimens were prepared and divided into 6 groups according to the type of curing unit and the thickness of resin disks interposed between the cement surface and light sourc...

  12. Hardening of a dual-cure resin cement using QTH and LED curing units

    Directory of Open Access Journals (Sweden)

    Maria Jacinta Moraes Coelho Santos

    2010-04-01

    Full Text Available OBJECTIVE: This study evaluated the surface hardness of a resin cement (RelyX ARC photoactivated through indirect composite resin (Cristobal disks of different thicknesses using either a light-emitting diode (LED or quartz tungsten halogen (QTH light source. MATERIAL AND METHODS: Eighteen resin cement specimens were prepared and divided into 6 groups according to the type of curing unit and the thickness of resin disks interposed between the cement surface and light source. Three indentations (50 g for 15 s were performed on the top and bottom surface of each specimen and a mean Vickers hardness number (VHN was calculated for each specimen. The data were analyzed using two-way ANOVA and Tukey-Kramer test was used for post-hoc pairwise comparisons. RESULTS: Increased indirect resin disk thickness resulted in decreased mean VHN values. Mean VHN values for the top surfaces of the resin cement specimens ranged from 23.2 to 46.1 (QTH and 32.3 to 41.7 (LED. The LED curing light source produced higher hardness values compared to the QTH light source for 2- and 3-mm-thick indirect resin disks. The differences were clinically, but not statistically significant. Increased indirect resin disk thickness also resulted in decreased mean VHN values for the bottom surfaces of the resin cement: 5.8 to 19.1 (QTH and 7.5 to 32.0 (LED. For the bottom surfaces, a statistically significant interaction was also found between the type of curing light source and the indirect resin disk thickness. CONCLUSION: Mean surface hardness values of resin cement specimens decreased with the increase of indirect resin disk thickness. The LED curing light source generally produced higher surface hardness values.

  13. Cure Behaviors of epoxy resin initiated by methylanilinium slats as latent cationic curing agent.

    Energy Technology Data Exchange (ETDEWEB)

    Park, S.J.; Kim, T.J.; Lee, C.J.; Lee, J.R. [Korea Research Institute of Chemical Technology, Taejeon (Korea); Park, J.K.; Shin, J.H. [Seoul National University, Seoul (Korea)

    2001-03-01

    The effect of novel N-crotyl-N, N-dimethyl-4-methylanilinium hexafluoroantimonate (CMH) curing agent as a thermal latent initiator on thermal behaviors, rheological properties, and thermal stability of diglycidylether of bisphenol A (DGEBA) epoxy cationic system was investigated. From DSC measurements of DGEBA/CMH system, it was shown that this system exhibits an excellent thermal latent characteristic at a given temperature. The conversion and conversion rate of DGEBA/CMH system increased with increasing the concentration of initiator, due to high activity of CMH. Rheological properties of the system were investigated under isothermal condition using a rheometer. The gelation time was obtained from the analysis of storage modulus (G'), loss modulus (G''), and damping factor (tan {delta}). As a result, the reduction of gelation time was affected by high curing temperature and concentration of CMH, resulting in high degree of network formation in cationic polymerization, due to difference of activity. The thermal stability of the cured epoxy resin was discussed in terms of the activation energy for decomposition and thermal factors determined from TGA measurements. (author). 29 refs., 3 tabs., 10 figs.

  14. ACID RESISTANCE OF FLYASH BASED GEOPOLYMER MORTAR UNDER AMBIENT CURING AND HEAT CURING

    Directory of Open Access Journals (Sweden)

    V.Sreevidya

    2012-02-01

    Full Text Available An Experimental study was conducted to assess the Acid resistance of flyash based geopolymer mortar specimens of size 50x50x50mm with a ratio of flyash to sand as 1:3.The ratio between solution(Sodiumhydroxide and Sodium silicate solution to flyash were 0.376,0.386,0.396 and 0.416. After casting the specimens were subjected to both ambient curing and heat curing. In heat curing the specimens were kept continuously at 60oC for 24 hrs. Durability of specimens was assessed by immersing them in 5% of sulfuric acid and 5%hydrochloric acid for a period of 14 weeks. Evaluation of its resistance in terms of change in weight, compressive strength and visual appearance at regular intervals was carried out. After exposure in the acid solutions for 14 weeks, the samples showed very low weight loss. Results obtained from the present study indicate that Geopolymers are highly resistance to sulfuric acid and hydrochloric acid.

  15. Curing mechanism of alkaline phenolic resin with organic ester

    Institute of Scientific and Technical Information of China (English)

    Huang Renhe; Wang Yanmin; Zhang Baoping

    2014-01-01

    To study the curing mechanism of alkaline phenolic resin with organic ester, three esters were chosen to react with three systems - alkaline phenolic resin, potassium hydroxide aqueous solution containing phenol, and potassium hydroxide aqueous solution. The variations of pH, heat release and gel pH during the reactions were monitored and measured. Infrared spectroscopy (IR) and thermal gravity analysis (TG) techniques were used to characterize the curing reaction. It was found that organic ester is only partial y hydrolyzed and resin can be cured through organic ester hydrolysis process as wel as the reaction with redundant organic ester. The sequential curing mechanism of alkaline phenolic resin cured by organic ester was identified as fol ows: a portion of organic ester is firstly hydrolyzed owing to the effect of the strong alkaline; the gel is then formed after the pH decreases to about 10.8-10.88, meanwhile, the redundant organic ester (i.e. non-hydrolysis ester) starts the curing reaction with the resin. It has also been found that the curing rate depends on the hydrolysis velocity of organic ester. The faster the hydrolysis speed of the ester, the faster the curing rate of the resin.

  16. Shortest exposure time possible with LED curing lights

    NARCIS (Netherlands)

    Busemann, I.; Lipke, C.; Schattenberg, A.V.M.B.; Willershausen, B.; Ernst, C.P.

    2011-01-01

    PURPOSE: To investigate the shortest exposure time of different light emitting diode (LED)-curing devices for different resin composites in a clinically relevant laboratory model. METHODS: Nine LED curing devices (Bluephase, Bluephase 16i, Bluephase G2, Bluephase 20i/Ivoclar Vivadent, DEMI/sds Kerr,

  17. Improved method facilitates debulking and curing of phenolic impregnated asbestos

    Science.gov (United States)

    Gaines, P.

    1966-01-01

    Workpieces covered with phenolic impregnated asbestos tape and then wrapped with a specified thickness of nylon yarn under pressure, are debulked and cured in a standard oven. This method of debulking and curing is used in the fabrication of ablative chambers for the Gemini and Apollo attitude control engines.

  18. Shortest exposure time possible with LED curing lights

    NARCIS (Netherlands)

    Busemann, I.; Lipke, C.; Schattenberg, A.V.M.B.; Willershausen, B.; Ernst, C.P.

    2011-01-01

    PURPOSE: To investigate the shortest exposure time of different light emitting diode (LED)-curing devices for different resin composites in a clinically relevant laboratory model. METHODS: Nine LED curing devices (Bluephase, Bluephase 16i, Bluephase G2, Bluephase 20i/Ivoclar Vivadent, DEMI/sds Kerr,

  19. Isothermal epoxy-cure monitoring using nonlinear ultrasonics

    NARCIS (Netherlands)

    Koissin, V.; Demcenko, A.; Korneev, V.A.

    2014-01-01

    Isothermal curing of LY 1564SP resin in an aluminium-adhesive-aluminium laminate is investigated, using a nonlinear ultrasonic immersion technique, to prove its applicability for this type of dynamic material transformation. For verification and comparison, epoxy-cure kinetics and rheological behavi

  20. Conjugal transfer of group B streptococcal plasmids and comobilization of Escherichia coli-Streptococcus shuttle plasmids to Lactobacillus plantarum.

    OpenAIRE

    1988-01-01

    The antibiotic resistance group B streptococcal plasmids, pIP501 and pVA797, were conjugally transferred from Streptococcus faecalis to Lactobacillus plantarum. The Escherichia coli-Streptococcus shuttle plasmids, pVA838 and pSA3, were mobilized from S. sanguis to L. plantarum by pVA797 via cointegrate formation. pVA838 readily resolved from pVA797 and was present in L. plantarum as deletion derivatives. The pVA797::pSA3 cointegrate failed to resolve in L. plantarum.

  1. 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.

  2. Genetic Characterization of ExPEC-Like Virulence Plasmids among a Subset of NMEC.

    Directory of Open Access Journals (Sweden)

    Bryon A Nicholson

    Full Text Available Neonatal Meningitis Escherichia coli (NMEC is one of the most common causes of neonatal bacterial meningitis in the US and elsewhere resulting in mortality or neurologic deficits in survivors. Large plasmids have been shown experimentally to increase the virulence of NMEC in the rat model of neonatal meningitis. Here, 9 ExPEC-like plasmids were isolated from NMEC and sequenced to identify the core and accessory plasmid genes of ExPEC-like virulence plasmids in NMEC and create an expanded plasmid phylogeny. Results showed sequenced virulence plasmids carry a strongly conserved core of genes with predicted functions in five distinct categories including: virulence, metabolism, plasmid stability, mobile elements, and unknown genes. The major functions of virulence-associated and plasmid core genes serve to increase in vivo fitness by adding multiple iron uptake systems to the genetic repertoire to facilitate NMEC's survival in the host's low iron environment, and systems to enhance bacterial resistance to host innate immunity. Phylogenetic analysis based on these core plasmid genes showed that at least two lineages of ExPEC-like plasmids could be discerned. Further, virulence plasmids from Avian Pathogenic E. coli and NMEC plasmids could not be differentiated based solely on the genes of the core plasmid genome.

  3. Development of various curing effect of nominal strength Geopolymer concrete

    Directory of Open Access Journals (Sweden)

    S. Kumaravel

    2014-04-01

    Full Text Available Geopolymer concrete is an innovative method and is produced by complete elimination of ordinary Portland cement byproduced in fly ash. This study on different condition of curing in geopolymer concrete suitable for curing at ambient and heat-cured condition of temperature will widen its application to concrete structures. Low lime fly ash is used as the base material, which is reacted by alkaline solution and additional use of ground granulated blast furnace slag. Workability of fresh concrete and compressive strength of geopolymer concrete was investigated. The curing effect of geopolymer concrete is steam, hot air and ambient cubes specimens are tested in different days. Results are compared for various curing and strength of concrete.

  4. STUDY ON THE POST-CURE KINETICS OF EPOXY RESIN IRRADIATED BY AN ELECTRON BEAM

    Institute of Scientific and Technical Information of China (English)

    Jian-wen Bao; Ye Li; Xiang-yu Zhong; Xiang-bao Chen; Feng-mei Li

    2004-01-01

    The post-cure kinetics of electron beam (EB) curing of epoxy resin initiated by diaryiodinium was investigated.The post-cure reaction fits first order reaction kinetics. The reaction rate constant increases with increasing treatment temperature of post-cure. The reaction rate of post-cure is much lower than the rate of its reaction on electron beam treatment.

  5. 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 recombi

  6. Geminiviruses: a tale of a plasmid becoming a virus

    Directory of Open Access Journals (Sweden)

    Krupovic Mart

    2009-05-01

    Full Text Available Abstract Background Geminiviruses (family Geminiviridae are small single-stranded (ss DNA viruses infecting plants. Their virion morphology is unique in the known viral world – two incomplete T = 1 icosahedra are joined together to form twinned particles. Geminiviruses utilize a rolling-circle mode to replicate their genomes. A limited sequence similarity between the three conserved motifs of the rolling-circle replication initiation proteins (RCR Reps of geminiviruses and plasmids of Gram-positive bacteria allowed Koonin and Ilyina to propose that geminiviruses descend from bacterial replicons. Results Phylogenetic and clustering analyses of various RCR Reps suggest that Rep proteins of geminiviruses share a most recent common ancestor with Reps encoded on plasmids of phytoplasmas, parasitic wall-less bacteria replicating both in plant and insect cells and therefore occupying a common ecological niche with geminiviruses. Capsid protein of Satellite tobacco necrosis virus was found to be the best template for homology-based structural modeling of the geminiviral capsid protein. Good stereochemical quality of the generated models indicates that the geminiviral capsid protein shares the same structural fold, the viral jelly-roll, with the vast majority of icosahedral plant-infecting ssRNA viruses. Conclusion We propose a plasmid-to-virus transition scenario, where a phytoplasmal plasmid acquired a capsid-coding gene from a plant RNA virus to give rise to the ancestor of geminiviruses.

  7. 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 funct...

  8. Resistant plasmid profile analysis of multidrug resistant Escherichia ...

    African Journals Online (AJOL)

    Objectives: This study was carried out to determine the resistant plasmids of ... resistance pattern of micro-organisms to common an- tibiotics1 ... ment has necessitated the need for regular monitoring of antibiotics susceptibility trends to provide the basis for developing rational prescription programs, mak- ..... Paediatrics and.

  9. 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

  10. Use of plasmid DNA for induction of protective immunity

    DEFF Research Database (Denmark)

    Lorenzen, Niels

    2004-01-01

    Vaccines based on plasmid DNA have been tested for a number of fish pathogens but so far it is only in case of the rhabdoviruses, where the technology has been a real break through in vaccine research. Aspects of dose, time-course and mechanisms of protection, as well as practical use are discussed....

  11. Effects of maternal plasmid GHRH treatment on offspring growth

    Science.gov (United States)

    To differentiate prenatal effects of plasmid growth hormone-releasing hormone (GHRH) treatment from maternal effects mediated by lactation on long-term growth of offspring, a cross-fostering study was designed. Pregnant sows (n = 12) were untreated (n = 6), or received either a Wt-GHRH (n = 2), or H...

  12. [Chromatographic separation of plasmid DNA by anion-exchange cryogel].

    Science.gov (United States)

    Guo, Yantao; Shen, Shaochuan; Yun, Junxian; Yao, Kejian

    2012-08-01

    Plasmid DNA (pDNA) is used as an important vector for gene therapy, and its wide application is restricted by the purity and yield. To obtain high-purity pDNA, a chromatographic method based on anion-exchange supermacroporous cryogel was explored. The anion-exchange cryogel was prepared by grafting diethylaminoethyl-dextran to the epoxide groups of polyacrylamide-based matrix and pUC19 plasmid was used as a target to test the method. The plasmid was transferred into Escherichia coli DH5alpha, cultivated, harvested and lysed. The obtained culture was centrifuged and the supernatant was used as the plasmid feedstock, which was loaded into the anion-exchange cryogel bed for chromatographic separation. By optimizing the pH of running buffer and the elution conditions, high-purity pDNA was obtained by elution with 0.5 mol/L sodium chloride solution at pH 6.6. Compared to the traditional methods for purification of pDNA, animal source enzymes and toxic reagents were not involved in the present separation process, ensuring the safety of both the purification operations and the obtained pDNA.

  13. Plasmid containing a DNA ligase gene from Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

    McCarthy, D.; Griffin, K.; Setlow, J.K.

    1984-05-01

    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.

  14. Tragedy of the commons among antibiotic resistance plasmids.

    Science.gov (United States)

    Smith, Jeff

    2012-04-01

    As social interactions are increasingly recognized as important determinants of microbial fitness, sociobiology is being enlisted to better understand the evolution of clinically relevant microbes and, potentially, to influence their evolution to aid human health. Of special interest are situations in which there exists a "tragedy of the commons," where natural selection leads to a net reduction in fitness for all members of a population. Here, I demonstrate the existence of a tragedy of the commons among antibiotic resistance plasmids of bacteria. In serial transfer culture, plasmids evolved a greater ability to superinfect already-infected bacteria, increasing plasmid fitness when evolved genotypes were rare. Evolved plasmids, however, fell victim to their own success, reducing the density of their bacterial hosts when they became common and suffering reduced fitness through vertical transmission. Social interactions can thus be an important determinant of evolution for the molecular endosymbionts of bacteria. These results also identify an avenue of evolution that reduces proliferation of both antibiotic resistance genes and their bacterial hosts. © 2012 The Author(s). Evolution© 2012 The Society for the Study of Evolution.

  15. The replication origin of a repABC plasmid

    Directory of Open Access Journals (Sweden)

    Cevallos Miguel A

    2011-06-01

    Full Text Available Abstract Background repABC operons are present on large, low copy-number plasmids and on some secondary chromosomes in at least 19 α-proteobacterial genera, and are responsible for the replication and segregation properties of these replicons. These operons consist, with some variations, of three genes: repA, repB, and repC. RepA and RepB are involved in plasmid partitioning and in the negative regulation of their own transcription, and RepC is the limiting factor for replication. An antisense RNA encoded between the repB-repC genes modulates repC expression. Results To identify the minimal region of the Rhizobium etli p42d plasmid that is capable of autonomous replication, we amplified different regions of the repABC operon using PCR and cloned the regions into a suicide vector. The resulting vectors were then introduced into R. etli strains that did or did not contain p42d. The minimal replicon consisted of a repC open reading frame under the control of a constitutive promoter with a Shine-Dalgarno sequence that we designed. A sequence analysis of repC revealed the presence of a large A+T-rich region but no iterons or DnaA boxes. Silent mutations that modified the A+T content of this region eliminated the replication capability of the plasmid. The minimal replicon could not be introduced into R. etli strain containing p42d, but similar constructs that carried repC from Sinorhizobium meliloti pSymA or the linear chromosome of Agrobacterium tumefaciens replicated in the presence or absence of p42d, indicating that RepC is an incompatibility factor. A hybrid gene construct expressing a RepC protein with the first 362 amino acid residues from p42d RepC and the last 39 amino acid residues of RepC from SymA was able to replicate in the presence of p42d. Conclusions RepC is the only element encoded in the repABC operon of the R. etli p42d plasmid that is necessary and sufficient for plasmid replication and is probably the initiator protein. The ori

  16. Transfer of plasmid-mediated ampicillin resistance from Haemophilus to Neisseria gonorrhoeae requires an intervening organism.

    Science.gov (United States)

    McNicol, P J; Albritton, W L; Ronald, A R

    1986-01-01

    Haemophilus species have been implicated as the source of plasmid-mediated ampicillin resistance in Neisseria gonorrhoeae. Previous attempts to transfer conjugally the resistance plasmids from Haemophilus species to N. gonorrhoeae have met with limited success. Using both biparental and triparental mating systems, it was found that transfer will occur if the commensal Neisseria species, Neisseria cinerea, is used as a transfer intermediate. This organism stably maintains resistance plasmids of Haemophilus and facilitates transfer of these plasmids to N. gonorrhoeae, in a triparental mating system, at a transfer frequency of 10(-8). Both Haemophilus ducreyi and N. gonorrhoeae carry mobilizing plasmids capable of mediating conjugal transfer of the same resistance plasmids. However, restriction endonuclease mapping and DNA hybridization studies indicate that the mobilizing plasmids are distinctly different molecules. Limited homology is present within the transfer region of these plasmids.

  17. 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

  18. Participation of the lytic replicon in bacteriophage P1 plasmid maintenance.

    OpenAIRE

    1989-01-01

    P1 bacteriophage carries at least two replicons: a plasmid replicon and a viral lytic replicon. Since the isolated plasmid replicon can maintain itself stably at the low copy number characteristic of intact P1 prophage, it has been assumed that this replicon is responsible for driving prophage replication. We provide evidence that when replication from the plasmid replicon is prevented, prophage replication continues, albeit at a reduced rate. The residual plasmid replication is due to incomp...

  19. 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....

  20. Enhanced brain targeting efficiency of intranasally administered plasmid DNA: an alternative route for brain gene therapy.

    Science.gov (United States)

    Han, In-Kwon; Kim, Mi Young; Byun, Hyang-Min; Hwang, Tae Sun; Kim, Jung Mogg; Hwang, Kwang Woo; Park, Tae Gwan; Jung, Woon-Won; Chun, Taehoon; Jeong, Gil-Jae; Oh, Yu-Kyoung

    2007-01-01

    Recently, nasal administration has been studied as a noninvasive route for delivery of plasmid DNA encoding therapeutic or antigenic genes. Here, we examined the brain targeting efficiency and transport pathways of intranasally administered plasmid DNA. Quantitative polymerase chain reaction (PCR) measurements of plasmid DNA in blood and brain tissues revealed that intranasally administered pCMVbeta (7.2 kb) and pN2/CMVbeta (14.1 kb) showed systemic absorption and brain distribution. Following intranasal administration, the beta-galactosidase protein encoded by these plasmids was significantly expressed in brain tissues. Kinetic studies showed that intranasally administered plasmid DNA reached the brain with a 2,595-fold higher efficiency than intravenously administered plasmid DNA did, 10 min post-dose. Over 1 h post-dose, the brain targeting efficiencies were consistently higher for intranasally administered plasmid DNA than for intravenously administered DNA. To examine how plasmid DNA enters the brain and moves to the various regions, we examined tissues from nine brain regions, at 5 and 10 min after intranasal or intravenous administration of plasmid DNA. Intravenously administered plasmid DNA displayed similar levels of plasmid DNA in the nine different regions, whereas, intranasally administered plasmid DNA exhibited different levels of distribution among the regions, with the highest plasmid DNA levels in the olfactory bulb. Moreover, plasmid DNA was mainly detected in the endothelial cells, but not in glial cells. Our results suggest that intranasally applied plasmid DNA may reach the brain through a direct route, possibly via the olfactory bulb, and that the nasal route might be an alternative method for efficiently delivering plasmid DNA to the brain.

  1. Conservation of Plasmid-Encoded Traits among Bean-Nodulating Rhizobium Species

    OpenAIRE

    Brom, Susana; Girard, Lourdes; García-de los Santos, Alejandro; Sanjuan-Pinilla, Julio M.; Olivares, José; Sanjuan, Juan

    2002-01-01

    Rhizobium etli type strain CFN42 contains six plasmids. We analyzed the distribution of genetic markers from some of these plasmids in bean-nodulating strains belonging to different species (Rhizobium etli, Rhizobium gallicum, Rhizobium giardinii, Rhizobium leguminosarum, and Sinorhizobium fredii). Our results indicate that independent of geographic origin, R. etli strains usually share not only the pSym plasmid but also other plasmids containing symbiosis-related genes, with a similar organi...

  2. Effect of plasmid pKM101 in ultraviolet irradiated uvr+ and uvr- Escherichia coli.

    Science.gov (United States)

    Slezáriková, V; Sedliaková, M; Andreeva, I V; Rusina OYu; Skavronskaya, A G

    1992-11-16

    The effect of plasmid pKM101 on UV irradiated excision proficient and excision deficient cells was investigated. The plasmid increased the survival of excision proficient cells while partially inhibiting thymine dimer excision. The frequency of mutations was almost unchanged. In excision deficient cells the effect of the plasmid on survival was less pronounced while cell mutability was increased. Our data indicate that the mucAB genes (carried by the plasmid) influence the two types of cells in a different way.

  3. Role of the metalloprotease Vvp and the virulence plasmid pR99 of Vibrio vulnificus serovar E in surface colonization and fish virulence.

