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Sample records for plasmid transfer rate

  1. 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.

  2. 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.

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

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    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.

  4. 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

  5. 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...

  6. 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.

  7. Conjugative Plasmid Transfer in Xylella fastidiosa Is Dependent on tra and trb Operon Functions

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    Van Horn, Christopher R.

    2017-01-01

    ABSTRACT The insect-transmitted plant pathogen Xylella fastidiosa is capable of efficient horizontal gene transfer (HGT) and recombination. Natural transformation occurs at high rates in X. fastidiosa, but there also is evidence that certain strains of X. fastidiosa carry native plasmids equipped with transfer and mobilization genes, suggesting conjugation as an additional mechanism of HGT in some instances. Two operons, tra and trb, putatively encoding a conjugative type IV secretion system, are found in some but not all X. fastidiosa isolates, often on native plasmids. X. fastidiosa strains that carry the conjugative transfer genes can belong to different subspecies and frequently differ in host ranges. Using X. fastidiosa strain M23 (X. fastidiosa subsp. fastidiosa) or Dixon (X. fastidiosa subsp. multiplex) as the donor strain and Temecula (X. fastidiosa subsp. fastidiosa) as the recipient strain, plasmid transfer was characterized using the mobilizable broad-host-range vector pBBR5pemIK. Transfer of plasmid pBBR5pemIK was observed under in vitro conditions with both donor strains and was dependent on both tra and trb operon functions. A conjugative mechanism likely contributes to gene transfer between diverse strains of X. fastidiosa, possibly facilitating adaptation to new environments or different hosts. IMPORTANCE Xylella fastidiosa is an important plant pathogen worldwide, infecting a wide range of different plant species. The emergence of new diseases caused by X. fastidiosa, or host switching of existing strains, is thought to be primarily due to the high frequency of HGT and recombination in this pathogen. Transfer of plasmids by a conjugative mechanism enables movement of larger amounts of genetic material at one time, compared with other routes of gene transfer such as natural transformation. Establishing the prevalence and functionality of this mechanism in X. fastidiosa contributes to a better understanding of HGT, adaptation, and disease emergence

  8. Conjugative plasmid transfer in Xylella fastidiosa is dependent on tra and trb operon functions.

    Science.gov (United States)

    Burbank, Lindsey P; Van Horn, Christopher R

    2017-08-14

    The insect-transmitted plant pathogen Xylella fastidiosa is capable of efficient horizontal gene transfer (HGT) and recombination. Natural transformation occurs at high rates in X. fastidiosa, but there also is evidence that certain strains of X. fastidiosa carry native plasmids equipped with transfer and mobilization genes, suggesting conjugation as an additional mechanism of HGT in some instances. Two operons, tra and trb, putatively encoding a conjugative Type IV secretion system are found in some but not all X. fastidiosa isolates, often on native plasmids. X. fastidiosa strains that carry the conjugative transfer genes can belong to different subspecies, and frequently differ in host ranges. Using X. fastidiosa strains M23 (subspecies fastidiosa) or Dixon (subspecies multiplex) as the donor strain and Temecula (subspecies fastidiosa) as the recipient strain, plasmid transfer was characterized using the mobilizable broad host range vector pBBR5pemIK. Transfer of plasmid pBBR5pemIK was observed under in vitro conditions with both donor strains, and was dependent on both tra and trb operon functions. A conjugative mechanism likely contributes to gene transfer between diverse strains of X. fastidiosa, possibly facilitating adaptation to new environments or different hosts.IMPORTANCEXylella fastidiosa is an important plant pathogen world-wide, infecting a wide range of different plant species. Emergence of new diseases caused by X. fastidiosa, or host-switching of existing strains is thought to be primarily due to the high frequency of HGT and recombination in this pathogen. Transfer of plasmids by a conjugative mechanism enables movement of larger amounts of genetic material at one time compared with other routes of gene transfer such as natural transformation. Establishing the prevalence and functionality of this mechanism in X. fastidiosa contributes to a better understanding of HGT and adaptation, and disease emergence in this diverse pathogen. This is a work

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

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    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.

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

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    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.

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

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    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.

  12. Conjugal plasmid transfer (pAM beta 1) in Lactobacillus plantarum.

    OpenAIRE

    Shrago, A W; Chassy, B M; Dobrogosz, W J

    1986-01-01

    The streptococcal plasmid pAM beta 1 (erythromycin resistance) was transferred via conjugation from Streptococcus faecalis to Lactobacillus plantarum and was transferred among L. plantarum strains. Streptococcus sanguis Challis was transformed with pAM beta 1 isolated from these transconjugants, and transformants harboring intact pAM beta 1 could conjugate the plasmid back to L. plantarum.

  13. Factors that affect transfer of the IncI1 β-lactam resistance plasmid pESBL-283 between E. coli strains.

    Directory of Open Access Journals (Sweden)

    Nadine Händel

    Full Text Available The spread of antibiotic resistant bacteria worldwide presents a major health threat to human health care that results in therapy failure and increasing costs. The transfer of resistance conferring plasmids by conjugation is a major route by which resistance genes disseminate at the intra- and interspecies level. High similarities between resistance genes identified in foodborne and hospital-acquired pathogens suggest transmission of resistance conferring and transferrable mobile elements through the food chain, either as part of intact strains, or through transfer of plasmids from foodborne to human strains. To study the factors that affect the rate of plasmid transfer, the transmission of an extended-spectrum β-lactamase (ESBL plasmid from a foodborne Escherichia coli strain to the β-lactam sensitive E. coli MG1655 strain was documented as a function of simulated environmental factors. The foodborne E. coli isolate used as donor carried a CTX-M-1 harboring IncI1 plasmid that confers resistance to β-lactam antibiotics. Cell density, energy availability and growth rate were identified as factors that affect plasmid transfer efficiency. Transfer rates were highest in the absence of the antibiotic, with almost every acceptor cell picking up the plasmid. Raising the antibiotic concentrations above the minimum inhibitory concentration (MIC resulted in reduced transfer rates, but also selected for the plasmid carrying donor and recombinant strains. Based on the mutational pattern of transconjugant cells, a common mechanism is proposed which compensates for fitness costs due to plasmid carriage by reducing other cell functions. Reducing potential fitness costs due to maintenance and expression of the plasmid could contribute to persistence of resistance genes in the environment even without antibiotic pressure. Taken together, the results identify factors that drive the spread and persistence of resistance conferring plasmids in natural isolates

  14. Factors that affect transfer of the IncI1 β-lactam resistance plasmid pESBL-283 between E. coli strains.

    Science.gov (United States)

    Händel, Nadine; Otte, Sarah; Jonker, Martijs; Brul, Stanley; ter Kuile, Benno H

    2015-01-01

    The spread of antibiotic resistant bacteria worldwide presents a major health threat to human health care that results in therapy failure and increasing costs. The transfer of resistance conferring plasmids by conjugation is a major route by which resistance genes disseminate at the intra- and interspecies level. High similarities between resistance genes identified in foodborne and hospital-acquired pathogens suggest transmission of resistance conferring and transferrable mobile elements through the food chain, either as part of intact strains, or through transfer of plasmids from foodborne to human strains. To study the factors that affect the rate of plasmid transfer, the transmission of an extended-spectrum β-lactamase (ESBL) plasmid from a foodborne Escherichia coli strain to the β-lactam sensitive E. coli MG1655 strain was documented as a function of simulated environmental factors. The foodborne E. coli isolate used as donor carried a CTX-M-1 harboring IncI1 plasmid that confers resistance to β-lactam antibiotics. Cell density, energy availability and growth rate were identified as factors that affect plasmid transfer efficiency. Transfer rates were highest in the absence of the antibiotic, with almost every acceptor cell picking up the plasmid. Raising the antibiotic concentrations above the minimum inhibitory concentration (MIC) resulted in reduced transfer rates, but also selected for the plasmid carrying donor and recombinant strains. Based on the mutational pattern of transconjugant cells, a common mechanism is proposed which compensates for fitness costs due to plasmid carriage by reducing other cell functions. Reducing potential fitness costs due to maintenance and expression of the plasmid could contribute to persistence of resistance genes in the environment even without antibiotic pressure. Taken together, the results identify factors that drive the spread and persistence of resistance conferring plasmids in natural isolates and shows how these

  15. Proposed model for the high rate of rearrangement and rapid migration observed in some IncA/C plasmid lineages.

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    Meinersmann, R J; Lindsey, R L; Bono, J L; Smith, T P; Oakley, B B

    2013-08-01

    IncA/C plasmids are a class of plasmids from the Enterobacteriaceae that are relatively large (49 to >180 kbp), that are readily transferred by conjugation, and that carry multiple antimicrobial resistance genes. Reconstruction of the phylogeny of these plasmids has been difficult because of the high rate of remodeling by recombination-mediated horizontal gene transfer (HGT). We hypothesized that evaluation of nucleotide polymorphisms relative to the rate of HGT would help to develop a clock to show whether anthropic practices have had significant influences on the lineages of the plasmid. A system was developed to rapidly sequence up to 191 known open reading frames from each of 39 recently isolated IncA/C plasmids from a diverse panel of Salmonella enterica and Escherichia coli strains. With these data plus sequences from GenBank, we were able to distinguish six distinct lineages that had extremely low numbers of polymorphisms within each lineage, especially among the largest group designated as group 1. Two regions, each about half the plasmid in size, could be distinguished with a separate lineal pattern. The distribution of group 1 showed that it has migrated extremely rapidly with fewer polymorphisms than can be expected in 2,000 years. Remodeling by frequent HGT was evident, with a pattern that appeared to have the highest rate just upstream of the putative conjugation origin of transfer (oriT). It seems likely that when an IncA/C plasmid is transferred by conjugation there is an opportunity for plasmid remodeling adjacent to the oriT, which was also adjacent to a multiple antimicrobial resistance gene cassette.

  16. 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.

  17. 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....

  18. 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...

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

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    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

  20. TOL plasmid transfer during bacterial conjugation in vitro and rhizoremediation of oil compounds in vivo

    Energy Technology Data Exchange (ETDEWEB)

    Jussila, Minna M. [Department of Applied Chemistry and Microbiology, Viikki Biocenter, P.O. Box 56 (Viikinkaari 9), FI-00014 University of Helsinki, Helsinki (Finland)]. E-mail: minna.m.jussila@helsinki.fi; Zhao, Ji [Department of Applied Chemistry and Microbiology, Viikki Biocenter, P.O. Box 56 (Viikinkaari 9), FI-00014 University of Helsinki, Helsinki (Finland); Suominen, Leena [Department of Applied Chemistry and Microbiology, Viikki Biocenter, P.O. Box 56 (Viikinkaari 9), FI-00014 University of Helsinki, Helsinki (Finland); Lindstroem, Kristina [Department of Applied Chemistry and Microbiology, Viikki Biocenter, P.O. Box 56 (Viikinkaari 9), FI-00014 University of Helsinki, Helsinki (Finland)

    2007-03-15

    Molecular profiling methods for horizontal transfer of aromatics-degrading plasmids were developed and applied during rhizoremediation in vivo and conjugations in vitro. pWW0 was conjugated from Pseudomonas to Rhizobium. The xylE gene was detected both in Rhizobium galegae bv. officinalis and bv. orientalis, but it was neither stably maintained in orientalis nor functional in officinalis. TOL plasmids were a major group of catabolic plasmids among the bacterial strains isolated from the oil-contaminated rhizosphere of Galega orientalis. A new finding was that some Pseudomonas migulae and Pseudomonas oryzihabitans strains harbored a TOL plasmid with both pWW0- and pDK1-type xylE gene. P. oryzihabitans 29 had received the archetypal TOL plasmid pWW0 from Pseudomonas putida PaW85. As an application for environmental biotechnology, the biodegradation potential of oil-polluted soil and the success of bioremediation could be estimated by monitoring changes not only in the type and amount but also in transfer of degradation plasmids. - Horizontal transfer of degradation plasmids in the oil-contaminated rhizosphere reveals the dynamic nature of the intrinsic biodegradation potential.

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

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    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...

  2. Plasmid Transfer into the Homoacetogen Acetobacterium woodii by Electroporation and Conjugation

    OpenAIRE

    Strätz, Michael; Sauer, Uwe; Kuhn, Anita; Dürre, Peter

    1994-01-01

    Shuttle vectors (pMS3 and pMS4) which replicated in Escherichia coli and in gram-positive Acetobacterium woodii were constructed by ligating the replication origin of plasmid pAMβ1 with the E. coli cloning vector pUC19 and the tetM gene of streptococcal transposon Tn916. Electrotransformation of A. woodii was achieved at frequencies of 4.5 × 103 transformants per μg of plasmid DNA. For conjugal plasmid transfer, the mobilizable shuttle vector pKV12 was constructed by cloning the tetM gene int...

  3. Biofilm models for the food industry: hot spots for plasmid transfer?

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    Van Meervenne, Eva; De Weirdt, Rosemarie; Van Coillie, Els; Devlieghere, Frank; Herman, Lieve; Boon, Nico

    2014-04-01

    Biofilms represent a substantial problem in the food industry, with food spoilage, equipment failure, and public health aspects to consider. Besides, biofilms may be a hot spot for plasmid transfer, by which antibiotic resistance can be disseminated to potential foodborne pathogens. This study investigated biomass and plasmid transfer in dual-species (Pseudomonas putida and Escherichia coli) biofilm models relevant to the food industry. Two different configurations (flow-through and drip-flow) and two different inoculation procedures (donor-recipient and recipient-donor) were tested. The drip-flow configuration integrated stainless steel coupons in the setup while the flow-through configuration included a glass flow cell and silicone tubing. The highest biomass density [10 log (cells cm-²)] was obtained in the silicone tubing when first the recipient strain was inoculated. High plasmid transfer ratios, up to 1/10 (transconjugants/total bacteria), were found. Depending on the order of inoculation, a difference in transfer efficiency between the biofilm models could be found. The ease by which the multiresistance plasmid was transferred highlights the importance of biofilms in the food industry as hot spots for the acquisition of multiresistance plasmids. This can impede the treatment of foodborne illnesses if pathogens acquire this multiresistance in or from the biofilm.

  4. A new and improved host-independent plasmid system for RK2-based conjugal transfer.

    Directory of Open Access Journals (Sweden)

    Trine Aakvik Strand

    Full Text Available Bacterial conjugation is a process that is mediated either by a direct cell-to-cell junction or by formation of a bridge between the cells. It is often used to transfer DNA constructs designed in Escherichia coli to recipient bacteria, yeast, plants and mammalian cells. Plasmids bearing the RK2/RP4 origin of transfer (oriT are mostly mobilized using the E. coli S17-1/SM10 donor strains, in which transfer helper functions are provided from a chromosomally integrated RP4::Mu. We have observed that large plasmids were occasionally modified after conjugal transfer when using E. coli S17-1 as a donor. All modified plasmids had increased in size, which most probably was a result of co-transfer of DNA from the chromosomally located oriT. It has earlier also been demonstrated that the bacteriophage Mu is silently transferred to recipient cells by these donor strains, and both occurrences are very likely to lead to mutations within the recipient DNA. Here we report the construction of a new biological system addressing both the above mentioned problems in which the transfer helper functions are provided by a plasmid lacking a functional oriT. This system is compatible with all other replicons commonly used in conjugation experiments and further enables the use of diverse bacterial strains as donors. Plasmids containing large inserts were successfully conjugated and the plasmid modifications observed when E. coli S17-1 was used as donor were eliminated by the use of the new host-independent vector system.

  5. Transfer of plasmid-mediated resistance to tetracycline in pathogenic bacteria from fish and aquaculture environments.

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    Guglielmetti, Elena; Korhonen, Jenni M; Heikkinen, Jouni; Morelli, Lorenzo; von Wright, Atte

    2009-04-01

    The transferability of a large plasmid that harbors a tetracycline resistance gene tet(S), to fish and human pathogens was assessed using electrotransformation and conjugation. The plasmid, originally isolated from fish intestinal Lactococcus lactis ssp. lactis KYA-7, has potent antagonistic activity against the selected recipients (Lactococcus garvieae and Listeria monocytogenes), preventing conjugation. Therefore the tetracycline resistance determinant was transferred via electroporation to L. garvieae. A transformant clone was used as the donor in conjugation experiments with three different L. monocytogenes strains. To our knowledge, this is the first study showing the transfer of an antibiotic resistance plasmid from fish-associated lactic bacteria to L. monocytogenes, even if the donor L. garvieae was not the original host of the tetracycline resistance but experimentally created by electroporation. These results demonstrate that the antibiotic resistance genes in the fish intestinal bacteria have the potential to spread both to fish and human pathogens, posing a risk to aquaculture and consumer safety.

  6. Transfer of an indigenous plasmid of Rhizobium loti to other rhizobia and Agrobacterium tumefaciens.

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    Pankhurst, C E; Broughton, W J; Wieneke, U

    1983-08-01

    Rhizobium loti strains NZP2037 and NZP2213 were each found to contain a single large plasmid: pRlo2037a (240 MDal) and pRlo2213a (120 MDal), respectively. Plasmid DNA present in crude cell lysates of each strain and purified pRlo2037a DNA did not hybridize with pID1, a recombinant plasmid containing part of the nitrogen fixation (nif) region of R. meliloti, indicating that nif genes were not present on these plasmids. The transposon Tn5 was inserted into pRlo2037a and this plasmid was then transferred into R. leguminosarum, R. meliloti and Agrobacterium tumefaciens. All transconjugants failed to nodulate Lotus pedunculatus, suggesting that the ability to nodulate this legume was also not carried on pRlo2037a. Transfer of pRlo2037a to R. loti strain NZP2213 did not alter the Nod+ Fix- phenotype of this strain for L. pedunculatus. Determinants for flavolan resistance, believed to be necessary for effective nodulation of L. pedunculatus, were not carried on pRlo2037a. These data suggest that nodulation, nitrogen fixation and flavolan resistance genes are not present on the large plasmid in R. loti strain NZP2037.

  7. Transfer of IncP Plasmids to Extremely Acidophilic Thiobacillus thiooxidans

    OpenAIRE

    Jin, S. M.; Yan, W. M.; Wang, Z. N.

    1992-01-01

    The broad-host-range IncP plasmids RP4, R68.45, RP1::Tn501, and and pUB307 were transferred directly to extremely acidophilic Thiobacillus thiooxidans from Escherichia coli by conjugation at frequencies of 10-5 to 10-7 per recipient. The ability of T. thiooxidans to receive and express the antibiotic resistance markers was examined. The plasmid RP4 was transferred back to E. coli from T. thiooxidans at a frequency of 1.0 × 10-3 per recipient.

  8. Transfer of IncP Plasmids to Extremely Acidophilic Thiobacillus thiooxidans.

    Science.gov (United States)

    Jin, S M; Yan, W M; Wang, Z N

    1992-01-01

    The broad-host-range IncP plasmids RP4, R68.45, RP1::Tn501, and and pUB307 were transferred directly to extremely acidophilic Thiobacillus thiooxidans from Escherichia coli by conjugation at frequencies of 10 to 10 per recipient. The ability of T. thiooxidans to receive and express the antibiotic resistance markers was examined. The plasmid RP4 was transferred back to E. coli from T. thiooxidans at a frequency of 1.0 x 10 per recipient.

  9. 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

  10. Horizontal Transfer of Plasmid-Mediated Cephalosporin Resistance Genes in the Intestine of Houseflies (Musca domestica).

    Science.gov (United States)

    Fukuda, Akira; Usui, Masaru; Okubo, Torahiko; Tamura, Yutaka

    2016-06-01

    Houseflies are a mechanical vector for various types of bacteria, including antimicrobial-resistant bacteria (ARB). If the intestine of houseflies is a suitable site for the transfer of antimicrobial resistance genes (ARGs), houseflies could also serve as a biological vector for ARB. To clarify whether cephalosporin resistance genes are transferred efficiently in the housefly intestine, we compared with conjugation experiments in vivo (in the intestine) and in vitro by using Escherichia coli with eight combinations of four donor and two recipient strains harboring plasmid-mediated cephalosporin resistance genes and chromosomal-encoded rifampicin resistance genes, respectively. In the in vivo conjugation experiment, houseflies ingested donor strains for 6 hr and then recipient strains for 3 hr, and 24 hr later, the houseflies were surface sterilized and analyzed. In vitro conjugation experiments were conducted using the broth-mating method. In 3/8 combinations, the in vitro transfer frequency (Transconjugants/Donor) was ≥1.3 × 10(-4); the in vivo transfer rates of cephalosporin resistance genes ranged from 2.0 × 10(-4) to 5.7 × 10(-5). Moreover, cephalosporin resistance genes were transferred to other species of enteric bacteria of houseflies such as Achromobacter sp. and Pseudomonas fluorescens. These results suggest that houseflies are not only a mechanical vector for ARB but also a biological vector for the occurrence of new ARB through the horizontal transfer of ARGs in their intestine.

  11. Construction of disarmed Ti plasmids transferable between Escherichia coli and Agrobacterium species.

    Science.gov (United States)

    Kiyokawa, Kazuya; Yamamoto, Shinji; Sakuma, Kei; Tanaka, Katsuyuki; Moriguchi, Kazuki; Suzuki, Katsunori

    2009-04-01

    Agrobacterium-mediated plant transformation has been used widely, but there are plants that are recalcitrant to this type of transformation. This transformation method uses bacterial strains harboring a modified tumor-inducing (Ti) plasmid that lacks the transfer DNA (T-DNA) region (disarmed Ti plasmid). It is desirable to develop strains that can broaden the host range. A large number of Agrobacterium strains have not been tested yet to determine whether they can be used in transformation. In order to improve the disarming method and to obtain strains disarmed and ready for the plant transformation test, we developed a simple scheme to make certain Ti plasmids disarmed and simultaneously maintainable in Escherichia coli and mobilizable between E. coli and Agrobacterium. To establish the scheme in nopaline-type Ti plasmids, a neighboring segment to the left of the left border sequence, a neighboring segment to the right of the right border sequence of pTi-SAKURA, a cassette harboring the pSC101 replication gene between these two segments, the broad-host-range IncP-type oriT, and the gentamicin resistance gene were inserted into a suicide-type sacB-containing vector. Replacement of T-DNA with the cassette in pTiC58 and pTi-SAKURA occurred at a high frequency and with high accuracy when the tool plasmid was used. We confirmed that there was stable maintenance of the modified Ti plasmids in E. coli strain S17-1lambdapir and conjugal transfer from E. coli to Ti-less Agrobacterium strains and that the reconstituted Agrobacterium strains were competent to transfer DNA into plant cells. As the modified plasmid delivery system was simple and efficient, conversion of strains to the disarmed type was easy and should be applicable in studies to screen for useful strains.