    Science.gov (United States)

    Valiente, Esmeralda; Lee, Chung-Te; Hor, Lien-I; Fouz, Belén; Amaro, Carmen

    2008-02-01

    The virulence for eels of Vibrio vulnificus biotype 2 serovar E (VSE) is conferred by a plasmid that codifies ability to survive in eel serum and cause septicaemia. To find out whether the plasmid and the selected chromosomal gene vvp plays a role in the initial steps of infection, the VSE strain CECT4999, the cured strain CT218 and the Vvp-deficient mutant CT201 (obtained in this work by allelic exchange) were used in colonization and virulence experiments. The eel avirulent biotype 1 (BT1) strain YJ016, whose genome has been sequenced, was used for comparative purposes. The global results demonstrate that the plasmid does not play a significant role in surface colonization because (i) CECT4999 and CT218 were equally chemoattracted towards and adherent to eel mucus and gills, and (ii) CT218 persisted in gills from bath-infected eels 2 weeks post infection. In contrast, mutation in vvp gene reduced significantly chemoattraction and attachment to eel mucus and gills, as well as virulence degree by immersion challenge. Co-infection experiments by bath with CECT4999 and CT201 confirmed that Vvp was involved in eel colonization and persistence in gills, because CECT4999 was recovered at higher numbers compared with CT201 from both internal organs of moribund fish (ratio 4:1) and gills from survivors (ratio 50:1). Interestingly, YJ016 also showed chemoattraction and attachment to mucus, and complementation of CT201 with BT1-vvp gene restored both activities together with virulence degree by immersion challenge. Additional experiments with algae mucus and purified mucin gave similar results. In conclusion, the protease Vvp of V. vulnificus seems to play an essential role in colonization of mucosal surfaces present in aquatic environments. Among the V. vulnificus strains colonizing fish mucus, only those harbouring the plasmid could survive in blood and cause septicaemia.

  4. Presence and analysis of plasmids in human and animal associated Arcobacter species

    DEFF Research Database (Denmark)

    Douidah, Laid; De Zutter, Lieven; Van Nieuwerburgh, Filip;

    2014-01-01

    In this study, we report the screening of four Arcobacter species for the presence of small and large plasmids. Plasmids were present in 9.9% of the 273 examined strains. One Arcobacter cryaerophilus and four Arcobacter butzleri plasmids were selected for further sequencing. The size of three sma...

  5. Occurrence and persistence of indigenous transconjugants carrying conjugative plasmids in soil.

    Science.gov (United States)

    Inoue, Daisuke; Soda, Satoshi; Tsutsui, Hirofumi; Yamazaki, Yuji; Murashige, Katsushi; Sei, Kazunari; Fujita, Masanori; Ike, Michihiko

    2009-09-01

    The transfer of the self-transmissible plasmids, RP4 and pJP4, from introduced bacteria to indigenous bacteria was examined in soil and slurry microcosms. The introduced plasmids persisted in indigenous transconjugants despite the low survival of introduced donors. The potential of the transconjugants for growth and conjugation affects the persistence of introduced plasmids in soil.

  6. Novel plasmid conferring kanamycin and tetracycline resistance in turkey-derived Campylobacter jejuni strain 11601MD

    Science.gov (United States)

    In Campylobacter spp., resistance to the antibiotics kanamycin and tetracycline is frequently associated with plasmid-borne genes. However, relatively few plasmids of Campylobacter jejuni have been fully characterized to date. A novel plasmid (p11601MD; 44,095 bp.) harboring tet(O) was identified in...

  7. 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 fixing nodules was investigated by hybridizati

  8. Mosaic structure and regulation of conjugal transfer of the Escherichia coli plasmid pRK100

    NARCIS (Netherlands)

    Starcic Erjavec, Marjanca

    2003-01-01

    Plasmids are extrachromosomal DNA elements that can be found in prokaryotic as well as in eukaryotic cells. They can vary in size and genetic make-up. The plasmid pRK100, which is the study subject of this thesis, is a large (145 kb) natural conjugative plasmid, which was isolated from an uropathoge

  9. Conjugal transfer of a virulence plasmid in the opportunistic intracellular actinomycete Rhodococcus equi.

    Science.gov (United States)

    Tripathi, V N; Harding, W C; Willingham-Lane, J M; Hondalus, M K

    2012-12-01

    Rhodococcus equi is a facultative intracellular, Gram-positive, soilborne actinomycete which can cause severe pyogranulomatous pneumonia with abscessation in young horses (foals) and in immunocompromised people, such as persons with AIDS. All strains of R. equi isolated from foals and approximately a third isolated from humans contain a large, ~81-kb plasmid which is essential for the intramacrophage growth of the organism and for virulence in foals and murine in vivo model systems. We found that the entire virulence plasmid could be transferred from plasmid-containing strains of R. equi (donor) to plasmid-free R. equi strains (recipient) at a high frequency and that plasmid transmission reestablished the capacity for intracellular growth in macrophages. Plasmid transfer required living cells and cell-to-cell contact and was unaffected by the presence of DNase, factors pointing to conjugation as the major means of genetic transfer. Deletion of a putative relaxase-encoding gene, traA, located in the proposed conjugative region of the plasmid, abolished plasmid transfer. Reversion of the traA mutation restored plasmid transmissibility. Finally, plasmid transmission to other Rhodococcus species and some additional related organisms was demonstrated. This is the first study showing a virulence plasmid transfer in R. equi, and it establishes a mechanism by which the virulence plasmid can move among bacteria in the soil.

  10. The evolution of a conjugative plasmid and its ability to increase bacterial fitness

    Science.gov (United States)

    Dionisio, F; Conceição, I.C; Marques, A.C.R; Fernandes, L; Gordo, I

    2005-01-01

    Conjugative plasmids are extra-chromosomal DNA elements that are capable of horizontal transmission and are found in many natural isolated bacteria. Although plasmids may carry beneficial genes to their bacterial host, they may also cause a fitness cost. In this work, we studied the evolution of the R1 plasmid and we found that, in spite of the R1 plasmid conferring an initial cost to its host, after 420 generations the cost disappeared in all five independent evolution experiments. In fact, in two of these five experiments evolved conjugative plasmids actually conferred a fitness advantage to their hosts. Furthermore, the relative fitness of the ancestral clone bearing one of the evolved plasmids is significantly higher than both the plasmid-free ancestral cells and the evolved cells carrying the evolved plasmid. Given that the R1 plasmid may spread among different species of enterobacteria, we wondered what the effect of the evolved plasmid would be inside Salmonella enterica cells. We found that the evolved plasmid is also able to dramatically increase the relative fitness of these cells. Our results suggest that even if general usage of antibiotics is halted, conjugative plasmids that have been selected with antibiotics in previous years can still persist among bacterial populations or even invade new strains. PMID:17148179

  11. A Bipolar Spindle of Antiparallel ParM Filaments Drives Bacterial Plasmid Segregation

    DEFF Research Database (Denmark)

    Gayathri, P; Fujii, T; Møller-Jensen, Jakob;

    2012-01-01

    To ensure their stable inheritance by daughter cells during cell division, bacterial low copy-number plasmids make simple DNA segregating machines that use an elongating protein filament between sister plasmids. In the ParMRC system of Escherichia coli R1 plasmid, ParM, an actin-like protein, forms...

  12. Cure kinetics and chemorheology of EPDM/graphene oxide nanocomposites

    Energy Technology Data Exchange (ETDEWEB)

    Allahbakhsh, Ahmad [Department of Polymer Engineering, Islamic Azad University, South Tehran Branch, 17776-13651 Tehran (Iran, Islamic Republic of); Mazinani, Saeedeh, E-mail: s.mazinani@aut.ac.ir [Amirkabir Nanotechnology Research Institute (ANTRI), Amirkabir University of Technology, Tehran (Iran, Islamic Republic of); Kalaee, Mohammad Reza [Department of Polymer Engineering, Islamic Azad University, South Tehran Branch, 17776-13651 Tehran (Iran, Islamic Republic of); Sharif, Farhad [Department of Polymer Engineering and Color Technology, Amirkabir University of Technology, Tehran (Iran, Islamic Republic of)

    2013-07-10

    Graphical abstract: - Highlights: • Graphene oxide content and dispersion as effective parameters on cure kinetics. • Graphene oxide as an effective controlling factor of crosslink density. • Interaction of graphene oxide with curing system (ZnO) during curing process. - Abstract: In this study, the effect of graphene oxide on cure behavior of ethylene–propylene–diene rubber (EPDM) nanocomposite is studied. In this regard, the cure kinetics of nanocomposite is studied employing different empirical methods. The required activation energy of nth-order cure process shows about 160 kJ/mol increments upon 5 phr graphene oxide loading compared to 1 phr graphene oxide loading. However, the required activation energy is significantly reduced followed by incorporation of graphene oxide in nanocomposites compared to neat EPDM sample. Furthermore, the effect of graphene oxide on structural properties of nanocomposites during the cure process is studied using X-ray diffraction, scanning electron microscopy and Fourier transform infrared spectrometry techniques. As the results show, graphene oxide interestingly affects the structure of zinc oxide during the vulcanization process. This behavior could be probably related to high tendency of zinc oxide to react with oxidized surface of graphene oxide.

  13. Development of a resin curing model for UV nanoimprint.

    Science.gov (United States)

    Lee, Jin Woo; Cho, Dong-Woo

    2005-10-01

    UV nanoimprint lithography uses UV light as an energy source. It is performed at room temperature and low pressure, and has its own merits as compared to thermal nanoimprint. In this paper, a measurement system was developed to measure the degree of resin curing in UV nanoimprint to improve our understanding of the resin solidification phenomenon. A curing model was then established based on the measurement results. The measurement system measured the degree of cure in real time and was composed of a Fourier transform infrared spectroscopy system, a UV light source, and an optical guide. Also, new UV-curable resins that had low viscosity values were developed for the UV nanoimprint process, and imprint tests using these resins were performed successfully. The curing model considered the UV irradiation time, power, and curing temperature, which are important parameters in the UV nanoimprint process. The degree of cure had an exponential relation to UV irradiation time, power, and temperature; thus, the curing model was expressed as an exponential function of the UV irradiation time, power, and temperature. The developed model was verified for various UV-curable resins.

  14. Composite depth of cure using four polymerization techniques

    Directory of Open Access Journals (Sweden)

    Ericson Janolio de Camargo

    2009-10-01

    Full Text Available The light-curing technique is relevant to reduce the degree of polymerization shrinkage, improving clinical and esthetic success of composite resin restorations. OBJECTIVE: To evaluate in vitro the effect of four light-curing techniques on depth of cure of a composite resin. MATERIAL AND METHODS: Ten specimens of a composite resin were made in cylindrical cavities prepared in PVC plates (3.0 X 7.0 mm for each light-curing technique. Four photoactivation methods were investigated: stepped, ramped, pulse-delay and traditional. Specimens were longitudinally sectioned and polished for microhardness measurements (kg/mm², which were made at 0.1, 1.0, 2.0 and 4.0 mm from the irradiated surface. Data were subjected to ANOVA and Tukey's test. RESULTS: The effect of factors studied (curing method and distance from the surface and the interaction of these factors was statistically significant (p<0.05. The traditional method of cure provided higher microhardness values (69.6 ± 2.5 than the stepped (63.5 ± 3.1 and pulsed (63.9 ± 3.2 methods at all depths evaluated, but it did not differ from the ramped method (66.7± 4.4 at 0.1 and 1.0 mm of depth. CONCLUSION: All techniques employed provided satisfactory cure of the composite resin up to the depth of 2.0 mm from the irradiated surface.

  15. A 'cure' for Down syndrome: what do parents want?

    Science.gov (United States)

    Inglis, A; Lohn, Z; Austin, J C; Hippman, C

    2014-10-01

    Recent advancements in molecular genetics raise the possibility that therapeutics or a 'cure' for Down syndrome (DS) may become available. However, there are no data regarding how parents of children with DS perceive the possibility of mitigating specific manifestations such as the intellectual disability (ID) associated with DS, or curing the condition entirely. To explore these issues, we distributed a questionnaire to members of the Lower Mainland Down Syndrome Society in British Columbia, Canada. Questionnaires were completed by 101 parents (response rate=41%). A majority (61%) viewed the possibility of reversing ID in DS positively, but only 41% said that they would 'cure' their child of DS if it were possible. Twenty-seven percent of respondents said they would not 'cure' their child, and 32% were unsure if they would 'cure' their child. The most commonly cited motivation for opting for a 'cure' was to increase their child's independence. However, parental attitudes' towards a 'cure' for DS were complex, affected by ethical issues, perceived societal values, and pragmatic factors such as the age of the individual and long-term care-giving burden. These findings could be used by healthcare professionals supporting families who include a member with DS and to direct future research.

  16. Electron Beam Cured Epoxy Resin Composites for High Temperature Applications

    Science.gov (United States)

    Janke, Christopher J.; Dorsey, George F.; Havens, Stephen J.; Lopata, Vincent J.; Meador, Michael A.

    1997-01-01

    Electron beam curing of Polymer Matrix Composites (PMC's) is a nonthermal, nonautoclave curing process that has been demonstrated to be a cost effective and advantageous alternative to conventional thermal curing. Advantages of electron beam curing include: reduced manufacturing costs; significantly reduced curing times; improvements in part quality and performance; reduced environmental and health concerns; and improvement in material handling. In 1994 a Cooperative Research and Development Agreement (CRADA), sponsored by the Department of Energy Defense Programs and 10 industrial partners, was established to advance the electron beam curing of PMC technology. Over the last several years a significant amount of effort within the CRADA has been devoted to the development and optimization of resin systems and PMCs that match the performance of thermal cured composites. This highly successful materials development effort has resulted in a board family of high performance, electron beam curable cationic epoxy resin systems possessing a wide range of excellent processing and property profiles. Hundreds of resin systems, both toughened and untoughened, offering unlimited formulation and processing flexibility have been developed and evaluated in the CRADA program.

  17. Variation of Resin Properties Through the Thickness of Cured Samples

    Science.gov (United States)

    1984-01-01

    It is the purpose of this work to gain knowledge of the glassy materials used as matrices in composites and to study the homogeneity resulting from the curing process. An attempt is made to link the glass transition quantitatively with the presence of a given material. Expoxy resins containing various amounts of hardener (TGDDM/DDS system) were cured in a muffle furnace at 473 K for seven hours. The glass transition temperature, T sub g versus weight minus percent of hardener in the epoxy resin were measured. A limit was rapidly reached in T sub g at only two percent hardener. Thus, the glass transition of the fully cured epoxy-amine matrix seems not much different from the epoxide-epoxide cure. The T sub g versus cure-time for the epoxide-epoxide reaction was also studied. My 720 was cured by itself in an oil bath at 473 K for different lengths of time. The T sub g was found to increase exponentially with the cure time, and a maximum T sub g of about 450 K was reached after eleven hours. The reaction was found to be inhibited by running the sample under argon.

  18. Development of Geopolymer Concrete with Different Curing Conditions

    Directory of Open Access Journals (Sweden)

    M.F. Nuruddin Nuruddin

    2011-02-01

    Full Text Available Environmental issues resulted from cement production have become a major concern today. To develop a sustainable future it is encouraged to limit the use of this construction material that can affect the environment. Cement replacement material was proposed to partially replace cement portion in concrete. Geopolymer is a part of inorganic polymer material that has similar bonding function like cement in concrete. It consists of alkaline solutions and geological source material. Alkaline liquids used in this research are 8 M sodium hydroxide (NaOH solution and sodium silicate (Na2SiO3 solutions, while source materials are fly ash and microwave incinerated rice husk ash (MIRHA. Three different curing regimes, namely hot gunny curing, ambient curing, and external exposure curing, were applied to obtain suitable method that was suitable with cast in situ application. Geopolymer concrete samples were tested on their compressive strength and microstructure properties. It was found that external exposure curing had the highest compressive strength compared to other two curing methods. Scanning electron microscopy analysis also showed better improvement in interfacial transition zone for concrete sample with external exposure curing.

  19. Plasmid diversity and phylogenetic consistency in the Lyme disease agent Borrelia burgdorferi.

    Science.gov (United States)

    Casjens, Sherwood R; Gilcrease, Eddie B; Vujadinovic, Marija; Mongodin, Emmanuel F; Luft, Benjamin J; Schutzer, Steven E; Fraser, Claire M; Qiu, Wei-Gang

    2017-02-15

    Bacteria from the genus Borrelia are known to harbor numerous linear and circular plasmids. We report here a comparative analysis of the nucleotide sequences of 236 plasmids present in fourteen independent isolates of the Lyme disease agent B. burgdorferi. We have sequenced the genomes of 14 B. burgdorferi sensu stricto isolates that carry a total of 236 plasmids. These individual isolates carry between seven and 23 plasmids. Their chromosomes, the cp26 and cp32 circular plasmids, as well as the lp54 linear plasmid, are quite evolutionarily stable; however, the remaining plasmids have undergone numerous non-homologous and often duplicative recombination events. We identify 32 different putative plasmid compatibility types among the 236 plasmids, of which 15 are (usually) circular and 17 are linear. Because of past rearrangements, any given gene, even though it might be universally present in these isolates, is often found on different linear plasmid compatibility types in different isolates. For example, the arp gene and the vls cassette region are present on plasmids of four and five different compatibility types, respectively, in different isolates. A majority of the plasmid types have more than one organizationally different subtype, and the number of such variants ranges from one to eight among the 18 linear plasmid types. In spite of this substantial organizational diversity, the plasmids are not so variable that every isolate has a novel version of every plasmid (i.e., there appears to be a limited number of extant plasmid subtypes). Although there have been many past recombination events, both homologous and nonhomologous, among the plasmids, particular organizational variants of these plasmids correlate with particular chromosomal genotypes, suggesting that there has not been rapid horizontal transfer of whole linear plasmids among B. burgdorferi lineages. We argue that plasmid rearrangements are essentially non-revertable and are present at a frequency of

  20. Changes on degree of conversion of dual-cure luting light-cured with blue LED

    Science.gov (United States)

    Bandéca, M. C.; El-Mowafy, O.; Saade, E. G.; Rastelli, A. N. S.; Bagnato, V. S.; Porto-Neto, S. T.

    2009-05-01

    The indirect adhesive procedures constitute recently a substantial portion of contemporary esthetic restorative treatments. The resin cements have been used to bond tooth substrate and restorative materials. Due to recently introduction of the self-bonding resin luting cement based on a new monomer, filler and initiation technology has become important to study the degree of conversion of these new materials. In the present work the polymerization reaction and the filler content of dual-cured dental resin cements were studied by means of infra-red spectroscopy (FT-IR) and thermogravimetry (TG). Twenty specimens were made in a metallic mold (8 mm diameter × 1 mm thick) from each of 2 cements, Panavia® F2.0 (Kuraray) and RelyX™ Unicem Applicap (3M/ESPE). Each specimen was cured with blue LED with power density of 500 mW/cm2 for 30 s. Immediately after curing, 24 and 48 h, and 7 days DC was determined. For each time interval 5 specimens were pulverized, pressed with KBr and analyzed with FT-IR. The TG measurements were performed in Netzsch TG 209 under oxygen atmosphere and heating rate of 10°C/min from 25 to 700°C. A two-way ANOVA showed DC (%) mean values statistically significance differences between two cements ( p 0.05). The Relx-Y™ Unicem mean values were significantly higher than Panavia® F 2.0. The degree of conversion means values increasing with the storage time and the filler content showed similar for both resin cements.

  1. Experimental observation of internal water curing of concrete

    DEFF Research Database (Denmark)

    Lura, Pietro; Jensen, Ole Mejlhede

    2007-01-01

    Internal water curing has a significant effect on concrete. In addition to affecting hydration and moisture distribution, it influences most concrete properties, such as strength, shrinkage, cracking, and durability. The following paper is an overview of experimental methods to study internal water...... curing of concrete and its consequences. The special techniques needed to study internal water curing are dealt with along with the consequences of this process. Examples of applications are given and new measuring techniques that may potentially be applied to this field are addressed....

  2. Determination of plasmid copy number reveals the total plasmid DNA amount is greater than the chromosomal DNA amount in Bacillus thuringiensis YBT-1520.

    Directory of Open Access Journals (Sweden)

    Chunying Zhong

    Full Text Available Bacillus thuringiensis is the most widely used bacterial bio-insecticide, and most insecticidal crystal protein-coding genes are located on plasmids. Most strains of B. thuringiensis harbor numerous diverse plasmids, although the plasmid copy numbers (PCNs of all native plasmids in this host and the corresponding total plasmid DNA amount remains unknown. In this study, we determined the PCNs of 11 plasmids (ranging from 2 kb to 416 kb in a sequenced B. thuringiensis subsp. kurstaki strain YBT-1520 using real-time qPCR. PCNs were found to range from 1.38 to 172, and were negatively correlated to plasmid size. The amount of total plasmid DNA (∼8.7 Mbp was 1.62-fold greater than the amount of chromosomal DNA (∼5.4 Mbp at the mid-exponential growth stage (OD(600 = 2.0 of the organism. Furthermore, we selected three plasmids with different sizes and replication mechanisms to determine the PCNs over the entire life cycle. We found that the PCNs dynamically shifted at different stages, reaching their maximum during the mid-exponential growth or stationary phases and remaining stable and close to their minimum after the prespore formation stage. The PCN of pBMB2062, which is the smallest plasmid (2062 bp and has the highest PCN of those tested, varied in strain YBT-1520, HD-1, and HD-136 (172, 115, and 94, respectively. These findings provide insight into both the total plasmid DNA amount of B. thuringiensis and the strong ability of the species to harbor plasmids.

  3. Remarkable stability of an instability-prone lentiviral vector plasmid in Escherichia coli Stbl3.

    Science.gov (United States)

    Al-Allaf, Faisal A; Tolmachov, Oleg E; Zambetti, Lia Paola; Tchetchelnitski, Viktoria; Mehmet, Huseyin

    2013-02-01

    Large-scale production of plasmid DNA to prepare therapeutic gene vectors or DNA-based vaccines requires a suitable bacterial host, which can stably maintain the plasmid DNA during industrial cultivation. Plasmid loss during bacterial cell divisions and structural changes in the plasmid DNA can dramatically reduce the yield of the desired recombinant plasmid DNA. While generating an HIV-based gene vector containing a bicistronic expression cassette 5'-Olig2cDNA-IRES-dsRed2-3', we encountered plasmid DNA instability, which occurred in homologous recombination deficient recA1 Escherichia coli strain Stbl2 specifically during large-scale bacterial cultivation. Unexpectedly, the new recombinant plasmid was structurally changed or completely lost in 0.5 L liquid cultures but not in the preceding 5 mL cultures. Neither the employment of an array of alternative recA1 E. coli plasmid hosts, nor the lowering of the culture incubation temperature prevented the instability. However, after the introduction of this instability-prone plasmid into the recA13E. coli strain Stbl3, the transformed bacteria grew without being overrun by plasmid-free cells, reduction in the plasmid DNA yield or structural changes in plasmid DNA. Thus, E. coli strain Stbl3 conferred structural and maintenance stability to the otherwise instability-prone lentivirus-based recombinant plasmid, suggesting that this strain can be used for the faithful maintenance of similar stability-compromised plasmids in large-scale bacterial cultivations. In contrast to Stbl2, which is derived wholly from the wild type isolate E. coli K12, E. coli Stbl3 is a hybrid strain of mixed E. coli K12 and E. coli B parentage. Therefore, we speculate that genetic determinants for the benevolent properties of E. coli Stbl3 for safe plasmid propagation originate from its E. coli B ancestor.

  4. Micro-leakage of a Fissure Sealant Cured Using Quartz-tungsten-halogen and Plasma Arc Light Curing Units.

    Science.gov (United States)

    Bahrololoomi, Zahra; Soleimani, Ali Asghar; Jafari, Najmeh; Varkesh, Bentolhoda

    2014-01-01

    Background and aims. Newer curing units such as plasma arc can polymerize the sealants in much shorter curing times. The aim of this study was to compare the effect of two different curing units on the micro-leakage of a fissure sealant material. Materials and methods. Sixty two extracted premolars without caries were randomly divided into two groups of 31 samples. Occlusal surfaces of all teeth were cleansed. Then, teeth surfaces were etched by 37% phosphoric acid. After rinsing and drying, occlusal surfaces of teeth were sealed by a fissure sealant. The sealant was then cured using either a halogen light curing unit or a plasma arc curing light. After sealing, the teeth were thermocycled for 500 cycles. The teeth were then sectioned and examined for micro-leakage. Statistical analyses were performed with Mann-Whitney test. Results. There was no significant difference between two groups regarding micro-leakage (P = 0.42). Conclusion. Results showed that there was no significant difference between two different curing units. Therefore, plasma arc unit might be a useful alternative for sealant polymerization.