  12. TcpM: a novel relaxase that mediates transfer of large conjugative plasmids from Clostridium perfringens.

    Science.gov (United States)

    Wisniewski, Jessica A; Traore, Daouda A; Bannam, Trudi L; Lyras, Dena; Whisstock, James C; Rood, Julian I

    2016-03-01

    Conjugative transfer of toxin and antibiotic resistance plasmids in Clostridium perfringens is mediated by the tcp conjugation locus. Surprisingly, neither a relaxase gene nor an origin of transfer (oriT) has been identified on these plasmids, which are typified by the 47 kb tetracycline resistance plasmid pCW3. The tcpM gene (previously called intP) encodes a potential tyrosine recombinase that was postulated to be an atypical relaxase. Mutagenesis and complementation studies showed that TcpM was required for wild-type transfer of pCW3 and that a tyrosine residue, Y259, was essential for TcpM activity, which was consistent with the need for a relaxase-mediated hydrophilic attack at the oriT site. Other catalytic residues conserved in tyrosine recombinases were not required for TcpM activity, suggesting that TcpM was not a site-specific recombinase. Mobilization studies led to the identification of the oriT site, which was located in the 391 bp intergenic region upstream of tcpM. The oriT site was localized to a 150 bp region, and gel mobility shift studies showed that TcpM could bind to this region. Based on these studies we postulate that conjugative transfer of pCW3 involves the atypical relaxase TcpM binding to and processing the oriT site to initiate plasmid transfer.

  13. 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.

  14. Selective pressure affects transfer and establishment of a Lactobacillus plantarum resistance plasmid in the gastrointestinal environment

    DEFF Research Database (Denmark)

    Feld, Louise; Schjorring, S.; Hammer, Karin;

    2008-01-01

    Objectives and methods: A Lactobacillus plantarum strain recently isolated from French raw-milk cheese was tested for its ability to transfer a small plasmid pLFE1 harbouring the erythromycin resistance gene erm(B) to Enterococcus faecalis. Mating was studied in vitro and in different...

  15. Strain-Specific Transfer of Antibiotic Resistance from an Environmental Plasmid to Foodborne Pathogens

    Directory of Open Access Journals (Sweden)

    Eva Van Meervenne

    2012-01-01

    Full Text Available Pathogens resistant to multiple antibiotics are rapidly emerging, entailing important consequences for human health. This study investigated if the broad-host-range multiresistance plasmid pB10, isolated from a wastewater treatment plant, harbouring amoxicillin, streptomycin, sulfonamide, and tetracycline resistance genes, was transferable to the foodborne pathogens Salmonella spp. or E. coli O157:H7 and how this transfer alters the phenotype of the recipients. The transfer ratio was determined by both plating and flow cytometry. Antibiotic resistance profiles were determined for both recipients and transconjugants using the disk diffusion method. For 14 of the 15 recipient strains, transconjugants were detected. Based on plating, transfer ratios were between 6.8×10−9 and 3.0×10−2 while using flow cytometry, transfer ratios were between <1.0×10−5 and 1.9×10−2. With a few exceptions, the transconjugants showed phenotypically increased resistance, indicating that most of the transferred resistance genes were expressed. In summary, we showed that an environmental plasmid can be transferred into foodborne pathogenic bacteria at high transfer ratios. However, the transfer ratio seemed to be recipient strain dependent. Moreover, the newly acquired resistance genes could turn antibiotic susceptible strains into resistant ones, paving the way to compromise human health.

  16. 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.

  17. Transfer and Occurrence of Large Mercury Resistance Plasmids in River Epilithon

    OpenAIRE

    Bale, M J; Fry, J C; Day, M J

    1988-01-01

    In situ mating experiments were done in the River Taff, South Wales, United Kingdom, by using a natural mercury resistance plasmid (pQM1) isolated from a mixture of epilithic bacteria in vitro. The river temperature from March to November was found to influence transfer frequencies strongly (6.8 × 10−9 to 1.5 × 10−2 per recipient). A linear relationship existed between log10 transfer frequency and river temperature (6 to 21°C), a 2.6°C change in temperature giving a 10-fold change in transfer...

  18. High-frequency conjugal plasmid transfer from gram-negative Escherichia coli to various gram-positive coryneform bacteria.

    Science.gov (United States)

    Schäfer, A; Kalinowski, J; Simon, R; Seep-Feldhaus, A H; Pühler, A

    1990-03-01

    We report on the mobilization of shuttle plasmids from gram-negative Escherichia coli to gram-positive corynebacteria mediated by P-type transfer functions. Introduction of plasmids into corynebacteria was markedly enhanced after heat treatment of the recipient cells. High-frequency plasmid transfer was also observed when the restriction system of the recipient was mutated. On the basis of our data, we conclude that efficient DNA transfer from gram-negative to gram-positive bacteria, at least to coryneform bacteria, is conceivable in certain natural ecosystems.

  19. Origin-of-transfer sequences facilitate mobilisation of non-conjugative antimicrobial-resistance plasmids in Staphylococcus aureus

    Science.gov (United States)

    O'Brien, Frances G.; Yui Eto, Karina; Murphy, Riley J. T.; Fairhurst, Heather M.; Coombs, Geoffrey W.; Grubb, Warren B.; Ramsay, Joshua P.

    2015-01-01

    Staphylococcus aureus is a common cause of hospital, community and livestock-associated infections and is increasingly resistant to multiple antimicrobials. A significant proportion of antimicrobial-resistance genes are plasmid-borne, but only a minority of S. aureus plasmids encode proteins required for conjugative transfer or Mob relaxase proteins required for mobilisation. The pWBG749 family of S. aureus conjugative plasmids can facilitate the horizontal transfer of diverse antimicrobial-resistance plasmids that lack Mob genes. Here we reveal that these mobilisable plasmids carry copies of the pWBG749 origin-of-transfer (oriT) sequence and that these oriT sequences facilitate mobilisation by pWBG749. Sequences resembling the pWBG749 oriT were identified on half of all sequenced S. aureus plasmids, including the most prevalent large antimicrobial-resistance/virulence-gene plasmids, pIB485, pMW2 and pUSA300HOUMR. oriT sequences formed five subfamilies with distinct inverted-repeat-2 (IR2) sequences. pWBG749-family plasmids encoding each IR2 were identified and pWBG749 mobilisation was found to be specific for plasmids carrying matching IR2 sequences. Specificity of mobilisation was conferred by a putative ribbon-helix-helix-protein gene smpO. Several plasmids carried 2–3 oriT variants and pWBG749-mediated recombination occurred between distinct oriT sites during mobilisation. These observations suggest this relaxase-in trans mechanism of mobilisation by pWBG749-family plasmids is a common mechanism of plasmid dissemination in S. aureus. PMID:26243776

  20. Conjugative transfer of an IncA/C plasmid-borne blaCMY-2 gene through genetic re-arrangements with an IncX1 plasmid.

    Science.gov (United States)

    Wiesner, Magdalena; Fernández-Mora, Marcos; Cevallos, Miguel A; Zavala-Alvarado, Crispín; Zaidi, Mussaret B; Calva, Edmundo; Silva, Claudia

    2013-11-21

    Our observation that in the Mexican Salmonella Typhimurium population none of the ST19 and ST213 strains harbored both the Salmonella virulence plasmid (pSTV) and the prevalent IncA/C plasmid (pA/C) led us to hypothesize that restriction to horizontal transfer of these plasmids existed. We designed a conjugation scheme using ST213 strain YU39 as donor of the blaCMY-2 gene (conferring resistance to ceftriaxone; CRO) carried by pA/C, and two E. coli lab strains (DH5α and HB101) and two Typhimurium ST19 strains (SO1 and LT2) carrying pSTV as recipients. The aim of this study was to determine if the genetic background of the different recipient strains affected the transfer frequencies of pA/C. YU39 was able to transfer CRO resistance, via a novel conjugative mechanism, to all the recipient strains although at low frequencies (10-7 to 10-10). The presence of pSTV in the recipients had little effect on the conjugation frequency. The analysis of the transconjugants showed that three different phenomena were occurring associated to the transfer of blaCMY-2: 1) the co-integration of pA/C and pX1; 2) the transposition of the CMY region from pA/C to pX1; or 3) the rearrangement of pA/C. In addition, the co-lateral mobilization of a small (5 kb) ColE1-like plasmid was observed. The transconjugant plasmids involving pX1 re-arrangements (either via co-integration or ISEcp1-mediated transposition) obtained the capacity to conjugate at very high levels, similar to those found for pX1 (10-1). Two versions of the region containing blaCMY-2 were found to transpose to pX1: the large version was inserted into an intergenic region located where the "genetic load" operons are frequently inserted into pX1, while the short version was inserted into the stbDE operon involved in plasmid addiction system. This is the first study to report the acquisition of an extended spectrum cephalosporin (ESC)-resistance gene by an IncX1 plasmid. We showed that the transfer of the YU39 blaCMY-2 gene

  1. Conjugal plasmid transfer in Streptomyces resembles bacterial chromosome segregation by FtsK/SpoIIIE.

    Science.gov (United States)

    Vogelmann, Jutta; Ammelburg, Moritz; Finger, Constanze; Guezguez, Jamil; Linke, Dirk; Flötenmeyer, Matthias; Stierhof, York-Dieter; Wohlleben, Wolfgang; Muth, Günther

    2011-06-01

    Conjugation is a major route of horizontal gene transfer, the driving force in the evolution of bacterial genomes. Antibiotic producing soil bacteria of the genus Streptomyces transfer DNA in a unique process involving a single plasmid-encoded protein TraB and a double-stranded DNA molecule. However, the molecular function of TraB in directing DNA transfer from a donor into a recipient cell is unknown. Here, we show that TraB constitutes a novel conjugation system that is clearly distinguished from DNA transfer by a type IV secretion system. We demonstrate that TraB specifically recognizes and binds to repeated 8 bp motifs on the conjugative plasmid. The specific DNA recognition is mediated by helix α3 of the C-terminal winged-helix-turn-helix domain of TraB. We show that TraB assembles to a hexameric ring structure with a central ∼3.1 nm channel and forms pores in lipid bilayers. Structure, sequence similarity and DNA binding characteristics of TraB indicate that TraB is derived from an FtsK-like ancestor protein, suggesting that Streptomyces adapted the FtsK/SpoIIIE chromosome segregation system to transfer DNA between two distinct Streptomyces cells.

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

    Science.gov (United States)

    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

  3. The interplay between relatedness and horizontal gene transfer drives the evolution of plasmid-carried public goods.

    Science.gov (United States)

    Mc Ginty, Sorcha É; Lehmann, Laurent; Brown, Sam P; Rankin, Daniel J

    2013-06-22

    Plasmids carry a wide range of genes that are often involved in bacterial social behaviour. The question of why such genes are frequently mobile has received increasing attention. Here, we use an explicit population genetic approach to model the evolution of plasmid-borne bacterial public goods production. Our findings highlight the importance of both transmission and relatedness as factors driving the evolution of plasmid-borne public goods production. We partition the effects of plasmid transfer of social traits into those of infectivity and the effect of increased relatedness. Our results demonstrate that, owing to its effect on relatedness, plasmid mobility increases the invasion and stability of public goods, in a way not seen in individually beneficial traits. In addition, we show that plasmid transfer increases relatedness when public goods production is rare but this effect declines when production is common, with both scenarios leading to an increase in the frequency of plasmid-borne public goods. Plasmids remain important vectors for the spread of social genes involved in bacterial virulence thus an understanding of their dynamics is highly relevant from a public health perspective.

  4. 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.

  5. Production of Fibronectin Binding Protein A at the surface of Lactococcus lactis increases plasmid transfer in vitro and in vivo.

    Directory of Open Access Journals (Sweden)

    Daniela Pontes

    Full Text Available Lactococci are noninvasive lactic acid bacteria frequently used as protein delivery vectors and, more recently, as DNA delivery vehicles. We previously showed that Lactococcus lactis (LL expressing the Fibronectin-Binding Protein A of Staphylococcus aureus (LL-FnBPA+ showed higher internalization rates in vitro in Caco-2 cells than the native (wt lactococci and were able to deliver a eukaryotic Green Fluorescent Protein (GFP expression plasmid in 1% of human Caco-2 cells. Here, using the bovine beta-lactoglobulin (BLG, one of the major cow's milk allergen, and GFP we characterized the potential of LL-FnBPA+ as an in vivo DNA vaccine delivery vehicle. We first showed that the invasive strain LL-FnBPA+ carrying the plasmid pValac:BLG (LL-FnBPA+ BLG was more invasive than LL-BLG and showed the same invasivity as LL-FnBPA+. Then we demonstrated that the Caco-2 cells, co-incubated with LL-FnBPA+ BLG produced up to 30 times more BLG than the Caco-2 cells co-incubated with the non invasive LL-BLG. Using two different gene reporters, BLG and GFP, and two different methods of detection, EIA and fluorescence microscopy, we showed in vivo that: i in order to be effective, LL-FnBPA+ required a pre-coating with Fetal Calf Serum before oral administration; ii plasmid transfer occurred in enterocytes without regard to the strains used (invasive or not; iii the use of LL-FnBPA+ increased the number of mice producing BLG, but not the level of BLG produced. We thus confirmed the good potential of invasive recombinant lactic acid bacteria as DNA delivery vector in vivo.

  6. Transfer of drug-resistance plasmids by conjugation from nosocomial strains of Serratia marcescens to Escherichia coli in biological fluids of human origin.

    Science.gov (United States)

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

    1982-09-01

    Six independent isolates of multi-resistant Serratia marcescens associated with nosocomial infections were examined for their ability to transfer drug-resistance plasmids by conjugation to Escherichia coli in biological fluids of human origin, such as normal and pathological urine, faeces, blood plasma and ascitic fluid. Luria broth was used as a control. Positive transfer was found in all media assayed. The different patterns of linked transferable resistance found in the transconjugants corresponded to the phenotypic expression of five plasmids. The frequencies of transfer varied with plasmid types and media employed. The culture media did not affect the phenotypic expression of the plasmids.

  7. Conjugative plasmid transfer from Escherichia coli is a versatile approach for genetic transformation of thermophilic Bacillus and Geobacillus species.

    Science.gov (United States)

    Tominaga, Yurie; Ohshiro, Takashi; Suzuki, Hirokazu

    2016-05-01

    We previously demonstrated efficient transformation of the thermophile Geobacillus kaustophilus HTA426 using conjugative plasmid transfer from Escherichia coli BR408. To evaluate the versatility of this approach to thermophile transformation, this study examined genetic transformation of various thermophilic Bacillus and Geobacillus spp. using conjugative plasmid transfer from E. coli strains. E. coli BR408 successfully transferred the E. coli-Geobacillus shuttle plasmid pUCG18T to 16 of 18 thermophiles with transformation efficiencies between 4.1 × 10(-7) and 3.8 × 10(-2)/recipient. Other E. coli strains that are different from E. coli BR408 in intracellular DNA methylation also generated transformants from 9 to 15 of the 18 thermophiles, including one that E. coli BR408 could not transform, although the transformation efficiencies of these strains were generally lower than those of E. coli BR408. The conjugation was performed by simple incubation of an E. coli donor and a thermophile recipient without optimization of experimental conditions. Moreover, thermophile transformants were distinguished from abundant E. coli donor only by high temperature incubation. These observations suggest that conjugative plasmid transfer, particularly using E. coli BR408, is a facile and versatile approach for plasmid introduction into thermophilic Bacillus and Geobacillus spp., and potentially a variety of other thermophiles.

  8. Effects of Metals on Antibiotic Resistance and Conjugal Plasmid Transfer in Soil Bacterial Communities

    DEFF Research Database (Denmark)

    Song, Jianxiao

    Antibiotic resistance currently represents one of the biggest challenges for human health and in recent years the environmental dimension of antibiotic resistance has been increasingly recognized. The soil environment serves as an important reservoir of antibiotic resistance determinants....... In addition to direct selection of antibiotic resistance by antibiotics, metals may co-select for antibiotic resistance via different mechanisms causing environmental selection of antibiotic resistance in metal contaminated soils. Horizontal gene transfer of mobile genetic elements (MGEs) like plasmids...... is generally considered one of the most important co-selection mechanisms as multiple resistance genes can be located on the same MGE. This PhD thesis focused on the impact of metals (Cu and Zn) on the development of antibiotic resistance in bacterial communities in soils exposed to different degrees...

  9. The tra locus of streptomycete plasmid pIJ101 mediates efficient transfer of a circular but not a linear version of the same replicon.

    Science.gov (United States)

    Wang, Jing; Pettis, Gregg S

    2010-09-01

    Conjugal transfer of circular plasmids in Streptomyces involves a unique mechanism employing few plasmid-encoded loci and the transfer of double-stranded DNA by an as yet uncharacterized intercellular route. Efficient transfer of the circular streptomycete plasmid pIJ101 requires only two plasmid loci: the pIJ101 tra gene, and as a cis-acting function known as clt. Here, we compared the ability of the pIJ101 transfer apparatus to promote conjugal transfer of circular versus linear versions of the same replicon. While the pIJ101 tra locus readily transferred the circular form of the replicon, the linear version was transferred orders of magnitude less efficiently and all plasmids isolated from the transconjugants were circular, regardless of their original configuration in the donor. Additionally, relatively rare circularization of linear plasmids was detectable in the donor cells, which is consistent with the notion that this event was a prerequisite for transfer by TraB(pIJ101). Linear versions of this same replicon did transfer efficiently, in that configuration, from strains containing the conjugative linear plasmid SLP2. Our data indicate that functions necessary and sufficient for transfer of circular DNA were insufficient for transfer of a related linear DNA molecule. The results here suggest that the conjugation mechanisms of linear versus circular DNA in Streptomyces spp. are inherently different and/or that efficient transfer of linear DNA requires additional components.

  10. Transfer of the virulence-associated protein a-bearing plasmid between field strains of virulent and avirulent Rhodococcus equi.

    Science.gov (United States)

    Stoughton, W; Poole, T; Kuskie, K; Liu, M; Bishop, K; Morrissey, A; Takai, S; Cohen, N

    2013-01-01

    Virulent and avirulent isolates of Rhodococcus equi coexist in equine feces and the environment and are a source of infection for foals. The extent to which plasmid transfer occurs among field strains is ill-defined and this information is important for understanding the epidemiology of R. equi infections of foals. To estimate the frequency of transfer of the virulence plasmid between virulent and avirulent strains of R. equi derived from foals and their environment. None. In vitro study; 5 rifampin-susceptible, virulent R. equi isolates obtained from clinically affected foals or air samples from a farm with a history of recurrent R. equi foal pneumonia were each mixed with 5 rifampin-resistant, avirulent isolates derived from soil samples, using solid medium, at a ratio of 10 donor cells (virulent) per recipient cell. Presumed transconjugates were detected by plating on media with rifampin and colony immunoblotting to detect the presence of the virulence-associated protein A. Three presumed transconjugates were detected among 2,037 recipient colonies, indicating an overall estimated transfer frequency of 0.15% (95% CI, 0.03–0.43%). All 3 transconjugates were associated with a single donor and 2 recipient strains. Genotyping and multiplex PCR of presumed transconjugates demonstrated transfer of the virulence-associated protein A-bearing plasmid between virulent and avirulent R. equi. Transfer of the virulence plasmid occurs with relatively high frequency. These findings could impact strategies to control or prevent R. equi through environmental management.

  11. Transfer of the pheromone-inducible plasmid pCF10 among Enterococcus faecalis microorganisms colonizing the intestine of mini-pigs

    DEFF Research Database (Denmark)

    Licht, Tine Rask; Laugesen, D.; Jensen, Lars Bogø;

    2002-01-01

    A new animal model, the streptomycin-treated mini-pig, was developed in order to allow colonization of defined strains of Enterococcus faecalis in numbers sufficient to study plasmid transfer. Transfer of the pheromone-inducible pCF10 plasmid between streptomycin-resistant strains of E. faecalis OG...... until the end of the experiment. These observations showed that even in the absence of selective tetracycline pressure, plasmid pCF10 was transferred from ingested E. faecalis cells to other E. faecalis organisms already present in the intestinal environment and that the plasmid subsequently persisted...

  12. Plasmid cloning vectors for the conjugal transfer of DNA from Escherichia coli to Streptomyces spp.

    Science.gov (United States)

    Bierman, M; Logan, R; O'Brien, K; Seno, E T; Rao, R N; Schoner, B E

    1992-07-01

    We have constructed cloning vectors for the conjugal transfer of DNA from Escherichia coli to Streptomyces spp. All vectors contain the 760-bp oriT fragment from the IncP plasmid, RK2. Transfer functions need to be supplied in trans by the E. coli donor strain. We have incorporated into these vectors selectable antibiotic-resistance markers (AmR, ThR, SpR) that function in Streptomyces spp. and other features that should allow for: (i) integration via homologous recombination between cloned DNA and the Streptomyces spp. chromosome, (ii) autonomous replication, or (iii) site-specific integration at the bacteriophage phi C31 attachment site. Shuttle cosmids for constructing genomic libraries and bacteriophage P1 cloning vector capable of accepting approx. 100-kb fragments are also described. A simple mating procedure has been developed for the conjugal transfer of these vectors from E. coli to Streptomyces spp. that involves plating of the donor strain and either germinated spores or mycelial fragments of the recipient strain. We have shown that several of these vectors can be introduced into Streptomyces fradiae, a strain that is notoriously difficult to transform by PEG-mediated protoplast transformation.

  13. 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

  14. 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.

  15. Gene electro transfer of plasmid encoding vascular endothelial growth factor for enhanced expression and perfusion in the ischemic swine heart.

    Science.gov (United States)

    Hargrave, Barbara; Strange, Robert; Navare, Sagar; Stratton, Michael; Burcus, Nina; Murray, Len; Lundberg, Cathryn; Bulysheva, Anna; Li, Fanying; Heller, Richard

    2014-01-01

    Myocardial ischemia can damage heart muscle and reduce the heart's pumping efficiency. This study used an ischemic swine heart model to investigate the potential for gene electro transfer of a plasmid encoding vascular endothelial growth factor for improving perfusion and, thus, for reducing cardiomyopathy following acute coronary syndrome. Plasmid expression was significantly greater in gene electro transfer treated tissue compared to injection of plasmid encoding vascular endothelial growth factor alone. Higher gene expression was also seen in ischemic versus non-ischemic groups with parameters 20 Volts (ptransfer of plasmid encoding vascular endothelial growth factor had increased perfusion in the area at risk compared to control groups. Troponin and creatine kinase increased across all groups, suggesting equivalent ischemia in all groups prior to treatment. Echocardiography was used to assess ejection fraction, cardiac output, stroke volume, left ventricular end diastolic volume, and left ventricular end systolic volume. No statistically significant differences in these parameters were detected during a 2-week time period. However, directional trends of these variables were interesting and offer valuable information about the feasibility of gene electro transfer of vascular endothelial growth factor in the ischemic heart. The results demonstrate that gene electro transfer can be applied safely and can increase perfusion in an ischemic area. Additional study is needed to evaluate potential efficacy.