  5. Type 3 Fimbriae Encoded on Plasmids Are Expressed from a Unique Promoter without Affecting Host Motility, Facilitating an Exceptional Phenotype That Enhances Conjugal Plasmid Transfer

    DEFF Research Database (Denmark)

    Madsen, Jonas Stenlokke; Riber, Leise; Kot, Witold;

    2016-01-01

    on plasmids is systematically different, as MrkH, a c-di-GMP dependent transcriptional activator is not needed for strong expression of the fimbriae. MrkH is required for expression of type 3 fimbriae of the Klebsiella pneumoniae chromosome, wherefrom the fimbriae operon (mrkABCDF) of plasmids is believed...... to have originated. We find that mrkABCDFs of plasmids are highly expressed via a unique promoter that differs from the original Klebsiella promoter resulting in fundamental behavioral consequences. Plasmid associated mrkABCDFs did not influence the swimming behavior of the host, that hereby acquired...

  6. Coupling between the Basic Replicon and the Kis-Kid Maintenance System of Plasmid R1: Modulation by Kis Antitoxin Levels and Involvement in Control of Plasmid Replication

    Directory of Open Access Journals (Sweden)

    Juan López-Villarejo

    2015-02-01

    Full Text Available kis-kid, the auxiliary maintenance system of plasmid R1 and copB, the auxiliary copy number control gene of this plasmid, contribute to increase plasmid replication efficiency in cells with lower than average copy number. It is thought that Kis antitoxin levels decrease in these cells and that this acts as the switch that activates the Kid toxin; activated Kid toxin reduces copB-mRNA levels and this increases RepA levels that increases plasmid copy number. In support of this model we now report that: (i the Kis antitoxin levels do decrease in cells containing a mini-R1 plasmid carrying a repA mutation that reduces plasmid copy number; (ii kid-dependent replication rescue is abolished in cells in which the Kis antitoxin levels or the CopB levels are increased. Unexpectedly we found that this coordination significantly increases both the copy number of the repA mutant and of the wt mini-R1 plasmid. This indicates that the coordination between plasmid replication functions and kis-kid system contributes significantly to control plasmid R1 replication.

  7. Measuring Vapors To Monitor the State of Cure of a Resin

    Science.gov (United States)

    Cramer, Elliott; Perey, Daniel F.; Yost, William T.

    2006-01-01

    A proposed noninvasive method of monitoring the cure path and the state of cure of an epoxy or other resin involves measurement of the concentration( s) of one or more compound(s) in the vaporous effluent emitted during the curing process. The method is based on the following general ideas: (1) The concentrations of the effluent compounds in the vicinity of the curing resin are approximately proportional to the instantaneous rate of curing. (2) As curing proceeds at a given temperature, subsequent decreases in the concentrations are indicative of approaching completion of cure; that is, the lower are the concentrations, the more nearly complete is the cure.

  8. The effect of curing light and chemical catalyst on the degree of conversion of two dual cured resin luting cements.

    Science.gov (United States)

    Souza-Junior, Eduardo José; Prieto, Lúcia Trazzi; Soares, Giulliana Panfiglio; Dias, Carlos Tadeu dos Santos; Aguiar, Flávio Henrique Baggio; Paulillo, Luís Alexandre Maffei Sartini

    2012-01-01

    The aim of this study was to evaluate the influence of different curing lights and chemical catalysts on the degree of conversion of resin luting cements. A total of 60 disk-shaped specimens of RelyX ARC or Panavia F of diameter 5 mm and thickness 0.5 mm were prepared and the respective chemical catalyst (Scotchbond Multi-Purpose Plus or ED Primer) was added. The specimens were light-cured using different curing units (an argon ion laser, an LED or a quartz-tungsten-halogen light) through shade A2 composite disks of diameter 10 mm and thickness 2 mm. After 24 h of dry storage at 37°C, the degree of conversion of the resin luting cements was measured by Fourier-transformed infrared spectroscopy. For statistical analysis, ANOVA and the Tukey test were used, with p ≤ 0.05. Panavia F when used without catalyst and cured using the LED or the argon ion laser showed degree of conversion values significantly lower than RelyX ARC, with and without catalyst, and cured with any of the light sources. Therefore, the degree of conversion of Panavia F with ED Primer cured with the quartz-tungsten-halogen light was significantly different from that of RelyX ARC regardless of the use of the chemical catalyst and light curing source. In conclusion, RelyX ARC can be cured satisfactorily with the argon ion laser, LED or quartz-tungsten-halogen light with or without a chemical catalyst. To obtain a satisfactory degree of conversion, Panavia F luting cement should be used with ED Primer and cured with halogen light.

  9. Remarkable stability of an instability-prone lentiviral vector plasmid in Escherichia coli Stbl3

    OpenAIRE

    Al-Allaf, Faisal A.; Tolmachov, Oleg E.; Zambetti, Lia Paola; Tchetchelnitski, Viktoria; Mehmet, Huseyin

    2012-01-01

    Large-scale production of plasmid DNA to prepare therapeutic gene vectors or DNA-based vaccines requires a suitable bacterial host, which can stably maintain the plasmid DNA during industrial cultivation. Plasmid loss during bacterial cell divisions and structural changes in the plasmid DNA can dramatically reduce the yield of the desired recombinant plasmid DNA. While generating an HIV-based gene vector containing a bicistronic expression cassette 5′-Olig2cDNA-IRES-dsRed2-3′, we encountered ...

  10. An Improved Method for Including Upper Size Range Plasmids in Metamobilomes

    DEFF Research Database (Denmark)

    Norman, Anders; Riber, Leise; Luo, Wenting

    2014-01-01

    cloning vector (pBR322), and a 56 Kbp conjugative plasmid (pKJK10), to represent lower- and upper plasmid size ranges, respectively. Subjecting a mixture of these plasmids to the overall isolation protocol revealed a 34-fold over-amplification of pBR322 after MDA. To address this bias, we propose......, as gene functions associated with these plasmids, such as conjugation, was exclusively encoded in the data output generated through the modified protocol. Thus, with the suggested modification, access to a large uncharacterized pool of accessory elements that reside on medium-to-large plasmids has been...

  11. Large plasmids of Escherichia coli and Salmonella encode highly diverse arrays of accessory genes on common replicon families.

    Science.gov (United States)

    Williams, Laura E; Wireman, Joy; Hilliard, Valda C; Summers, Anne O

    2013-01-01

    Plasmids are important in evolution and adaptation of host bacteria, yet we lack a comprehensive picture of their own natural variation. We used replicon typing and RFLP analysis to assess diversity and distribution of plasmids in the ECOR, SARA, SARB and SARC reference collections of Escherichia coli and Salmonella. Plasmids, especially large (≥30 kb) plasmids, are abundant in these collections. Host species and genotype clearly impact plasmid prevalence; plasmids are more abundant in ECOR than SAR, but, within ECOR, subgroup B2 strains have the fewest large plasmids. The majority of large plasmids have unique RFLP patterns, suggesting high variation, even within dominant replicon families IncF and IncI1. We found only four conserved plasmid types within ECOR, none of which are widely distributed. Within SAR, conserved plasmid types are primarily serovar-specific, including a pSLT-like plasmid in 13 Typhimurium strains. Conservation of pSLT contrasts with variability of other plasmids, suggesting evolution of serovar-specific virulence plasmids is distinct from that of most enterobacterial plasmids. We sequenced a conserved serovar Heidelberg plasmid but did not detect virulence or antibiotic resistance genes. Our data illustrate the high degree of natural variation in large plasmids of E. coli and Salmonella, even among plasmids sharing backbone genes.

  12. Comparative Study of Alkali-Activated Fly Ash Manufactured Under Pulsed Microwave Curing and Thermal Oven Curing

    OpenAIRE

    Shi, Shi; Bai, Yun; Li, H.; Xu, D. L.; Basheer, P. A. Muhammed

    2014-01-01

    This paper compares the alkali-activated fly ash (AAFA) manufactured with thermal oven curing and pulsed microwave curing methods. Fly ash activated by 8M NaOH solution at a liquid to solid ratio of 0.3 was cured by thermal oven at 85°C and domestic microwave oven, respectively. Apart from compressive strength test, the temperature profiles of AAFA samples were captured by thermal camera. Reaction products of AAFA were characterised with XRD, FTIR and TG/DTG, while the microstructure of AAFA ...

  13. Cationic lipids delay the transfer of plasmid DNA to lysosomes.

    Science.gov (United States)

    Wattiaux, R; Jadot, M; Laurent, N; Dubois, F; Wattiaux-De Coninck, S

    1996-10-14

    Plasmid 35S DNA, naked or associated with different cationic lipid preparations was injected to rats. Subcellular distribution of radioactivity in the liver one hour after injection, was established by centrifugation methods. Results show that at that time, 35S DNA has reached lysosomes. On the contrary, when 35S DNA was complexed with lipids, radioactivity remains located in organelles whose distribution after differential and isopycnic centrifugation, is clearly distinct from that of arylsulfatase, lysosome marker enzyme. Injection of Triton WR 1339, a specific density perturbant of lysosomes, four days before 35S DNA injection causes a density decrease of radioactivity bearing structures, apparent one hour after naked 35S DNA injection but visible only after more than five hours, when 35S DNA associated with a cationic lipid is injected. These observations show that cationic lipids delay the transfer to lysosomes, of plasmid DNA taken up by the liver.

  14. Replisome Assembly at Bacterial Chromosomes and Iteron Plasmids

    Directory of Open Access Journals (Sweden)

    Katarzyna Ewa Wegrzyn

    2016-08-01

    Full Text Available The proper initiation and occurrence of DNA synthesis depends on the formation and rearrangements of nucleoprotein complexes within the origin of DNA replication. In this review article, we present the current knowledge on the molecular mechanism of replication complex assembly at the origin of bacterial chromosome and plasmid replicon containing direct repeats (iterons within the origin sequence. We describe recent findings on chromosomal and plasmid replication initiators, DnaA and Rep proteins, respectively, and their sequence-specific interactions with double and single stranded DNA. Also, we discuss the current understanding of the activities of DnaA and Rep proteins required for replisome assembly that is fundamental to the duplication and stability of genetic information in bacterial cells.

  15. Plasmids and packaging cell lines for use in phage display

    Science.gov (United States)

    Bradbury, Andrew M.

    2012-07-24

    The invention relates to a novel phagemid display system for packaging phagemid DNA into phagemid particles which completely avoids the use of helper phage. The system of the invention incorporates the use of bacterial packaging cell lines which have been transformed with helper plasmids containing all required phage proteins but not the packaging signals. The absence of packaging signals in these helper plasmids prevents their DNA from being packaged in the bacterial cell, which provides a number of significant advantages over the use of both standard and modified helper phage. Packaged phagemids expressing a protein or peptide of interest, in fusion with a phage coat protein such as g3p, are generated simply by transfecting phagemid into the packaging cell line.

  16. A Rebeccamycin Analog Provides Plasmid-Encoded Niche Defense.

    Science.gov (United States)

    Van Arnam, Ethan B; Ruzzini, Antonio C; Sit, Clarissa S; Currie, Cameron R; Clardy, Jon

    2015-11-18

    Bacterial symbionts of fungus-growing ants occupy a highly specialized ecological niche and face the constant existential threat of displacement by another strain of ant-adapted bacteria. As part of a systematic study of the small molecules underlying this fraternal competition, we discovered an analog of the antitumor agent rebeccamycin, a member of the increasingly important indolocarbazole family. While several gene clusters consistent with this molecule's newly reported modification had previously been identified in metagenomic studies, the metabolite itself has been cryptic. The biosynthetic gene cluster for 9-methoxyrebeccamycin is encoded on a plasmid in a manner reminiscent of plasmid-derived peptide antimicrobials that commonly mediate antagonism among closely related Gram-negative bacteria.

  17. Dataset of plasmid DNA extraction using different magnetic nanoparticles (MNPs

    Directory of Open Access Journals (Sweden)

    H. Rahnama

    2016-12-01

    MNPs were characterized by energy dispersive spectroscopy (EDS and transmission electron microscopy (TEM. Finally, the overall efficiency of different MNPs (Fe3O4, Fe3O4/SiO2, Fe3O4/SiO2/TiO2 in plasmid DNA isolation was compared using gel electrophoresis analysis. The data supplied in this article supports the accompanying publication “Comparative study of three magnetic nano-particles (FeSO4, FeSO4/SiO2, FeSO4/SiO2/TiO2 in plasmid DNA extraction” (H. Rahnama, A. Sattarzadeh, F. Kazemi, N. Ahmadi, F. Sanjarian, Z. Zand, 2016 [1].

  18. Liquid-Crystalline Mesophases of Plasmid DNA in Bacteria

    Science.gov (United States)

    Reich, Ziv; Wachtel, Ellen J.; Minsky, Abraham

    1994-06-01

    Bacterial plasmids may often reach a copy number larger than 1000 per cell, corresponding to a total amount of DNA that may exceed the amount of DNA within the bacterial chromosome. This observation highlights the problem of cellular accommodation of large amounts of closed-circular nucleic acids, whose interwound conformation offers negligible DNA compaction. As determined by x-ray scattering experiments conducted on intact bacteria, supercoiled plasmids segregate within the cells into dense clusters characterized by a long-range order. In vitro studies performed at physiological DNA concentrations indicated that interwound DNA spontaneously forms liquid crystalline phases whose macroscopic structural properties are determined by the features of the molecular supercoiling. Because these features respond to cellular factors, DNA supercoiling may provide a sensitive regulatory link between cellular parameters and the packaging modes of interwound DNA in vivo.

  19. Current trends in separation of plasmid DNA vaccines: a review.

    Science.gov (United States)

    Ghanem, Ashraf; Healey, Robert; Adly, Frady G

    2013-01-14

    Plasmid DNA (pDNA)-based vaccines offer more rapid avenues for development and production if compared to those of conventional virus-based vaccines. They do not rely on time- or labour-intensive cell culture processes and allow greater flexibility in shipping and storage. Stimulating antibodies and cell-mediated components of the immune system are considered as some of the major advantages associated with the use of pDNA vaccines. This review summarizes the current trends in the purification of pDNA vaccines for practical and analytical applications. Special attention is paid to chromatographic techniques aimed at reducing the steps of final purification, post primary isolation and intermediate recovery, in order to reduce the number of steps necessary to reach a purified end product from the crude plasmid.

  20. Polymerase chain reaction-based gene removal from plasmids

    Directory of Open Access Journals (Sweden)

    Vishnu Vardhan Krishnamurthy

    2015-09-01

    Full Text Available This data article contains supplementary figures and methods to the research article entitled, “Multiplex gene removal by two-step polymerase chain reactions” (Krishnamurthy et al., Anal. Biochem., 2015, doi:http://dx.doi.org/10.1016/j.ab.2015.03.033, which presents a restriction-enzyme free method to remove multiple DNA segments from plasmids. Restriction-free cloning methods have dramatically improved the flexibility and speed of genetic manipulation compared to conventional assays based on restriction enzyme digestion (Lale and Valla, 2014. DNA Cloning and Assembly Methods, vol. 1116. Here, we show the basic scheme and characterize the success rate for single and multiplex gene removal from plasmids. In addition, we optimize experimental conditions, including the amount of template, multiple primers mixing, and buffers for DpnI treatment, used in the one-pot reaction for multiplex gene removal.

  1. Curing of Epoxy Resin Induced by Femtosecond Laser Pulse

    Institute of Scientific and Technical Information of China (English)

    LI Yubin; ZHANG Zuoguang

    2005-01-01

    The possibility of curing of epoxy resin induced by femtosecond laser beam was explored through choosing different initiators . Absorption spectroscopy, infrared spectroscopy (IR), stereomicroscopy and scanning electron microscopy (SEM) were applied to analyze the structure of epoxy resin systems after irradiation with a femtosecond laser beam. The experimental results show that the epoxy resin systems containing diaryliodonium salts can be cured by irradiation of Jemtosecond laser pulse, while the systems containing benzoin can not be cured. It is found that diaryliodonium salts decompose under the irradiation of femtosecond laser pulse through multi ( two ) -photon absorption, initiating the ring-opening polymerization of epoxy resin. And the appearance of cured area has a sheet structure consisting of many tiny lamellar structures.

  2. Cure monitoring using ultrasonic guided waves in wires.

    Science.gov (United States)

    Vogt, T; Lowe, M; Cawley, P

    2003-09-01

    The possibility of using ultrasonic guided waves for monitoring the cure process of epoxy resins is investigated. The two techniques presented use a wire waveguide which is partly embedded in the resin. The first technique is based on the measurement of attenuation due to leakage of bulk waves into the resin surrounding the waveguide. The second technique measures the reflection of the guided wave that occurs at the point where the waveguide enters the resin. Both the attenuation and the reflection coefficient change significantly during cure, and the numerical methods to relate these to the material properties of the curing resin are presented in this paper. The results from the modeling are experimentally verified and show good agreement. The applicability of each testing method is discussed, and typical cure-monitoring curves are presented.

  3. effect of light curing unit characteristics on light intensity output ...

    African Journals Online (AJOL)

    2013-09-09

    Sep 9, 2013 ... Objective: To determine the characteristics of light curing units (LCUs) in dental clinics in Nairobi and ... generation which has dogged the new generation of. LEDs without ..... Knezevic A, Tarle Z, Meniga A, Sutalo J, Pichler G.

  4. Cure monitoring using ultrasonic guided waves in wires

    Science.gov (United States)

    Vogt, T.; Lowe, M.; Cawley, P.

    2003-09-01

    The possibility of using ultrasonic guided waves for monitoring the cure process of epoxy resins is investigated. The two techniques presented use a wire waveguide which is partly embedded in the resin. The first technique is based on the measurement of attenuation due to leakage of bulk waves into the resin surrounding the waveguide. The second technique measures the reflection of the guided wave that occurs at the point where the waveguide enters the resin. Both the attenuation and the reflection coefficient change significantly during cure, and the numerical methods to relate these to the material properties of the curing resin are presented in this paper. The results from the modeling are experimentally verified and show good agreement. The applicability of each testing method is discussed, and typical cure-monitoring curves are presented.

  5. Editorial: Short-course adjuvant trastuzumab will increase cure rates ...

    African Journals Online (AJOL)

    Editorial: Short-course adjuvant trastuzumab will increase cure rates in patients with human epidermal growth factor receptor 2-positive breast cancer. ... Journal Home > Vol 107, No 8 (2017) >. Log in or Register to get ... from 32 Countries:.

  6. GENERAL ENVIRONMENTAL CORPORATION; CURE ELECTROCOAGULATION TECHNOLOGY: INNOVATIVE TECHNOLOGY EVALUATION REPORT

    Science.gov (United States)

    The CURE electrocoagulation technology was demonstrated under the Superfund Innovative Technology Evaluation (SITE) program at the U.S. Department of Energy (DOE) Rocky Flats Environmental Technology Site (RFETS), where water from the solar evaporation ponds (SEPs) was contaminat...

  7. Chemistry and technology of Cured and Smoked Meat Products

    Directory of Open Access Journals (Sweden)

    V. K. Mathur

    1967-05-01

    Full Text Available Recent developments in the chemistry and technology of cured and smoked meat products are reviewed. Various factors physical, chemical and microbiological affecting water holding capacity of cured meats and cured colour formation and its stability during storage are discussed. Methods of preparation of various types of cured and smoked meat products have been briefly described. Developments in the use of synthetic casings and skinless sausages have been mentioned. Smoke constituents, their role and physico-chemical aspects of mechanism and functions of smoking are discussed. Advancements in the methods of smoke generation, conventional smoking and some kilns, electrostatic smoking and smoking by use of liquid smokes have been described. Some of the methods of preparation of liquid smokes are also given.

  8. effect of light intensity on the cure characteristics of photo ...

    African Journals Online (AJOL)

    2012-05-05

    May 5, 2012 ... Design: A laboratory based cross sectional study. Setting: Public and ... set out to determine the light intensity emitted by light curing units ... polymerised with it. MATERIAL AND METHODS ..... J. of Oral Rehab. 2005; 32:.

  9. Factors affecting dry-cured ham consumer acceptability.

    Science.gov (United States)

    Morales, R; Guerrero, L; Aguiar, A P S; Guàrdia, M D; Gou, P

    2013-11-01

    The objectives of the present study were (1) to compare the relative importance of price, processing time, texture and intramuscular fat in purchase intention of dry-cured ham through conjoint analysis, (2) to evaluate the effect of dry-cured ham appearance on consumer expectations, and (3) to describe the consumer sensory preferences of dry-cured ham using external preference mapping. Texture and processing time influenced the consumer preferences in conjoint analysis. Red colour intensity, colour uniformity, external fat and white film presence/absence influenced consumer expectations. The consumer disliked hams with bitter and metallic flavour and with excessive saltiness and piquantness. Differences between expected and experienced acceptability were found, which indicates that the visual preference of consumers does not allow them to select a dry-cured ham that satisfies their sensory preferences of flavour and texture.

  10. Development of hard materials by radiation curing technology

    Energy Technology Data Exchange (ETDEWEB)

    Salleh, N.G. E-mail: nik_ghazali@mint.gov.my; Glaesel, H.J.; Mehnert, R

    2002-03-01

    For studying nanoglobular modification effects in radiation cured polymeric composites, we prepared polymerization active silico-organic nanoparticles. With their polymerization active ligands, these nanoparticles form crosslinks by modifying the viscoelastic properties in radiation cured polymeric nanocomposites. In this process, there was a polymerization activity imparted to the particle surfaces of nanopowders, thus applying the physico-chemical modification scheme of a heterogeneous copolymerization to novel scratch and abrasion resistant coatings. By varying the nanoparticle-monomer formulation and the curing method, additional property can be achieved. In this works, we also investigated the influence of various factors such as addition of photoinitiators and other additives into the formulations. The coating materials were applied to the substrate by using different type of coaters. These materials were cured by ultraviolet light and electron beam irradiation. Properties of coatings were characterized using Universal scratch tester and Taber abrasion tester.

  11. Characterization of two novel plasmids from Geobacillus sp. 610 and 1121 strains.

    Science.gov (United States)

    Kananavičiūtė, Rūta; Butaitė, Elena; Citavičius, Donaldas

    2014-01-01

    We describe two cryptic low molecular weight plasmids, pGTD7 (3279bp) and pGTG5 (1540bp), isolated from Geobacillus sp. 610 and 1121 strains, respectively. Homology analysis of the replication protein (Rep) sequences and detection of ssDNA indicate that both of them replicate via rolling circle mechanism. As revealed by sequence similarities of dso region and Rep protein, plasmid pGTD7 belongs to pC194/pUB110 plasmid family. The replicon of pGTD7 was proved to be functional in another Geobacillus host. For this purpose, a construct pUCK7, containing a replicon of the analyzed plasmid, was created and transferred to G. stearothermophilus NUB3621R strain by electroporation. Plasmid pGTG5, based on Rep protein sequence similarity, was found to be related mostly to some poorly characterized bacterial plasmids. Rep proteins encoded by these plasmids contain conservative motifs that are most similar to those of Microviridae phages. This feature suggests that pGTG5, together with other plasmids containing the same motifs, could constitute a new family of bacterial plasmids. To date, pGTG5 is the smallest plasmid identified in bacteria belonging to the genus Geobacillus. The two plasmids described in this study can be used for the construction of new vectors suitable for biotechnologically important bacteria of the genus Geobacillus.