  16. Transfer and loss of naturally-occuring plasmids among isolates of Rhizobium leguminosarum bv. viciae in heavy metal contaminated soils

    NARCIS (Netherlands)

    Lakzian, A.; Murphy, P.J.; Giller, K.E.

    2007-01-01

    Plasmid transfer among isolates of Rhizobium leguminosarum bv. viciae in heavy metal contaminated soils from a long-term experiment in Braunschweig, Germany, was investigated under laboratory conditions. Three replicate samples each of four sterilized soils with total Zn contents of 54, 104, 208 and

  17. Plasmid Biopharmaceuticals.

    Science.gov (United States)

    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.

  18. Plasmid Transfer of Plasminogen K1-5 Reduces Subcutaneous Hepatoma Growth by Affecting Inflammatory Factors

    Directory of Open Access Journals (Sweden)

    Lea A. Koch

    2014-01-01

    Full Text Available There is evidence that plasminogen K1-5 (PlgK1-5 directly affects tumour cells and inflammation. Therefore, we analysed if PlgK1-5 has immediate effects on hepatoma cells and inflammatory factors in vitro and in vivo. In vitro, effects of plasmid encoding PlgK1-5 (pK1-5 on Hepa129, Hepa1-6, and HuH7 cell viability, apoptosis, and proliferation as well as VEGF and TNF-alpha expression and STAT3-phosphorylation were investigated. In vivo, tumour growth, proliferation, vessel density, and effects on vascular endothelial growth factor (VEGF and tumour necrosis factor alpha (TNF-alpha expression were examined following treatment with pK1-5. In vivo, pK1-5 halved cell viability; cell death was increased by up to 15% compared to the corresponding controls. Proliferation was not affected. VEGF, TNF-alpha, and STAT3-phosphorylation were affected following treatment with pK1-5. In vivo, ten days after treatment initiation, pK1-5 reduced subcutaneous tumour growth by 32% and mitosis by up to 77% compared to the controls. Vessel density was reduced by 50%. TNF-alpha levels in tumour and liver tissue were increased, whereas VEGF levels in tumours and livers were reduced after pK1-5 treatment. Taken together, plasmid gene transfer of PlgK1-5 inhibits hepatoma (cell growth not only by reducing vessel density but also by inducing apoptosis, inhibiting proliferation, and triggering inflammation.

  19. Dissemination of a clone carrying a fosA3-harbouring plasmid mediates high fosfomycin resistance rate of KPC-producing Klebsiella pneumoniae in China.

    Science.gov (United States)

    Jiang, Yan; Shen, Ping; Wei, Zeqing; Liu, Lilin; He, Fang; Shi, Keren; Wang, Yanfei; Wang, Haiping; Yu, Yunsong

    2015-01-01

    Fosfomycin has been proposed as an adjunct to other active agents for treating KPC-producing Klebsiella pneumoniae infections. This study aimed to investigate the prevalence of fosfomycin resistance and plasmid-mediated resistance determinants among KPC-producing K. pneumoniae isolates from clinical samples in China. In total, 278 KPC-producing and 80 extended-spectrum β-lactamase (ESBL)-producing (non-KPC-producing) clinical K. pneumoniae isolates were collected in 12 hospitals from 2010 to 2013. Fosfomycin susceptibility testing was carried out using the agar dilution method. Phylogenetic clonal patterns were revealed by pulsed-field gel electrophoresis (PFGE). Isolates were screened for plasmid-mediated fosfomycin resistance genes (fosA, fosA3 and fosC2) by PCR amplification. A plasmid was completely sequenced by next-generation sequencing. The fosfomycin resistance rate in KPC-producers (60.8%; 169/278) was significantly higher than in ESBL-producers (12.5%; 10/80). In addition, 94 KPC-producing isolates were positive for fosA3 and most of them were clonally related. A 23939-bp plasmid (pFOS18) co-harbouring fosA3 and bla(KPC-2) was completely sequenced, revealing that the fosA3 gene was flanked by two copies of IS26; however, bla(KPC-2) was located on a Tn3-Tn4401 integration structure. Although the fosA3 and blaKPC-2 genes are located on different transposon systems, they are able to spread together worldwide through plasmid transfer. Dissemination of the clone carrying the fosA3-harbouring plasmid mediates the high fosfomycin resistance rate of KPC-producing K. pneumoniae in China. Fosfomycin as an alternative option for treating infections caused by KPC-producing K. pneumoniae should not be recommended in hospitals in which fosfomycin-resistant clonal dissemination is emerging.

  20. Novel R-plasmid conjugal transfer inhibitory and antibacterial activities of phenolic compounds from Mallotus philippensis (Lam.) Mull. Arg.

    Science.gov (United States)

    Oyedemi, Blessing O M; Shinde, Vaibhav; Shinde, Kamlesh; Kakalou, Dionysia; Stapleton, Paul D; Gibbons, Simon

    2016-06-01

    Antimicrobial resistance severely limits the therapeutic options for many clinically important bacteria. In Gram-negative bacteria, multidrug resistance is commonly facilitated by plasmids that have the ability to accumulate and transfer refractory genes amongst bacterial populations. The aim of this study was to isolate and identify bioactive compounds from the medicinal plant Mallotus philippensis (Lam.) Mull. Arg. with both direct antibacterial properties and the capacity to inhibit plasmid conjugal transfer. A chloroform-soluble extract of M. philippensis was subjected to bioassay-guided fractionation using chromatographic and spectrometric techniques that led to the isolation of the known compounds rottlerin [5,7-dihydroxy-2,2-dimethyl-6-(2,4,6-trihydroxy-3-methyl-5-acetylbenzyl)-8-cinnamoyl-1,2-chromene] and the red compound (8-cinnamoyl-5,7-dihydroxy-2,2,6-trimethylchromene). Both compounds were characterised and elucidated using one-dimensional and two-dimensional nuclear magnetic resonance (NMR). Rottlerin and the red compound showed potent activities against a panel of clinically relevant Gram-positive bacteria, including meticillin-resistant Staphylococcus aureus (MRSA). No significant direct activities were observed against Gram-negative bacteria. However, both rottlerin and the red compound strongly inhibited conjugal transfer of the plasmids pKM101, TP114, pUB307 and R6K amongst Escherichia coli at a subinhibitory concentration of 100mg/L. Interestingly, despite the planar nature of the compounds, binding to plasmid DNA could not be demonstrated by a DNA electrophoretic mobility shift assay. These results show that rottlerin and the red compound are potential candidates for antibacterial drug lead development. Further studies are needed to elucidate the mode of inhibition of the conjugal transfer of plasmids.

  1. Gene Transfer into the Lung by Nanoparticle Dextran-Spermine/Plasmid DNA Complexes

    Directory of Open Access Journals (Sweden)

    Syahril Abdullah

    2010-01-01

    Full Text Available A novel cationic polymer, dextran-spermine (D-SPM, has been found to mediate gene expression in a wide variety of cell lines and in vivo through systemic delivery. Here, we extended the observations by determining the optimal conditions for gene expression of D-SPM/plasmid DNA (D-SPM/pDNA in cell lines and in the lungs of BALB/c mice via instillation delivery. In vitro studies showed that D-SPM could partially protect pDNA from degradation by nuclease and exhibited optimal gene transfer efficiency at D-SPM to pDNA weight-mixing ratio of 12. In the lungs of mice, the levels of gene expression generated by D-SPM/pDNA are highly dependent on the weight-mixing ratio of D-SPM to pDNA, amount of pDNA in the complex, and the assay time postdelivery. Readministration of the complex at day 1 following the first dosing showed no significant effect on the retention and duration of gene expression. The study also showed that there was a clear trend of increasing size of the complexes as the amount of pDNA was increased, where the sizes of the D-SPM/pDNA complexes were within the nanometer range.

  2. Gene electro transfer of plasmid encoding vascular endothelial growth factor for enhanced expression and perfusion in the ischemic swine heart.

    Directory of Open Access Journals (Sweden)

    Barbara Hargrave

    Full Text Available Myocardial ischemia can damage heart muscle and reduce the heart's pumping efficiency. This study used an ischemic swine heart model to investigate the potential for gene electro transfer of a plasmid encoding vascular endothelial growth factor for improving perfusion and, thus, for reducing cardiomyopathy following acute coronary syndrome. Plasmid expression was significantly greater in gene electro transfer treated tissue compared to injection of plasmid encoding vascular endothelial growth factor alone. Higher gene expression was also seen in ischemic versus non-ischemic groups with parameters 20 Volts (p<0.03, 40 Volts (p<0.05, and 90 Volts (p<0.05, but not with 60 Volts (p<0.09 while maintaining a pulse width of 20 milliseconds. The group with gene electro transfer of plasmid encoding vascular endothelial growth factor had increased perfusion in the area at risk compared to control groups. Troponin and creatine kinase increased across all groups, suggesting equivalent ischemia in all groups prior to treatment. Echocardiography was used to assess ejection fraction, cardiac output, stroke volume, left ventricular end diastolic volume, and left ventricular end systolic volume. No statistically significant differences in these parameters were detected during a 2-week time period. However, directional trends of these variables were interesting and offer valuable information about the feasibility of gene electro transfer of vascular endothelial growth factor in the ischemic heart. The results demonstrate that gene electro transfer can be applied safely and can increase perfusion in an ischemic area. Additional study is needed to evaluate potential efficacy.

  3. 人源志贺菌毒力大质粒体外转移及致病性分析%Transfer of strong virulent plasmid of human Shigella in vitro and pathogenicity of plasmid-transferred

    Institute of Scientific and Technical Information of China (English)

    刘红英; 杨霞; 陈陆; 王新卫; 许兰菊; 王川庆

    2011-01-01

    目的 为了解志贺菌毒力大质粒能否在体外转移及其转移后是否仍具有致病性.方法 将人源福氏志贺菌及其毒力大质粒分别与鸡源鲍氏志贺菌混合,并在37℃培养箱和4℃冰箱中混合培养24h;用不同剂量的毒力大质粒转移菌株通过口服和腹腔注射分别人工感染雏鸡和小白鼠.结果 人源福氏志贺菌的毒力大质成功转移到丢失自身毒力大质粒的鸡源鲍氏志贺菌中;转移菌株对雏鸡和小白鼠均有致病性,且转移菌株的培养特性也有所改变.结论 当志贺菌的毒力大质粒裸露出来且具有适宜的条件时,可以在体外转移到相近的病原菌中,且新生成的转移菌株具有一定稳定性和致病性;志贺菌的高致病性需要有毒力大质粒和菌体的其他特殊致病因子同时存在才能完成.%Objective To understand whether the strong virulent plasmid of Shigella can transfer in vitro and still possess pathogenicity after the transfer. Methods Human Shigella flexneri and the large virulent plasmid of Shigella were separately mixed with chicken Shigella boydii, and then cultured at 371 and 4t refrigerator for 24hours. Chickens and mice were experimentally infected either orally or intraperitoneally with transferred-strain of large plasmid in different dosages. Results The strong virulent plasmid of human Shigella was successfully transferred into chicken Shigella boydii which had lost the large virulence plasmid. Chicken and mice inoculated with transferred-strain showed pathogenicity of the strains transferred. The culture characteristics of transferred-strain was changed. Conclusion The strong virulent plasmid of Shigella in a suitable environment can be transferred in to pathogenic bacteria in the close category in vitro and the new transferred-strain possessed certain stability and pathogenicity. The high pathogenicity of Shigella would occurred if there were strong virulent plasmid and other special

  4. Conjugative plasmid transfer in Xylella fastidiosa is dependent on tra and trb operon functions

    Science.gov (United States)

    The insect-transmitted plant pathogen Xylella fastidiosa is capable of efficient horizontal gene transfer and recombination, leading to diversity between strains and the categorization of X. fastidiosa into multiple subspecies. Although natural transformation is shown to occur at high rates in X. fa...

  5. Experimental Study of Plasmid TGF-β1 DNA Gene Transfer with Lipofectamine into Rabbit Corneal Epithelial Cells In Vitro

    Institute of Scientific and Technical Information of China (English)

    黄琼; 胡燕华; 姜发纲; 陈宏

    2002-01-01

    To investigate whether the TGF-β1 plasmid DNA carried by lipofectamine could be introduced into cultured rabbit corneal epithelial cells, specific expression of the plasmid pMAM TGF-β1in the cultured corneal epithelial cells was studied. Two days after 12 h of transfection of pMAMTGF-β1 mediated by lipofectamine into the cultured corneal epithelial cells, the TGF-β1 protein expression specific for pMAMTGF-β1 in the cells was detected by means of immunohistochemical staining and the positive rate was 23. 37 %. The results suggested that foreign plasmid DNA could be effectively delivered into cultured rabbit corneal epithelial cells by means of lipofectamine, and this will provide a promising method of studying TGF-β1 on the mechanism of physiology and pathology concerned with corneal epithelial cells.

  6. Broad host range plasmid-based gene transfer system in the cyanobacterium Gloeobacter violaceus which lacks thylakoids

    Institute of Scientific and Technical Information of China (English)

    GUO Haitao; XU Xudong

    2004-01-01

    Gloeobacter violaceus, a cyanobacterium lack of thylakoids, is refractory to genetic manipulations because its cells are enveloped by a thick gelatinous sheath and in colonial form.In this study, a large number of single cells were obtained by repeated pumping with a syringe with the gelatinous sheath removed.And an exogenous broad host range plasmid pKT210 was conjugatively transferred into G.violaceus.Analyses with dot-blot hybridization and restriction mapping showed that the exogenous plasmid pKT210 had been introduced into G.violaceus and stably maintained with no alteration in its structure.pKT210 extracted from G.violaceus exconjugants could be transformed into the mcr- mrr- E.coli strain DH10B but not the mcr+ mrr+ strain DH5α, which suggests that a methylase system may be present in G.violaceus.

  7. Transfer of bcl-xs plasmid is effective in preventing and inhibiting rat hepatocellular carcinoma induced by N-nitrosomorpholine.

    Science.gov (United States)

    Baba, M; Iishi, H; Tatsuta, M

    2001-08-01

    To examine the effect of the bcl-xs gene on the sequence from hepatic precancerous lesions, foci and neoplastic nodules, to hepatocellular carcinomas, Sprague-Dawley rats were given water containing 175 mg/l N-nitrosomorpholine (NNM) for 8 weeks. At weeks 1, 4 and 7, the left lobe of the rat liver was exposed and injected with the bcl-xs plasmid (pCR3.1-rat bcl-xs cDNA) or pCR3.1 encapsulated in cationic empty liposomes each at a dose of 80 microg plasmid/kg body weight. One minute later, low-field-strength, long-duration electric pulses were applied to the left lobe using a pincette electrode with circular poles 1 cm in diameter. The in vivo electroporation procedure significantly increased the transfer of the reporter gene chloramphenicol acetyl transferase (CAT) plasmid via empty liposomes. Thus, CAT mRNA was expressed not only at the sites of electrode contact but at sites 0.5-1.0 cm away from the electrode, and expression also increased with increasing doses of plasmid, meaning that in vivo electroporation enabled the expression of plasmid DNA throughout an extensive area of the rat liver. By week 11, the neoplastic nodules were significantly fewer and smaller in the bcl-xs group than in the pCR3.1 group at the two sites, one with and the other without electrode contact. No hepatocellular carcinomas were found in the rats that had received the bcl-xs plasmid, whereas these tumors were observed in 30% of the rats given pCR3.1. Moreover, overexpression of the bcl-xs protein was detected, and apoptotic activity was significantly increased in the neoplastic nodules, foci and hepatocytes adjacent to the hepatic lesions. These results indicate that the bcl-xs plasmid inhibits the occurrence and growth of rat hepatocellular carcinoma and may thus be effective for the prevention and treatment of human liver tumors.

  8. Inhibition of resistance plasmid transfer in Escherichia coli by ionophores, chlortetracycline, bacitracin, and ionophore/antimicrobial combinations.

    Science.gov (United States)

    Mathers, Jeremy J; Clark, Steven R; Hausmann, Denny; Tillman, Paul; Benning, Valerie R; Gordon, Shelly K

    2004-01-01

    Medicinal feed additives bacitracin, chlortetracycline (CTC), laidlomycin, lasalocid, and salinomycin inhibited the transfer of multiresistance-conferring plasmid pBR325 (Tet(r) Amp(r) Cp(r), 6.0 kb) into selected gram-negative strains with the use of an in vitro model. High concentrations of ampicillin-sensitive competence-pretreated Escherichia coli HB 101 cells were exposed to 10% (v/v) of 1:10 dimethyl sulfoxide/agent : water containing test mixtures for 0.5 hr prior to plasmid addition and transforming conditions. Transformation was inhibited for all antimicrobials and showed a positive association wich higher concentration. Additional testing of ionophore compounds separately and in combination with bacitracin, chlortetracycline, lincomycin, roxarsone, tylosin, and virginiamycin at representative feed concentrations demonstrated 80.6% to >99.9% inhibition (P < 0.001) of resistance transfer. Bacitracin alone inhibited transformation within the range of 50-500 ppm. No increase in resistance transfer was observed when poultry-derived and reference gram-negative isolates having low or no transformation efficiency were additionally tested. The results suggest that these compounds, at relevant concentrations used in animal feed, may interfere with cell envelope-associated DNA uptake channels or other transformation competence mechanisms. Through these mechanisms, ionophores and cell membrane-interactive feed agents such as CTC and bacitracin may act to inhibit resistance transfer mechanisms within poultry and livestock.

  9. 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...

  10. Intergenerational transfers and the social discount rate

    Energy Technology Data Exchange (ETDEWEB)

    Howarth, R.B.; Norgaard, R.B.

    1992-08-01

    This paper investigates the relationship between intergenerational asset transfers and the choice of the discount rate for use in cost-benefit analysis in a model of a competitive overlapping generations economy constrained by a socially managed exhaustible resource. Provided that there are no distortions in capital markets and that all agents hold perfect foresight, cost-benefit techniques will result in a Pareto efficient resource allocation if the discount rate is set equal to the market rate of interest. But since the path of the interest rate depends on the level of intergenerational transfers, cost-benefit techniques do not ensure a socially desirable distribution of welfare between generations; a social optimum will result only if intergenerational transfers are properly chosen and enforced. Decentralized private altruism may result in intergenerational transfers that both present and future individuals would agree are too small if members of the present generation attach positive weight to the general welfare of future generations, not simply their personal descendants. In a world where intergenerational transfers are non-optimal, second-best policy-making may imply a constrained optimum that is inefficient. Together, these findings suggest that cost-benefit analysis is at best a partial criterion to policy formulation that should be used only in conjunction with ethical principles that define the proper distribution of welfare between present and future generations.

  11. Unusual distance dependences of electron transfer rates.

    Science.gov (United States)

    Kuss-Petermann, Martin; Wenger, Oliver S

    2016-07-28

    Usually the rates for electron transfer (kET) decrease with increasing donor-acceptor distance, but Marcus theory predicts a regime in which kET is expected to increase when the transfer distance gets longer. Until recently, experimental evidence for such counter-intuitive behavior had been very limited, and consequently this effect is much less well-known than the Gaussian free energy dependence of electron transfer rates leading to the so-called inverted driving-force effect. This article presents the theoretical concepts that lead to the prediction of electron transfer rate maxima at large donor-acceptor distances, and it discusses conditions that are expected to favor experimental observations of such behavior. It continues with a consideration of specific recent examples in which electron transfer rates were observed to increase with increasing donor-acceptor distance, and it closes with a discussion of the importance of this effect in the context of light-to-chemical energy conversion.

  12. Transposon characterization of vancomycin-resistant Enterococcus faecium (VREF) and dissemination of resistance associated with transferable plasmids

    DEFF Research Database (Denmark)

    Migura, Lourdes Garcia; Liebana, Ernesto; Jensen, Lars Bogø

    2007-01-01

    diversity demonstrated by PFGE (n = 62). Conjugation experiments were carried out to assess the transfer of vancomycin resistance. Co-transfer of vanA together with erm(B) positioned on the same conjugative plasmid containing a replicon similar to pRE25 was demonstrated and also the presence of different......Objectives: VanA glycopeptide resistance has persisted on broiler farms in the UK despite the absence of the antimicrobial selective pressure, avoparcin. This study aimed to investigate the contribution of horizontal gene transfer of Tn 1546 versus clonal spread in the dissemination...... of the resistance. Methods and results: One hundred and one vancomycin-resistant Enterococcus faecium isolated from 19 unrelated farms have been investigated. Tn 1546 characterization by long PCR and Clal-digestions of amplicons showed a very low diversity of Tn types (n = 4) in comparison to the high genotypic...

  13. Improvement of in vivo transfer of plasmid DNA in muscle : Comparison of electroporation versus ultrasound

    NARCIS (Netherlands)

    Kusumanto, Yoka H.; Mulder, Nanno H.; Dam, Wendy A.; Losen, Mario H.; Meijer, Coby; Hospers, Geke A. P.

    2007-01-01

    Plasmid-based gene delivery to muscle is a treatment strategy for many diseases with potential advantages above viral-based gene delivery methods, however, with a relative low transfection efficiency. We compared two physical methods-electroporation and ultrasound-that facilitate DNA uptake into cel

  14. 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...

  15. A novel peptide delivers plasmids across blood-brain barrier into neuronal cells as a single-component transfer vector.

    Directory of Open Access Journals (Sweden)

    Ailing Fu

    Full Text Available There is no data up to now to show that peptide can deliver plasmid into brain as a single-component transfer vector. Here we show that a novel peptide, RDP (consisted of 39 amino acids, can be exploited as an efficient plasmid vector for brain-targeting delivery. The plasmids containing Lac Z reporter gene (pVAX-Lac Z and BDNF gene (pVAX-BDNF are complexed with RDP and intravenously injected into mice. The results of gel retardation assay show that RDP enables to bind DNA in a dose-dependent manner, and the X-Gal staining identity that Lac Z is specifically expressed in the brain. Also, the results of Western blot and immunofluorescence staining of BDNF indicate that pVAX-BDNF complexed with RDP can be delivered into brain, and show neuroprotective properties in experimental Parkinson's disease (PD model. The results demonstrate that RDP enables to bind and deliver DNA into the brain, resulting in specific gene expression in the neuronal cells. This strategy provides a novel, simple and effective approach for non-viral gene therapy of brain diseases.