  12. Bacillus stearothermophilus contains a plasmid-borne gene for alpha-amylase.

    Science.gov (United States)

    Mielenz, J R

    1983-01-01

    The gene for thermostable alpha-amylase from the thermophilic bacterium Bacillus stearothermophilus has been cloned and expressed in Escherichia coli. Each alpha-amylase-producing colony contained at least a 9.7-kilobase-pair (kb) chimeric plasmid composed of the vector pBR322 and a common 5.4-kb HindIII fragment of DNA. B. stearothermophilus contains four plasmids with sizes from 12 kb to over 108 kb. Restriction endonuclease analysis of these naturally occurring plasmids showed they also contain a 5.4-kb HindIII fragment of DNA. Cloning experiments with the four plasmids yielded alpha-amylase-producing E. coli that contained the same 9.7-kb chimeric plasmid. Restriction endonuclease analysis and further recombinant DNA experiments identified a 26-kb plasmid that contains the gene for alpha-amylase. A spontaneous mutant of B. stearothermophilus unable to produce alpha-amylase was missing the 26-kb plasmid but contained a 20-kb plasmid. A 6-kb deletion within the region of the 5.4-kb HindIII fragment yielded the 20-kb plasmid unable to code for alpha-amylase. A nick-translated probe for the alpha-amylase coding region did not hybridize to either plasmid or total cellular DNA from this mutant strain of B. stearothermophilus. These results demonstrate the gene for alpha-amylase is located exclusively on a 26-kb plasmid in B. stearothermophilus with no genetic counterpart present on the chromosome. Images PMID:6193526

  13. Presence and analysis of plasmids in human and animal associated arcobacter species.

    Science.gov (United States)

    Douidah, Laid; De Zutter, Lieven; Van Nieuwerburgh, Filip; Deforce, Dieter; Ingmer, Hanne; Vandenberg, Olivier; Van den Abeele, Anne-Marie; Houf, Kurt

    2014-01-01

    In this study, we report the screening of four Arcobacter species for the presence of small and large plasmids. Plasmids were present in 9.9% of the 273 examined strains. One Arcobacter cryaerophilus and four Arcobacter butzleri plasmids were selected for further sequencing. The size of three small plasmids isolated from A. butzleri and the one from A. cryaerophilus strains ranged between 4.8 and 5.1 kb, and the size of the large plasmid, isolated from A. butzleri, was 27.4 kbp. The G+C content of all plasmids ranged between 25.4% and 26.2%. A total of 95% of the large plasmid sequence represents coding information, which contrasts to the 20 to 30% for the small plasmids. Some of the open reading frames showed a high homology to putative conserved domains found in other related organisms, such as replication, mobilization and genes involved in type IV secretion system. The large plasmid carried 35 coding sequences, including seven genes in a contiguous region of 11.6 kbp that encodes an orthologous type IV secretion system found in the Wolinella succinogenes genome, Helicobacter pylori and Campylobacter jejuni plasmids, which makes this plasmid interesting for further exploration.

  14. Presence and analysis of plasmids in human and animal associated arcobacter species.

    Directory of Open Access Journals (Sweden)

    Laid Douidah

    Full Text Available In this study, we report the screening of four Arcobacter species for the presence of small and large plasmids. Plasmids were present in 9.9% of the 273 examined strains. One Arcobacter cryaerophilus and four Arcobacter butzleri plasmids were selected for further sequencing. The size of three small plasmids isolated from A. butzleri and the one from A. cryaerophilus strains ranged between 4.8 and 5.1 kb, and the size of the large plasmid, isolated from A. butzleri, was 27.4 kbp. The G+C content of all plasmids ranged between 25.4% and 26.2%. A total of 95% of the large plasmid sequence represents coding information, which contrasts to the 20 to 30% for the small plasmids. Some of the open reading frames showed a high homology to putative conserved domains found in other related organisms, such as replication, mobilization and genes involved in type IV secretion system. The large plasmid carried 35 coding sequences, including seven genes in a contiguous region of 11.6 kbp that encodes an orthologous type IV secretion system found in the Wolinella succinogenes genome, Helicobacter pylori and Campylobacter jejuni plasmids, which makes this plasmid interesting for further exploration.

  15. Efficient transformation of Bacillus thuringiensis requires nonmethylated plasmid DNA.

    OpenAIRE

    Macaluso, A; Mettus, A M

    1991-01-01

    The transformation efficiency of Bacillus thuringiensis depends upon the source of plasmid DNA. DNA isolated from B. thuringiensis, Bacillus megaterium, or a Dam- Dcm- Escherichia coli strain efficiently transformed several B. thuringiensis strains, B. thuringiensis strains were grouped according to which B. thuringiensis backgrounds were suitable sources of DNA for transformation of other B. thuringiensis strains, suggesting that B. thuringiensis strains differ in DNA modification and restri...

  16. Plasmid Isolation in Legionella pneumophila and Legionella-like Organisms.

    Science.gov (United States)

    1980-08-22

    834. 14. Macrina, F. L., D. J. Kopecko, K. R. Jones, D. J. Ayers, and S. M. McCowen. 1978. A multiple plasmic-containing Escherichi coli strain...smaller 20 Mdal cryptic plasmid and was used as a control marker with the screening procedure. Escherichia coli V517 was supplied by E. M. Lederberg...Tris-borate buffer. This purified preparation was suitable for electrophoresis. Molecular weight estimates. Escherichia coli V517 was employed as an

  17. Growth dependence of conjugation explains limited plasmid invasion in biofilms: an individual‐based modelling study

    DEFF Research Database (Denmark)

    Merkey, Brian; Lardon, Laurent; Seoane, Jose Miguel;

    2011-01-01

    . By extending an individual‐based model of microbial growth and interactions to include the dynamics of plasmid carriage and transfer by individual cells, we were able to conduct in silico tests of this and other hypotheses on the dynamics of conjugal plasmid transfer in biofilms. For a generic model plasmid...... and scan speed) and spatial reach (EPS yield, conjugal pilus length) are more important for successful plasmid invasion than the recipients' growth rate or the probability of segregational loss. While this study identifies one factor that can limit plasmid invasion in biofilms, the new individual......Plasmid invasion in biofilms is often surprisingly limited in spite of the close contact of cells in a biofilm. We hypothesized that this poor plasmid spread into deeper biofilm layers is caused by a dependence of conjugation on the growth rate (relative to the maximum growth rate) of the donor...

  18. 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.

  19. Scaling-up recombinant plasmid DNA for clinical trial: current concern, solution and status.

    Science.gov (United States)

    Ismail, Ruzila; Allaudin, Zeenathul Nazariah; Lila, Mohd-Azmi Mohd

    2012-09-07

    Gene therapy and vaccines are rapidly developing field in which recombinant nucleic acids are introduced in mammalian cells for enhancement, restoration, initiation or silencing biochemical function. Beside simplicity in manipulation and rapid manufacture process, plasmid DNA-based vaccines have inherent features that make them promising vaccine candidates in a variety of diseases. This present review focuses on the safety concern of the genetic elements of plasmid such as propagation and expression units as well as their host genome for the production of recombinant plasmid DNA. The highlighted issues will be beneficial in characterizing and manufacturing plasmid DNA for save clinical use. Manipulation of regulatory units of plasmid will have impact towards addressing the safety concerns raised in human vaccine applications. The gene revolution with plasmid DNA by alteration of their plasmid and production host genetics will be promising for safe delivery and obtaining efficient outcomes.

  20. Partition-associated incompatibility caused by random assortment of pure plasmid clusters

    DEFF Research Database (Denmark)

    Ebersbach, Gitte; Sherratt, David J; Gerdes, Kenn;

    2005-01-01

    Summary Bacterial plasmids and chromosomes encode centromere-like partition loci that actively segregate DNA before cell division. The molecular mechanism behind DNA segregation in bacteria is largely unknown. Here we analyse the mechanism of partition-associated incompatibility for plasmid pB171......-lived pairing of plasmids. Instead, pure R1 and F foci were positioned along the length of the cell, and in a random order. Thus, our results raise the possibility that partition-mediated plasmid incompatibility is not caused by pairing of heterologous plasmids but instead by random positioning of pure plasmid...... clusters along the long axis of the cell. The strength of the incompatibility was correlated with the capability of the plasmids to compete for the mid-cell position....

  1. In-situ cure monitoring of epoxy resin systems

    OpenAIRE

    Powell, Graham

    1998-01-01

    This thesis was submitted for the degree of Doctor of Philosophy and awarded by Brunel University. This thesis describes the work carried out at Brunel University to develop novel optical fibre sensors capable of monitoring the cure state of an epoxy/amine resin system. The sensors were of simple construction, consisting of an optical fibre from which the silicone cladding layer had been removed over a short length. This stripped length was embedded into the curing resin system. The sensor...

  2. Some electroanalytical investigations into the cure chemistry of industrial sealants

    OpenAIRE

    Raftery, Declan Patrick

    1996-01-01

    This thesis represents a study of the cure chemistry of three contrasting adhesive technologies, applying a range of analytical approaches to gain further insight into the complex chemistry of adhesives. An introduction is given in chapter one into the general chemistry of adhesives and their analysis, with particular emphasis on anaerobic adhesives and the crucial role played by transition metals in the cure chemistry. In order to elucidate the role played by tertiary amines and saccahri...

  3. Cure Characteristics of Tricyanate Ester High Temperature Composite Resins (PREPRINT)

    Science.gov (United States)

    2011-02-09

    isothermal DSC data were analyzed via the Kissinger [17], Ozawa [18], and Friedman [19] methods, as described in the recent analysis of dicyanate ester...7 present the graphical analysis that underlies the Kissinger [17], Ozawa [18], and Friedman [19] methods, respectively, of non-isothermal DSC cure...where β is the heating rate and Tp the absolute temperature at the peak cure rate, is proportional to the activation energy. For the Ozawa method

  4. The Prevention and Cure of Cervical Vertebra Disease

    Institute of Scientific and Technical Information of China (English)

    1994-01-01

    CERVICAL vertebra problems are common and usually occur in middleaged and elderly people, especially those who often bend over. The symptoms patients might encounter are dizziness, headache, neck and shoulder pain and numbness, and even viscera trouble. The following exercises are a simple and convenient way to prevent and cure cervical vertebra problems. If you continue doing the exercises, you can prevent and cure them. Here are the exercises step by step:

  5. Identification of two replicons in phage-plasmid P4.

    Science.gov (United States)

    Tocchetti, A; Serina, S; Terzano, S; Dehò, G; Ghisotti, D

    1998-06-05

    DNA replication of phage-plasmid P4 proceeds bidirectionally from the ori1 site (previously named ori), but requires a second cis-acting region, crr. Replication depends on the product of the P4 alpha gene, a protein with primase and helicase activity, that binds both ori1 and crr. A negative regulator of P4 DNA replication, the Cnr protein, is required for copy number control of plasmid P4. Using a plasmid complementation test for replication, we found that two replicons, both dependent on the alpha gene product, coexist in P4. The first replicon is made by the cnr and alpha genes and the ori1 and crr sites. The second is limited to the alpha and crr region. Thus, in the absence of the ori1 region, replication can initiate at a different site. By deletion mapping, a cis-acting region, ori2, essential for replication of the alpha-crr replicon was mapped within a 270-bp fragment in the first half of the alpha gene. The ori2 site was found to be dispensable in a replicon that contains ori1. A construct that besides crr and alpha carries also the cnr gene was unable to replicate, suggesting that Cnr not only controls replication from ori1, but also silences ori2.

  6. CURING PROCESS OF PHOTOPOLYMER RESIN BOND DIAMOND TOOLS

    Institute of Scientific and Technical Information of China (English)

    GAO Tao; PENG Wei; YAO Chunyan

    2007-01-01

    Analytical simulation and corresponding proof-test are adopted to study the principle of the curing process of photopolymer resin diamond tools. The influence of the diamond as abrasives in photopolymer resin owing to the absorptivity of the diamond for the UV light on the photopolymer resin curing process is discussed. Based on the above, a kind of diamond tool-dicing blade is selected to analyze the curing process of photopolymer bond diamond tools. An analytical model of curing process is developed and a correlation curve between the depth of polymerization of the photopolymer resin diamond tools and the exposure time to represent the curing process of photopolymer bond dicing blade. A test is done to proof-test the validity of the analytical model and the correlation curve. The simulated data fit the experimental results, which demonstrates the analytical models and numerical algorithm are of high reliability. The analytical simulation method could possibly be used to optimize the curing cycle and improve the quality of the photopolymers resin bond diamond tools.

  7. Characterization of dental composite curing kinetics using dynamic light scattering

    Science.gov (United States)

    Wells-Gray, Elaine M.; Kirkpatrick, Sean J.; Sakaguchi, Ron L.

    2009-02-01

    Polymer resin composites are a class of widely used restorative dental materials that undergo a complex polymerization curing process that has been the subject of substantial research. This study uses speckle correlation methods based on dynamic light scattering as a tool to monitor the rate and extent of dental composite polymerization during and after photo-curing. Thin disc-shaped samples (composite consisting of 50:50 BisGMA/TEGDMA resin, quartz silica filler particles, and camphorquinone as photo-initiator. A 633 nm HeNe laser beam was used to probe the top surface of the sample via a backscattered speckle pattern, while the bottom surface was illuminated with a halogen curing lamp (peak wavelength=470nm) to initiate the polymerization reaction. The speckle patterns were recorded with a CCD camera, and stored as a 'speckle cube' for post processing. Correlation values of the intensity fluctuation were calculated on a pixel-by-pixel basis for pairs of subsequent speckle images and then ensemble averaged. Results show a sharp decrease in correlation at the onset of curing, indicating a large amount of double bond conversion and movement within the composite. Correlation values then quickly increase, eventually reaching a plateau near unity, indicating cessation of molecular rearrangement. The kinetic behavior demonstrated by our correlation curves are in good agreement with curing data found in the literature, and demonstrate the usefulness of this technique for monitoring dental composite curing.

  8. Depth of cure of bulk-fill flowable composite resins.

    Science.gov (United States)

    Pedalino, Inaam; Hartup, Grant R; Vandewalle, Kraig S

    2015-01-01

    In recent years, manufacturers have introduced flowable composite resins that reportedly can be placed in increments of 4 mm or greater. The purpose of this study was to evaluate the depth of cure of bulk-fill flowable composite resins (SureFil SDR Flow, Grandio Flow, and Venus Bulk Fill) and a conventional flowable composite resin (Revolution Formula 2). Depth of cure was measured in terms of bottom-maximum Knoop hardness number (KHN) ratios and the International Organization for Standardization (ISO) 4049 scrape technique. Shades A2 and A3 of SureFil SDR Flow, Grandio Flow, and Revolution Formula 2 were tested. Venus Bulk Fill was tested in its only available shade (universal). Specimens in thicknesses of 2, 3, 4, 5, and 6 mm were polymerized for 20 or 40 seconds, and a hardness tester was used to determine the hardness ratios for each shade at each thickness. For the scraping technique, after specimens were exposed to the curing light, unpolymerized composite resin was removed with a plastic instrument, the polymerized composite was measured, and the length was divided by 2 per ISO guidelines. According to the KHN ratios and the scrape test, Venus Bulk Fill predictably exceeded the manufacturer's claim of a 4-mm depth of cure at both 20 and 40 seconds of curing time. The overall results for depth of cure showed that Venus Bulk Fill ≥ SureFil SDR Flow ≥ Grandio Flow ≥ Revolution Formula 2.

  9. Monitoring the Cure State of Thermosetting Resins by Ultrasound

    Directory of Open Access Journals (Sweden)

    Alfonso Maffezzoli

    2013-09-01

    Full Text Available The propagation of low intensity ultrasound in a curing resin, acting as a high frequency oscillatory excitation, has been recently proposed as an ultrasonic dynamic mechanical analysis (UDMA for cure monitoring. The technique measures sound velocity and attenuation, which are very sensitive to changes in the viscoelastic characteristics of the curing resin, since the velocity is related to the resin storage modulus and density, while the attenuation is related to the energy dissipation and scattering in the curing resin. The paper reviews the results obtained by the authors’ research group in the last decade by means of in-house made ultrasonic set-ups for both contact and air-coupled ultrasonic experiments. The basics of the ultrasonic wave propagation in polymers and examples of measurements of the time-evolution of ultrasonic longitudinal modulus and chemical conversion of different thermosetting resins are presented. The effect of temperature on the cure kinetics, the comparison with rheological, low frequency dynamic mechanical and calorimetric results, and the correlation between ultrasonic modulus and crosslinking density will be also discussed. The paper highlights the reliability of ultrasonic wave propagation for monitoring the physical changes taking place during curing and the potential for online monitoring during polymer and polymer matrix composite processing.

  10. Competing ParA structures space bacterial plasmids equally over the nucleoid.

    Directory of Open Access Journals (Sweden)

    Robert Ietswaart

    2014-12-01

    Full Text Available Low copy number plasmids in bacteria require segregation for stable inheritance through cell division. This is often achieved by a parABC locus, comprising an ATPase ParA, DNA-binding protein ParB and a parC region, encoding ParB-binding sites. These minimal components space plasmids equally over the nucleoid, yet the underlying mechanism is not understood. Here we investigate a model where ParA-ATP can dynamically associate to the nucleoid and is hydrolyzed by plasmid-associated ParB, thereby creating nucleoid-bound, self-organizing ParA concentration gradients. We show mathematically that differences between competing ParA concentrations on either side of a plasmid can specify regular plasmid positioning. Such positioning can be achieved regardless of the exact mechanism of plasmid movement, including plasmid diffusion with ParA-mediated immobilization or directed plasmid motion induced by ParB/parC-stimulated ParA structure disassembly. However, we find experimentally that parABC from Escherichia coli plasmid pB171 increases plasmid mobility, inconsistent with diffusion/immobilization. Instead our observations favor directed plasmid motion. Our model predicts less oscillatory ParA dynamics than previously believed, a prediction we verify experimentally. We also show that ParA localization and plasmid positioning depend on the underlying nucleoid morphology, indicating that the chromosomal architecture constrains ParA structure formation. Our directed motion model unifies previously contradictory models for plasmid segregation and provides a robust mechanistic basis for self-organized plasmid spacing that may be widely applicable.

  11. Influence of curing time, overlay material and thickness on three light-curing composites used for luting indirect composite restorations.

    Science.gov (United States)

    D'Arcangelo, Camillo; De Angelis, Francesco; Vadini, Mirco; Carluccio, Fabio; Vitalone, Laura Merla; D'Amario, Maurizio

    2012-08-01

    To assess the microhardness of three resin composites employed in the adhesive luting of indirect composite restorations and examine the influence of the overlay material and thickness as well as the curing time on polymerization rate. Three commercially available resin composites were selected: Enamel Plus HRI (Micerium) (ENA), Saremco ELS (Saremco Dental) (SAR), Esthet-X HD (Dentsply/DeTrey) (EST-X). Post-polymerized cylinders of 6 different thicknesses were produced and used as overlays: 2 mm, 3 mm, 3.5 mm, 4 mm, 5 mm, and 6 mm. Two-mm-thick disks were produced and employed as underlays. A standardized amount of composite paste was placed between the underlay and the overlay surfaces which were maintained at a fixed distance of 0.5 mm. Light curing of the luting composite layer was performed through the overlays for 40, 80, or 120 s. For each specimen, the composite to be cured, the cured overlay, and the underlay were made out of the same batch of resin composite. All specimens were assigned to three experimental groups on the basis of the resin composite used, and to subgroups on the basis of the overlay thickness and the curing time, resulting in 54 experimental subgroups (n = 5). Forty-five additional specimens, 15 for each material under investigation, were produced and subjected to 40, 80, or 120 s of light curing using a microscope glass as an overlay; they were assigned to 9 control subgroups (n = 5). Three Vicker's hardness (VH) indentations were performed on each specimen. Means and standard deviations were calculated. Data were statistically analyzed using 3-way ANOVA. Within the same material, VH values lower than 55% of control were not considered acceptable. The used material, the overlay thickness, and the curing time significantly influenced VH values. In the ENA group, acceptable hardness values were achieved with 3.5-mm or thinner overlays after 120 or 80 s curing time (VH 41.75 and 39.32, respectively), and with 2-mm overlays after 40 s (VH 54

  12. Mechanistic basis of plasmid-specific DNA binding of the F plasmid regulatory protein, TraM.

    Science.gov (United States)

    Peng, Yun; Lu, Jun; Wong, Joyce J W; Edwards, Ross A; Frost, Laura S; Mark Glover, J N

    2014-11-11

    The conjugative transfer of bacterial F plasmids relies on TraM, a plasmid-encoded protein that recognizes multiple DNA sites to recruit the plasmid to the conjugative pore. In spite of the high degree of amino acid sequence conservation between TraM proteins, many of these proteins have markedly different DNA binding specificities that ensure the selective recruitment of a plasmid to its cognate pore. Here we present the structure of F TraM RHH (ribbon-helix-helix) domain bound to its sbmA site. The structure indicates that a pair of TraM tetramers cooperatively binds an underwound sbmA site containing 12 base pairs per turn. The sbmA is composed of 4 copies of a 5-base-pair motif, each of which is recognized by an RHH domain. The structure reveals that a single conservative amino acid difference in the RHH β-ribbon between F and pED208 TraM changes its specificity for its cognate 5-base-pair sequence motif. Specificity is also dictated by the positioning of 2-base-pair spacer elements within sbmA; in F sbmA, the spacers are positioned between motifs 1 and 2 and between motifs 3 and 4, whereas in pED208 sbmA, there is a single spacer between motifs 2 and 3. We also demonstrate that a pair of F TraM tetramers can cooperatively bind its sbmC site with an affinity similar to that of sbmA in spite of a lack of sequence similarity between these DNA elements. These results provide a basis for the prediction of the DNA binding properties of the family of TraM proteins.

  13. Characterization of Multidrug-Resistant Escherichia coli by Plasmid Replicon Typing and Pulsed-Field Gel Electrophoresis

    Science.gov (United States)

    Background: Characterization of plasmids has particular clinical significance because genes encoding important traits such as antimicrobial resistance are frequently present in plasmids. Plasmid replicon typing is a multiplex PCR based method that can be used to classify 18 of the 26 known plasmid t...

  14. Changes in the temperature of a dental light-cured composite resin by different light-curing units

    Science.gov (United States)

    Rastelli, A. N. S.; Jacomassi, D. P.; Bagnato, V. S.

    2008-08-01

    The purpose of this study was to evaluate the temperature increase during the polymerization process through the use of three different light-curing units with different irradiation times. One argon laser (Innova, Coherent), one halogen (Optilight 501, Demetron), and one blue LED (LEC 1000, MM Optics) LCU with 500 mW/cm2 during 5, 10, 20, 30, 40, 50, and 60 s of irradiation times were used in this study. The composite resin used was a microhybrid Filtek Z-250 (3M/ESPE) at color A2. The samples were made in a metallic mold 2 mm in thickness and 4 mm in diameter and previously light-cured during 40 s. A thermocouple (Model 120 202 EAJ, Fenwal Electronic, Milford, MA, USA) was introduced in the composite resin to measure the temperature increase during the curing process. The highest temperature increase was recorded with a Curing Light 2500 halogen LCU (5 and 31°C after 5 and 60 s, respectively), while the lowest temperature increase was recorded for the Innova LCU based on an argon laser (2 and 11°C after 5 and 60 s, respectively). The temperature recorded for LCU based on a blue LED was 3 and 22°C after 5 and 60 s, respectively. There was a quantifiable amount of heat generated during the visible light curing of a composite resin. The amount of heat generated was influenced by the characteristics of the light-curing units used and the irradiation times.