  16. A novel peptide delivers plasmids across blood-brain barrier into neuronal cells as a single-component transfer vector.

    Science.gov (United States)

    Fu, Ailing; Zhang, Miaomiao; Gao, Feiyan; Xu, Xingran; Chen, Zhangbao

    2013-01-01

    There is no data up to now to show that peptide can deliver plasmid into brain as a single-component transfer vector. Here we show that a novel peptide, RDP (consisted of 39 amino acids), can be exploited as an efficient plasmid vector for brain-targeting delivery. The plasmids containing Lac Z reporter gene (pVAX-Lac Z) and BDNF gene (pVAX-BDNF) are complexed with RDP and intravenously injected into mice. The results of gel retardation assay show that RDP enables to bind DNA in a dose-dependent manner, and the X-Gal staining identity that Lac Z is specifically expressed in the brain. Also, the results of Western blot and immunofluorescence staining of BDNF indicate that pVAX-BDNF complexed with RDP can be delivered into brain, and show neuroprotective properties in experimental Parkinson's disease (PD) model. The results demonstrate that RDP enables to bind and deliver DNA into the brain, resulting in specific gene expression in the neuronal cells. This strategy provides a novel, simple and effective approach for non-viral gene therapy of brain diseases.

  17. Differences in the carriage and the ability to utilize the serotype associated virulence plasmid in strains of Salmonella enterica serotype Typhimurium investigated by use of a self-transferable virulence plasmid, pOG669.

    Science.gov (United States)

    Olsen, John E; Brown, Derek J; Thomsen, Line E; Platt, David J; Chadfield, Mark S

    2004-06-01

    Most strains of Salmonella enterica subspecies enterica serotype typhimurium (S. typhimurium) naturally harbour a virulence plasmid which carries the salmonella plasmid virulence (spv) genes. However, isolates belonging to certain phage types are generally found without the plasmid. We have utilized a self-transferable virulence plasmid, pOG669 to investigate the effect of introduction of spv genes into strains of such phage types. The use of the co-integrate plasmid, pOG669, was validated on a diverse collection of strains. pOG669 was transferred into strains of serotypes that are normally associated with the possession of virulence plasmids. All strains maintained the wild type level of virulence in a mouse model, except that introduction of pOG669 restored normal virulence levels in an avirulent, plasmid free strain of S. dublin and resulted in a decrease in virulence in a strain of S. dublin from clonal line Du3. S. gallinarum did not become virulent in mice, but pOG669 was functionally interchangeable with the wild type plasmid when strains were tested in a chicken model. Strains of serotypes not normally associated with the carriage of a virulence plasmid did not increase in virulence upon the introduction of pOG669. An IncX plasmid pOG670 that was included as control was incompatible with the virulence plasmid in a strain of S. dublin, demonstrating that the common virulence plasmid of this serotype is of a different incompatibility group than other virulence plasmids. Strains of S. typhimurium from phage types that do not normally carry a virulence plasmid responded differently to attempts to introduce pOG669. No transconjugants were observed with the strains of DT5 and DT21. The introduction of pOG669 did not alter the virulence of JEO3942(DT10), DT35 and JEO3949(DT66) significantly, while DT1 and DT27 became more virulent. DT27 became as virulent as wild type C5, while logVC(10) of DT1 only increased from 4.1 to 5.7. The ability to express spv-genes was

  18. Transfer Rate Models for Gnutella Signaling Traffic

    OpenAIRE

    2006-01-01

    This paper reports on transfer rate models for the Gnutella signaling protocol. New results on message-level and IP-level rates are presented. The models are based on traffic captured at the Blekinge Institute of Technology (BTH) campus in Sweden and offer several levels of granularity: message type, application layer and network layer. The aim is to obtain parsimonous models suitable for analysis and simulation of P2P workload. IEEE Explorer

  19. Mass Action Models Describing Extant Horizontal Transfer of Plasmids: Inferences and Parameter Sensitivities

    DEFF Research Database (Denmark)

    Smets, Barth F.; Lardon, Laurent

    2009-01-01

    of the outcomes to the various plasmid dynamic parameters. For our analysis, we developed a set of user-friendly MatLab® routines, which are deposited in the public domain. We hope that the availability of these routines will encourage the computationally untrained microbiologist to make use of these mathematical...... models. Finally, further permutations, as well as limitations of these mass action models in view of the structured complexity of most microbial systems are addressed....

  20. Encapsulation of plasmid DNA in calcium phosphate nanoparticles: stem cell uptake and gene transfer efficiency

    Directory of Open Access Journals (Sweden)

    Cao X

    2011-12-01

    Full Text Available Xia Cao*, Wenwen Deng*, Yuan Wei*, Weiyan Su, Yan Yang, Yawei Wei, Jiangnan Yu, Ximing XuDepartment of Pharmaceutics, School of Pharmacy, and Center for Nano Drug/Gene Delivery and Tissue Engineering, Jiangsu University, Jingkou District, Zhenjiang, People's Republic of China*These authors contributed equally to this workBackground: The purpose of this study was to develop calcium phosphate nanocomposite particles encapsulating plasmid DNA (CP-pDNA nanoparticles as a nonviral vector for gene delivery.Methods: CP-pDNA nanoparticles employing plasmid transforming growth factor beta 1 (TGF-β1 were prepared and characterized. The transfection efficiency and cell viability of the CP-pDNA nanoparticles were evaluated in mesenchymal stem cells, which were identified by immunofluorescence staining. Cytotoxicity of plasmid TGF-β1 and calcium phosphate to mesenchymal stem cells were evaluated by MTT assay.Results: The integrity of TGF-β1 encapsulated in the CP-pDNA nanoparticles was maintained. The well dispersed CP-pDNA nanoparticles exhibited an ultralow particle size (20–50 nm and significantly lower cytotoxicity than Lipofectamine™ 2000. Immunofluorescence staining revealed that the cultured cells in this study were probably mesenchymal stem cells. The cellular uptake and transfection efficiency of the CP-pDNA nanoparticles into the mesenchymal stem cells were higher than that of needle-like calcium phosphate nanoparticles and a standard calcium phosphate transfection kit. Furthermore, live cell imaging and confocal laser microscopy vividly showed the transportation process of the CP-pDNA nanoparticles in mesenchymal stem cells. The results of a cytotoxicity assay found that both plasmid TGF-β1 and calcium phosphate were not toxic to mesenchymal stem cells.Conclusion: CP-pDNA nanoparticles can be developed into an effective alternative as a nonviral gene delivery system that is highly efficient and has low cytotoxicity.Keywords: calcium

  1. In vivo transfer of an incFIB plasmid harbouring a class 1 integron with gene cassettes dfrA1-aadA1

    NARCIS (Netherlands)

    Essen-Zandbergen, van A.; Smith, H.; Veldman, K.T.; Mevius, D.J.

    2009-01-01

    Transfer of resistance genes from bacteria from food producing animals to human pathogens is a potential risk to human health. The aim of this study was to determine in vivo transfer of a plasmid harbouring a class 1 integron containing gene cassettes dfrA1-aadA1 from Salmonella to Escherichia coli

  2. The multidrug resistance IncA/C transferable plasmid encodes a novel domain-swapped dimeric protein-disulfide isomerase.

    Science.gov (United States)

    Premkumar, Lakshmanane; Kurth, Fabian; Neyer, Simon; Schembri, Mark A; Martin, Jennifer L

    2014-01-31

    The multidrug resistance-encoding IncA/C conjugative plasmids disseminate antibiotic resistance genes among clinically relevant enteric bacteria. A plasmid-encoded disulfide isomerase is associated with conjugation. Sequence analysis of several IncA/C plasmids and IncA/C-related integrative and conjugative elements (ICE) from commensal and pathogenic bacteria identified a conserved DsbC/DsbG homolog (DsbP). The crystal structure of DsbP reveals an N-terminal domain, a linker region, and a C-terminal catalytic domain. A DsbP homodimer is formed through domain swapping of two DsbP N-terminal domains. The catalytic domain incorporates a thioredoxin-fold with characteristic CXXC and cis-Pro motifs. Overall, the structure and redox properties of DsbP diverge from the Escherichia coli DsbC and DsbG disulfide isomerases. Specifically, the V-shaped dimer of DsbP is inverted compared with EcDsbC and EcDsbG. In addition, the redox potential of DsbP (-161 mV) is more reducing than EcDsbC (-130 mV) and EcDsbG (-126 mV). Other catalytic properties of DsbP more closely resemble those of EcDsbG than EcDsbC. These catalytic differences are in part a consequence of the unusual active site motif of DsbP (CAVC); substitution to the EcDsbC-like (CGYC) motif converts the catalytic properties to those of EcDsbC. Structural comparison of the 12 independent subunit structures of DsbP that we determined revealed that conformational changes in the linker region contribute to mobility of the catalytic domain, providing mechanistic insight into DsbP function. In summary, our data reveal that the conserved plasmid-encoded DsbP protein is a bona fide disulfide isomerase and suggest that a dedicated oxidative folding enzyme is important for conjugative plasmid transfer.

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

    Directory of Open Access Journals (Sweden)

    Timothy J Johnson

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

  4. 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...

  5. Factors that affect transfer of the IncI1 β-lactam resistance plasmid pESBL-283 between E. coli strains

    NARCIS (Netherlands)

    Händel, N.; Otte, S.; Jonker, M.J.; Brul, S.; ter Kuile, B.H.

    2015-01-01

    The spread of antibiotic resistant bacteria worldwide presents a major health threat to human health care that results in therapy failure and increasing costs. The transfer of resistance conferring plasmids by conjugation is a major route by which resistance genes disseminate at the intra- and inter

  6. Vibrationally Resolved Electron Transfer Rates in Solution

    Science.gov (United States)

    Spears, Kenneth G.

    2002-03-01

    We have re-examined our earlier report of electron transfer in the [Co(Cp)_2|V(CO)_6] radical-pair using ultrafast infrared transient absorption spectroscopy in room temperature solutions. The radical-pair is created from the [Co(Cp)_2^+|V(CO)_6^-] ion-pair by ultrafast visible charge-transfer excitation. Transient absorption experiments with ps time constants. A small ET component with a 75 ps time constant is due to some separation and reformation of the radical-pairs. Transient absorption experiments monitoring the recovery of the ion-pair state shows that both fast components are due to ET rather than some other vibrational relaxation (VR) process in the radical state. We analyze the visible charge-transfer band and assign the two fast ET decay times to two ion-pair contact geometries with absorption origins different by about 1250 ± 350 cm-1. For excitation at 800 and 700 nm the 700 fs ET lifetime depends on the vibrational quantum state of the nontotally symmetric CO stretch in the V(CO)6 radical, where the lifetime decreases by 10% for the first vibrational quantum and 45% for the second quantum. There is no quantum effect for the second ion-pair geometry with a 5 ps ET lifetime. Standard ET rate models cannot explain the rate dependence upon vibrational quantum state for a nontotally symmetric vibration, and it may arise from a breakdown of the Condon approximation. We also report rates for IVR of CO stretching modes and for VR of low frequency vibrations. At excitation wavelengths of 620 and 555 nm there is sufficient internal vibrational energy in low-frequency vibrations to cause geometric inter-conversion between energetically similar Jahn-Teller geometries in the V(CO)6 radical. This process creates a 200 fs rise time for the V(CO)6 radical species to assume a stable geometry, which requires VR of low frequency vibrations to the solvent. These results demonstrate that earlier ET measurements from our group on the same molecule had insufficient time

  7. Comparison of 2,4-Dichlorophenoxyacetic Acid Degradation and Plasmid Transfer in Soil Resulting from Bioaugmentation with Two Different pJP4 Donors

    Science.gov (United States)

    Newby, D. T.; Gentry, T. J.; Pepper, I. L.

    2000-01-01

    A pilot field study was conducted to assess the impact of bioaugmentation with two plasmid pJP4-bearing microorganisms: the natural host, Ralstonia eutropha JMP134, and a laboratory-generated strain amenable to donor counterselection, Escherichia coli D11. The R. eutropha strain contained chromosomal genes necessary for mineralization of 2,4-dichlorophenoxyacetic acid (2,4-D), while the E. coli strain did not. The soil system was contaminated with 2,4-D alone or was cocontaminated with 2,4-D and Cd. Plasmid transfer to indigenous populations, plasmid persistence in soil, and degradation of 2,4-D were monitored over a 63-day period in the bioreactors. To assess the impact of contaminant reexposure, aliquots of bioreactor soil were reamended with additional 2,4-D. Both introduced donors remained culturable and transferred plasmid pJP4 to indigenous recipients, although to different extents. Isolated transconjugants were members of the Burkholderia and Ralstonia genera, suggesting multiple, if not successive, plasmid transfers. Upon a second exposure to 2,4-D, enhanced degradation was observed for all treatments, suggesting microbial adaptation to 2,4-D. Upon reexposure, degradation was most rapid for the E. coli D11-inoculated treatments. Cd did not significantly impact 2,4-D degradation or transconjugant formation. This study demonstrated that the choice of donor microorganism might be a key factor to consider for bioaugmentation efforts. In addition, the establishment of an array of stable indigenous plasmid hosts at sites with potential for reexposure or long-term contamination may be particularly useful. PMID:10919798

  8. Conjugal plasmid transfer in Streptomyces resembles bacterial chromosome segregation by FtsK/SpoIIIE

    OpenAIRE

    Vogelmann, Jutta; Ammelburg, Moritz; Finger, Constanze; Guezguez, Jamil; LINKE, Dirk; Flötenmeyer, Matthias; Stierhof, York-Dieter; Wohlleben, Wolfgang; Muth, Günther

    2011-01-01

    Most bacteria share virulence and resistance genes by transferring single-stranded DNA through a type IV secretion system. Streptomycetes, however, exchange dsDNA, using a system found to closely resemble machineries for prokaryotic chromosome segregation or DNA translocation during spore formation.

  9. 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.

  10. Plasmid-mediated transfer of CTX-M-55 extended-spectrum beta-lactamase among different strains of Salmonella and Shigella spp. in the Republic of Korea.

    Science.gov (United States)

    Kim, Jin Seok; Kim, Soojin; Park, Jungsun; Shin, Eunkyung; Yun, Young-Sun; Lee, Deog-Yong; Kwak, Hyo-Sun; Seong, Won Keun; Chung, Gyung Tae; Kim, Junyoung

    2017-09-01

    We screened 10 CTX-M-55-producing Shigella and Salmonella isolates from a national surveillance in Korea. The blaCTX-M-55 was located on the IncI1 (n=5), IncA/C (n=4) and IncZ (n=1) plasmids, downstream of ISEcp1, IS26-ISEcp1 and ISEcp1-IS5 sequences, respectively. These results indicate that CTX-M-55 has disseminated to other bacteria by lateral plasmid transfer. Copyright © 2017. Published by Elsevier Inc.

  11. Sequence-based analysis of pQBR103; a representative of a unique, transfer-proficient mega plasmid resident in the microbial community of sugar beet.

    Science.gov (United States)

    Tett, Adrian; Spiers, Andrew J; Crossman, Lisa C; Ager, Duane; Ciric, Lena; Dow, J Maxwell; Fry, John C; Harris, David; Lilley, Andrew; Oliver, Anna; Parkhill, Julian; Quail, Michael A; Rainey, Paul B; Saunders, Nigel J; Seeger, Kathy; Snyder, Lori A S; Squares, Rob; Thomas, Christopher M; Turner, Sarah L; Zhang, Xue-Xian; Field, Dawn; Bailey, Mark J

    2007-08-01

    The plasmid pQBR103 was found within Pseudomonas populations colonizing the leaf and root surfaces of sugar beet plants growing at Wytham, Oxfordshire, UK. At 425 kb it is the largest self-transmissible plasmid yet sequenced from the phytosphere. It is known to enhance the competitive fitness of its host, and parts of the plasmid are known to be actively transcribed in the plant environment. Analysis of the complete sequence of this plasmid predicts a coding sequence (CDS)-rich genome containing 478 CDSs and an exceptional degree of genetic novelty; 80% of predicted coding sequences cannot be ascribed a function and 60% are orphans. Of those to which function could be assigned, 40% bore greatest similarity to sequences from Pseudomonas spp, and the majority of the remainder showed similarity to other gamma-proteobacterial genera and plasmids. pQBR103 has identifiable regions presumed responsible for replication and partitioning, but despite being tra+ lacks the full complement of any previously described conjugal transfer functions. The DNA sequence provided few insights into the functional significance of plant-induced transcriptional regions, but suggests that 14% of CDSs may be expressed (11 CDSs with functional annotation and 54 without), further highlighting the ecological importance of these novel CDSs. Comparative analysis indicates that pQBR103 shares significant regions of sequence with other plasmids isolated from sugar beet plants grown at the same geographic location. These plasmid sequences indicate there is more novelty in the mobile DNA pool accessible to phytosphere pseudomonas than is currently appreciated or understood.

  12. Genetic environments of the transferable plasmid-mediated blaCTX-M-3 gene in Serratia marcescens isolates.

    Science.gov (United States)

    Chu, Pei-Yu; Peng, Chien-Fang

    2014-01-01

    In this study, genetic environments of the transferable plasmid-mediated blaCTX-M-3 gene were characterized among 14 isolates of cefotaxime-resistant Serratia marcescens using PCR and BLAST DNA sequence analysis. A total of 3 types of genetic architectures in the regions surrounding this blaCTX-M-3 gene were identified. Type I architecture was characterized by the presence of a complete insertion sequence of tnpA-ISEcp1, identified as interrupting a reverse IS26 sequence in the upstream region of the blaCTX-M-3 gene. A reverse-directional orf477 fragment was located downstream of the blaCTX-M-3 gene, which was in the same direction of the mucA gene. A common region containing the orf513 element was located upstream of the mucA gene. Moreover, a copy of the 3'-CS2 element was located immediately upstream of the orf513 element. A novel complex class 1 integron was characterized by the presence of the dfrA19 gene, which was flanked by two copies of class 1 integrons. This is the first report to describe the dfrA19 gene within a novel complex class 1 integron in S. marcescens isolates from Taiwan. This novel complex class 1 integron structure was located distantly upstream of the blaCTX-M-3 gene.

  13. 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

  14. Conditional Cash Transfers and School Dropout Rates

    Science.gov (United States)

    Dearden, Lorraine; Emmerson, Carl; Frayne, Christine; Meghir, Costas

    2009-01-01

    This paper evaluates a United Kingdom pilot study designed to test whether a means-tested conditional cash transfer paid to 16- to 18-year-olds for staying in full-time education is an effective way of reducing the proportion of school dropouts. The transfer's impact is substantial: In the first year, full-time education participation rates…

  15. Conditional Cash Transfers and School Dropout Rates

    Science.gov (United States)

    Dearden, Lorraine; Emmerson, Carl; Frayne, Christine; Meghir, Costas

    2009-01-01

    This paper evaluates a United Kingdom pilot study designed to test whether a means-tested conditional cash transfer paid to 16- to 18-year-olds for staying in full-time education is an effective way of reducing the proportion of school dropouts. The transfer's impact is substantial: In the first year, full-time education participation rates…

  16. In Vivo Acquisition of Carbapenemase Gene blaKPC-2 in Multiple Species of Enterobacteriaceae through Horizontal Transfer of Insertion Sequence or Plasmid

    Directory of Open Access Journals (Sweden)

    Baixing Ding

    2016-10-01

    Full Text Available Objectives: Current worldwide spread of carbapenem resistance in Enterobacteriaceae constitutes a critical public health threat. This study aims to investigate how carbapenem resistance is acquired in Enterobacteriaceae in patients during antimicrobial therapy. Methods: Clinical strains from the same anatomical site of the same patients that converted from carbapenem-susceptible to resistant during antimicrobial therapy and showed identical or similar PFGE patterns were identified. The similarly sized plasmids carried by the susceptible and resistant strains, the latter containing the carbapenemase genes, were sequenced and analyzed. Results: Paired strains were identified from four patients: three had neurosurgical conditions while the other had acute exacerbation of COPD. Two pairs of Klebsiella pneumoniae (KP1-S/R and KP2-S/R, S and R indicating susceptible and resistant strains, respectively, one pair of Morganella morganii (MM-S/R and one pair of Enterobacter aerogenes (EA-S/R were collected. All four carbapenem-resistant strains carried plasmids harboring blaKPC-2. Compared with the similarly sized plasmids in KP1-S and KP2-S, an insertion sequence that includes ISKpn6-like, blaKPC-2 and ISKpn8 was noted in pKP1-R and pKP2-R. Strains MM-R and EA-R had blaKPC-2-carrying plasmids not resembling plasmids in strains MM-S and EA-S suggesting their new acquisition while on therapy. Conclusions: Enterobacteriaceae can acquire carbapenem resistance during antimicrobial therapy through horizontal transfer of an insertion sequence or plasmid.

  17. In vivo Acquisition of Carbapenemase Gene blaKPC-2 in Multiple Species of Enterobacteriaceae through Horizontal Transfer of Insertion Sequence or Plasmid

    Science.gov (United States)

    Ding, Baixing; Shen, Zhen; Hu, Fupin; Ye, Meiping; Xu, Xiaogang; Guo, Qinglan; Wang, Minggui

    2016-01-01

    Objectives: Current worldwide spread of carbapenem resistance in Enterobacteriaceae constitutes a critical public health threat. This study aims to investigate how carbapenem resistance is acquired in Enterobacteriaceae in patients during antimicrobial therapy. Methods: Clinical strains from the same anatomical site of the same patients that converted from carbapenem-susceptible to resistant during antimicrobial therapy and showed identical or similar PFGE patterns were identified. The similarly sized plasmids carried by the susceptible and resistant strains, the latter containing the carbapenemase genes, were sequenced and analyzed. Results: Paired strains were identified from four patients: three had neurosurgical conditions while the other had acute exacerbation of COPD. Two pairs of Klebsiella pneumoniae (KP1-S/R and KP2-S/R, S and R indicating susceptible and resistant strains, respectively), one pair of Morganella morganii (MM-S/R) and one pair of Enterobacter aerogenes (EA-S/R) were collected. All four carbapenem-resistant strains carried plasmids harboring blaKPC−2. Compared with the similarly sized plasmids in KP1-S and KP2-S, an insertion sequence that includes ISKpn6-like, blaKPC−2 and ISKpn8 was noted in pKP1-R and pKP2-R. Strains MM-R and EA-R had blaKPC−2-carrying plasmids not resembling plasmids in strains MM-S and EA-S suggesting their new acquisition while on therapy. Conclusions: Enterobacteriaceae can acquire carbapenem resistance during antimicrobial therapy through horizontal transfer of an insertion sequence or plasmid. PMID:27818649

  18. The master regulator of IncA/C plasmids is recognized by the Salmonella Genomic island SGI1 as a signal for excision and conjugal transfer.