  15. Degree of cure-dependent modelling for polymer curing processes at small-strain. Part I: consistent reformulation

    Science.gov (United States)

    Hossain, M.; Steinmann, P.

    2014-04-01

    A physically-based small strain curing model has been developed and discussed in our previous contribution (Hossain et al. in Comput Mech 43:769-779, 2009a) which was extended later for finite strain elasticity and viscoelasticity including shrinkage in Hossain et al. (Comput Mech 44(5):621-630, 2009b) and in Hossain et al. (Comput Mech 46(3):363-375, 2010), respectively. The previously proposed constitutive models for curing processes are based on the temporal evolution of the material parameters, namely the shear modulus and the relaxation time (in the case of viscoelasticity). In the current paper, a thermodynamically consistent small strain constitutive model is formulated that is directly based on the degree of cure, a key parameter in the curing (reaction) kinetics. The new formulation is also in line with the earlier proposed hypoelastic approach. The curing process of polymers is a complex phenomenon involving a series of chemical reactions which transform a viscoelastic fluid into a viscoelastic solid during which the temperature, the chemistry and the mechanics are coupled. Part I of this work will deal with an isothermal viscoelastic formulation including shrinkage effects whereas the following Part II will give emphasis on the thermomechanical coupled approach. Some representative numerical examples conclude the paper and show the capability of the newly proposed constitutive formulation to capture major phenomena observed during the curing processes of polymers.

  16. Type 3 Fimbriae Encoded on Plasmids Are Expressed from a Unique Promoter without Affecting Host Motility, Facilitating an Exceptional Phenotype That Enhances Conjugal Plasmid Transfer

    Science.gov (United States)

    Madsen, Jonas Stenløkke; Riber, Leise; Kot, Witold; Basfeld, Alrun; Burmølle, Mette; Hansen, Lars Hestbjerg; Sørensen, Søren Johannes

    2016-01-01

    Horizontal gene transfer (HGT), the transmission of genetic material to a recipient that is not the progeny of the donor, is fundamental in bacterial evolution. HGT is often mediated by mobile genetic elements such as conjugative plasmids, which may be in conflict with the chromosomal elements of the genome because they are independent replicons that may petition their own evolutionary strategy. Here we study differences between type 3 fimbriae encoded on wild type plasmids and in chromosomes. Using known and newly characterized plasmids we show that the expression of type 3 fimbriae encoded on plasmids is systematically different, as MrkH, a c-di-GMP dependent transcriptional activator is not needed for strong expression of the fimbriae. MrkH is required for expression of type 3 fimbriae of the Klebsiella pneumoniae chromosome, wherefrom the fimbriae operon (mrkABCDF) of plasmids is believed to have originated. We find that mrkABCDFs of plasmids are highly expressed via a unique promoter that differs from the original Klebsiella promoter resulting in fundamental behavioral consequences. Plasmid associated mrkABCDFs did not influence the swimming behavior of the host, that hereby acquired an exceptional phenotype being able to both actively swim (planktonic behavior) and express biofilm associated fimbriae (sessile behavior). We show that this exceptional phenotype enhances the conjugal transfer of the plasmid. PMID:27627107

  17. Type 3 fimbriae encoded on plasmids are expressed from a unique promoter without affecting host motility, facilitating an exceptional phenotype that enhances conjugal plasmid transfer

    DEFF Research Database (Denmark)

    Madsen, Jonas Stenløkke; Riber, Leise; Kot, Witold Piotr;

    2016-01-01

    of the genome because they are independent replicons that may petition their own evolutionary strategy. Here we study differences between type 3 fimbriae encoded on wild type plasmids and in chromosomes. Using known and newly characterized plasmids we show that the expression of type 3 fimbriae encoded...

  18. Conjugative transferability of the A/C plasmids from Salmonella enterica isolates that possess or lack blaCMY in the A/C plasmid backbone

    Science.gov (United States)

    The objective of this study was to gain a better understanding of the conjugative transfer of antimicrobial resistance plasmids from 205 Salmonella enterica strains, isolated from cattle to E. coli or Salmonella recipients. PCR-based replicon typing (PBRT) was used to type incompatibility plasmid r...

  19. Molecular cloning with bifunctional plasmid vectors in Bacillus subtilis: isolation of a spontaneous mutant of Bacillus subtilis with enhanced transformability for Escherichia coli-propagated chimeric plasmid DNA.

    OpenAIRE

    Ostroff, G. R.; Pène, J. J.

    1983-01-01

    Hybrid plasmid DNA cloned in Escherichia coli undergoes deletions when returned to competent Bacillus subtilis, even in defined restriction and modification mutants of strain 168. We have isolated a mutant of B. subtilis MI112 which is stably transformed at high frequency by chimeric plasmid DNA propagated in E. coli.

  20. Expansion of a plasmid classification system for Gram-positive bacteria and determination of the diversity of plasmids in Staphylococcus aureus strains of human, animal, and food origins

    DEFF Research Database (Denmark)

    Lozano, C.; Garcia-Migura, L.; Aspiroz, C.

    2012-01-01

    An expansion of a previously described plasmid classification was performed and used to reveal the plasmid content of a collection of 92 Staphylococcus aureus strains of different origins. rep genes of other genera were detected in Staphylococcus. S1 pulsed-field gel electrophoresis (PFGE) hybrid...

  1. Analysis of plasmid diversity in 96 Rhodococcus equi strains isolated in Normandy (France) and sequencing of the 87-kb type I virulence plasmid.

    Science.gov (United States)

    Duquesne, Fabien; Hébert, Laurent; Sévin, Corinne; Breuil, Marie-France; Tapprest, Jackie; Laugier, Claire; Petry, Sandrine

    2010-10-01

    To characterize the potential epidemiological relationship between the origin of Rhodococcus equi strains and the type of their virulence plasmids, we performed a comparative analysis of virulence plasmid types encountered in 96 R. equi strains isolated from (1) autopsied horses, (2) organic samples (horse faeces, manure and straw) and (3) environmental samples. Our results revealed no clear epidemiological link between virulence plasmid type and the origin of R. equi strains isolated from horse-related environments. To understand this result, we determined the nucleotide sequence of the second most frequently isolated virulence plasmid type: a 87-kb type I (pVAPA116) plasmid and compared it with the previously sequenced (and most commonly encountered) 85-kb type I (pVAPA1037) plasmid. Our results show that the divergence between these two plasmids is mainly due to the presence of three allelic exchange loci, resulting in the deletion of two genes and the insertion of three genes in pVAPA116 compared with pVAPA1037. In conclusion, it appears that the divergence between the two sequenced rhodococcal virulence plasmids is not associated with the vap pathogenicity island and may result from an evolutionary process driven by a mobility-related invertase/resolvase invA-like gene. © 2010 ANSES. Journal compilation © 2010 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd.

  2. A Site-Specific Integrative Plasmid Found in Pseudomonas aeruginosa Clinical Isolate HS87 along with A Plasmid Carrying an Aminoglycoside-Resistant Gene.

    Directory of Open Access Journals (Sweden)

    Dexi Bi

    Full Text Available Plasmids play critical roles in bacterial fitness and evolution of Pseudomonas aeruginosa. Here two plasmids found in a drug-resistant P. aeruginosa clinical isolate HS87 were completely sequenced. The pHS87b plasmid (11.2 kb carries phage-related genes and function-unknown genes. Notably, pHS87b encodes an integrase and has an adjacent tRNAThr-associated attachment site. A corresponding integrated form of pHS87b at the tRNAThr locus was identified on the chromosome of P. aeruginosa, showing that pHS87b is able to site-specifically integrate into the 3'-end of the tRNAThr gene. The pHS87a plasmid (26.8 kb displays a plastic structure containing a putative replication module, stability factors and a variable region. The RepA of pHS87a shows significant similarity to the replication proteins of pPT23A-family plasmids. pHS87a carries a transposon Tn6049, a truncated insertion sequence ΔIS1071 and a Tn402-like class 1 integron which contains an aacA4 cassette that may confer aminoglycoside resistance. Thus, pHS87b is a site-specific integrative plasmid whereas pHS87a is a plastic antibiotic resistance plasmid. The two native plasmids may promote the fitness and evolution of P. aeruginosa.

  3. UV-cured methacrylic-silica hybrids: Effect of oxygen inhibition on photo-curing kinetics

    Energy Technology Data Exchange (ETDEWEB)

    Corcione, C. Esposito; Striani, R.; Frigione, M., E-mail: mariaenrica.frigione@unisalento.it

    2014-01-20

    Highlights: • The kinetic behavior of novel photopolymerizable organic–inorganic hybrid system was studied as a function of the composition and of the atmosphere for reactions. • The UV-curing reaction of the hybrid mixture was found fast and complete. • The combined presence of thiol monomer and nanostructured silica allows to reduce the effect of inhibition of oxygen towards the radical photopolymerization. - Abstract: The kinetic behavior of innovative photopolymerizable UV-cured methacrylic–silica hybrid formulations, previously developed, was studied and compared to that of a reference control system. The organic–inorganic (O–I) hybrids proposed in this study are obtained from organic precursors with a high siloxane content mixed with tetraethoxysilane (TEOS) in such a way to produce co-continuous silica nano-domains dispersed within a cross-linked organic phase, as a result of the hydrolysis and condensation reactions. The kinetics of the radical photopolymerization mechanism induced by UV-radiations, in presence of a suitable photoinitiator, was studied by calorimetric, FTIR and Raman spectroscopic analyses, by varying the composition of the mixtures and the atmosphere for reactions. The well known effect of oxygen on the kinetic mechanism of the free radical photopolymerization of the methacrylic–siloxane based monomers was found to be strongly reduced in the hybrid system, especially when a proper thiol was used. The experimental calorimetric data were fitted using a simple kinetic model for radical photopolymerization reactions, obtaining a good agreement between the experimental data and the theoretical model. From the comparison of the kinetic constants calculated for control and hybrid systems, it was possible to assess the effect of the composition, as well as of the atmosphere used during the photo-polymerization process, on the kinetic of photopolymerization reaction.

  4. Comparison of Shear Bond Strength and Estimation of Adhesive Remnant Index between Light-cure Composite and Dual-cure Composite: An in vitro Study.

    Science.gov (United States)

    Verma, Geeta; Trehan, Mridula; Sharma, Sunil

    2013-09-01

    To measure and compare the shear bond strength and adhesive remnant index of light-cure composite. (Enlight, Ormco.) and dual-cure composite (Phase II dual cure, Reliance Ortho). Sixty extracted human premolar teeth were divided into two groups: group I (blue): conventional light cure composite resin. (Enlight, Ormco.) and group II (green): dual cure composite resin. (Phase II dual cure, Reliance Ortho.) with 30 teeth in each group. These samples were tested on the universal testing machine to measure the shear bond strength. Student t-test showed that the mean shear bond strength of the conventional light cure group (8.54 MPa - 10.42 MPa) was significantly lower than dual cure group (10.45 MPa -12.17 MPa). These findings indicate that the shear bond strength of dual-cure composite resin (Phase II dual cure, Reliance Ortho) is comparatively higher than conventional light-cure composite resin (Enlight, Ormco). In the majority of the samples, adhesive remnant index (ARI) scores were 4 and 5 in both the groups whereas score 1 is attained by the least number of samples in both the groups. How to cite this article: Verma G, Trehan M, Sharma S. Comparison of Shear Bond Strength and Estimation of Adhesive Remnant Index between Light-cure Composite and Dual-cure Composite: An in vitro Study. Int J Clin Pediatr Dent 2013;6(3):166-170.

  5. Influence of argon laser curing on resin bond strength.

    Science.gov (United States)

    Hinoura, K; Miyazaki, M; Onose, H

    1993-04-01

    Light cured resin composites are usually cured with halogen lamps whose light output decreases with time and distance to the resin surface. This study compared bond strengths of resins to tooth structure cured with either an argon laser or a conventional halogen light. The enamel and dentin of bovine incisors were ground on the buccal surface with wet #600 grit SiC paper. A 4 x 2 mm mold was placed on the tooth surface and Scotchbond 2/Silux and Clearfil Photobond/Photo Clearfil A were placed into the molds and cured using a Quick Light or an argon laser for exposure times of 10, 20, and 30 seconds, and distances of 0.0, 0.5, 1.0, and 1.5 mm from the resin surface. The intensity of the Quick Light was measured as 510 mW/cm2 at 470 +/- 15 nm and the intensity of the argon laser was adjusted to 510 mW/cm2 before curing. Shear bond tests at a crosshead speed of 1.0 mm/min were performed after 24 hours of storage in water. The bond strengths obtained with the halogen lamp and the laser were not significantly different at the same exposure times and at 0.0 or 0.5 mm from the resin surface. The laser cured bond strengths did not decrease with increasing distance whereas there was a significant decrease in halogen bond strengths at distances greater than 0.5 mm for both resins. The use of the laser might provide a clinical advantage in cases where the curing light source cannot be brought into proximity to the surface of the resin.

  6. SENSORY CHARACTERISTICS OF NATIVE CHICKEN QUEEN PINEAPPLE-CURED HAM

    Directory of Open Access Journals (Sweden)

    Dr. Lilibeth A. Roxas

    2015-01-01

    Full Text Available The potential of Native Chicken to be processed into palatable ham was conducted making use of Queen Pineapple (QP crude extract as one of the curing ingredients. Primarily, the main goal is to develop a protocol in the manufacture of processed native chicken ham and determine the organoleptic quality of native chicken ham product. The age of the bird and maturity of the fruit were considered for the best organoleptic quality of chicken ham. In this study, the combine injection and dry cure (CIDC method of the conventional formula was adopted. The desired amount of QP crude extract was first determined for the pump pickle. Curing salt was used for the control while different volume of pineapple crude extract was used in two treatments. The protocols for processing native chicken were developed using slaughter native chicken, and QP crude extract as curing ingredient for ham making. Color, flavor, juiciness and tenderness were among the desirable characteristics considered in this study. The sensory evaluation by trained panelists on QP-cured ham samples demonstrated comparable results. All the cooked meat samples were apparently acceptable to the sensory panel. The mean scores for flavor, juiciness and tenderness of meat samples have slight differences; however, they are not statistically significant. Indeed, native chicken can be processed into palatable ham with queen pineapple (Formosa variety extract that served as curing ingredient, flavor enhancer and tenderizer. Native Chicken QP-Cured ham is a commendable value-added product for both native chicken and queen pineapple by-products (butterball size.

  7. Insights into dynamics of mobile genetic elements in hyperthermophilic environments from five new Thermococcus plasmids.

    Directory of Open Access Journals (Sweden)

    Mart Krupovic

    Full Text Available Mobilome of hyperthermophilic archaea dwelling in deep-sea hydrothermal vents is poorly characterized. To gain insight into genetic diversity and dynamics of mobile genetic elements in these environments we have sequenced five new plasmids from different Thermococcus strains that have been isolated from geographically remote hydrothermal vents. The plasmids were ascribed to two subfamilies, pTN2-like and pEXT9a-like. Gene content and phylogenetic analyses illuminated a robust connection between pTN2-like plasmids and Pyrococcus abyssi virus 1 (PAV1, with roughly half of the viral genome being composed of genes that have homologues in plasmids. Unexpectedly, pEXT9a-like plasmids were found to be closely related to the previously sequenced plasmid pMETVU01 from Methanocaldococcus vulcanius M7. Our data suggests that the latter observation is most compatible with an unprecedented horizontal transfer of a pEXT9a-like plasmid from Thermococcales to Methanococcales. Gene content analysis revealed that thermococcal plasmids encode Hfq-like proteins and toxin-antitoxin (TA systems of two different families, VapBC and RelBE. Notably, although abundant in archaeal genomes, to our knowledge, TA and hfq-like genes have not been previously found in archaeal plasmids or viruses. Finally, the plasmids described here might prove to be useful in developing new genetic tools for hyperthermophiles.

  8. RK2 plasmid dynamics in Caulobacter crescentus cells--two modes of DNA replication initiation.

    Science.gov (United States)

    Wegrzyn, Katarzyna; Witosinska, Monika; Schweiger, Pawel; Bury, Katarzyna; Jenal, Urs; Konieczny, Igor

    2013-06-01

    Undisturbed plasmid dynamics is required for the stable maintenance of plasmid DNA in bacterial cells. In this work, we analysed subcellular localization, DNA synthesis and nucleoprotein complex formation of plasmid RK2 during the cell cycle of Caulobacter crescentus. Our microscopic observations showed asymmetrical distribution of plasmid RK2 foci between the two compartments of Caulobacter predivisional cells, resulting in asymmetrical allocation of plasmids to progeny cells. Moreover, using a quantitative PCR (qPCR) method, we estimated that multiple plasmid particles form a single fluorescent focus and that the number of plasmids per focus is approximately equal in both swarmer and predivisional Caulobacter cells. Analysis of the dynamics of TrfA-oriV complex formation during the Caulobacter cell cycle revealed that TrfA binds oriV primarily during the G1 phase, however, plasmid DNA synthesis occurs during the S and G2 phases of the Caulobacter cell cycle. Both in vitro and in vivo analysis of RK2 replication initiation in C. crescentus cells demonstrated that it is independent of the Caulobacter DnaA protein in the presence of the longer version of TrfA protein, TrfA-44. However, in vivo stability tests of plasmid RK2 derivatives suggested that a DnaA-dependent mode of plasmid replication initiation is also possible.

  9. Polar Fixation of Plasmids during Recombinant Protein Production in Bacillus megaterium Results in Population Heterogeneity.

    Science.gov (United States)

    Münch, Karin M; Müller, Johannes; Wienecke, Sarah; Bergmann, Simone; Heyber, Steffi; Biedendieck, Rebekka; Münch, Richard; Jahn, Dieter

    2015-09-01

    During the past 2 decades, Bacillus megaterium has been systematically developed for the gram-per-liter scale production of recombinant proteins. The plasmid-based expression systems employed use a xylose-controlled promoter. Protein production analyses at the single-cell level using green fluorescent protein as a model product revealed cell culture heterogeneity characterized by a significant proportion of less productive bacteria. Due to the enormous size of B. megaterium, such bistable behavior seen in subpopulations was readily analyzed by time lapse microscopy and flow cytometry. Cell culture heterogeneity was not caused simply by plasmid loss: instead, an asymmetric distribution of plasmids during cell division was detected during the exponential-growth phase. Multicopy plasmids are generally randomly distributed between daughter cells. However, in vivo and in vitro experiments demonstrated that under conditions of strong protein production, plasmids are retained at one of the cell poles. Furthermore, it was found that cells with accumulated plasmids and high protein production ceased cell division. As a consequence, the overall protein production of the culture was achieved mainly by the subpopulation with a sufficient plasmid copy number. Based on our experimental data, we propose a model whereby the distribution of multicopy plasmids is controlled by polar fixation under protein production conditions. Thereby, cell lines with fluctuating plasmid abundance arise, which results in population heterogeneity. Our results provide initial insights into the mechanism of cellular heterogeneity during plasmid-based recombinant protein production in a Bacillus species.

  10. Insights into dynamics of mobile genetic elements in hyperthermophilic environments from five new Thermococcus plasmids.

    Science.gov (United States)

    Krupovic, Mart; Gonnet, Mathieu; Hania, Wajdi Ben; Forterre, Patrick; Erauso, Gaël

    2013-01-01

    Mobilome of hyperthermophilic archaea dwelling in deep-sea hydrothermal vents is poorly characterized. To gain insight into genetic diversity and dynamics of mobile genetic elements in these environments we have sequenced five new plasmids from different Thermococcus strains that have been isolated from geographically remote hydrothermal vents. The plasmids were ascribed to two subfamilies, pTN2-like and pEXT9a-like. Gene content and phylogenetic analyses illuminated a robust connection between pTN2-like plasmids and Pyrococcus abyssi virus 1 (PAV1), with roughly half of the viral genome being composed of genes that have homologues in plasmids. Unexpectedly, pEXT9a-like plasmids were found to be closely related to the previously sequenced plasmid pMETVU01 from Methanocaldococcus vulcanius M7. Our data suggests that the latter observation is most compatible with an unprecedented horizontal transfer of a pEXT9a-like plasmid from Thermococcales to Methanococcales. Gene content analysis revealed that thermococcal plasmids encode Hfq-like proteins and toxin-antitoxin (TA) systems of two different families, VapBC and RelBE. Notably, although abundant in archaeal genomes, to our knowledge, TA and hfq-like genes have not been previously found in archaeal plasmids or viruses. Finally, the plasmids described here might prove to be useful in developing new genetic tools for hyperthermophiles.

  11. Exploring Antibiotic Resistance Genes and Metal Resistance Genes in Plasmid Metagenomes from Wastewater Treatment Plants

    Directory of Open Access Journals (Sweden)

    An-Dong eLi

    2015-09-01

    Full Text Available Plasmids operate as independent genetic elements in microorganism communities. Through horizontal gene transfer, they can provide their host microorganisms with important functions such as antibiotic resistance and heavy metal resistance. In this study, six metagenomic libraries were constructed with plasmid DNA extracted from influent, activated sludge and digested sludge of two wastewater treatment plants. Compared with the metagenomes of the total DNA extracted from the same sectors of the wastewater treatment plant, the plasmid metagenomes had significantly higher annotation rates, indicating that the functional genes on plasmids are commonly shared by those studied microorganisms. Meanwhile, the plasmid metagenomes also encoded many more genes related to defense mechanisms, including ARGs. Searching against an antibiotic resistance genes (ARGs database and a metal resistance genes (MRGs database revealed a broad-spectrum of antibiotic (323 out of a total 618 subtypes and metal resistance genes (23 out of a total 23 types on these plasmid metagenomes. The influent plasmid metagenomes contained many more resistance genes (both ARGs and MRGs than the activated sludge and the digested sludge metagenomes. Sixteen novel plasmids with a complete circular structure that carried these resistance genes were assembled from the plasmid metagenomes. The results of this study demonstrated that the plasmids in wastewater treatment plants could be important reservoirs for resistance genes, and may play a significant role in the horizontal transfer of these genes.

  12. Exploring antibiotic resistance genes and metal resistance genes in plasmid metagenomes from wastewater treatment plants.

    Science.gov (United States)

    Li, An-Dong; Li, Li-Guan; Zhang, Tong

    2015-01-01

    Plasmids operate as independent genetic elements in microorganism communities. Through horizontal gene transfer (HGT), they can provide their host microorganisms with important functions such as antibiotic resistance and heavy metal resistance. In this study, six metagenomic libraries were constructed with plasmid DNA extracted from influent, activated sludge (AS) and digested sludge (DS) of two wastewater treatment plants (WWTPs). Compared with the metagenomes of the total DNA extracted from the same sectors of the wastewater treatment plant, the plasmid metagenomes had significantly higher annotation rates, indicating that the functional genes on plasmids are commonly shared by those studied microorganisms. Meanwhile, the plasmid metagenomes also encoded many more genes related to defense mechanisms, including ARGs. Searching against an antibiotic resistance genes (ARGs) database and a metal resistance genes (MRGs) database revealed a broad-spectrum of antibiotic (323 out of a total 618 subtypes) and MRGs (23 out of a total 23 types) on these plasmid metagenomes. The influent plasmid metagenomes contained many more resistance genes (both ARGs and MRGs) than the AS and the DS metagenomes. Sixteen novel plasmids with a complete circular structure that carried these resistance genes were assembled from the plasmid metagenomes. The results of this study demonstrated that the plasmids in WWTPs could be important reservoirs for resistance genes, and may play a significant role in the horizontal transfer of these genes.