    Science.gov (United States)

    Kiss, János; Papp, Péter Pál; Szabó, Mónika; Farkas, Tibor; Murányi, Gábor; Szakállas, Erik; Olasz, Ferenc

    2015-10-15

    The genomic island SGI1 and its variants, the important vehicles of multi-resistance in Salmonella strains, are integrative elements mobilized exclusively by the conjugative IncA/C plasmids. Integration and excision of the island are carried out by the SGI1-encoded site-specific recombinase Int and the recombination directionality factor Xis. Chromosomal integration ensures the stable maintenance and vertical transmission of SGI1, while excision is the initial step of horizontal transfer, followed by conjugation and integration into the recipient. We report here that SGI1 not only exploits the conjugal apparatus of the IncA/C plasmids but also utilizes the regulatory mechanisms of the conjugation system for the exact timing and activation of excision to ensure efficient horizontal transfer. This study demonstrates that the FlhDC-family activator AcaCD, which regulates the conjugation machinery of the IncA/C plasmids, serves as a signal of helper entry through binding to SGI1 xis promoter and activating SGI1 excision. Promoters of int and xis genes have been identified and the binding site of the activator has been located by footprinting and deletion analyses. We prove that expression of xis is activator-dependent while int is constitutively expressed, and this regulatory mechanism is presumably responsible for the efficient transfer and stable maintenance of SGI1. © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

  19. Factors Differentiating Hospital Transfers from Long-term Care Facilities with High and Low Transfer Rates.

    Science.gov (United States)

    Teresi, Jeanne A.; And Others

    1991-01-01

    Examined differences among nursing homes in rates at which they transfer patients to hospitals. Data from nursing staff and charts on 286 most recent transfers from 10 nursing homes revealed that, from relatively equivalent patient populations, high-rate facilities transferred more chronically ill, physically frail patients and patients with…

  20. Transfer of a gonococcal beta-lactamase plasmid to conjugation-deficient Neisseria cinerea strains by transformation.

    Science.gov (United States)

    Genco, C A; Clark, V L

    1988-12-01

    We have previously shown that some strains of Neisseria cinerea can serve as recipients in conjugation (Con+) with Neisseria gonorrhoeae while others cannot (Con-). To determine if a replication defect contributes to the inability of certain strains of N. cinerea to serve as recipients in conjugation, we attempted to introduce a naturally occurring gonococcal beta-lactamase plasmid into N. cinerea by transformation. Various methods were employed, and all proved unsuccessful. Since specific sequences are required for DNA uptake in transformation of N. gonorrhoeae, we constructed a number of hybrid plasmids containing N. cinerea chromosomal DNA inserted into the N. gonorrhoeae/Escherichia coli beta-lactamase shuttle vector, pLES2. When nine randomly selected plasmids with inserts were used to transform an N. cinerea strain which did not accept the gonococcal beta-lactamase plasmid by conjugation, transformants were observed with four of the hybrid plasmids. The presence of one of the hybrid plasmids, pCAG9, in transformants was confirmed by agarose gel electrophoresis, Southern hybridization, and beta-lactamase production. When an N. gonorrhoeae donor strain containing pCAG9 was used in conjugation with several N. cinerea strains, only those strains that were previously shown to act as recipients could accept and maintain pCAG9. The ability of pCAG9 and the other three hybrid plasmids to transform Con- strains demonstrates that the beta-lactamase plasmid can replicate in Con- strains, and, therefore, the Con- phenotype is due to a block in some other stage of the conjugation process.

  1. A plasmid RK2-based broad-host-range cloning vector useful for transfer of metagenomic libraries to a variety of bacterial species.

    Science.gov (United States)

    Aakvik, Trine; Degnes, Kristin Fløgstad; Dahlsrud, Rannveig; Schmidt, Frank; Dam, Ragnar; Yu, Lihua; Völker, Uwe; Ellingsen, Trond Erling; Valla, Svein

    2009-06-01

    The majority of microorganisms in natural environments are difficult to cultivate, but their genes can be studied via metagenome libraries. To enhance the chances that these genes become expressed we here report the construction of a broad-host-range plasmid vector (pRS44) for fosmid and bacterial artificial chromosome (BAC) cloning. pRS44 can be efficiently transferred to numerous hosts by conjugation. It replicates in such hosts via the plasmid RK2 origin of replication, while in Escherichia coli it replicates via the plasmid F origin. The vector was found to be remarkably stable due to the insertion of an additional stability element (parDE). The copy number of pRS44 is adjustable, allowing for easy modifications of gene expression levels. A fosmid metagenomic library consisting of 20 000 clones and BAC clones with insert sizes up to 200 kb were constructed. The 16S rRNA gene analysis of the fosmid library DNA confirmed that it represents a variety of microbial species. The entire fosmid library and the selected BAC clones were transferred to Pseudomonas fluorescens and Xanthomonas campestris (fosmids only), and heterologous proteins from the fosmid library were confirmed to be expressed in P. fluorescens. To our knowledge no other reported vector system has a comparable potential for functional screening across species barriers.

  2. Large IncHI2-plasmids encode extended-spectrum β-lactamases (ESBLs) in Enterobacter spp. bloodstream isolates, and support ESBL-transfer to Escherichia coli.

    Science.gov (United States)

    Nilsen, E; Haldorsen, B C; Sundsfjord, A; Simonsen, G S; Ingebretsen, A; Naseer, U; Samuelsen, O

    2013-11-01

    We investigated the prevalence of extended-spectrum β-lactamases (ESBLs) in Enterobacter spp. bloodstream isolates from 19 hospital laboratories in Norway during 2011. A total of 62/230 (27%) isolates were resistant to third-generation cephalosporins and four (1.7%) were ESBL-positive; blaCTX -M-15 (n = 3) and blaSHV -12 (n = 1). This is comparable to the prevalence of ESBLs in clinical isolates of Escherichia coli and Klebsiella pneumoniae in Norway during the same period. All ESBL-positive isolates were multidrug resistant (MDR) and harboured plasmid-mediated quinolone resistance. Three isolates supported transfer of large IncHI2-plasmids harbouring ESBL- and MDR-encoding genes to E. coli recipients by in vitro conjugation.

  3. [Characterization of the transfer-related tra region of the conjugative plasmid p19 from a Bacillus subtilis soil strain].

    Science.gov (United States)

    Poluéktova, E U; Gagarina, E Iu; Nezametdinova, V Z; Shilovskiĭ, I P; Rodionova, S A; Prozorov, A A

    2010-01-01

    The nucleotide sequences of three DNA fragments (total size 30574 bp) of the plasmid p19 from the Bacillus subtilis 19 soil strain have been determined. Thirty open reading frames (ORFs) have been identified in these fragments. oriT of the plasmid has also been identified. As shown by the search for homologs of hypothetical protein products of these ORFs in databases, such homology exists for 18 ORFs. The protein products of nine ORFs can be assumed to have specific functions. Several ORFs were inactivated via insertional mutagenesis, and the conjugation capacity of the mutant plasmids was estimated. According to the data on homology of protein products and the results of ORF inactivation, regions of a total size of about 20 kb from the DNA fragments sequenced by us were inferred to belong to the tra region of p19. As follows from the analysis of the identified ORFs of the p19 tra region, it differs from the earlier described tra regions of other plasmids, irrespective of a certain similarity with the corresponding regions of plasmids of gram-positive bacteria from the genera Bacillus, Clostridium, and Listeria.

  4. Evidence for the horizontal transfer of an integrase gene from a fusellovirus to a pRN-like plasmid within a single strain of Sulfolobus and the implications for plasmid survival

    DEFF Research Database (Denmark)

    Peng, Xu

    2008-01-01

    seven ORFs, three of which encode an atypical RepA, a PlrA and a CopG protein. A fourth ORF exhibits a high nucleotide sequence identity to the SSV4 integrase gene, which suggests that it has been transferred to the plasmid from SSV4. A single point mutation within an otherwise identical 500 bp region...... infectivity. The virus and plasmid carry genomes of 15 135 and 6970 bp, respectively. For SSV4, 33 predicted ORFs are compactly organized with a strong preference for UGA stop codons, three-quarters of which overlap with either the Shine-Dalgarno motif or the start codon of the following gene. pXZ1 carries...... of the integrase gene occurs in the viral attachment site (attP), which corresponds to the anticodon region of the targeted tRNA gene in the host chromosome. This point mutation confers on pXZ1 the ability to integrate into the tRNA(Glu)[CUC] gene, which differs from the integration site of SSV4, t...

  5. Determination of Royalty Rates in the International Technology Transfer Contracts

    OpenAIRE

    Kapitsa, Yu.; Aralova, N.

    2015-01-01

    The existing approaches used in determination of the royalty rates for technology transfer contracts and based on the experience of research institutions of the National Academy of Sciences of Ukraine, research organizations and universities in Europe and USA were reviewed. The analysis of the existing rates has been made as well as recommendations on determination of the royalty rates for technology transfer contracts between research institutions and foreign and domestic partners have been ...

  6. Determination of Royalty Rates in the International Technology Transfer Contracts

    Directory of Open Access Journals (Sweden)

    Kapitsa, Yu.

    2015-03-01

    Full Text Available The existing approaches used in determination of the royalty rates for technology transfer contracts and based on the experience of research institutions of the National Academy of Sciences of Ukraine, research organizations and universities in Europe and USA were reviewed. The analysis of the existing rates has been made as well as recommendations on determination of the royalty rates for technology transfer contracts between research institutions and foreign and domestic partners have been worked out.

  7. Escherichia coli DNA helicase I catalyzes a sequence-specific cleavage/ligation reaction at the F plasmid origin of transfer.

    Science.gov (United States)

    Sherman, J A; Matson, S W

    1994-10-21

    Recent studies have shown that the Escherichia coli F plasmid-encoded traI gene product (TraIp), also known as DNA helicase I, catalyzes the formation of the site- and strand-specific nick that initiates F plasmid DNA transfer. Scission of the phosphodiester bond at the nic site within the origin of transfer (oriT) is accompanied by the covalent attachment of TraIp to the 5'-phosphate of the nicked DNA strand. This mechanism suggests that TraIp may also be capable of catalyzing a DNA ligation reaction using the energy stored in the protein-DNA intermediate. To test this possibility, an in vitro assay was designed that utilized short single-stranded DNA oligonucleotides of different lengths derived from the region within oriT that spanned the nic site. Purified TraIp was capable of efficiently cleaving single-stranded DNA that contained a nic site, and upon cleavage, the protein became covalently linked to the 5'-end of the nic site. When TraIp was incubated with two oligonucleotides of different length that contained the nic site, there was formation of novel recombinant products resulting from a TraIp-catalyzed cleavage/ligation reaction. Furthermore, the cleavage and ligation reactions were both sequence-specific. These data suggest that TraIp plays an important role in the initiation and termination of conjugative DNA transfer.

  8. Implementasi Manajemen Transfer Rate pada Proses HDFS Berbasis SDN

    Directory of Open Access Journals (Sweden)

    Narendra Hanif Wicaksana

    2017-01-01

    Full Text Available Pada kluster Hadoop perpindahan data akan sering terjadi, karena data yang disimpan akan tersebar ke dalam Datanode terutama pada saat melakukan proses penyimpanan ke dalam HDFS. Lalu lintas jaringan data mempengaruhi performa kinerja klaster Hadoop secara keseluruhan. Permasalahan ketersediaan bandwith dan juga congestion yang disebabkan lalu lintas data lain dapat mempengaruhi proses penyimpanan data ke dalam HDFS. SDN memiiliki fungsi untuk melakukan pengaturan manajemen transter rate sehingga dapat mengkategorikan lalu lintas data dan juga menyediakan nilai transfer rate dengan menggunkan mekanisme queue. Memanfaatkan arsitektur jaringan SDN, pada klaster Hadoop dilakukan manajemen transfer rate untuk dapat mengoptimalkan proses perpindahan data pada saat penyimpanan ke HDFS. Manajemen transfer rate dilakukan dengan cara memanfaatkan fitur queue pada switch OpenFlow. Tiap queue digunakan untuk mengkategorikan lalu lintas data pada jaringan klaster Hadoop. Nilai transfer rate untuk lalu lintas data HDFS dipisahkan dan diberikan nilai transfer rate yang lebih tinggi. Berdasarkan hasil uji coba dengan melakukan manajemen transfer rate waktu proses penyimpanan data ke HDFS tidak terpengaruh walaupun pada saat proses penyimpanan data terdapat lalu lintas data lain yang mengakibatkan congestion.

  9. 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

  10. 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.

  11. 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.

  12. Transfer component skill deficit rates among Veterans who use wheelchairs

    Directory of Open Access Journals (Sweden)

    Alicia M. Koontz, PhD

    2016-03-01

    Full Text Available The purpose of this study was to quantify the deficit rates for transfer component skills in a Veteran cohort and explore the relationship between deficit rates and subject characteristics. Seventy-four men and 18 women performed up to four transfers independently from their wheelchair to a mat table while a therapist evaluated their transfer techniques using the Transfer Assessment Instrument. The highest deficit rates concerned the improper use of handgrips (63%. Other common problems included not setting the wheelchair up at the proper angle (50% and not removing the armrest (58%. Veterans over 60 yr old and Veterans with moderate shoulder pain were more likely to set up their wheelchairs inappropriately than younger Veterans (p = 0.003 and Veterans with mild shoulder pain (p = 0.004. Women were less likely to remove their armrests than men (p = 0.03. Subjects with disabilities other than spinal cord injury were less inclined to set themselves up for a safe and easy transfer than the subjects with spinal cord injury (p transfer skills among Veterans and will inform the development of future transfer training programs both within and outside of the Department of Veterans Affairs.

  13. Conjugal transfer of the Sinorhizobium meliloti 1021 symbiotic plasmid is governed through the concerted action of one- and two-component signal transduction regulators.

    Science.gov (United States)

    Nogales, Joaquina; Blanca-Ordóñez, Helena; Olivares, José; Sanjuán, Juan

    2013-03-01

    Conjugal transfer of Sinorhizobium meliloti and Rhizobium etli symbiotic plasmids are repressed by the transcriptional regulator RctA. Here we report on new key players in the signal transduction cascade towards S. meliloti pSym conjugation. We have identified S. meliloti pSymA gene SMa0974 as an orthologue of the R. etli rctB gene which is required to antagonize repression by RctA. In S. meliloti two additional genes, rctR and rctC participate in control of rctB expression. rctR (SMa0955) encodes a protein of the GntR family of transcriptional regulators involved in repression of rctB. A rctR mutant promotes pSymA conjugal transfer and displays increased transcription of tra, virB and rctB genes even in presence of wild-type rctA gene. Among genes repressed by RctR, rctC (SMa0961) encodes a response regulator required to activate rctB transcription and therefore for derepression of plasmid conjugative functions. We conclude that in both R. etli and S. meliloti pSym conjugal transfer is derepressed via rctB, however the regulatory cascades to achieve activation of rctB are probably different. Upstream of rctB, the S. meliloti pSym conjugal transfer is regulated through the concerted action of genes representing one- (rctR) and two-component (rctC) signal transduction systems in response to yet unidentified signals.

  14. 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...

  15. Rates of mass, momentum, and energy transfer at the magnetopause

    Science.gov (United States)

    Hill, T. W.

    1979-01-01

    Empirical estimates of the global rates of transfer of solar wind mass, tangential momentum, and energy at the Earth's magnetopause are presented for comparison against model estimates based on the four principal mechanisms that have been proposed to explain such transfer. The comparisons, although not quite conclusive, strongly favor a model that incorporates some combination of direct magnetic connection and anomalous cross field diffusion. An additional global constraint, the rate at which magnetic flux is cycled through the magnetospheric convection system, strongly suggests that direct magnetic connection plays a significant if not dominant role in the solar wind/magnetosphere interaction.

  16. A convenient cancer vaccine therapy with in vivo transfer of interleukin 12 expression plasmid using gene gun technology after priming with irradiated carcinoma cells.

    Science.gov (United States)

    Nishitani, Masa-aki; Sakai, Tohru; Ishii, Kazunari; Zhang, Manxin; Nakano, Yoko; Nitta, Yoshio; Miyazaki, Jun-ichi; Kanayama, Hiro-omi; Kagawa, Susumu; Himeno, Kunisuke

    2002-02-01

    We studied interleukin (IL)-12 gene therapy using a gene gun as a new autologous vaccination strategy for cancer. In the first experiment, BALB/c mice were inoculated with syngeneic murine renal cancer cells (Renca) intradermally in the abdomen. This was followed by an injection of IL-12 expression plasmid using the gene gun. About 40% of the mice exhibited rejection of the tumor after the treatment and these mice also acquired immunological resistance against a secondary challenge with Renca cells. Based on these results, we examined whether antitumor activity can be potentiated when mice undergo combination treatment with intradermal inoculation of irradiated Renca cells and transfection with IL-12 gene. Inoculation of irradiated Renca cells alone was partially effective in inducing antitumor immunity, whereas the combined treatment remarkably intensified this effect. Moreover, this combined treatment inhibited tumor establishment and enhanced survival of the mice with tumor infiltration by CD4(+) and CD8(+) T cells, even when the treatment was started after tumor-implantation at a distant site. This antitumor effect was antigen specific and we confirmed the induction of antitumor cytotoxic T cells by this treatment. These results show that local cutaneous transfer of IL-12 expression plasmid using gene gun technology enhances systemic and specific antitumor immunity primed by irradiated tumor cells.

  17. 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.

  18. Testing Theories of Transfer Using Error Rate Learning Curves.

    Science.gov (United States)

    Koedinger, Kenneth R; Yudelson, Michael V; Pavlik, Philip I

    2016-07-01

    We analyze naturally occurring datasets from student use of educational technologies to explore a long-standing question of the scope of transfer of learning. We contrast a faculty theory of broad transfer with a component theory of more constrained transfer. To test these theories, we develop statistical models of them. These models use latent variables to represent mental functions that are changed while learning to cause a reduction in error rates for new tasks. Strong versions of these models provide a common explanation for the variance in task difficulty and transfer. Weak versions decouple difficulty and transfer explanations by describing task difficulty with parameters for each unique task. We evaluate these models in terms of both their prediction accuracy on held-out data and their power in explaining task difficulty and learning transfer. In comparisons across eight datasets, we find that the component models provide both better predictions and better explanations than the faculty models. Weak model variations tend to improve generalization across students, but hurt generalization across items and make a sacrifice to explanatory power. More generally, the approach could be used to identify malleable components of cognitive functions, such as spatial reasoning or executive functions. Copyright © 2016 Cognitive Science Society, Inc.

  19. 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

  20. Horizontal transfer of tet(M) and erm(B) resistance plasmids from food strains of Lactobacillus plantarum to Enterococcus faecalis JH2-2 in the gastrointestinal tract of gnotobiotic rats

    DEFF Research Database (Denmark)

    Jacobsen, Louise; Wilcks, Andrea; Hammer, Karin

    2007-01-01

    Two wild-type strains of Lactobacillus plantarum previously isolated from fermented dry sausages were analysed for their ability to transfer antibiotic resistance plasmids in the gastrointestinal tract. For this purpose, we used gnotobiotic rats as an in vivo model. Rats were initially inoculated...

  1. Theoretical Model of the Relationship between Single Embryo Transfer Rate and Multiple Pregnancy Rate in Japan

    OpenAIRE

    Syuichi Ooki

    2012-01-01

    The purpose of the present study was to examine the effect of single embryo transfer (SET) in assisted reproductive technology (ART) on the reduction of the multiple pregnancy rate. We also estimated the monozygotic (MZ) twinning rates according to the SET diffusion indirectly. A reverse sigmoid curve was assumed and examined using nationwide data of SET from 2007 to 2009 in Japan. The multiple pregnancy rate decreased almost linearly where the SET pregnancy rate was between about 40% and 80%...

  2. Horizontal transfer of tet(M) and erm(B) resistance plasmids from food strains of Lactobacillus plantarum to Enterococcus faecalis JH2-2 in the gastrointestinal tract of gnotobiotic rats

    DEFF Research Database (Denmark)

    Jacobsen, Louise; Wilcks, Andrea; Hammer, Karin;

    2007-01-01

    Two wild-type strains of Lactobacillus plantarum previously isolated from fermented dry sausages were analysed for their ability to transfer antibiotic resistance plasmids in the gastrointestinal tract. For this purpose, we used gnotobiotic rats as an in vivo model. Rats were initially inoculated...... with the recipient Enterococcus faecalis JH2-2 at a concentration of 10(10) CFU mL(-1). After a week, either of the two donors L. plantarum DG 522 (harbouring a tet(M)-containing plasmid of c. 40 kb) or L. plantarum DG 507 [harbouring a tet(M)-containing plasmid of c. 10 kb and an erm(B)-containing plasmid of c. 8......(2) CFU g(-1) faeces towards the end of the experiment. For erm(B)-TCs, the number was significantly higher and increased to c. 10(3) CFU g(-1) faeces. To our knowledge, this is the first study showing in vivo transfer of wild-type antibiotic resistance plasmids from L. plantarum to E. faecalis....

  3. 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

  4. [Electron transfer between globular proteins. Dependence of the rate of transfer on distance].

    Science.gov (United States)

    Lakhno, V D; Chuev, G N; Ustinin, M N; Komarov, V M

    1998-01-01

    Based on the assumption that electron transfer between globular proteins occurs by a collective excitation of polaron type, the dependence of the rate of this process on the distance between the donor and acceptor centers with regard to their detailed electron structure was calculated. The electron structure of the heme was calculated by the quantum-chemical MNDO-PM3 method. The results were compared with experimental data on interprotein and intraglobular electron transfer. It is shown that, in the framework of this model, the electron transfer is not exponential and does not require a particular transfer pathway since the whole protein macromolecule is involved in the formation of the electron excited state.

  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. mcr-1.2, a New mcr Variant Carried on a Transferable Plasmid from a Colistin-Resistant KPC Carbapenemase-Producing Klebsiella pneumoniae Strain of Sequence Type 512.

    Science.gov (United States)

    Di Pilato, Vincenzo; Arena, Fabio; Tascini, Carlo; Cannatelli, Antonio; Henrici De Angelis, Lucia; Fortunato, Simona; Giani, Tommaso; Menichetti, Francesco; Rossolini, Gian Maria

    2016-09-01

    A novel mcr variant, named mcr-1.2, encoding a Gln3-to-Leu functional variant of MCR-1, was detected in a KPC-3-producing ST512 Klebsiella pneumoniae isolate collected in Italy from a surveillance rectal swab from a leukemic child. The mcr-1.2 gene was carried on a transferable IncX4 plasmid whose structure was very similar to that of mcr-1-bearing plasmids previously found in Escherichia coli and K. pneumoniae strains from geographically distant sites (Estonia, China, and South Africa).

  7. Transfer of spontaneously hatching or hatched blastocyst yields better pregnancy rates than expanded blastocyst transfer

    Directory of Open Access Journals (Sweden)

    Natachandra M Chimote

    2013-01-01

    Full Text Available Context: Blastocyst stage embryo transfer (ET has become routine practice in recent years. However, probably due to limitations of assisted hatching techniques, expanded blastocyst transfer (EBT is still the preferred mode. Inexplicably, not much consideration has been given to spontaneously hatching/hatched blastocyst transfer (SHBT. Aim: This study aimed to investigate developmental potential of spontaneously hatching/hatched blastocyst against EBT in in vitro fertilization (IVF cycles. Settings and Design: Prospective study of 146 women undergoing their first IVF- ET cycle. SUBJECTS AND Methods: On the basis of blastocyst status, women were classified into SHBT and EBT groups. Intracytoplasmic sperm injection cycles were excluded to remove male factor bias. Implantation rate (IR, clinical pregnancy rate, and live birth rate were the main outcome measures. Statistical Analysis: Graph-pad Prism 5 statistical package. Results: SHBT group showed significantly higher blastocyst formation rate (53.3 ± 17.5 vs. 43.1 ± 14.5%, P = 0.0098, top-quality blastocysts (71.8 vs. 53.7%, P = 0.0436, IR (43.6 vs. 27.9%, P = 0.0408, pregnancy rate (59.4 vs. 45.1%, P = 0.0173, and live birth rate (36.8 vs. 22.8%, P = 0.003 compared to EBT group. Multiple pregnancy rates remained comparable between the two groups. Implantation correlated strongly with top-quality blastocysts (Pearson, r = 0.4441 in SHBT group, while the correlation was nonsignificant in EBT group. Conclusion: Extending culture of expanded blastocysts by a few hours to allow transfer of spontaneously hatching/hatched blastocysts gives higher implantation and pregnancy rates with no added risk of multiple gestations. Spontaneously hatching/hatched blastocysts have a better potential to implant and develop into a positive pregnancy.

  8. Sequence-based analysis of pQBR103; a representative of a unique, transfer-proficient mega plasmid resident in the microbial community of sugar beet

    OpenAIRE

    Tett, Adrian; Spiers, Andrew J.; Crossman, Lisa C.; Ager, Duane; Ciric, Lena; Dow, J. Maxwell; Fry, John C.; Harris,David; Lilley, Andrew; Oliver, Anna; Parkhill, Julian; Quail, Michael A; Paul. B. Rainey; Saunders, Nigel J.; Seeger, Kathy

    2007-01-01

    The plasmid pQBR103 was found within Pseudomonas populations colonizing the leaf and root surfaces of sugar beet plants growing at Wytham, Oxfordshire, UK. At 425 kb it is the largest self-transmissible plasmid yet sequenced from the phytosphere. It is known to enhance the competitive fitness of its host, and parts of the plasmid are known to be actively transcribed in the plant environment. Analysis of the complete sequence of this plasmid predicts a coding sequence (CDS)-rich genome contain...

  9. Electron transfer rates and equilibrium within cytochrome c oxidase

    DEFF Research Database (Denmark)

    Farver, O; Einarsdóttir, O; Pecht, I

    2000-01-01

    identical within experimental error and independent of the enzyme concentration. This demonstrates that a fast intramolecular electron equilibration is taking place between CuA and heme a. The rate constants for CuA --> heme a ET and the reverse (heme a --> CuA) process were found to be 13 000 s-1 and 3700......Intramolecular electron transfer (ET) between the CuA center and heme a in bovine cytochrome c oxidase was investigated by pulse radiolysis. CuA, the initial electron acceptor, was reduced by 1-methyl nicotinamide radicals in a diffusion-controlled reaction, as monitored by absorption changes...... at 830 nm. After the initial reduction phase, the 830 nm absorption was partially restored, corresponding to reoxidation of the CuA center. Concomitantly, the absorption at 445 nm and 605 nm increased, indicating reduction of heme a. The rate constants for heme a reduction and CuA reoxidation were...

  10. Effect of pheromone induction on transfer of the Enterococcus faecalis plasmid pCF10 in intestinal mucus ex vivo

    DEFF Research Database (Denmark)

    Licht, Tine Rask; Hammerum, Anette Marie; Jensen, Lars Bogø;

    2001-01-01

    The effect of synthetic sex pheromone on pheromone-inducible conjugation between the isogenic Enterococcus faecalis strains OG1RF and OG1SS was investigated in (i) Todd-Hewitt broth medium and (ii) intestinal mucus isolated from germ-free rats. In broth, the presence of synthetic pheromone cCF10...... in the experiment. We suggest that due to differences in diffusion rates and medium-binding of the pheromones, the effect of the synthetic cCF10 was immediately dominated by the effect of pheromones produced by the recipient E. faecalis strain in broth, while this happened later in mucus....

  11. Transfer entropy analysis of maternal and fetal heart rate coupling.

    Science.gov (United States)

    Marzbanrad, Faezeh; Kimura, Yoshitaka; Endo, Miyuki; Palaniswami, Marimuthu; Khandoker, Ahsan H

    2015-01-01

    Although evidence of the short term relationship between maternal and fetal heart rates has been found in previous model-based studies, knowledge about the mechanism and patterns of the coupling during gestation is still limited. In this study, a model-free method based on Transfer Entropy (TE) was applied to quantify the maternal-fetal heart rate couplings in both directions. Furthermore, analysis of the lag at which TE was maximum and its changes throughout gestation, provided more information about the mechanism of coupling and its latency. Experimental results based on fetal electrocardiograms (fECGs) and maternal ECG showed the evidence of coupling for 62 out of 65 healthy mothers and fetuses in each direction, by statistically validating against the surrogate pairs. The fetuses were divided into three gestational age groups: early (16-25 weeks), mid (26-31 weeks) and late (32-41 weeks) gestation. The maximum TE from maternal to fetal heart rate significantly increased from early to mid gestation, while the coupling delay on both directions decreased significantly from mid to late gestation. These changes occur concomitant with the maturation of the fetal sensory and autonomic nervous systems with advancing gestational age. In conclusion, the application of TE with delays revealed detailed information about the changes in fetal-maternal heart rate coupling strength and latency throughout gestation, which could provide novel clinical markers of fetal development and well-being.

  12. Frequency of conjugative transfer of plasmid-encoded ISEcp1 - blaCTX-M-15 and aac(6'-lb-cr genes in Enterobacteriaceae at a tertiary care center in Lebanon - role of transferases

    Directory of Open Access Journals (Sweden)

    Araj George F

    2010-07-01

    Full Text Available Abstract Background The frequency of transfer of genes encoding resistance to antimicrobial agents was determined by conjugation in ESBL-producing and/or fluoroquinolone or aminoglycoside resistant Enterobacteriaceae clinical isolates at a tertiary care center in Lebanon. In addition, the role of tra genes encoding transferases in mediating conjugation was assessed. Methods Conjugation experiments were done on 53 ESBL-producing and/or fluoroquinolone resistant E. coli and K. pneumoniae and ESBL-producing S. sonnei isolates. Antimicrobial susceptibility testing on parent and transconjugant isolates, and PCR amplifications on plasmid extracts of the resistance-encoding genes: blaCTX-M-15 with the ISEcp1 insertion sequence, the aac(6'-lb-cr and qnrS genes, as well as tra encoding transferases genes were done. Random amplified polymorphic DNA (RAPD analysis was performed to demonstrate whether conjugative isolates are clonal and whether they are linked epidemiologically to a particular source. Results Antimicrobial susceptibility testing on transconjugants revealed that 26 out of 53 (49% ESBL-producing Enterobacteriaceae were able to transfer antimicrobial resistance to the recipients. Transfer of high-level resistance to the transconjugants encoded by the blaCTX-M-15 gene downstream the ISEcp1 insertion sequence against 3rd generation cephalosporins, and of low-level resistance against ciprofloxacin, and variable levels of resistance against aminoglycosides encoded by aac(6'-lb-cr gene, were observed in transconjugants. tra encoding transferase genes were detected exclusively in conjugative isolates. Conclusion In conclusion, the frequency of transfer of antimicrobial resistance in non clonal Enterobacteriaceae at the tertiary care center by conjugation was 49%. Conjugation occurred in isolates expressing the tra encoding transferase genes. Multiple conjugative strains harboring the plasmid encoded antimicrobial resistant genes were circulating in

  13. Genetic and functional characterization of a yet-unclassified rhizobial Dtr (DNA-transfer-and-replication) region from a ubiquitous plasmid conjugal system present in Sinorhizobium meliloti, in Sinorhizobium medicae, and in other nonrhizobial Gram-negative bacteria.

    Science.gov (United States)

    Giusti, María de los Ángeles; Pistorio, Mariano; Lozano, Mauricio J; Tejerizo, Gonzalo A Torres; Salas, María Eugenia; Martini, María Carla; López, José Luis; Draghi, Walter O; Del Papa, María Florencia; Pérez-Mendoza, Daniel; Sanjuán, Juan; Lagares, Antonio

    2012-05-01

    Rhizobia are Gram-negative bacteria that live in soils and associate with leguminous plants to establish nitrogen-fixing symbioses. The ability of these bacteria to undergo horizontal gene transfer (HGT) is thought to be one of the main features to explain both the origin of their symbiotic life-style and the plasticity and dynamics of their genomes. In our laboratory we have previously characterized at the species level the non-pSym plasmid mobilome in Sinorhizobium meliloti, the symbiont of Medicago spp., and have found a high incidence of conjugal activity in many plasmids (Pistorio et al., 2008). In this work we characterized the Dtr (DNA-transfer-and-replication) region of one of those plasmids, pSmeLPU88b. This mobilization region was found to represent a previously unclassified Dtr type in rhizobia (hereafter type-IV), highly ubiquitous in S. meliloti and found in other genera of Gram-negative bacteria as well; including Agrobacterium, Ochrobactrum, and Chelativorans. The oriT of the type-IV Dtr described here could be located by function within a DNA fragment of 278 bp, between the divergent genes parA and mobC. The phylogenetic analysis of the cognate relaxase MobZ indicated that this protein groups close to the previously defined MOB(P3) and MOB(P4) type of enzymes, but is located in a separate and novel cluster that we have designated MOB(P0). Noteworthy, MOB(P0) and MOB(P4) relaxases were frequently associated with plasmids present in rhizospheric soil bacteria. A comparison of the nod-gene locations with the phylogenetic topology of the rhizobial relaxases revealed that the symbiotic genes are found on diverse plasmids bearing any of the four Dtr types, thus indicating that pSym plasmids are not specifically associated with any particular mobilization system. Finally, we demonstrated that the type-IV Dtr promoted the mobilization of plasmids from S. meliloti to Sinorhizobium medicae as well as from these rhizobia to other bacteria by means of their own

  14. The qacC Gene Has Recently Spread between Rolling Circle Plasmids of Staphylococcus, Indicative of a Novel Gene Transfer Mechanism

    DEFF Research Database (Denmark)

    Wassenaar, Trudy M; Ussery, David W; Ingmer, Hanne

    2016-01-01

    identity of these plasmids compared to the strict nucleotide conservation of their qacC means that this gene has recently spread. In the absence of insertion sequences or other genetic elements explaining the mobility, we sought for an explanation of mobilization by sequence comparison. Publically...... available sequences of qac genes, their flanking genes and the replication gene that is invariably present in RC-plasmids were compared to reconstruct the evolutionary history of these plasmids and to explain the recent spread of qacC. Here we propose a new model that explains how qacC is mobilized...... in RC-plasmids, which has also been employed by other genes, such as lnuA (conferring lincomycin resistance). The proposed gene mobility has aided to the wide spread of clinically relevant resistance genes in Staphylococcus populations....

  15. 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 ...

  16. 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

  17. 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...

  18. Impacts of organic carbon availability and recipient bacteria characteristics on the potential for TOL plasmid genetic bioaugmentation in soil slurries.

    Science.gov (United States)

    Ikuma, Kaoru; Holzem, Ryan M; Gunsch, Claudia K

    2012-09-01

    The effectiveness of genetic bioaugmentation relies on efficient plasmid transfer between donor and recipient cells as well as the plasmid's phenotype in the recipient cell. In the present study, the effects of varying organic carbon substrates, initial recipient-to-donor cell density ratios, and mixtures of known recipient bacterial strains on the conjugation and function of a TOL plasmid were tested in sterile soil slurry batch reactors. The presence of soil organic carbon was sufficient in ensuring TOL plasmid transconjugant occurrence (up to 2.1±0.5%) for most recipient strains in soil slurry batch mating experiments. The addition of glucose had limited effects on transconjugant occurrence; however, glucose amendment increased the specific toluene degradation rates of some Enterobacteriaceae transconjugants in soil slurry. Initial cell density ratios and mixtures of recipient strains had smaller impacts on plasmid conjugation and resulting phenotype functionality. These observations suggest that genetic bioaugmentation may be improved by minimal altering of environmental conditions.

  19. Transfer of Wild-Type Plasmids Harbouring Tetracycline or Erythromycin Resistance Genes from Native Strains of Lactobacillus plantarum to other Bacteria in a Gastrointestinal Environment

    DEFF Research Database (Denmark)

    Feld, Louise

    2008-01-01

    Overførsel af oprindelige plasmider indeholdende tetracyklin eller erythromycin resistensgener fra naturligt forekommende Lactobacillus plantarum stammer til andre bakterier i et tarmmiljø Denne rapport omhandler overførsel af plasmider vha. konjugation fra naturlige laktobaciller til andre...... fokus på tetracyklin og erythromycin samt på laktobaciller, konjugation og rolling-circle replikerende plasmider, idet de praktiske studier specielt har koncentreret sig om dette. 6 Manuskript I, havde til formål at teste to naturligt forekommende Lactobacillus plantarum stammers (DG 507 og DG 522....... Det blev vist at pLFE1 har et bredt værtsspektrum, idet der blev observeret transkonjuganter fra mating mellem donorstammen L. plantarum M345 og recipienter af Lactobacillus rhamnosus, Lactococcus lactis, Listeria innocua, E. faecalis og Listeria monocytogenes....

  20. 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...

  1. Elective transfer of two embryos: Reduction of multiple gestations while maintaining high pregnancy rates

    OpenAIRE

    Dowling-Lacey, Donna; Jones, Estella; Mayer, Jacob; Bocca, Silvina; Stadtmauer, Laurel; Oehninger, Sergio

    2006-01-01

    Purpose: To determine if the elective transfer of two embryos reduced the incidence of multiple gestations while maintaining high pregnancy rates. Methods: IVF patients and recipients of oocyte donation with an elective day-3 transfer of 2 or 3 embryos were studied. Result(s): In IVF, the elective transfer of 2 embryos resulted in similar pregnancy rate but significantly reduced the overall incidence of multiple gestations (20% versus 39%) when compared to the elective transfer of 3 embryos. ...

  2. Type 3 fimbriae, encoded by the conjugative plasmid pOLA52, enhance biofilm formation and transfer frequencies in Enterobacteriaceae strains

    DEFF Research Database (Denmark)

    Burmølle, Mette; Bahl, Martin Iain; Jensen, Lars Bogø

    2008-01-01

    biofilm-deficient mutants, which all mapped within a putative operon with high homology to the mrkABCDF operon of Klebsiella pneumoniae, where these genes are responsible for type 3 fimbriae expression, attachment to surfaces and biofilm formation. Biofilm formation in microtitre plates and in urinary......, resulting in enhanced conjugation frequencies and biofilm formation of the plasmid-harbouring strain....

  3. The qacC Gene Has Recently Spread between Rolling Circle Plasmids of Staphylococcus, Indicative of a Novel Gene Transfer Mechanism

    DEFF Research Database (Denmark)

    Wassenaar, Trudy M; Ussery, David W; Ingmer, Hanne

    2016-01-01

    identity of these plasmids compared to the strict nucleotide conservation of their qacC means that this gene has recently spread. In the absence of insertion sequences or other genetic elements explaining the mobility, we sought for an explanation of mobilization by sequence comparison. Publically...

  4. Measurements of file transfer rates over dedicated long-haul connections

    Energy Technology Data Exchange (ETDEWEB)

    Rao, Nageswara S [ORNL; Settlemyer, Bradley W [ORNL; Imam, Neena [ORNL; Hinkel, Gregory Carl [ORNL

    2016-01-01

    Wide-area file transfers are an integral part of several High-Performance Computing (HPC) scenarios. Dedicated network connections with high capacity, low loss rate and low competing traffic, are increasingly being provisioned over current HPC infrastructures to support such transfers. To gain insights into these file transfers, we collected transfer rate measurements for Lustre and xfs file systems between dedicated multi-core servers over emulated 10 Gbps connections with round trip times (rtt) in 0-366 ms range. Memory transfer throughput over these connections is measured using iperf, and file IO throughput on host systems is measured using xddprof. We consider two file system configurations: Lustre over IB network and xfs over SSD connected to PCI bus. Files are transferred using xdd across these connections, and the transfer rates are measured, which indicate the need to jointly optimize the connection and host file IO parameters to achieve peak transfer rates. In particular, these measurements indicate that (i) peak file transfer rate is lower than peak connection and host IO throughput, in some cases by as much as 50% or lower, (ii) xdd request sizes that achieve peak throughput for host file IO do not necessarily lead to peak file transfer rates, and (iii) parallelism in host IO and TCP transport does not always improve the file transfer rates.

  5. Chemotherapy of Bacterial Plasmids

    Science.gov (United States)

    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

  6. 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 ...

  7. Engineered electron-transfer chain in photosystem 1 based photocathodes outperforms electron-transfer rates in natural photosynthesis.

    Science.gov (United States)

    Kothe, Tim; Pöller, Sascha; Zhao, Fangyuan; Fortgang, Philippe; Rögner, Matthias; Schuhmann, Wolfgang; Plumeré, Nicolas

    2014-08-25

    Photosystem 1 (PS1) triggers the most energetic light-induced charge-separation step in nature and the in vivo electron-transfer rates approach 50 e(-)  s(-1)  PS1(-1). Photoelectrochemical devices based on this building block have to date underperformed with respect to their semiconductor counterparts or to natural photosynthesis in terms of electron-transfer rates. We present a rational design of a redox hydrogel film to contact PS1 to an electrode for photocurrent generation. We exploit the pH-dependent properties of a poly(vinyl)imidazole Os(bispyridine)2Cl polymer to tune the redox hydrogel film for maximum electron-transfer rates under optimal conditions for PS1 activity. The PS1-containing redox hydrogel film displays electron-transfer rates of up to 335±14 e(-)  s(-1)  PS1(-1), which considerably exceeds the rates observed in natural photosynthesis or in other semiartificial systems. Under O2 supersaturation, photocurrents of 322±19 μA cm(-2) were achieved. The photocurrents are only limited by mass transport of the terminal electron acceptor (O2). This implies that even higher electron-transfer rates may be achieved with PS1-based systems in general. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Spread of CTX-M-15 extended-spectrum β-lactamase-producing Escherichia coli isolates through household contact and plasmid transfer.

    Science.gov (United States)

    Kojima, Yasuko; Harada, Sohei; Aoki, Kotaro; Ishii, Yoshikazu; Sawa, Tomoka; Hasegawa, Kei; Saji, Tsutomu; Yamaguchi, Keizo; Tateda, Kazuhiro

    2014-05-01

    We document the household spread of extended-spectrum β-lactamase-producing Escherichia coli. One isolate belonged to sequence type 1193 and caused urinary tract infection in a 4-month-old female, and the other isolate belonged to sequence type 131 and colonized three family members, including the index patient. These isolates carried similar Inc-I1-Iγ plasmids, harboring blaCTX-M-15.

  9. Spread of CTX-M-15 Extended-Spectrum β-Lactamase-Producing Escherichia coli Isolates through Household Contact and Plasmid Transfer

    OpenAIRE

    Kojima, Yasuko; Harada, Sohei; Aoki, Kotaro; ISHII, Yoshikazu; Sawa, Tomoka; Hasegawa, Kei; Saji, Tsutomu; Yamaguchi, Keizo; Tateda, Kazuhiro

    2014-01-01

    We document the household spread of extended-spectrum β-lactamase-producing Escherichia coli. One isolate belonged to sequence type 1193 and caused urinary tract infection in a 4-month-old female, and the other isolate belonged to sequence type 131 and colonized three family members, including the index patient. These isolates carried similar Inc-I1-Iγ plasmids, harboring blaCTX-M-15.

  10. Acquisition through horizontal gene transfer of plasmid pSMA198 by Streptococcus macedonicus ACA-DC 198 points towards the dairy origin of the species.

    Directory of Open Access Journals (Sweden)

    Konstantinos Papadimitriou

    Full Text Available Streptococcus macedonicus is an intriguing streptococcal species whose most frequent source of isolation is fermented foods similarly to Streptococcus thermophilus. However, S. macedonicus is closely related to commensal opportunistic pathogens of the Streptococcus bovis/Streptococcus equinus complex.We analyzed the pSMA198 plasmid isolated from the dairy strain Streptococcus macedonicus ACA-DC 198 in order to provide novel clues about the main ecological niche of this bacterium. pSMA198 belongs to the narrow host range pCI305/pWV02 family found primarily in lactococci and to the best of our knowledge it is the first such plasmid to be reported in streptococci. Comparative analysis of the pSMA198 sequence revealed a high degree of similarity with plasmids isolated from Lactococcus lactis strains deriving from milk or its products. Phylogenetic analysis of the pSMA198 Rep showed that the vast majority of closely related proteins derive from lactococcal dairy isolates. Additionally, cloning of the pSMA198 ori in L. lactis revealed a 100% stability of replication over 100 generations. Both pSMA198 and the chromosome of S. macedonicus exhibit a high percentage of potential pseudogenes, indicating that they have co-evolved under the same gene decay processes. We identified chromosomal regions in S. macedonicus that may have originated from pSMA198, also supporting a long co-existence of the two replicons. pSMA198 was also found in divergent biotypes of S. macedonicus and in strains isolated from dispersed geographic locations (e.g. Greece and Switzerland showing that pSMA198's acquisition is not a recent event.Here we propose that S. macedonicus acquired plasmid pSMA198 from L. lactis via an ancestral genetic exchange event that took place most probably in milk or dairy products. We provide important evidence that point towards the dairy origin of this species.