  13. Conservation of plasmids among Escherichia coli K1 isolates of diverse origins.

    Science.gov (United States)

    Mercer, A A; Morelli, G; Heuzenroeder, M; Kamke, M; Achtman, M

    1984-12-01

    Escherichia coli K1 isolates of various O types were previously assigned to different clonal groups. Members of the two clones defined by membrane pattern 9 (MP9) and serotypes O18:K1 and O1:K1 had been found to be very similar to each other. The plasmid contents of these bacteria confirmed this conclusion. Both groups carried a self-transmissible plasmid of the FI incompatibility group that coded for colicin production and a major outer membrane protein called the plasmid-coded protein (PCP). The size of this plasmid varied from 76 to 96 megadaltons, but restriction endonuclease digestion and DNA heteroduplex analysis revealed that these plasmids were highly related. O18:K1 bacteria of MP6 had previously been determined to represent a subclone, related to but different from O18:K1 MP9 bacteria. These MP6 bacteria carried a different, smaller IncFI plasmid which did not code for colicin production or the PCP protein. This smaller plasmid was primarily related to the larger plasmid within the regions of DNA encoding incompatibility, replication, and conjugation. O1:K1 bacteria of MP5 contained other unrelated plasmids in agreement with the previous conclusion that they are unrelated to O1:K1 bacteria of MP9. The bacteria examined had been isolated from two continents over a time span of 38 years, and the results attest to conservative inheritance of plasmids within bacteria of common descent.

  14. Conservation of plasmids among Escherichia coli K1 isolates of diverse origins.

    Science.gov (United States)

    Mercer, A A; Morelli, G; Heuzenroeder, M; Kamke, M; Achtman, M

    1984-01-01

    Escherichia coli K1 isolates of various O types were previously assigned to different clonal groups. Members of the two clones defined by membrane pattern 9 (MP9) and serotypes O18:K1 and O1:K1 had been found to be very similar to each other. The plasmid contents of these bacteria confirmed this conclusion. Both groups carried a self-transmissible plasmid of the FI incompatibility group that coded for colicin production and a major outer membrane protein called the plasmid-coded protein (PCP). The size of this plasmid varied from 76 to 96 megadaltons, but restriction endonuclease digestion and DNA heteroduplex analysis revealed that these plasmids were highly related. O18:K1 bacteria of MP6 had previously been determined to represent a subclone, related to but different from O18:K1 MP9 bacteria. These MP6 bacteria carried a different, smaller IncFI plasmid which did not code for colicin production or the PCP protein. This smaller plasmid was primarily related to the larger plasmid within the regions of DNA encoding incompatibility, replication, and conjugation. O1:K1 bacteria of MP5 contained other unrelated plasmids in agreement with the previous conclusion that they are unrelated to O1:K1 bacteria of MP9. The bacteria examined had been isolated from two continents over a time span of 38 years, and the results attest to conservative inheritance of plasmids within bacteria of common descent. Images PMID:6094355

  15. 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.

  16. Plasmids of the pRM/pRF family occur in diverse Rickettsia species.

    Science.gov (United States)

    Baldridge, Gerald D; Burkhardt, Nicole Y; Felsheim, Roderick F; Kurtti, Timothy J; Munderloh, Ulrike G

    2008-02-01

    The recent discoveries of the pRF and pRM plasmids of Rickettsia felis and R. monacensis have contravened the long-held dogma that plasmids are not present in the bacterial genus Rickettsia (Rickettsiales; Rickettsiaceae). We report the existence of plasmids in R. helvetica, R. peacockii, R. amblyommii, and R. massiliae isolates from ixodid ticks and in an R. hoogstraalii isolate from an argasid tick. R. peacockii and four isolates of R. amblyommii from widely separated geographic locations contained plasmids that comigrated with pRM during pulsed-field gel electrophoresis and larger plasmids with mobilities similar to that of pRF. The R. peacockii plasmids were lost during long-term serial passage in cultured cells. R. montanensis did not contain a plasmid. Southern blots showed that sequences similar to those of a DnaA-like replication initiator protein, a small heat shock protein 2, and the Sca12 cell surface antigen genes on pRM and pRF were present on all of the plasmids except for that of R. massiliae, which lacked the heat shock gene and was the smallest of the plasmids. The R. hoogstraalii plasmid was most similar to pRM and contained apparent homologs of proline/betaine transporter and SpoT stringent response genes on pRM and pRF that were absent from the other plasmids. The R. hoogstraalii, R. helvetica, and R. amblyommii plasmids contained homologs of a pRM-carried gene similar to a Nitrobacter sp. helicase RecD/TraA gene, but none of the plasmids hybridized with a probe derived from a pRM-encoded gene similar to a Burkholderia sp. transposon resolvase gene.

  17. Effect of excessive cadmium chloride on the plasmids of E. coli HB 101 in vivo

    Institute of Scientific and Technical Information of China (English)

    1999-01-01

    After Escherichia coli HB101 with plasmid pWH58, pWH98, or pTBa5 were cultered respectively in amp LB broth which contained 50 mg/L CdCl2 constantly for 24 h, these plasmids were isolated from E. coli, and the effect of excessive CdCl2 on the E. coli HB101 and plasmid DNA was studied by surveying the growth of E. coli HB101 and plasmid, argarose gel electrophoresis and analysis of restriction fragment length polymorphism (RFLP) of plasmids, and plasmid transformation. The results showed that 50 mg/L CdCl2 treatment lagged the growth of E. coli HB101 for at least 4h, but after grown for 24h there were not significant differences in the growths of E. coli HB101s and the productions of plasmids between the treatment and control. These results implified that E. coli HB101 have induced adaptability to cadmium stress and excessive CdCl2 did not inhibit the replication and amp+ gene's expression of plasmid DNA in vivo of E. coli significantly. 50 mg/L CdCl2 treatment for 24 hours might cause the sequence's change of plasmid DNA, but could not lead to the random breakage of plasmid DNA strands. Moreover, after 50 mg/L of CdCl2 treatment in vivo the transformation activities of plasmid did not altered, implied excessive CdCl2 could not affect the superhelical structure of plasmid and also not break the loop of plasmid DNA evidently.

  18. Dual-Replicon Plasmids for Elimination of Resistant Plasmids%双复制子质粒消除耐药质粒的研究

    Institute of Scientific and Technical Information of China (English)

    莫冰; 余克花; 黎帆; 李蓉; 宋矿余

    2014-01-01

    Objective To observe the eliminating effect of dual-replicon plasmids on resistant plasmids.Methods The dual-replicon plasmids pKT230-oriV were constructed and transferred into bacteria with resistant plasmids pRK290 through the transformation and conjugational trans-fer pathways.The eliminating effect of dual-replicon plasmids on resistant plasmids was ob-served.Results The bacteria were cultured for 5 generations after the transfer of dual-replicon plasmids,and resistant plasmids pRK290 in bacteria were eliminated.Conclusion The conjuga-tional transfer of dual-replicon plasmids is a pathway to eliminate resistant plasmids.%目的:观察双复制子质粒对耐药质粒的消除作用。方法构建双复制子质粒 pKT230-oriV,并通过转化或接合转移途径转入带有 pRK290质粒的细菌中,观察双复制子质粒对耐药质粒的消除作用。结果转入双复制子质粒菌培养5代后,细菌中的 pRK290耐药质粒被消除。结论双复制子利用接合转移途径是消除耐药质粒的一个途径。

  19. 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.

  20. Plasmid DNA Supercoiling and Gyrase Activity in Escherichia coli Wild-Type and rpoS Stationary-Phase Cells

    Science.gov (United States)

    Reyes-Domínguez, Yazmid; Contreras-Ferrat, Gabriel; Ramírez-Santos, Jesús; Membrillo-Hernández, Jorge; Gómez-Eichelmann, M. Carmen

    2003-01-01

    Stationary-phase cells displayed a distribution of relaxed plasmids and had the ability to recover plasmid supercoiling as soon as nutrients became available. Preexisting gyrase molecules in these cells were responsible for this recovery. Stationary-phase rpoS cells showed a bimodal distribution of plasmids and failed to supercoil plasmids after the addition of nutrients, suggesting that rpoS plays a role in the regulation of plasmid topology during the stationary phase. PMID:12533486

  1. Caul and method for bonding and curing intricate composite structures

    Science.gov (United States)

    Willden, Kurtis S. (Inventor); Goodno, Kenneth N. (Inventor)

    1993-01-01

    The invention disclosed here is a method for forming and curing an intricate structure of criss-crossing composite stringers and frames that are bonded to a skin panel. A structure constructed in accordance with the invention would be well-suited for use as a portion of an aircraft fuselage, a boat hull, or the like. The method is preferably practiced by applying uncured composite stringers to an uncured composite sheet panel. This is followed by placing cured frames crosswise over the stringers. The frames have openings at the locations where they intersect with the stringers which enables the frames to come into direct contact with the skin along most of their length. During the forming and curing process, the stringers are covered with a plurality of cauls, and the entire assembly of skin panel, stringers, frames and cauls is subjected to a vacuum bagging and curing process. The cauls serve to maintain both part shape and to control the flow of resin within the stringers as they are cured. Further, they probably eliminate the need for intermediate protective materials between the vacuum bag and the stringers.

  2. Photothermal radiometry monitoring of light curing in resins

    Energy Technology Data Exchange (ETDEWEB)

    Zambrano-Arjona, M A [Applied Physics Department, Cinvestav-Unidad Merida, A.P. 73 Cordemex, Merida, 97310 (Mexico); Medina-Esquivel, R [Cinvestav-Unidad Queretaro, Libramiento Norponiente 2000 Fracc. Real de Juriquilla, CP 76230, Queretaro (Mexico); Alvarado-Gil, J J [Applied Physics Department, Cinvestav-Unidad Merida, A.P. 73 Cordemex, Merida, 97310 (Mexico)

    2007-10-07

    Real time measurement of thermal diffusivity during the evolution of the light curing process in dental resins is reported using photothermal radiometry. The curing is induced by a non-modulated blue light beam, and at the same time, a modulated red laser beam is sent onto the sample, generating a train of thermal waves that produce modulated infrared radiation. The monitoring of this radiation permits to follow the time evolution of the process. The methodology is applied to two different commercially available light curing resin-based composites. In all cases thermal diffusivity follows a first order kinetics with similar stabilization characteristic times. Analysis of this kinetics permits to exhibit the close relationship of increase in thermal diffusivity with the decrease in monomer concentration and extension of the polymerization in the resin, induced by the curing light. It is also shown that the configuration in which the resin is illuminated by the modulated laser can be the basis for the development of an in situ technique for the determination of the degree of curing.

  3. Thermal stability and curing kinetics of polycarbosilane fibers

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Thermal stability and curing kinetics of polycarbosilane (PCS) fibers were studied by thermogravimetry (TG), Fourier transform infrared spectroscopy(FT-IR). Curing is an essential step in the preparation of SiC fibers and the properties of SiC fibers are affected greatly by curing conditions. TG measurement performed in air shows that mass gain starts at approximately 200℃ and PCS fibers are sensitive to oxygen. Curing with oxygen, which results in crosslinking on the surface, enabled PCS fibers to retain its shape during high-temperature pyrolysis. The curing of PCS fibers is oxidation of Si-H and Si-CH3, then Si-O-Si and Si-O-C bonds are formed. This is a first order reaction, with activation energy of 79.27 kJ/mol, and the pre-exponential factor is calculated as 3.07 × 106.The kinetics model was obtained and the experimental data of PCS fibers show good agreement with the kinetics model.

  4. Fiber-Matrix Interface Studies on Electron Beam Cured Composites

    Energy Technology Data Exchange (ETDEWEB)

    Drazel, L.T.; Janke, C.J.; Yarborough, K.D.

    1999-05-23

    The recently completed Department of Energy (DOE) and industry sponsored Cooperative Research and Development Agreement (CRADA) entitled, ''Electron Beam Curing of Polymer Matrix Composites,'' determined that the interlaminar shear strength properties of the best electron beam cured IM7/epoxy composites were 19-28% lower than autoclave cured IM7/epoxy composites (i.e. IM7/977-2 and IM7/977-3). Low interlaminar shear strength is widely acknowledged as the key barrier to the successful acceptance and implementation of electron beam cured composites in the aircraft/aerospace industry. The objective of this work was to improve the interlaminar shear strength properties of electron beam cured composites by formulating and evaluating several different fiber sizings or coating materials. The researchers have recently achieved some promising results by having discovered that the application of epoxy-based, electron beam compatible sizings or coatings onto surface-treated, unsized IM7 carbon fibers improved the composite interlaminar shear strength properties by as much as 55% versus composites fabricated from surface-treated, unsized IM7 fibers. In addition, by applying these same epoxy-based sizings or coatings onto surface-treated, unsized IM7 fibers it was possible to achieve an 11% increase in the composite interlaminar shear strength compared to composites made from surface-treated, GP-sized IM7 fibers. Work is continuing in this area of research to further improve these properties.

  5. Characterization of Moisture Diffusion in Cured Concrete Slabs at Early Ages

    OpenAIRE

    Xiao Zhang; Hongduo Zhao

    2015-01-01

    The objective of this paper is to investigate the characterization of moisture diffusion inside early-age concrete slabs subjected to curing. Time-dependent relative humidity (RH) distributions of three mixture proportions subjected to three different curing methods (i.e., air curing, water curing, and membrane-forming compounds curing) and sealed condition were measured for 28 days. A one-dimensional nonlinear moisture diffusion partial differential equation (PDE) based on Fick’s second law,...

  6. Prediction of 28-day Compressive Strength of Concrete from Early Strength and Accelerated Curing Parameters

    OpenAIRE

    T.R. Neelakantan; S. Ramasundaram; Shanmugavel, R.; R. Vinoth

    2013-01-01

    Predicting 28-day compressive strength of concrete is an important research task for many years. In this study, concrete specimens were cured in two phases, initially at room temperature for a maximum of 30 h and later at a higher temperature for accelerated curing for a maximum of 3 h. Using the early strength obtained after the two-phase curing and the curing parameters, regression equations were developed to predict the 28-day compressive strength. For the accelerated curing (higher temper...

  7. Comparative evaluation of surface porosities in conventional heat polymerized acrylic resin cured by water bath and microwave energy with microwavable acrylic resin cured by microwave energy

    Directory of Open Access Journals (Sweden)

    Sunint Singh

    2013-01-01

    Full Text Available Background: Conventional heat cure poly methyl methacrylate (PMMA is the most commonly used denture base resin despite having some short comings. Lengthy polymerization time being one of them and in order to overcome this fact microwave curing method was recommended. Unavailability of specially designed microwavable acrylic resin made it unpopular. Therefore, in this study, conventional heat cure PMMA was polymerized by microwave energy. Aim and Objectives: This study was designed to evaluate the surface porosities in PMMA cured by conventional water bath and microwave energy and compare it with microwavable acrylic resin cured by microwave energy. Materials and Methods: Wax samples were obtained by pouring molten wax into a metal mold of 25 mm × 12 mm × 3 mm dimensions. These samples were divided into three groups namely C, CM, and M. Group C denotes conventional heat cure PMMA cured by water bath method, CM denotes conventional heat cure PMMA cured by microwave energy, M denotes specially designed microwavable acrylic denture base resin cured by microwave energy. After polymerization, each sample was scanned in three pre-marked areas for surface porosities using the optical microscope. As per the literature available, this instrument is being used for the first time to measure the porosity in acrylic resin. It is a reliable method of measuring area of surface pores. Portion of the sample being scanned is displayed on the computer and with the help of software area of each pore was measured and data were analyzed. Results: Conventional heat cure PMMA samples cured by microwave energy showed maximum porosities than the samples cured by conventional water bath method and microwavable acrylic resin cured by microwave energy. Higher percentage of porosities was statistically significant, but well within the range to be clinically acceptable. Conclusion: Within the limitations of this in-vitro study, conventional heat cure PMMA can be cured by

  8. [Plasmid characteristics of naphthalene and salicylate biodegradation in Pseudomonas putida].

    Science.gov (United States)

    Zakharian, R A; Bakunin, K A; Gasparian, N S; Kocharian, Sh M; Arakelov, G M

    1980-01-01

    The object of this work was to study the physico-chemical and biological properties of DNAs of the biodegradation plasmids NAH and SAL. A comparative analysis of the physico-chemical parameters for these DNAs made it possible to detect a number of identical properties in them: the same sedimentation profile for covalently-closed circular DNA forms, 68--70 S; the molecular weight of ca. 50 MD; a roughly equal number of fragments (up to 23) was found when the DNAs of NAH and SAL were restricted by EcoRI endonuclease. The transformation of the plasmidless strain PpGI was done.

  9. pTAR-Encoded Proteins in Plasmid Partitioning

    OpenAIRE

    Kalnin, Kirill; Stegalkina, Svetlana; Yarmolinsky, Michael

    2000-01-01

    Partition cassettes, essential for the segregational stability of low-copy-number bacterial plasmids, typically encode two autoregulated proteins and an adjacent cis-acting centromere analog to which one or perhaps both proteins bind. The diminutive partition region of pTAR of Agrobacterium spp. was reported to be exceptional, encoding only a single protein, ParA (D. R. Gallie and C. I. Kado, J. Mol. Biol. 193:465–478, 1987). However, resequencing of the region revealed two small downstream g...

  10. Recombinogenic engineering of conjugative plasmids with fluorescent marker cassettes

    DEFF Research Database (Denmark)

    Reisner, A.; Molin, Søren; Zechner, E.L.

    2002-01-01

    functions using very short regions of homology. Initial manipulation of the IncFII target plasmids R1 and R1drd19 indicated that the linear targeting DNA should be devoid of all extraneous homologies to. the target molecule for optimal insertion specificity. Indeed, a simple recombination assay proved...... that in the presence of additional homologous regions in the targeting DNA, strand exchanges occurred exclusively within the longest regions of homology. A versatile panel of vectors was created to facilitate convenient PCR amplification of targeting DNAs containing various combinations of different antibiotic...

  11. Brazilian purpuric fever caused by Haemophilus influenzae biogroup aegyptius strains lacking the 3031 plasmid.

    Science.gov (United States)

    Tondella, M L; Quinn, F D; Perkins, B A

    1995-01-01

    Brazilian purpuric fever (BPF) is a life-threatening pediatric infection caused by Haemophilus influenzae biogroup aegyptius (Hae), an organism formerly associated with only self-limited purulent conjunctivitis. Strains of Hae causing BPF have a 24-MDa plasmid with a specific AccI restriction pattern designated 3031. This plasmid was thought to code for a virulence factor because it had been detected only among Hae strains isolated from BPF cases or their contacts. From 3 typical BPF cases recently identified in São Paulo State, sterile-site Hae isolates were obtained; these isolates were similar to earlier BPF-associated Hae except they did not possess a 3031 plasmid. HindIII restricted chromosomal DNA from these strains was probed with purified 3031 plasmid DNA under high-stringency conditions. There was no evidence that 3031 plasmid DNA had become chromosomally integrated. It appears that the 3031 plasmid does not code for BPF-specific virulence factors.

  12. IncA/C plasmids: An emerging threat to human and animal health?

    Science.gov (United States)

    Johnson, Timothy J; Lang, Kevin S

    2012-01-01

    Incompatibility group IncA/C plasmids are large, low copy, theta-replicating plasmids that have been described in the literature for over 40 years. However, they have only recently been intensively studied on the genomic level because of their associations with the emergence of multidrug resistance in enteric pathogens of humans and animals. These plasmids are unique among other enterobacterial plasmids in many aspects, including their modular structure and gene content. While the IncA/C plasmid genome structure has now been well defined, many questions remain pertaining to their basic biological mechanisms of dissemination and regulation. Here, we discuss the history of IncA/C plasmids in light of our recent understanding of their population distribution, genomics, and effects on host bacteria.

  13. Long- term manure exposure increases soil bacterial community potential for plasmid uptake

    DEFF Research Database (Denmark)

    Musovic, Sanin; Klümper, Uli; Dechesne, Arnaud;

    2014-01-01

    Microbial communities derived from soils subject to different agronomic treatments were challenged with three broad host range plasmids, RP4, pIPO2tet and pRO101, via solid surface filter matings to assess their permissiveness. Approximately 1 in 10 000 soil bacterial cells could receive and main......Microbial communities derived from soils subject to different agronomic treatments were challenged with three broad host range plasmids, RP4, pIPO2tet and pRO101, via solid surface filter matings to assess their permissiveness. Approximately 1 in 10 000 soil bacterial cells could receive...... and maintain the plasmids. The community permissiveness increased up to 100% in communities derived from manured soil. While the plasmid transfer frequency was significantly influenced by both the type of plasmid and the agronomic treatment, the diversity of the transconjugal pools was purely plasmid dependent...

  14. Novel archaeal plasmid pAH1 and its interactions with the lipothrixvirus AFV1

    DEFF Research Database (Denmark)

    Basta, Tamara; Smyth, John; Forterre, Patrick

    2009-01-01

    to establish a system for studying plasmid-virus interactions we characterized the genome of pAH1 which closely resembles those of the Sulfolobus conjugative plasmids pARN3 and pARN4. pAH1 integrates site specifically into, and excises from, the host chromosome indicating a dynamic interaction with the latter....... Although nucleotide sequence comparisons revealed extensive intergenomic exchange during the evolution of archaeal conjugative plasmids, pAH1 was shown to be stably maintained suggesting that the host system is suitable for studying plasmid-virus interactions. AFV1 infection and propagation leads to a loss...... of the circular form of pAH1 and this effect correlates positively with the increase in the intracellular quantity of AFV1 DNA. We infer that the virus inhibits plasmid replication since no pAH1 degradation was observed. This mechanism of archaeal viral inhibition of plasmid propagation is not observed...

  15. Strategies and approaches in plasmidome studies—uncovering plasmid diversity disregarding of linear elements?

    Science.gov (United States)

    Dib, Julián R.; Wagenknecht, Martin; Farías, María E.; Meinhardt, Friedhelm

    2015-01-01

    The term plasmid was originally coined for circular, extrachromosomal genetic elements. Today, plasmids are widely recognized not only as important factors facilitating genome restructuring but also as vehicles for the dissemination of beneficial characters within bacterial communities. Plasmid diversity has been uncovered by means of culture-dependent or -independent approaches, such as endogenous or exogenous plasmid isolation as well as PCR-based detection or transposon-aided capture, respectively. High-throughput-sequencing made possible to cover total plasmid populations in a given environment, i.e., the plasmidome, and allowed to address the quality and significance of self-replicating genetic elements. Since such efforts were and still are rather restricted to circular molecules, here we put equal emphasis on the linear plasmids which—despite their frequent occurrence in a large number of bacteria—are largely neglected in prevalent plasmidome conceptions. PMID:26074886

  16. Influence of light curing units on failure of directcomposite restorations

    Directory of Open Access Journals (Sweden)

    Sameer Jadhav

    2011-01-01

    Full Text Available Light polymerizable tooth colored restorative materials are most widely preferred for advantages such as esthetics, improved physical properties and operator′s control over the working time. Since the introduction of these light polymerizable restorative materials, there has been a concern about the depth of appropriate cure throughout the restoration. Photopolymerization of the composite is of fundamental importance because adequate polymerization is a crucial factor for optimization of the physical and mechanical properties and clinical results of the composite material. Inadequate polymerization results in greater deterioration at the margins of the restoration, decreased bond strength between the tooth and the restoration, greater cytotoxicity, and reduced hardness. Therefore, the dentist must use a light curing unit that delivers adequate and sufficient energy to optimize composite polymerization. Varying light intensity affects the degree of conversion of monomer to polymer and depth of cure.