  11. Why close a bacterial genome? The plasmid of Alteromonas macleodii HOT1A3 is a vector for inter-specific transfer of a flexible genomic island

    Directory of Open Access Journals (Sweden)

    Eduard eFadeev

    2016-03-01

    Full Text Available Genome sequencing is rapidly becoming a staple technique in environmental and clinical microbiology, yet computational challenges still remain, leading to many draft genomes which are typically fragmented into many contigs. We sequenced and completely assembled the genome of a marine heterotrophic bacterium, Alteromonas macleodii HOT1A3, and compared its full genome to several draft genomes obtained using different reference-based and de-novo methods. In general, the de-novo assemblies clearly outperformed the reference-based or hybrid ones, covering>99% of the genes and representing essentially all of the gene functions. However, only the fully closed genome (~4.5Mbp allowed us to identify the presence of a large, 148 kbp plasmid, pAM1A3. While HOT1A3 belongs to Alteromonas macleodii, typically found in surface waters (surface ecotype, this plasmid consists of an almost complete flexible genomic island, containing many genes involved in metal resistance previously identified in the genomes of Alteromonas mediterranea (deep ecotype. Indeed, similar to A. mediterranea, A. macleodii HOT1A3 grows at concentrations of zinc, mercury and copper that are inhibitory for other A. macleodii strains. The presence of a plasmid encoding almost an entire flexible genomic island suggests that wholesale genomic exchange between heterotrophic marine bacteria belonging to related but ecologically different populations is not uncommon.

  12. 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.

  13. Role of Rom protein in copy number control of plasmid pBR322 at different growth rates in Escherichia coli K-12

    DEFF Research Database (Denmark)

    Tove, Atlung; Christensen, Bjarke Bak; Hansen, Flemming G.

    1999-01-01

    The copy number per cell mass of plasmid pBR322 and a rom(-) derivative was measured as a function of generation time. In fast growing cells the copy number per cell mass was virtually identical for rom(+) and rom(-) derivatives. However, the copy number of pBR322 only increased 3- to 4-fold from...... a 20- to 80-min generation time, whereas the copy number of the torn derivative increased 7- to 10-fold. The copy number stayed constant for the rom(+) and rom(-) plasmids at generation times longer than 80-100 min. Thus, the presence of the rom gene decreased the copy number of plasmid pBR322...... effect on the copy number of pBR322 rom(-) at fast growth, but it decreased its copy number at slow growth to the same level as found for pBR322, i.e., complemented the pBR322 rom(-) plasmid. The pACYC184 plasmid and its rom(+) derivatives showed copy numbers similar to those of pBR322 rom(-) and pBR322...

  14. Ectopic pregnancy rates with day 3 versus day 5 embryo transfer: a retrospective analysis

    OpenAIRE

    Jun Sunny H; Milki Amin A

    2003-01-01

    Abstract Background Blastocyst transfer may theoretically decrease the incidence of ectopic pregnancy following IVF-ET in view of the decreased uterine contractility reported on day 5. The purpose of our study is to specifically compare the tubal pregnancy rates between day 3 and day 5 transfers. Methods A retrospective analysis of all clinical pregnancies conceived in our IVF program since 1998 was performed. The ectopic pregnancy rates were compared for day 3 and day 5 transfers. Results Th...

  15. 78 FR 13661 - Energy Transfer Fuel, LP; Notice of Petition for Rate Approval

    Science.gov (United States)

    2013-02-28

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF ENERGY Federal Energy Regulatory Commission Energy Transfer Fuel, LP; Notice of Petition for Rate Approval Take notice that on February 15, 2013, Energy Transfer Fuel, LP filed for approval of rates for...

  16. Transfer Rate Edited experiment for the selective detection of Chemical Exchange via Saturation Transfer (TRE-CEST).

    Science.gov (United States)

    Friedman, Joshua I; Xia, Ding; Regatte, Ravinder R; Jerschow, Alexej

    2015-07-01

    Chemical Exchange Saturation Transfer (CEST) magnetic resonance experiments have become valuable tools in magnetic resonance for the detection of low concentration solutes with far greater sensitivity than direct detection methods. Accurate measures of rates of chemical exchange provided by CEST are of particular interest to biomedical imaging communities where variations in chemical exchange can be related to subtle variations in biomarker concentration, temperature and pH within tissues using MRI. Despite their name, however, traditional CEST methods are not truly selective for chemical exchange and instead detect all forms of magnetization transfer including through-space NOE. This ambiguity crowds CEST spectra and greatly complicates subsequent data analysis. We have developed a Transfer Rate Edited CEST experiment (TRE-CEST) that uses two different types of solute labeling in order to selectively amplify signals of rapidly exchanging proton species while simultaneously suppressing 'slower' NOE-dominated magnetization transfer processes. This approach is demonstrated in the context of both NMR and MRI, where it is used to detect the labile amide protons of proteins undergoing chemical exchange (at rates⩾30s(-1)) while simultaneously eliminating signals originating from slower (∼5s(-1)) NOE-mediated magnetization transfer processes. TRE-CEST greatly expands the utility of CEST experiments in complex systems, and in-vivo, in particular, where it is expected to improve the quantification of chemical exchange and magnetization transfer rates while enabling new forms of imaging contrast.

  17. Geometry effect on energy transfer rate in a coupled-quantum-well structure: nonlinear regime

    Science.gov (United States)

    Salavati-fard, T.; Vazifehshenas, T.

    2014-12-01

    We study theoretically the effect of geometry on the energy transfer rate at nonlinear regime in a coupled-quantum-well system using the balance equation approach. To investigate comparatively the effect of both symmetric and asymmetric geometry, different structures are considered. The random phase approximation dynamic dielectric function is employed to include the contributions from both quasiparticle and plasmon excitations. Also, the short-range exchange interaction is taken into account through the Hubbard approximation. Our numerical results show that the energy transfer rate increases by increasing the well thicknesses in symmetric structures. Furthermore, by increasing spatial asymmetry, the energy transfer rate decreases for the electron temperature range of interest. From numerical calculations, it is obtained that the nonlinear energy transfer rate is proportional to the square of electron drift velocity in all structures and also, found that the influence of Hubbard local field correction on the energy transfer rate gets weaker by increasing the strength of applied electric field.

  18. [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.

  19. Acquisition through Horizontal Gene Transfer of Plasmid pSMA198 by Streptococcus macedonicus ACA-DC 198 Points towards the Dairy Origin of the Species

    Science.gov (United States)

    Papadimitriou, Konstantinos; Anastasiou, Rania; Maistrou, Eleni; Plakas, Thomas; Papandreou, Nikos C.; Hamodrakas, Stavros J.; Ferreira, Stéphanie; Supply, Philip; Renault, Pierre; Pot, Bruno; Tsakalidou, Effie

    2015-01-01

    Background Streptococcus macedonicus is an intriguing streptococcal species whose most frequent source of isolation is fermented foods similarly to Streptococcus thermophilus. However, S. macedonicus is closely related to commensal opportunistic pathogens of the Streptococcus bovis/Streptococcus equinus complex. Methodology/Principal Findings We analyzed the pSMA198 plasmid isolated from the dairy strain Streptococcus macedonicus ACA-DC 198 in order to provide novel clues about the main ecological niche of this bacterium. pSMA198 belongs to the narrow host range pCI305/pWV02 family found primarily in lactococci and to the best of our knowledge it is the first such plasmid to be reported in streptococci. Comparative analysis of the pSMA198 sequence revealed a high degree of similarity with plasmids isolated from Lactococcus lactis strains deriving from milk or its products. Phylogenetic analysis of the pSMA198 Rep showed that the vast majority of closely related proteins derive from lactococcal dairy isolates. Additionally, cloning of the pSMA198 ori in L. lactis revealed a 100% stability of replication over 100 generations. Both pSMA198 and the chromosome of S. macedonicus exhibit a high percentage of potential pseudogenes, indicating that they have co-evolved under the same gene decay processes. We identified chromosomal regions in S. macedonicus that may have originated from pSMA198, also supporting a long co-existence of the two replicons. pSMA198 was also found in divergent biotypes of S. macedonicus and in strains isolated from dispersed geographic locations (e.g. Greece and Switzerland) showing that pSMA198’s acquisition is not a recent event. Conclusions/Significance Here we propose that S. macedonicus acquired plasmid pSMA198 from L. lactis via an ancestral genetic exchange event that took place most probably in milk or dairy products. We provide important evidence that point towards the dairy origin of this species. PMID:25584532

  20. Persistence of a pKPN3-like CTX-M-15-encoding IncFIIK plasmid in a Klebsiella pneumonia ST17 host during two years of intestinal colonization.

    Directory of Open Access Journals (Sweden)

    Iren Høyland Löhr

    Full Text Available To characterize the CTX-M-15-encoding plasmid in a Klebsiella pneumoniae ST17 strain, responsible for an outbreak at a Norwegian neonatal intensive care unit and subsequent colonization of affected children for up to two years. To identify plasmid-mediated features relevant for the outbreak dynamics, and to investigate the plasmids capability of horizontal transfer, its segregational stability and plasmid-mediated fitness costs.Plasmid profiling was performed by S1-nuclease PFGE, PCR-based replicon typing and Southern blot-hybridization. The complete sequence of the CTX-M-15-encoding plasmid was obtained by 454 sequencing. Plasmid self-transferability was investigated by broth- and filter mating, segregational stability was explored by serial passage, and plasmid-conferred fitness costs were examined in pairwise head-to-head competitions and by growth rate comparisons.CTX-M-15 was encoded by a ~180 kb IncFIIK plasmid in K. pneumoniae ST17. S1-nuclease PFGE profiles of the first and the last CTX-M-15-producing K. pneumoniae isolates, recovered from the four children colonized the longest, suggested that the plasmid was stably maintained during intestinal carriage of up to two years. The DNA sequence of the pKPN3-like plasmid, pKp848CTX, uncovered a Tn3-like antibiotic resistance region and multiple heavy metal- and thermoresistance determinants. Plasmid pKp848CTX could not be transferred to Escherichia coli in vitro and we found no evidence to support horizontal plasmid transfer in vivo. Segregational plasmid loss ranging from 0.83% to 17.5% was demonstrated in evolved populations in vitro, but only minor fitness costs were associated with plasmid-carriage.Plasmid pKp848CTX encodes phenotypic traits, which may have had an impact on the fitness and survival of the K. pneumoniae ST17 strain in the outbreak setting. The antibiotic resistance plasmid pKp848CTX was stably maintained during two years of intestinal colonization, conferring negligible

  1. Factors affecting conception rates in cattle following embryo transfer

    African Journals Online (AJOL)

    ACSS

    ... such as lack of trained personnel and equipment for ovulation and packaging ... Temperatures range from 10 to 270 C, .... conception rates are within the range of ... efficiency of heat detection, early .... Faculty of Veterinary Medicine, Debre.

  2. Absorption and desorption mass transfer rates in chemically enhanced reactive systems. Part II : Reverse kinetic rate parameters

    NARCIS (Netherlands)

    Hamborg, Espen S.; Versteeg, Geert F.

    2012-01-01

    The forward and reverse kinetic rate parameters have been determined for CO2 absorption and desorption mass transfer processes in aqueous 2.0 M MDEA solutions at temperatures of 298.15, 313.15, and 333.15 K and the loading of CO2 ranging from 0 to 0.8. The derived kinetic rate parameters have been b

  3. The day of embryo transfer affects delivery rate, birth weights, female-to-male ratio, and monozygotic twin rate

    Directory of Open Access Journals (Sweden)

    Valentina Sotiroska

    2015-12-01

    Conclusion: The study suggests that if a blastocyst can be obtained in patients of advanced age (≧36 years, it improves their baby take-home rates. Younger patients (aged < 36 years should undergo elective single blastocyst transfers to reduce multiple pregnancy rates.

  4. 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.

  5. 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...

  6. Analysis of Factors Influencing Pregnancy Rate in Frozen-thawed Embryo Transfer

    Institute of Scientific and Technical Information of China (English)

    Lu LI; Xiao-xi SUN; Jun-ling CHEN; Xiao-hong GAO; Yong-wei WANG; Jie-wei TAO; Li-nan CHENG

    2004-01-01

    Objective To analyse factors influencing the outcome of frozen-thawed embryo transfer (FET)Method A retrospective analysis was performed in our center on 129 thawing cycles from March 2001 to April 2003. The related parameters were compared between conceived and non-conceived cycles.Results There were totally 129 clinical pregnancies in these transfers (pregnancy rate: 27.1%). Frozen-thawed embryos were transferred to natural cycles and CC cycling and hormone replacement treatment had equal success. Groups of IVF and ICSI did not differ significantly in pregnancy rates (P>0. 05). The pregnancy rates for one, two, three and four pre-embryos transfer were 0, 20.0%,44.1% and 75.0%,respectively (P<0. 05). There were statistical differences between pregnancy group or non- pregnancy group in the endometrial thickness, CES, CES/No. Of embryo. A higher pregnancy rate was observed in embryo transfers which had at least one 4-cell grade I embryo (d 2)(P<0.01). Conclusions The most important factors influencing the implantation rate and pregnancy rate of frozen-thawed embryo transfer are age, endometrium thickness, and the number, morphology and growth rate of transferred frozen embryos of women participants.

  7. Analyzing Service Rates for File Transfers in Peer-to-peer File Sharing Systems

    Institute of Scientific and Technical Information of China (English)

    WANG Kai; PAN Li; LI Jian-hua

    2008-01-01

    When examining the file transfer performance in a peer-to-peer file sharing system, a fundamental problem is how to describe the service rate for a file transfer.In this paper, the problem is examined by analyzing the distribution of server-like nodes' upstream-bandwidth among their concurrent transfers.A sufficient condition for the service rate, what a receiver obtains for downloading a file, to asymptotically be uniform is presented.On the aggregate service rate for transferring a file in a system, a sufficient condition for it to asymptotically follow a Zipf distribution is presented.These asymptotic equalities are both in the mean square sense.These analyses and the sufficient conditions provide a mathematic base for modeling file transfer processes in peer-to-peer file sharing systems.

  8. Heart-rate recommendations: transfer between running and cycling exercise?

    Science.gov (United States)

    Roecker, K; Striegel, H; Dickhuth, H-H

    2003-04-01

    With the expanding use of portable heart rate (HR) monitors in endurance sports, HR is increasingly used as a marker for exercise intensity. Hereby, HR at the so-called individual anaerobic threshold (IAT) is one possible reference point. However, once determined, it is often attempted to apply HR recommendations from one type of ergometry to different kinds of exercises. We examined whether HR at IAT and at 4 mmol x l -1 blood lactate is predictable from cycling to running and vice versa. Data of 371 subjects (304 male, 67 female) were analyzed. All subjects underwent an incremental test on a treadmill (TR, starting speed 6 or 8 km x h -1, increments 2 km x h -1 every 3 min) and on a bicycle ergometer (BE, start at 50 Watt, increments 25 or 50 Watt every 3 min). IAT was determined at a net increase of lactate concentration of 1 - 5 mmol x l -1 above lactate concentration at lactate threshold for running (as in: Med Sci Sports Exerc 1998, 30 (10); 1552 - 1557) and 1.0 mmol x l -1 for cycling. A maximum time span of three weeks was allowed between the tests. We found that heart rate at IAT or at 4 mmol x l -1 blood lactate did not correlate between cycling and running. A sports specific test seems to be a prerequisite for reliable heart rate recommendations.

  9. Stimulation of chondrocytes in vitro by gene transfer with plasmids coding for epidermal growth factor (hEGF) and basic fibroblast growth factor (bFGF)

    DEFF Research Database (Denmark)

    Schmal, H; Mehlhorn, A T; Zwingmann, J

    2005-01-01

    Human epidermal growth factor (hEGF) and basic fibroblast growth factor (bFGF) influence critical characteristics of chondrocytes. The effects on metabolism and differentiation were evaluated following transfection using specific plasmids coding for both cytokines. Chondrocytes were isolated from...... of recombinant hEGF and bFGF resulted in a significant increase in cell proliferation and glucosaminoglycan production. Chondrocytes were transfected with vectors coding for either hEGF or bFGF and the production of these proteins was measured in supernatants by ELISA. Expression kinetics showed different...... patterns: hEGF was detectable 2.5 days following transfection and peaked at day 5.5, whereas bFGF-production reached its maximum 1.5 days after transfection, declining thereafter. Chondrocytes endogenously produced significant amounts of bFGF within 5 days following isolation. Proliferation of h...

  10. 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

  11. Maximum orbit plane change with heat-transfer-rate considerations

    Science.gov (United States)

    Lee, J. Y.; Hull, D. G.

    1990-01-01

    Two aerodynamic maneuvers are considered for maximizing the plane change of a circular orbit: gliding flight with a maximum thrust segment to regain lost energy (aeroglide) and constant altitude cruise with the thrust being used to cancel the drag and maintain a high energy level (aerocruise). In both cases, the stagnation heating rate is limited. For aeroglide, the controls are the angle of attack, the bank angle, the time at which the burn begins, and the length of the burn. For aerocruise, the maneuver is divided into three segments: descent, cruise, and ascent. During descent the thrust is zero, and the controls are the angle of attack and the bank angle. During cruise, the only control is the assumed-constant angle of attack. During ascent, a maximum thrust segment is used to restore lost energy, and the controls are the angle of attack and bank angle. The optimization problems are solved with a nonlinear programming code known as GRG2. Numerical results for the Maneuverable Re-entry Research Vehicle with a heating-rate limit of 100 Btu/ft(2)-s show that aerocruise gives a maximum plane change of 2 deg, which is only 1 deg larger than that of aeroglide. On the other hand, even though aerocruise requires two thrust levels, the cruise characteristics of constant altitude, velocity, thrust, and angle of attack are easy to control.

  12. DETERMINATION OF HETEROGENEOUS ELECTRON TRANSFER RATE CONSTANTS AT MICROFABRICATED IRIDIUM ELECTRODES. (R825511C022)

    Science.gov (United States)

    There has been an increasing use of both solid metal and microfabricated iridium electrodes as substrates for various types of electroanalysis. However, investigations to determine heterogeneous electron transfer rate constants on iridium, especially at an electron beam evapor...

  13. Water and acrylamide monomer transfer rates from a settling basin to groundwaters.

    Science.gov (United States)

    Binet, Stéphane; Bru, Kathy; Klinka, Thomas; Touzé, Solène; Motelica-Heino, Mickael

    2015-05-01

    The aim of this paper was to estimate the potential leakage of acrylamide monomer, used for flocculation in a settling basin, towards the groundwaters. Surface-groundwater interactions were conceptualized with a groundwater transport model, using a transfer rate to describe the clogged properties of the interface. The change in the transfer rate as a function of the spreading of the clogged layer in the settling basin was characterized with respect to time. It is shown that the water and the Acrylamide transfer rate are not controlled by the spreading of the clogged layer until this layer fully covers the interface. When the clogged layer spreads out, the transfer rate remains in the same order of magnitude until the area covered reaches 80 %. The main flux takes place through bank seepage. In these early stage conditions of a working settling basin, the acrylamide flux towards groundwaters remains constant, at close to 10 g/year (±5).

  14. Reducing twin pregnancy rates after IVF--elective single embryo transfer (eSET).

    LENUS (Irish Health Repository)

    Milne, P

    2010-01-01

    Multiple pregnancy is a major complication of IVF and is associated with increased maternal, fetal and neonatal morbidity. Elective single embryo transfer (eSET) during IVF, rather than the more standard transfer of two embryos (double embryo transfer or DET), has been shown to significantly reduce the multiple pregnancy rate associated with IVF, while maintaining acceptable pregnancy rates. Couples undergoing IVF in 2008 who met good prognostic criteria had eSET performed. Pregnancy and twinning rates were compared with those for similar couples in 2007 who had DET. Couples unsuccessful with a fresh cycle of treatment had subsequent frozen embryo transfer cycles with DET. The cumulative pregnancy rate was similar for each group. However there were no multiple pregnancies in the eSET group, compared to 4 twins of 5 pregnancies in the DET group. 96% of eligible couples agreed to eSET. ESET is successful in and acceptable to good prognosis Irish couples undergoing IVF.

  15. DETERMINATION OF HETEROGENEOUS ELECTRON TRANSFER RATE CONSTANTS AT MICROFABRICATED IRIDIUM ELECTRODES. (R825511C022)

    Science.gov (United States)

    There has been an increasing use of both solid metal and microfabricated iridium electrodes as substrates for various types of electroanalysis. However, investigations to determine heterogeneous electron transfer rate constants on iridium, especially at an electron beam evapor...

  16. Effect of age of equine embryos and method of transfer on pregnancy rate.

    Science.gov (United States)

    Iuliano, M F; Squires, E L; Cook, V M

    1985-01-01

    A 2 X 2 cross-classified experiment was conducted to investigate the effect of age of equine embryo (7 vs 8 d postovulation) and method of transfer (surgical vs nonsurgical) on pregnancy rates at 50 d of gestation. Embryos were recovered 7 or 8 d postovulation using a Foley catheter and 3 liters of modified Dulbecco's phosphate-buffered saline (PBS). Upon identification, the embryos were placed in millipore-filtered PBS containing 20% heat-inactivated steer serum and maintained at room temperature until transferred. At the time of recovery, embryos were randomly assigned to be transferred either nonsurgically using a sterile insemination pipette or surgically via a flank incision. For nonsurgical transfer, the embryo was deposited into the uterine body; whereas, in surgical transfer, the embryo was placed in the uterine horn ipsilateral to the corpus luteum. Recovery rates for embryos collected on d 7 (75.5%) or 8 (81.9%) were similar (P greater than .05). Age of embryo did not affect (P greater than .05) pregnancy rate. At 50 d, pregnancy rates were 60 and 57% for mares receiving d 7 or 8 embryos. However, more (P less than .05) pregnancies were obtained after transfer of embryos surgically (72%) than nonsurgically (45%). More (P less than .05) pregnancies were obtained after transfer of d 8 embryos surgically (75%) compared with nonsurgically (40%). Within method of transfer, pregnancy rates were similar (P less than .05) for surgical transfer of d 7 and 8 embryos (69 and 75%), but tended (P less than .25) to be higher for nonsurgical transfer of d 7 embryos (50%) compared with d 8 embryos (40%).(ABSTRACT TRUNCATED AT 250 WORDS)

  17. Combination of the somatic cell nuclear transfer method and RNAi technology for the production of a prion gene-knockdown calf using plasmid vectors harboring the U6 or tRNA promoter.