  17. The Effect of Temperature Curing on Geopolymer Concrete

    Directory of Open Access Journals (Sweden)

    Triwulan

    2017-01-01

    Full Text Available This study presents curing temperature influences to behaviors of geopolymer concrete. The concrete was fly ash based, with the ratio of fly ash to alkaline activator was 65%:35%. Ratio coarse aggregate to fine aggregate was 60%:40%, ratios Na2SiO3/NaOH were 1.5:2 and 2.5. A polycarboxilate superplasticizer with ratio was 2% to fly ash weight. Form of the test specimens were cylinder with diameter of 10 cm and 20 cm high. Steam curing conducted were at 40°C, 60°C and 80°C for 24 hours. The control specimens were treated at normal moist curing. Tests conducted were compressive strength, tensile strength, porosity and elasticity modulus. The results indicated that at the elevated temperature the compressive strength increases as well as tensile strength but decreases the closed porosity of specimens. The elasticity modulus had a similar tendency.

  18. Modified Castor Oil as an Epoxy Resin Curing Agent

    Directory of Open Access Journals (Sweden)

    B. P. Patel

    2004-01-01

    Full Text Available A castor oil, an agricultural product, is an unsaturated material. Hence the castor oil is brominated by bromine liquid. The brominated castor oil (BCO was then reacted with excess of aliphatic diamines viz; ethylene diamine, 1,3-propane diamine and 1,6-hexane diamine. The resultant amino functionalized castor oil (ACO samples were then characterized by elemental analysis, IR spectral study and number of amino groups. All the three ACO samples were then employed for the curing of commercial Brominated epoxy resin. The curing of epoxy resin by ACO was monitored on differential scanning calorimeter (DSC and based on the DSC study their glass fibre reinforced composites (GRC were fabricated. The cured samples (i.e. unreinforced were also subjected to thermo gravimetric analysis (TGA. The chemical, mechanical and electrical properties of the glass fibre reinforced composites (GRC were also evaluated.

  19. Characterization and comparative overview of complete sequences of the first plasmids of Pandoraea across clinical and non-clinical strains

    Directory of Open Access Journals (Sweden)

    Delicia Yong

    2016-10-01

    Full Text Available To date, information on plasmid analysis in Pandoraea spp. is scarce. To address the gap of knowledge on this, the complete sequences of eight plasmids from Pandoraea spp. namely Pandoraea faecigallinarum DSM 23572 (pPF72-1, pPF72-2, Pandoraea oxalativorans DSM 23570 (pPO70-1, pPO70-2, pPO70-3, pPO70-4, Pandoraea vervacti NS15 (pPV15 and Pandoraea apista DSM 16535 (pPA35 were studied for the first time in this study. The information on plasmid sequences in Pandoraea spp. is useful because these plasmid sequences did not match to any known plasmid sequence deposited in public databases. Replication genes were not identified in some plasmids, a situation that has led to the possibility of host interaction involvement. Some plasmids were also void of par genes and intriguingly, repA gene was also not discovered in these plasmids. This further leads to the hypothesis of host-plasmid interaction. Plasmid stabilization/stability protein-encoding genes were observed in some plasmids but were not established for participating in plasmid segregation. Toxin-antitoxin systems MazEF, VapBC, RelBE, YgiT-MqsR, HigBA and ParDE were identified across the plasmids and their presence would improve plasmid maintenance. Conjugation genes were identified portraying the conjugation ability amongst Pandoraea plasmids. Additionally, we found a shared region amongst some of the plasmids that consists of conjugation genes. The identification of genes involved in replication, segregation, toxin-antitoxin systems and conjugation, would aid the design of drugs to prevent the survival or transmission of plasmids carrying pathogenic properties. Additionally, genes conferring virulence and antibiotic resistance were identified among the plasmids. The observed features in the plasmids shed light on the Pandoraea spp. as opportunistic pathogens.

  20. Plasmid profiles and antibiotic susceptibility patterns of Staphylococcus aureus isolates from Nigeria.

    Science.gov (United States)

    Olukoya, D K; Asielue, J O; Olasupo, N A; Ikea, J K

    1995-06-01

    In an investigation into the problems of infections due to Staphylococcus aureus in Nigeria, 100 strains were isolated from various hospitals in Lagos. The strains were screened for the presence of plasmids and for susceptibility to antimicrobial agents. Plasmids were extracted by modification of the method of Takahashi and Nagono[1]. The plasmids were diverse in nature. The strains were found to be highly resistant to commonly prescribed antibiotics.

  1. 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

    on various other plasmids belonging to IncA/C and IncL/M, while blaTEM-52c was found on IncN-like as well as on IncR plasmids. In the majority of cases (n = 21) the blaTEM-52 gene was located on a Tn3 transposon. Seven out of 10 blaTEM-52 plasmids tested in conjugation experiments were shown to be capable...

  2. Key features of mcr-1-bearing plasmids from Escherichia coli isolated from humans and food.

    Science.gov (United States)

    Zurfluh, Katrin; Nüesch-Inderbinen, Magdalena; Klumpp, Jochen; Poirel, Laurent; Nordmann, Patrice; Stephan, Roger

    2017-01-01

    Mcr-1-harboring Enterobacteriaceae are reported worldwide since their first discovery in 2015. However, a limited number of studies are available that compared full-length plasmid sequences of human and animal origins. In this study, mcr-1-bearing plasmids from seven Escherichia coli isolates recovered from patients (n = 3), poultry meat (n = 2) and turkey meat (n = 2) in Switzerland were further analyzed and compared. Isolates were characterized by multilocus sequence typing (MLST). The mcr-1-bearing plasmids were transferred by transformation into reference strain E. coli DH5α and MCR-1-producing transformants were selected on LB-agar supplemented with 2 mg/L colistin. Purified plasmids were then sequenced and compared. MLST revealed six distinct STs, illustrating the high clonal diversity among mcr-1-positive E. coli isolates of different origins. Two different mcr-1-positive plasmids were identified from a single E. coli ST48 human isolate. All other isolates possessed a single mcr-1 harboring plasmid. Transferable IncI2 (size ca. 60-61 kb) and IncX4 (size ca. 33-35 kb) type plasmids each bearing mcr-1 were found associated with human and food isolates. None of the mcr-1-positive IncI2 and IncX4 plasmids possessed any additional resistance determinants. Surprisingly, all but one of the sequenced mcr-1-positive plasmids lacked the ISApl1 element, which is a key element mediating acquisition of mcr-1 into various plasmid backbones. There is strong evidence that the food chain may be an important transmission route for mcr-1-bearing plasmids. Our data suggest that some "epidemic" plasmids rather than specific E. coli clones might be responsible for the spread of the mcr-1 gene along the food chain.

  3. Plasmid Profiles of Virulent Rhodococcus equi Strains Isolated from Infected Foals in Poland.

    Science.gov (United States)

    Kalinowski, Marcin; Grądzki, Zbigniew; Jarosz, Łukasz; Kato, Kiyoko; Hieda, Yu; Kakuda, Tsutomu; Takai, Shinji

    2016-01-01

    Rhodococcus equi is an important bacterial pathogen in foals up to 6 months old, widespread in horse farms all over the world. It was found that only virulent R. equi strains expressing 15-17 kDa virulence-associated protein (VapA) and having large virulence plasmid of 85-90 kb containing vapA gene are pathogenic for horses. To date, 12 plasmid types have been reported in VapA positive strains from horses. There are no data concerning plasmid types of Polish field R. equi strains isolated from horses and horse farm environment. The aim of the study is to determine plasmid profiles of virulent R. equi strains isolated in Poland from dead foals as well as from soil samples taken from horse breeding farms. Plasmid profiles of 10 clinical strains derived from 8 farms and 11 environmental strains from 3 farms, confirmed as virulent by PCR, were compared with 12 reference strains containing the known plasmid size and type. Plasmid DNAs were analysed by digestion with the restriction endonucleases BamHI, EcoRI, EcoT22I, and HindIII for detailed comparison and estimation of plasmid sizes. The results of RFLP analysis revealed that all except one isolates used in the study are classified as VapA 85 kb type I plasmid. One strain harboured VapA 87 kb type I plasmid. This is the first report of plasmid types of Polish field R. equi strains. The results of our preliminary investigations on horse farms located in central and eastern Poland indicate that the virulent R. equi strains thus far isolated from diseased foals and horse farms environment represent a highly uniform plasmid pattern.

  4. Occurrence of Plasmids in the Aromatic Degrading Bacterioplankton of the Baltic Sea

    OpenAIRE

    Ain Heinaru; Jaanis Juhanson; Eve Vedler; Eeva Heinaru; Jekaterina Jutkina

    2011-01-01

    Plasmids are mobile genetic elements that provide their hosts with many beneficial traits including in some cases the ability to degrade different aromatic compounds. To fulfill the knowledge gap regarding catabolic plasmids of the Baltic Sea water, a total of 209 biodegrading bacterial strains were isolated and screened for the presence of these mobile genetic elements. We found that both large and small plasmids are common in the cultivable Baltic Sea bacterioplankton and are particularly p...

  5. Genetic Characterization of Plasmid-Associated Benzalkonium Chloride Resistance Determinants in a Listeria monocytogenes Strain from the 1998-1999 Outbreak ▿

    Science.gov (United States)

    Elhanafi, Driss; Dutta, Vikrant; Kathariou, Sophia

    2010-01-01

    Quaternary ammonium compounds such as benzalkonium chloride (BC) are widely used as disinfectants in both food processing and medical environments. BC-resistant strains of Listeria monocytogenes have been implicated in multistate outbreaks of listeriosis and have been frequently isolated from food processing plants. However, the genetic basis for BC resistance in L. monocytogenes remains poorly understood. In this study, we have characterized a plasmid (pLM80)-associated BC resistance cassette in L. monocytogenes H7550, a strain implicated in the 1998-1999 multistate outbreak involving contaminated hot dogs. The BC resistance cassette (bcrABC) restored resistance to BC (MIC, 40 μg/ml) in a plasmid-cured derivative of H7550. All three genes of the cassette were essential for imparting BC resistance. The transcription of H7550 BC resistance genes was increased under sublethal (10 μg/ml) BC exposure and was higher at reduced temperatures (4, 8, or 25°C) than at 37°C. The level of transcription was higher at 10 μg/ml than at 20 or 40 μg/ml. In silico analysis suggested that the BC resistance cassette was harbored by an IS1216 composite transposon along with other genes whose functions are yet to be determined. The findings from this study will further our understanding of the adaptations of this organism to disinfectants such as BC and may contribute to the elucidation of possible BC resistance dissemination in L. monocytogenes. PMID:20971860

  6. Temperature and curing time affect composite sorption and solubility

    Directory of Open Access Journals (Sweden)

    Fabrício Luscino Alves de Castro

    2013-04-01

    Full Text Available Objective: This study evaluated the effect of temperature and curing time on composite sorption and solubility. Material and Methods: Seventy five specimens (8×2 mm were prepared using a commercial composite resin (ICE, SDI. Three temperatures (10°C, 25°C and 60°C and five curing times (5 s, 10 s, 20 s, 40 s and 60 s were evaluated. The specimens were weighed on an analytical balance three times: A: before storage (M1; B: 7 days after storage (M2; C: 7 days after storage plus 1 day of drying (M3. The storage solution consisted of 75% alcohol/25% water. Sorption and solubility were calculated using these three weights and specimen dimensions. The data were analyzed using the Kruskal-Wallis and Mann-Whitney U Tests (α=5%. Results: The results showed that time, temperature and their interaction influenced the sorption and solubility of the composite (p0.05. The 60°C composite temperature led to lower values of sorption for all curing times when compared with the 10°C temperature (p0.05. Solubility was similar at 40 s and 60 s for all temperatures (p>0.05, but was higher at 10°C than at 60°C for all curing times (p0.05. Conclusions: In conclusion, higher temperatures or longer curing times led to lower sorption and solubility values for the composite tested; however, this trend was only significant in specific combinations of temperature and curing times.

  7. Hardness Evaluation of Composite Resins Cured with QTH and LED

    Directory of Open Access Journals (Sweden)

    Behnaz Esmaeili

    2014-03-01

    Full Text Available Background and aims. Today light cured composites are widely used. Physical and mechanical properties of composites are related to the degree of conversion. Light curing unit (LCU is an important factor for composite polymerization. Aim of this study is evaluation of composite resins hardness using halogen and LED light curing units. Materials and methods. In this study, 30 samples of Filtek Z250 and C-Fill composite resins were provided. Samples were light cured with Ultralume2, Valo and Astralis7. Vickers hardness number (VHN was measured in 0, 1, 2 mm depth. Statistical analysis used: Data were analysed by SPSS software and compared with each other by T-test, one-way and twoway ANOVA and Post-hoc Tukey test. Results. In Filtek Z250, at top surface, VHN of Ultralume2 was higher than VHN of Valo (P = 0.02 and Astralis7 (P = 0.04, but in depth of 1, 2 mm, VHN of Ultralume2 and Astralis7 were almost the same and both LCUs were more than Valo which the difference between Ultralume2 and Valo was significant in depth of 1mm (0.05 and 2mm (0.02. In C-Fill composite, at top surface, Astralis7 showed higher VHN, but in depth of 2 mm, performance of all devices were rather similar. Conclusion. In Z250, which contains camphorquinone initiator, light cure LED Ultra-lume2 with narrow wavelength showed higher hardness number than Valo. In C-fill, in top surface, Astralis7 with more exposure time, resulted higher VHN. But In depth of 2 mm, various light curing devices had rather similar hardness number.

  8. Effects of medium composition on the production of plasmid DNA vector potentially for human gene therapy

    Institute of Scientific and Technical Information of China (English)

    XU Zhi-nan; SHEN Wen-he; CHEN Hao; CEN Pei-lin

    2005-01-01

    Plasmid vector is increasingly applied to gene therapy or gene vaccine. The production of plasmid pCMV-AP3 for cancer gene therapy was conducted in a modified MBL medium using a recombinant E. coli BL21 system. The effects of different MMBL components on plasmid yield, cell mass and specific plasmid DNA productivity were evaluated on shake-flask scale. The results showed that glucose was the optimal carbon source. High plasmid yield (58.3 mg/L) was obtained when 5.0 g/L glucose was added to MMBL. Glycerol could be chosen as a complementary carbon source because of the highest specific plasmid productivity (37.9 mg DNA/g DCW). After tests of different levels of nitrogen source and inorganic phosphate, a modified MMBL medium was formulated for optimal plasmid production. Further study showed that the initial acetate addition (less than 4.0 g/L) in MMBL improved plasmid production significantly, although it inhibited cell growth. The results will be useful for large-scale plasmid production using recombinant E. coli system.

  9. Modular construction of plasmids by parallel assembly of linear vector components.

    Science.gov (United States)

    Gao, XinZheng; Yan, Pu; Shen, Wentao; Li, Xiaoying; Zhou, Peng; Li, Yuenan

    2013-06-15

    Construction of plasmids is the basic and pivotal technology in molecular biology. The common method for constructing plasmids is to cut DNA fragments by restriction enzymes and then join the resulting fragments using ligase. We present here a modified Golden Gate cloning method for modular construction of plasmids. Unlike the original Golden Gate cloning system for cloning from entry vector to expression vector, this method can be used to construct plasmids immediately from linear DNA fragments. After polymerase chain reaction (PCR) amplification for flanking with BsaI sites, multiple linear DNA components (modules) can be parallel assembled into a circle plasmid by a single restriction-ligation reaction using the method. This method is flexible to construct different types of plasmids because the modules can be freely selected and assembled in any combination. This method was applied successfully to construct a prokaryotic expression plasmid from four modules and a plant expression plasmid from five modules (fragments). The results suggest that this method provides a simple and flexible platform for modular construction of plasmids.

  10. The evolution of collective restraint: policing and obedience among non-conjugative plasmids.

    Science.gov (United States)

    Kentzoglanakis, Kyriakos; García López, Diana; Brown, Sam P; Goldstein, Richard A

    2013-04-01

    The repression of competition by mechanisms of policing is now recognized as a major force in the maintenance of cooperation. General models on the evolution of policing have focused on the interplay between individual competitiveness and mutual policing, demonstrating a positive relationship between within-group diversity and levels of policing. We expand this perspective by investigating what is possibly the simplest example of reproductive policing: copy number control (CNC) among non-conjugative plasmids, a class of extra-chromosomal vertically transmitted molecular symbionts of bacteria. Through the formulation and analysis of a multi-scale dynamical model, we show that the establishment of stable reproductive restraint among plasmids requires the co-evolution of two fundamental plasmid traits: policing, through the production of plasmid-coded trans-acting replication inhibitors, and obedience, expressed as the binding affinity of plasmid-specific targets to those inhibitors. We explain the intrinsic replication instabilities that arise in the absence of policing and we show how these instabilities are resolved by the evolution of copy number control. Increasing levels of policing and obedience lead to improvements in group performance due to tighter control of local population size (plasmid copy number), delivering benefits both to plasmids, by reducing the risk of segregational loss and to the plasmid-host partnership, by increasing the rate of cell reproduction, and therefore plasmid vertical transmission.

  11. Pheromone-responsive conjugative vancomycin resistance plasmids in Enterococcus faecalis isolates from humans and chicken feces.

    Science.gov (United States)

    Lim, Suk-Kyung; Tanimoto, Koichi; Tomita, Haruyoshi; Ike, Yasuyoshi

    2006-10-01

    The drug resistances and plasmid contents of a total of 85 vancomycin-resistant enterococcus (VRE) strains that had been isolated in Korea were examined. Fifty-four of the strains originated from samples of chicken feces, and 31 were isolated from hospital patients in Korea. Enterococcus faecalis KV1 and KV2, which had been isolated from a patient and a sample of chicken feces, respectively, were found to carry the plasmids pSL1 and pSL2, respectively. The plasmids transferred resistances to vancomycin, gentamicin, kanamycin, streptomycin, and erythromycin to E. faecalis strains at a high frequency of about 10(-3) per donor cell during 4 hours of broth mating. E. faecalis strains containing each of the pSL plasmids formed clumps after 2 hours of incubation in broth containing E. faecalis FA2-2 culture filtrate (i.e., the E. faecalis sex pheromone), and the plasmid subsequently transferred to the recipient strain in a 10-min short mating in broth, indicating that the plasmids are responsive to E. faecalis pheromones. The pSL plasmids did not respond to any of synthetic pheromones for the previously characterized plasmids. The pheromone specific for pSL plasmids has been designated cSL1. Southern hybridization analysis showed that specific FspI fragments from each of the pSL plasmids hybridized with the aggregation substance gene (asa1) of the pheromone-responsive plasmid pAD1, indicating that the plasmids had a gene homologous to asa1. The restriction maps of the plasmids were identical, and the size of the plasmids was estimated to be 128.1 kb. The plasmids carried five drug resistance determinants for vanA, ermB, aph(3'), aph(6'), and aac(6')/aph(2'), which encode resistance to vancomycin, erythromycin, kanamycin, streptomycin, and gentamicin/kanamycin, respectively. Nucleotide sequence analyses of the drug resistance determinants and their flanking regions are described in this report. The results described provide evidence for the exchange of genetic information

  12. Postsymbiotic plasmid acquisition and evolution of the repA1-replicon in Buchnera aphidicola

    Science.gov (United States)

    Van Ham, Roeland C. H. J.; González-Candelas, Fernando; Silva, Francisco J.; Sabater, Beatriz; Moya, Andrés; Latorre, Amparo

    2000-01-01

    Buchnera aphidicola is an obligate, strictly vertically transmitted, bacterial symbiont of aphids. It supplies its host with essential amino acids, nutrients required by aphids but deficient in their diet of plant phloem sap. Several lineages of Buchnera show adaptation to their nutritional role in the form of plasmid-mediated amplification of key-genes involved in the biosynthesis of tryptophan (trpEG) and leucine (leuABCD). Phylogenetic analyses of these plasmid-encoded functions have thus far suggested the absence of horizontal plasmid exchange among lineages of Buchnera. Here, we describe three new Buchnera plasmids, obtained from species of the aphid host families Lachnidae and Pemphigidae. All three plasmids belong to the repA1 family of Buchnera plasmids, which is characterized by the presence of a repA1-replicon responsible for replication initiation. A comprehensive analysis of this family of plasmids unexpectedly revealed significantly incongruent phylogenies for different plasmid and chromosomally encoded loci. We infer from these incongruencies a case of horizontal plasmid transfer in Buchnera. This process may have been mediated by secondary endosymbionts, which occasionally undergo horizontal transmission in aphids. PMID:10984505

  13. Enhancing yields of low and single copy number plasmid DNAs from Escherichia coli cells.

    Science.gov (United States)

    Wood, Whitney N; Smith, Kyle D; Ream, Jennifer A; Kevin Lewis, L

    2017-02-01

    Many plasmids used for gene cloning and heterologous protein expression in Escherichia coli cells are low copy number or single copy number plasmids. The extraction of these types of plasmids from small bacterial cell cultures produces low DNA yields. In this study, we have quantitated yields of low copy and single copy number plasmid DNAs after growth of cells in four widely used broths (SB, SOC, TB, and 2xYT) and compared results to those obtained with LB, the most common E. coli cell growth medium. TB (terrific broth) consistently generated the greatest amount of plasmid DNA, in agreement with its ability to produce higher cell titers. The superiority of TB was primarily due to its high levels of yeast extract (24g/L) and was independent of glycerol, a unique component of this broth. Interestingly, simply preparing LB with similarly high levels of yeast extract (LB24 broth) resulted in plasmid yields that were equivalent to those of TB. By contrast, increasing ampicillin concentration to enhance plasmid retention did not improve plasmid DNA recovery. These experiments demonstrate that yields of low and single copy number plasmid DNAs from minipreps can be strongly enhanced using simple and inexpensive media. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. Rapid plasmid library screening using RecA-coated biotinylated probes.

    Science.gov (United States)

    Rigas, B; Welcher, A A; Ward, D C; Weissman, S M

    1986-12-01

    A method for the rapid physical isolation of recombinant plasmids of interest from a mixture of plasmids such as a plasmid cDNA library is presented. This method utilizes the ability of RecA protein to form stable complexes between linear single-stranded and circular double-stranded DNA molecules sharing sequence homology, and procedures allowing isolation of biotinylated nucleic acid. Biotinylated linear DNA probes coated with RecA have been used to screen reconstituted plasmid libraries consisting of two plasmid species, one homologous and the other heterologous to the probe. When the link between biotin and the nucleotide base could be cleaved by reducing agents, the complex was purified by streptavidin-agarose chromatography and the recovered plasmid was propagated in Escherichia coli. When the link was not cleavable the complex was bound to avidin in solution and purified by cupric iminodiacetic acid-agarose chromatography. The complex was then dissociated and the plasmids were propagated in E. coli. With either protocol, homologous plasmid recovery was between 10% and 20%, and enrichment was between 10(4)- and 10(5)-fold. Potential applications and extensions of this method, such as plasmid, cosmid, and phage library screening and facilitation of physical mapping of macroregions of mammalian genomes are presented and discussed.

  15. 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.

  16. Microneedle-mediated transcutaneous immunization with plasmid DNA coated on cationic PLGA nanoparticles

    Science.gov (United States)

    Kumar, Amit; Wonganan, Piyanuch; Sandoval, Michael A.; Li, Xinran; Zhu, Saijie; Cui, Zhengrong

    2012-01-01

    Previously, it was shown that microneedle-mediated transcutaneous immunization with plasmid DNA can potentially induce a stronger immune response than intramuscular injection of the same plasmid DNA. In the present study, we showed that the immune responses induced by transcutaneous immunization by applying plasmid DNA onto a skin area pretreated with solid microneedles were significantly enhanced by coating the plasmid DNA on the surface of cationic nanoparticles. In addition, the net surface charge of the DNA-coated nanoparticles significantly affected their in vitro skin permeation and their ability to induce immune responses in vivo. Transcutaneous immunization with plasmid DNA-coated net positively charged anoparticles elicited a stronger immune response than with plasmid DNA-coated net negatively charged nanoparticles or by intramuscular immunization with plasmid DNA alone. Transcutaneous immunization with plasmid DNA-coated net positively charged nanoparticles induced comparable immune responses as intramuscular injection of them, but transcutaneous immunization was able to induce specific mucosal immunity and a more balanced T helper type 1 and type 2 response. The ability of the net positively charged DNA-coated nanoparticles to induce a strong immune response through microneedle-mediated transcutaneous immunization may be attributed to their ability to increase the expression of the antigen gene encoded by the plasmid and to more effectively stimulate the maturation of antigen-presenting cells. PMID:22921518

  17. 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.

  18. Mechanism of acquisition of chromosomal markers by plasmids in Haemophilus influenzae.

    OpenAIRE

    Setlow, J K; Cabrera-Juárez, E; Griffin, K

    1984-01-01

    The hybrid plasmid pNov1 readily acquired genetic information from the chromosome of wild-type, but not rec-2, cells. Most of the recombination had taken place 1 h after entrance of the plasmid into the cell, as judged by transformation of rec-2 by lysates made from wild-type cells exposed to pNov1. Measurement of physical transfer from radioactively labeled cellular DNA to plasmids recombining in wild-type cells failed, since there was little more radioactivity in plasmids from such cells th...

  19. Influence of Plasmid Type on the Replication of Rhodococcus equi in Host Macrophages.

    Science.gov (United States)

    Willingham-Lane, Jennifer M; Berghaus, Londa J; Giguère, Steeve; Hondalus, Mary K

    2016-01-01

    The soil-dwelling, saprophytic actinomycete Rhodococcus equi is a multihost, facultative intracellular pathogen of macrophages. When inhaled by susceptible foals, it causes severe bronchopneumonia. It is also a pathogen of pigs, which may develop submaxillary lymphadenitis upon exposure. R. equi isolates obtained from foals and pigs possess conjugative plasmids housing a pathogenicity island (PAI) containing a novel family of genes of unknown function called the virulence-associated protein or vap family. The PAI regions of the equine and swine plasmids differ in vap gene composition, with equine isolates possessing six vap genes, including the major virulence determinant vapA, while the PAIs of swine isolates house vapB and five other unique vap genes. Possession of the pVAPA-type virulence plasmid by equine isolates bestows the capacity for intramacrophage replication essential for disease development in vivo. Swine isolates of R. equi are largely unstudied. Here, we show that R. equi isolates from pigs, carrying pVAPB-type plasmids, are able to replicate in a plasmid-dependent manner in macrophages obtained from a variety of species (murine, swine, and equine) and anatomical locations. Similarly, equine isolates carrying pVAPA-type plasmids are capable of replication in swine macrophages. Plasmid swapping between equine and swine strains through conjugation did not alter the intracellular replication capacity of the parental strain, indicating that coevolution of the plasmid and chromosome is not crucial for this attribute. These results demonstrate that while distinct plasmid types exist among R. equi isolates obtained from equine and swine sources, this tropism is not determined by host species-specific intramacrophage replication capabilities. IMPORTANCE This work greatly advances our understanding of the opportunistic pathogen Rhodococcus equi, a disease agent of animals and immunocompromised people. Clinical isolates from diseased foals carry a

  20. A DNA polymerase mutation that suppresses the segregation bias of an ARS plasmid in Saccharomyces cerevisiae.

    Science.gov (United States)

    Houtteman, S W; Elder, R T

    1993-03-01

    Yeast autonomously replicating sequence (ARS) plasmids exhibit an unusual segregation pattern during mitosis. While the nucleus divides equally into mother and daughter cells, all copies of the ARS plasmid will often remain in the mother cell. A screen was designed to isolate mutations that suppress this segregation bias. A plasmid with a weak ARS (wARS) that displayed an extremely high segregation bias was constructed. When cells were grown under selection for the wARS plasmid, the resulting colonies grew slowly and had abnormal morphology. A spontaneous recessive mutation that restored normal colony morphology was identified. This mutation suppressed plasmid segregation bias, as indicated by the increased stability of the wARS plasmid in the mutant cells even though the plasmid was present at a lower copy number. An ARS1 plasmid was also more stable in mutant cells than in wild-type cells. The wild-type allele for this mutant gene was cloned and identified as POL delta (CDC2). This gene encodes DNA polymerase delta, which is essential for DNA replication. These results indicate that DNA polymerase delta plays some role in causing the segregation bias of ARS plasmids.

  1. High instability of a nematicidal Cry toxin plasmid in Bacillus thuringiensis.

    Science.gov (United States)

    Sheppard, Anna E; Nakad, Rania; Saebelfeld, Manja; Masche, Anna C; Dierking, Katja; Schulenburg, Hinrich

    2016-01-01

    In bacterial pathogens, virulence factors are often carried on plasmids and other mobile genetic elements, and as such, plasmid evolution is central in understanding pathogenicity. Bacillus thuringiensis is an invertebrate pathogen that uses plasmid-encoded crystal (Cry) toxins to establish infections inside the host. Our study aimed to quantify stability of two Cry toxin-encoding plasmids, BTI_23p and BTI_16p, under standard laboratory culturing conditions. These two plasmids are part of the genome of the B. thuringiensis strain MYBT18679, which is of particular interest because of its high pathogenicity towards nematodes. One of the plasmids, BTI_23p, was found to be highly unstable, with substantial loss occurring within a single growth cycle. Nevertheless, longer term experimental evolution in the absence of a host revealed maintenance of the plasmid at low levels in the bacterial populations. BTI_23p encodes two nematicidal Cry toxins, Cry21Aa2 and Cry14Aa1. Consistent with previous findings, loss of the plasmid abolished pathogenicity towards the nematode Caenorhabditis elegans, which could be rescued by addition of Cry21Aa2-expressing Escherichia coli. These results implicate BTI_23p as a plasmid that is required for successful infection, yet unstable when present at high frequency in the population, consistent with the role of Cry toxins as public goods.

  2. Cloning, sequencing, and sequence analysis of two novel plasmids from the thermophilic anaerobic bacterium Anaerocellum thermophilum

    DEFF Research Database (Denmark)

    Clausen, Anders; Mikkelsen, Marie Just; Schrøder, I.

    2004-01-01

    The nucleotide sequence of two novel plasmids isolated from the extreme thermophilic anaerobic bacterium Anaerocellum thermophilum DSM6725 (A. thermophilum), growing optimally at 70degreesC, has been determined. pBAS2 was found to be a 3653 bp plasmid with a GC content of 43%, and the sequence...... was found, but no single stranded intermediates, characteristic of rolling circle replication, were found on Southern blots. The larger plasmid, pBAL, was found to be a 8294 bp plasmid with a GC content of 39%. It revealed 17 ORFs, of which three showed similarity at the amino acid (aa) level to known...

  3. Construction of Recombinant Plasmid Containing S. Mutans F-ATPase β Subunit Gene

    Institute of Scientific and Technical Information of China (English)

    YU Dan-ni; JIANG Li

    2005-01-01

    objective: construct a homologous recombinant plasmid which was expected to be transformed into S. mutans Methods: a region at the 5' terminus of the S. mutans F-ATPase β subunit gene was amplified by PCR, the PCR product was inserted into vector pVA891, yielding recombinant plasmid. Results: the DNA sequence of the recombinant plasmid was identified correct in whole by restriction endonuclease and DNA sequence techniques. Conclusion: the recombinant plasmid of S. mutans DNA was cloned in effect ,it may assist in construction of homologues recombinant mutant.

  4. The evolution of collective restraint: policing and obedience among non-conjugative plasmids.

    Directory of Open Access Journals (Sweden)

    Kyriakos Kentzoglanakis

    2013-04-01

    Full Text Available The repression of competition by mechanisms of policing is now recognized as a major force in the maintenance of cooperation. General models on the evolution of policing have focused on the interplay between individual competitiveness and mutual policing, demonstrating a positive relationship between within-group diversity and levels of policing. We expand this perspective by investigating what is possibly the simplest example of reproductive policing: copy number control (CNC among non-conjugative plasmids, a class of extra-chromosomal vertically transmitted molecular symbionts of bacteria. Through the formulation and analysis of a multi-scale dynamical model, we show that the establishment of stable reproductive restraint among plasmids requires the co-evolution of two fundamental plasmid traits: policing, through the production of plasmid-coded trans-acting replication inhibitors, and obedience, expressed as the binding affinity of plasmid-specific targets to those inhibitors. We explain the intrinsic replication instabilities that arise in the absence of policing and we show how these instabilities are resolved by the evolution of copy number control. Increasing levels of policing and obedience lead to improvements in group performance due to tighter control of local population size (plasmid copy number, delivering benefits both to plasmids, by reducing the risk of segregational loss and to the plasmid-host partnership, by increasing the rate of cell reproduction, and therefore plasmid vertical transmission.

  5. Plasmid profile in oral Fusobacterium nucleatum from humans and Cebus apella monkeys

    Directory of Open Access Journals (Sweden)

    Paula Marcia O.

    2003-01-01

    Full Text Available Fusobacterium nucleatum is a strict anaerobe and is indigenous of the human oral cavity. This organism is commonly recovered from different monomicrobial and mixed infections in humans and animals. In this study, the plasmid profile, the plasmid stability and the penicillin-resistance association in oral F. nucleatum isolated from periodontal patients, healthy subjects and Cebus apella monkeys were evaluated. Forty-five F. nucleatum strains from patients, 38 from healthy subjects and seven from C. apella were identified and analyzed. Plasmid extraction was performed in all the isolated strains. These elements were found in 26.7% strains from patients and one strain from C. apella. Strains from healthy subjects did not show any plasmid. Most of strains showed two plasmid bands ranging from 4 to 16 Kb, but digestions with endonucleases showed that they belonged to a single plasmid. The plasmid profile was similar and stable in human and monkey strains. Also, plasmids were classified into three groups according to size. Two strains were positive to beta-lactamase production and no plasmid DNA-hybridization with a beta-lactamase gene probe was observed, suggesting a chromosomal resistance.

  6. Quantifying and visualizing the transfer of exogenous plasmids to environmental microbial communities

    DEFF Research Database (Denmark)

    Dechesne, Arnaud

    2015-01-01

    of a community to take up exogenous plasmid should, however, be an important element affecting the fate of mobile genetic elements released in the environment. We have devised a method to evaluate the permissiveness of a bacterial community towards exogenous plasmids, both quantitatively (how many bacteria can......, our findings highlight the high potential for exogenous plasmids to be transferred to soil microbial communities and indicate that community permissiveness – as affected by environmental conditions- needs to be considered to predict the fate of plasmids in the environment....

  7. Structural similarity and distribution of small cryptic plasmids of Lactobacillus curvatus and L. sake.

    Science.gov (United States)

    Vogel, R F; Lohmann, M; Weller, A N; Hugas, M; Hammes, W P

    1991-11-15

    Plasmid profiles of strains of Lactobacillus curvatus and L. sake isolated from meat or sauerkraut were analysed to investigate plasmid homology and distribution in relation to the ecology of these organisms in fermenting foods. A hybridisation probe was constructed by cloning of pLc2, a cryptic, 2.6-kbp plasmid from L. curvatus LTH683, into the Escherichia coli plasmid pRV50. In Southern hybridisations with the digoxygenine labeled pLc2 probe, pLc2-related small plasmids were frequently detected in meat-borne strains of L. casei subsp. pseudoplantarum, L. curvatus, L. sake, L. alimentarius, L. farciminis and L. halotolerans and in L. curvatus and L. sake isolated from sauerkraut. Among 27 Lactobacillus type strains originally isolated from habitats other than meat this type of homology was detected only with plasmids of L. buchneri and L. mali. Restriction-enzyme mapping of six small cryptic plasmids from L. curvatus and L. sake revealed strong structural homology but no similarity to previously characterized plasmids of lactobacilli. The presence of a variable region in addition to a conserved one and the occurrence of deletions during cloning of pLc2 suggest that vectors derived from these plasmids are likely to be structurally unstable.

  8. Synthesis and curing of alkyd enamels based on ricinoleic acid

    Directory of Open Access Journals (Sweden)

    Jovičić Mirjana C.

    2010-01-01

    Full Text Available A combination of an alkyd resin with a melamine-formaldehyde resin gives a cured enamel film with the flexibility of the alkyd constituent and the high chemical resistance and hardness of the melamine resin at the same time. The melamine resin is a minor constituent and plays the role of a crosslinking agent. In this paper, alkyd resins of high hydroxyl numbers based on trimethylolpropane, ricinoleic acid and phthalic anhydride were synthesized. Two alkyds having 30 and 40 wt% of ricinoleic acid were formulated by calculation on alkyd constant. Alkyds were characterized by FTIR and by the determination of acid and hydroxyl numbers. Then synthesized alkyds were made into baking enamels by mixing with melamine-formaldehyde resins (weight ratio of 70:30 based on dried mass. Two types of commercial melamine resins were used: threeisobutoxymethyl melamine-formaldehyde resin (TIMMF and hexamethoxymethyl melamine resin (HMMMF. Prepared alkyd/melamine resin mixtures were cured in a differential scanning calorimeter (DSC under non-isothermal mode. Apparent degree of curing as a function of temperature was calculated from the curing enthalpies. Kinetic parameters of curing were calculated using Freeman-Carroll method. TIMMF resin is more reactive with synthesized alkyds than HMMMF resin what was expected. Alkyd resin with 30 wt% of ricinoleic acid is slightly more reactive than alkyd with 40 wt% of ricinoleic acid, probably because it has the high contents of free hydroxyl and acid groups. The gel content, Tg, thermal stability, hardness, elasticity and impact resistance of coated films cured at 150°C for 60 min were measured. Cured films show good thermal stability since the onset of films thermal degradation determined by thermogravimetric analysis (TGA is observed at the temperatures from 281 to 329°C. Films based on alkyd 30 are more thermal stable than those from alkyd 40, with the same melamine resin. The type of alkyd resin has no significant

  9. Grey Disorder in Field Flue—Cured Tobacco

    Institute of Scientific and Technical Information of China (English)

    ZHANGXIN; ZHAOQIBO; 等

    1999-01-01

    A field study was carried out on the grey disorder in flue-cured tobacco( Nicotiana tabacum L.) leaves.The results showed that low Eh and high available Fe content in the soils were responsible for the grey tobacco and dead roots.Grey disorder leaves produced low quality lamina,which was low in N,nicotine,reduced sugar and K contents,In this study,soil available Mn and leaf Mn contents were in a relatively low level and should not be an important factor in the occurrence of grey disorder in flue-cured tobacco.

  10. Magnetoactive elastomeric composites: Cure, tensile, electrical and magnetic properties

    Indian Academy of Sciences (India)

    K Sasikumar; G Suresh; K A Thomas; Reji John; V Natarajan; T Mukundan; R M R Vishnubhatla

    2006-11-01

    Magnetically active elastomer materials were prepared by incorporating nickel powder in synthetic elastomeric matrices, polychloroprene and nitrile rubber. Cure characteristics, mechanical, electrical and magnetic properties were experimentally determined for different volume fractions of magnetoactive filler. The cure time decreases sharply for initial filler loading and the decrease is marginal for additional loading of filler. The tensile strength and modulus at 100% strain was found to increase with increase in the volume fraction of nickel due to reinforcement action. The magnetic impedance and a.c. conductivity are found to increase with increase in volume fraction of nickel as well as frequency.

  11. Realtime 3D stress measurement in curing epoxy packaging

    DEFF Research Database (Denmark)

    Richter, Jacob; Hyldgård, A.; Birkelund, Karen;

    2007-01-01

    This paper presents a novel method to characterize stress in microsystem packaging. A circular p-type piezoresistor is implemented on a (001) silicon chip. We use the circular stress sensor to determine the packaging induced stress in a polystyrene tube filled with epoxy. The epoxy curing process...... is monitored by stress measurements. From the stress measurements we conclude that the epoxy cures in 8 hours at room temperature. We find the difference in in-plane normal stresses to be sigmaxx-sigmayy=6.7 MPa and (sigmaxx+sigmayy-0.4sigmazz)=232 MPa....

  12. Preliminary toxicological study of Sylgard 184 curing agent

    Energy Technology Data Exchange (ETDEWEB)

    Smith, D.M.; London, J.E.; Drake, G.A.; Thomas, R.G.

    1978-06-01

    The acute oral LD/sub 50//sup 30/ values for mice and rats receiving Sylgard 184 curing agent were greater than 5 g/kg. According to classical guidelines, the compound would be considered slightly toxic or practically nontoxic in both species. Skin application studies in the rabbit demonstrated the compound to be mildly irritating. Eye irritation studies, also in the rabbit, showed that Sylgard 184 curing agent was a mild but transitory irritant. The sensitization study in guinea pigs did not show the resin to be deleterious.

  13. Viral hepatitis B and C. Cure or treatment?

    Directory of Open Access Journals (Sweden)

    Dimitrios A. Kountouras

    2014-12-01

    Full Text Available HBV and HCV infections are among the most important global health problems; both represent also the leading cause of cirrhosis and HCC worldwide. HBV treatment cannot be considered cure but effective viral suppression can be achieved and remains the current principal goal of therapy. Talking about HCV treatment today equals to talking about total cure of the patient, with treatments of very high SVR rates, shorter if not shortest duration, minimal risk for resistance, pangenotypic and practically with no serious adverse events, no fibrosis or previous treatment status limitations, but also with a very high cost.

  14. Cure Monitoring and Control with Combined Dielectric/Temperature Probes.

    Science.gov (United States)

    1983-01-10

    DGEBA with expressions include an additional menthane diamine at 90*C were used parameter, 8, which accounts for to make the plots (the actual data...parameters to match Fig. 2 data. DGEBA cured with menthane diamine at Frequencies are 1,10,100 and 1000 Rz 900C. -5- ally with time (following the to slow down...mum. An example of such a result ure 2 shows plots correspondizg to is presented in Section 3.1 below. the model curves of Fig. 1 for DGEBA cured

  15. Optimization of the curing process of a sandwich panel

    Science.gov (United States)

    Phyo Maung, Pyi; Tatarnikov, O.; Malysheva, G.

    2016-10-01

    This study presented finite element modelling and experimental measurements of temperatures during the autoclave curing of the T-50 aircraft wing sandwich panel. This panel consists of upper and lower carbon fibre based laminates and an aluminium foil honeycomb. The finite element modelling was performed using the Femap-Nastran product. During processing, the temperature at various points on the surface of the panel was measured using the thermocouples. The finite element method simulated the thermal conditions and determined the temperatures in the different parts of the panel for a full cycle of the curing process. A comparison of the calculated and experimental data shows that their difference does not exceed 6%.

  16. Emergent Cure Chemistry in the Development of Aerospace Materials

    Science.gov (United States)

    2015-03-25

    R4 R3-SHR3-S-CH2-CH2-R4Product Anti-Markovnikov • Thiol-ene chemistry is a hybrid of free radical and condensation chemistry . It combines the speed...Charts 3. DATES COVERED (From - To) March 2015-March 2015 4. TITLE AND SUBTITLE Emergent cure chemistry in the development of aerospace materials...cure chemistry in the development of aerospace materials Joseph M. Mabry, Ph.D. Air Force Research Laboratory joseph.mabry@us.af.mil (661) 275-5857

  17. Nutidig mental sundhedsfremme i et historisk Mind-Cure perspektiv

    OpenAIRE

    Wistoft, Karen

    2014-01-01

    Mental sundhedsfremme og dertil knyttede metoder er i stærk vækst i velfærdssamfundet. Mental sundhed er dog ikke et nyt fænomen. Ved begyndelsen af det tyvende århundrede fremhævede Mind-Cure-bevægelsen i USA den helbredende kraft af positive følelser og overbevisninger. William James (1842-1910) anså Mind-Cure som en amerikansk pragmatisk drejning af den tids sundhedsfremme hen imod en mere optimistisk orientering i livet, en menneskelig karakter, der kunne opøves. James var fortaler for, a...

  18. Effects of feeding high protein or conventional canola meal on dry cured and conventionally cured bacon.

    Science.gov (United States)

    Little, K L; Bohrer, B M; Stein, H H; Boler, D D

    2015-05-01

    Objectives were to compare belly, bacon processing, bacon slice, and sensory characteristics from pigs fed high protein canola meal (CM-HP) or conventional canola meal (CM-CV). Soybean meal was replaced with 0 (control), 33, 66, or 100% of both types of canola meal. Left side bellies from 70 carcasses were randomly assigned to conventional or dry cure treatment and matching right side bellies were assigned the opposite treatment. Secondary objectives were to test the existence of bilateral symmetry on fresh belly characteristics and fatty acid profiles of right and left side bellies originating from the same carcass. Bellies from pigs fed CM-HP were slightly lighter and thinner than bellies from pigs fed CM-CV, yet bacon processing, bacon slice, and sensory characteristics were unaffected by dietary treatment and did not differ from the control. Furthermore, testing the existence of bilateral symmetry on fresh belly characteristics revealed that bellies originating from the right side of the carcasses were slightly (P≤0.05) wider, thicker, heavier and firmer than bellies from the left side of the carcass.

  19. pTAR-encoded proteins in plasmid partitioning.

    Science.gov (United States)

    Kalnin, K; Stegalkina, S; Yarmolinsky, M

    2000-04-01

    Partition cassettes, essential for the segregational stability of low-copy-number bacterial plasmids, typically encode two autoregulated proteins and an adjacent cis-acting centromere analog to which one or perhaps both proteins bind. The diminutive partition region of pTAR of Agrobacterium spp. was reported to be exceptional, encoding only a single protein, ParA (D. R. Gallie and C. I. Kado, J. Mol. Biol. 193:465-478, 1987). However, resequencing of the region revealed two small downstream genes, parB and orf-84, of which only parB was found to be essential for partitioning in A. tumefaciens. Purified ParA exhibited a weak ATPase activity that was modestly increased by nonspecific DNA. ParB bound in vitro to repeated sequences present in a region, parS, that possesses centromere and operator functions and within which we identified the primary transcription start site by primer extension. In certain respects the Par proteins behave normally in the foreign host Escherichia coli. In E. coli, as in A. tumefaciens, ParB repressed the partition operon; ParA, inactive alone, augmented this repression. Functional similarities between the partition system of pTAR and those of other plasmids and bacteria are prominent, despite differences in size, organization, and amino acid sequence.

  20. Adsorption behavior of plasmid DNA onto perfusion chromatographic matrix

    Institute of Scientific and Technical Information of China (English)

    Miladys LIMONTA; Lourdes ZUMALACARREGUI; Dayana SOLER

    2012-01-01

    Anion exchange chromatography is the most popular chromatographic method for plasmid separa-tion.POROS RI 50 is a perfusion chromatographic support which is a reversed phase matrix and is an alterna-tive to conventional ones due to its mass transfer properties.The adsorption and elution of the pIDKE2 plasmidonto reversed phase POROS RI 50 was studied.Langmuir isotherm model was adjusted in order to get the max-imum adsorption capacity and the dissociation constant for POROS RI 50-plasmid DNA (pDNA) system.Break-through curves were obtained for volumetric flows between 0.69-3.33mL/min,given dynamic capacity up to2.3 times higher than those reported for ionic exchange matrix used during the purification process of plasmidswith similar size to that of pIDKE2.The efficiency was less than 45% for the flow conditions and initial concen-tration studied,which means that the support will not be operated under saturation circumstances.