    Science.gov (United States)

    Wongsrikeao, Pimprapar; Sutou, Shizuyo; Kunishi, Miho; Dong, Ya Juan; Bai, Xuejin; Otoi, Takeshige

    2011-01-01

    By combining RNAi technology with SCNT method, we attempted to produce transgenic calves with knocked down bPRNP for technological assessments. The respective utilities of type II (tRNA) and type III (hU6) Pol III promoters in mediating plasmid vector-based RNAi for the production of a bPRNP-knockdown calf were compared. Plasmid harboring DNA for siRNA expression was introduced stably into the genome of primary cultured bovine cells. By inserting the transgenic cell into an enucleated bovine egg, SCNT embryos were produced. The ability for SCNT embryos to develop to blastocysts was higher in hU6 based vector groups (44-53%) than in a tRNA group (32%). In all, 30 hU6-embryos and 12 tRNA-embryos were transferred to 11 recipients. Only tRNA-embryos were able to impregnate recipients (6 out of 11 transfers), resulting in four aborted fetuses, one stillbirth, and one live-born calf. The expression of EGFP, a marker, was detected in all six. The bPRNP transcript levels in the nervous tissues (brain, cerebellum, spinal bulb, and spinal cord) from the calf, which was killed 20 days after birth, were reduced to 35% of those of the control calf on average, as determined by qRT-PCR. The PrPC levels, as estimated by western blot were reduced to 86% on average in the nervous tissues. These findings suggest that SCNT technology remains immature, that the tRNA promoter is useful, and that RNAi can significantly reduce PRNP mRNA levels, but insufficient reduction of PrPC levels exists in cattle under these conditions.

  18. Influence of fluid dynamic conditions on enzymatic hydrolysis of lignocellulosic biomass: Effect of mass transfer rate.

    Science.gov (United States)

    Wojtusik, Mateusz; Zurita, Mauricio; Villar, Juan C; Ladero, Miguel; Garcia-Ochoa, Felix

    2016-09-01

    The effect of fluid dynamic conditions on enzymatic hydrolysis of acid pretreated corn stover (PCS) has been assessed. Runs were performed in stirred tanks at several stirrer speed values, under typical conditions of temperature (50°C), pH (4.8) and solid charge (20% w/w). A complex mixture of cellulases, xylanases and mannanases was employed for PCS saccharification. At low stirring speeds (mass transfer coefficients and rates, when compared to chemical hydrolysis rates, lead to results that clearly show low mass transfer rates, being this phenomenon the controlling step of the overall process rate. However, for stirrer speed from 300rpm upwards, the overall process rate is controlled by hydrolysis reactions. The ratio between mass transfer and overall chemical reaction rates changes with time depending on the conditions of each run.

  19. Lithium Ion Coupled Electron-Transfer Rates in Superconcentrated Electrolytes: Exploring the Bottlenecks for Fast Charge-Transfer Rates with LiMn2O4 Cathode Materials.

    Science.gov (United States)

    Nikitina, Victoria A; Zakharkin, Maxim V; Vassiliev, Sergey Yu; Yashina, Lada V; Antipov, Evgeny V; Stevenson, Keith J

    2017-09-19

    The charge-transfer kinetics of lithium ion intercalation into LixMn2O4 cathode materials was examined in dilute and concentrated aqueous and carbonate LiTFSI solutions using electrochemical methods. Distinctive trends in ion intercalation rates were observed between water-based and ethylene carbonate/diethyl carbonate solutions. The influence of the solution concentration on the rate of lithium ion transfer in aqueous media can be tentatively attributed to the process associated with Mn dissolution, whereas in carbonate solutions the rate is influenced by the formation of a concentration-dependent solid electrolyte interface (SEI). Some indications of SEI layer formation at electrode surfaces in carbonate solutions after cycling are detected by X-ray photoelectron spectroscopy. The general consequences related to the application of superconcentrated electrolytes for use in advanced energy storage cathodes are outlined and discussed.

  20. Effects of Rate-Limited Mass Transfer on Modeling Vapor Intrusion with Aerobic Biodegradation.

    Science.gov (United States)

    Chen, Yiming; Hou, Deyi; Lu, Chunhui; Spain, Jim C; Luo, Jian

    2016-09-06

    Most of the models for simulating vapor intrusion accept the local equilibrium assumption for multiphase concentration distributions, that is, concentrations in solid, liquid and vapor phases are in equilibrium. For simulating vapor transport with aerobic biodegradation controlled by counter-diffusion processes, the local equilibrium assumption combined with dual-Monod kinetics and biomass decay may yield near-instantaneous behavior at steady state. The present research investigates how predicted concentration profiles and fluxes change as interphase mass transfer resistances are increased for vapor intrusion with aerobic biodegradation. Our modeling results indicate that the attenuation coefficients for cases with and without mass transfer limitations can be significantly different by orders of magnitude. Rate-limited mass transfer may lead to larger overlaps of contaminant vapor and oxygen concentrations, which cannot be simulated by instantaneous reaction models with local equilibrium mass transfer. In addition, the contaminant flux with rate-limited mass transfer is much smaller than that with local equilibrium mass transfer, indicating that local equilibrium mass transfer assumption may significantly overestimate the biodegradation rate and capacity for mitigating vapor intrusion through the unsaturated zone. Our results indicate a strong research need for field tests to examine the validity of local equilibrium mass transfer, a widely accepted assumption in modeling vapor intrusion.

  1. Information transfer through stochastic transmission of a linear combination of rates.

    Science.gov (United States)

    Smyrnakis, Ioannis; Smirnakis, Stelios

    2013-09-01

    In this work, the Shannon information transfer rate due to the transmission of a linear combination of the firing rates of a number of afferent neurons is examined. The transmission of this linear combination (transfer statistic) takes place through a stochastic firing process, while a rate code is assumed. Constraints are imposed on the transmission process by the requirement that the coefficient of variation for the transfer statistic is small and by the relative variance of the individual terms in the calculation of the statistic. In the regime of no noise or signal correlations among the input neurons, simulations suggest that information transfer for fixed overall input is favored when there are few high-firing neurons, as opposed to more lower-firing neurons. Signal correlations among low-firing neurons can result in aggregates of high firing rates, improving in this way information transfer and calculational robustness. Under reasonable rate code assumptions, information transfer rates obtained are of the order 3 to 10 bit/sec.

  2. Thickness and local field effects on energy transfer rate in coupled quantum wells system: Linear regime

    Energy Technology Data Exchange (ETDEWEB)

    Vazifehshenas, T.; Bahrami, B. [Department of Physics, Shahid Beheshti University, G.C., Evin, 1983963113 Tehran (Iran, Islamic Republic of); Salavati-fard, T., E-mail: taha@udel.edu [Department of Physics and Astronomy, University of Delaware, Newark, DE 19716 (United States)

    2012-12-15

    We investigate theoretically the dependence of energy transfer rate in Double-Quantum-Well system on the well thickness by using the balance equation formalism. Also, by including the local field correction in our calculations through the zero- and finite-temperature Hubbard approximations, we study the effect of the short-range interactions on the energy transfer phenomenon. Calculations consider both the static and dynamic screening approximations. Our numerical results predict that the energy transfer rate increases considerably by increasing the layers' thicknesses and by taking into account the short-range interactions, as well.

  3. 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...

  4. Mass Transfer in a closed stirred gas/liquid contactor: Part 1: The mass transfer rate kLS

    NARCIS (Netherlands)

    Koetsier, W.T.; Thoenes, D.; Frankena, J.F.

    1973-01-01

    Liquid phase mass transfer rates kLS for the absorption of oxygen in tap water and in aqueous ionic solutions have been determined in two closed stirred tank contactors for a power input between 3 and 70 W/kg and (impeller diameter)f(tank diameter) ratios DifT of 0.3, 0.35 and 0.4. The contactors

  5. IncA/C plasmids harboured in serious multidrug-resistant Vibrio cholerae serogroup O139 strains in China.

    Science.gov (United States)

    Wang, Ruibai; Yu, Dong; Zhu, Lianhui; Li, Jie; Yue, Junjie; Kan, Biao

    2015-03-01

    Vibrio cholerae serogroup O139 emerged in 1992 and is one of two major serogroups to have caused cholera epidemics. After 1998, serious multidrug-resistant (MDR) O139 strains quickly became common in China, showing a multidrug resistance profile to eight antibiotics. It is a great threat to public health, and elucidation of its mechanisms of resistance will provide a helpful guide for the clinical treatment and prevention of cholera. In this study, mega-plasmids from MDR V. cholerae O139 strains were identified by pulsed-field gel electrophoresis (PFGE) without enzyme digestion. One plasmid was isolated and sequenced, belonging to the IncA/C family. Ten antibiotic resistance genes were found in the MDR regions, including a blaTEM-20 gene, and these genes endowed the host with resistance to seven antibiotics. This kind of plasmid was positive in 71.2% (198/278) of toxigenic O139 strains, and the rate of plasmid positivity was consistent with the yearly change in MDR rates of these strains. This study reveals an important role of the IncA/C family plasmid in the spread of multiple antibiotic resistance of epidemic V. cholerae serogroup O139 strains, which has recombined with plasmids from different bacterial species and transferred among V. cholerae strains. Copyright © 2014 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.

  6. Role of Plasmid in Production of Acetobacter Xylinum Biofilms

    Directory of Open Access Journals (Sweden)

    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.

  7. 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.

  8. Rotational Energy Transfer and Collisional Induced Raman Linewidths in N2 Gas. 1; Energy Transfer Rates

    Science.gov (United States)

    Huo, Winifred M.; Green, Sheldon; Langhoff, Stephen R. (Technical Monitor)

    1995-01-01

    Rotationally inelastic transitions of N2 have been studied in the coupled state (CS) and infinite-order-sudden (IOS) approximations, using the N2-N2 rigidrotor potential of van der Avoird et al. For benchmarking purposes, close coupling (CC) calculations have also been carried out over a limited energy range. The CC and CS cross sections have been obtained both with and without identical molecule exchange symmetry, whereas exchange was neglected in the IOS calculations. The CS results track the CC cross sections rather well; between 113 - 219 cm(exp -1) the average deviation is 14%. Comparison between the CS and IOS cross sections at the high energy end of the CS calculation, 500 - 680 cm(exp -1), shows that IOS is sensitive to the amount of inelasticity and the results for large DELTA J transitions are subject to larger errors. It is found that the state-to-state cross sections with even and odd exchange symmetry agree to better than 2% and are well represented as a sum of direct and exchange cross sections for distinguishable molecules, an indication of the applicability of a classical treatment for this system. This result, however, does not apply to partial cross sections for given total J, but arises from a near cancellation in summing over partial waves. In order to use rigid-rotor results for the calculation of effective rotational excitation rates of N2 in the v=1 vibrational level colliding with bath N2 molecules in the v=0 level, it is assumed that exchange scattering between molecules in different vibrational levels is negligible and direct scattering is independent of Y. Good agreement with room temperature experimental data is obtained. The effective rates determined using the IOS and energy corrected sudden (ECS) approximations are also in reasonable agreement with experiment, with the ECS results being somewhat better. The problem with a degeneracy factor in earlier cross section expressions for collisions between identical molecules is pointed out

  9. A Very Efficient Transfer Function Bounding Technique on Bit Error Rate for Viterbi Decoded, Rate 1/N Convolutional Codes

    Science.gov (United States)

    Lee, P. J.

    1984-01-01

    For rate 1/N convolutional codes, a recursive algorithm for finding the transfer function bound on bit error rate (BER) at the output of a Viterbi decoder is described. This technique is very fast and requires very little storage since all the unnecessary operations are eliminated. Using this technique, we find and plot bounds on the BER performance of known codes of rate 1/2 with K 18, rate 1/3 with K 14. When more than one reported code with the same parameter is known, we select the code that minimizes the required signal to noise ratio for a desired bit error rate of 0.000001. This criterion of determining goodness of a code had previously been found to be more useful than the maximum free distance criterion and was used in the code search procedures of very short constraint length codes. This very efficient technique can also be used for searches of longer constraint length codes.

  10. Chlamydial plasmids and bacteriophages.

    Science.gov (United States)

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

    2015-01-01

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

  11. Increasing vaginal progesterone gel supplementation after frozen-thawed embryo transfer significantly increases the delivery rate

    DEFF Research Database (Denmark)

    Alsbjerg, Birgit; Polyzos, Nikolaos P; Elbaek, Helle Olesen

    2013-01-01

    The aim of this study was to evaluate the reproductive outcome in patients receiving frozen-thawed embryo transfer before and after doubling of the vaginal progesterone gel supplementation. The study was a retrospective study performed in The Fertility Clinic, Skive Regional Hospital, Denmark....... A total of 346 infertility patients with oligoamenorrhoea undergoing frozen-thawed embryo transfer after priming with oestradiol and vaginal progesterone gel were included. The vaginal progesterone dose was changed from 90 mg (Crinone) once a day to twice a day and the reproductive outcome during the two...... rate (8.7% versus 20.5%, respectively; P=0.002). Doubling of the vaginal progesterone gel supplementation during frozen-thawed embryo transfer cycles decreased the early pregnancy loss rate, resulting in a significantly higher delivery rate. This study evaluated the reproductive outcome of 346 women...

  12. 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.

  13. Heat and water rate transfer processes in the human respiratory tract at various altitudes.

    Science.gov (United States)

    Kandjov, I M

    2001-02-01

    The process of the respiratory air conditioning as a process of heat and mass exchange at the interface inspired air-airways surface was studied. Using a model of airways (Olson et al., 1970) where the segments of the respiratory tract are like cylinders with a fixed length and diameter, the corresponding heat transfer equations, in the paper are founded basic rate exchange parameters-convective heat transfer coefficient h(c)(W m(-2) degrees C(-1)) and evaporative heat transfer coefficient h(e)(W m(-2)hPa(-1)). The rate transfer parameters assumed as sources with known heat power are connected to airflow rate in different airways segments. Relationships expressing warming rate of inspired air due to convection, warming rate of inspired air due to evaporation, water diffused in the inspired air from the airways wall, i.e. a system of air conditioning parameters, was composed. The altitude dynamics of the relations is studied. Every rate conditioning parameter is an increasing function of altitude. The process of diffusion in the peripheral bronchial generations as a basic transfer process is analysed. The following phenomenon is in effect: the diffusion coefficient increases with altitude and causes a compensation of simultaneous decreasing of O(2)and CO(2)densities in atmospheric air. Due to this compensation, the diffusion in the peripheral generations with altitude is approximately constant. The elements of the human anatomy optimality as well as the established dynamics are discussed and assumed. The square form of the airways after the trachea expressed in terms of transfer supposes (in view of maximum contact surface), that a maximum heat and water exchange is achieved, i.e. high degree of air condition at fixed environmental parameters and respiration regime.

  14. Mexico’s conditional cash transfer programme increases cesarean section rates among the rural poor

    Science.gov (United States)

    2010-01-01

    Background: Caesarean section rates are increasing in Mexico and Latin America. This study evaluates the impact of a large-scale, conditional cash transfer programme in Mexico on caesarean section rates. The programme provides cash transfers to participating low income, rural households in Mexico conditional on accepting health care and nutrition supplements. Methods: The primary analyses uses retrospective reports from 979 women in poor rural communities participating in an effectiveness study and randomly assigned to incorporation into the programme in 1998 or 1999 across seven Mexican states. Using multivariate and instrumental variable analyses, we estimate the impact of the programme on caesarean sections and predict the adjusted mean rates by clinical setting. Programme participation is measured by beneficiary status, programme months and cash transfers. Results: More than two-thirds of poor rural women delivered in a health facility. Beneficiary status is associated with a 5.1 percentage point increase in caesarean rates; this impact increases to 7.5 percentage points for beneficiaries enrolled in the programme for ≥6 months before delivery. Beneficiaries had significantly higher caesarean delivery rates in social security facilities (24.0 compared with 5.6% among non-beneficiaries) and in other government facilities (19.3 compared with 9.5%). Conclusion: The Oportunidades conditional cash transfer programme is associated with higher caesarean section rates in social security and government health facilities. This effect appears to be driven by the increases in disposable income from the cash transfer. These findings are relevant to other countries implementing conditional cash transfer programmes and health care requirements. PMID:19933778

  15. Mexico's conditional cash transfer programme increases cesarean section rates among the rural poor.

    Science.gov (United States)

    Barber, Sarah L

    2010-08-01

    Caesarean section rates are increasing in Mexico and Latin America. This study evaluates the impact of a large-scale, conditional cash transfer programme in Mexico on caesarean section rates. The programme provides cash transfers to participating low income, rural households in Mexico conditional on accepting health care and nutrition supplements. The primary analyses uses retrospective reports from 979 women in poor rural communities participating in an effectiveness study and randomly assigned to incorporation into the programme in 1998 or 1999 across seven Mexican states. Using multivariate and instrumental variable analyses, we estimate the impact of the programme on caesarean sections and predict the adjusted mean rates by clinical setting. Programme participation is measured by beneficiary status, programme months and cash transfers. More than two-thirds of poor rural women delivered in a health facility. Beneficiary status is associated with a 5.1 percentage point increase in caesarean rates; this impact increases to 7.5 percentage points for beneficiaries enrolled in the programme for >or=6 months before delivery. Beneficiaries had significantly higher caesarean delivery rates in social security facilities (24.0 compared with 5.6% among non-beneficiaries) and in other government facilities (19.3 compared with 9.5%). The Oportunidades conditional cash transfer programme is associated with higher caesarean section rates in social security and government health facilities. This effect appears to be driven by the increases in disposable income from the cash transfer. These findings are relevant to other countries implementing conditional cash transfer programmes and health care requirements.

  16. Comparison of pregnancy rate between fresh embryo transfers and frozen-thawed embryo transfers following ICSI treatment

    Directory of Open Access Journals (Sweden)

    Zahra Basirat

    2016-01-01

    Full Text Available Background: The use of assisted reproductive technology (ART is increasing in the world. The rate, efficacy and safety of ART are very different among countries. There is an increase in the use of intra cytoplasmic sperm injection (ICSI, single fresh embryo transfer (ET and frozen-thawed embryo transfer (FET. Objective: The objective of this study was to compare pregnancy rate in fresh ET and FET. Materials and Methods: In this retrospective cross-sectional study 1014 ICSI-ET cycles (426 fresh ET and 588 FET from 753 women undergoing ICSI treatment referred to Fatemezahra Infertility and Reproductive Health Research Center in Babol, Iran from 2008 to 2013 were reviewed. Results: There were no significant differences between biochemical pregnancy rate (23% versus 18.8%, OR 1.301; 95% CI .95-1.774, gestational sac (95.6% versus 100% in FET, OR 0.60; 95% CI 0.54-0.67, and fetal heart activity (87.2% versus 93.6% OR .46; 95% CI .16-1.32 in fresh ET and FET cycles, respectively. P< 0.05 was considered statistically significant for all measures. Conclusion: Although, the result showed no significantly difference between the fresh ET and the FET cycles, however the embryos are able to be stored for subsequent ART. Therefore, we recommend FET cycles as an option alongside the fresh ET.

  17. Transfer of human frozen-thawed embryos with further cleavage during culture increases pregnancy rates

    Directory of Open Access Journals (Sweden)

    Bharat V Joshi

    2010-01-01

    Full Text Available Aim: To compare the pregnancy rate following transfer of frozen-thawed embryos with or without overnight culture after thawing. Settings and Design: This is a retrospective analysis of frozen-thawed embryo transfer (FET cycles performed between January 2006 and December 2008. Materials and Methods: Out of 518 thaw cycles, 504 resulted in embryo transfers (ETs. Of the total FET cycles, 415 were performed after an overnight culture of embryos (group A; and in 89 cycles, ET was performed within 2 hours of embryo thawing (group B. Statistical Analysis: The data were statistically analyzed using chi-square test. Results: We observed that with FET, women ≤30 years of age had a significantly higher (P=0.003 pregnancy rate (PR=28.9% as compared to women >30 years of age (17.5%. A significantly higher (P<0.001FNx08 pregnancy rate was also observed in women receiving 3 frozen-thawed embryos (29% as compared to those who received less than 3 embryos (10.7%. The difference in PR between group A (PR=24.3% and group B (PR=20.3% was not statistically significant. However, within group A, ET with cleaved embryos showed significantly ( P≤0.01 higher pregnancy rate compared to the uncleaved embryos, depending on the number of cleaved embryos transferred. Conclusion: No significant difference was noticed between FETs made with transfer of embryos with overnight culture and those without culture. However, within the cultured group, transfer of embryos cleaved during overnight culture gave significantly higher PR than transfers without any cleavage.

  18. Rate of heat transfer in polypropylene tubes in solar water heaters

    Energy Technology Data Exchange (ETDEWEB)

    Razavi, J.; Mahmoodi, M. [Sharif University of Technology, Tehran (Iran). Chemical Engineering Dept.; Riazi, M.R. [Kuwait Univ., Safat (Kuwait). Chemical Engineering Dept.

    2003-06-01

    A heat transfer rate was determined for polypropylene tubes in solar water heaters for the Reynolds number range 800-5600. Experiments were conducted in ambient temperatures of 34 to 37 {sup o}C. Data were correlated in the form of Nusselt numbers as: Nu=0.0015 Re{sup 0.75}Pr{sup 1/3} with correlation coefficient of 0.95. Such data can be used to predict heat transfer rates in a polypropylene solar heater in Tehran where the experiments were performed. An application of the results is shown in an example. (author)

  19. Learned cardiac control with heart rate biofeedback transfers to emotional reactions.

    Directory of Open Access Journals (Sweden)

    Nathalie Peira

    Full Text Available Emotions involve subjective feelings, action tendencies and physiological reactions. Earlier findings suggest that biofeedback might provide a way to regulate the physiological components of emotions. The present study investigates if learned heart rate regulation with biofeedback transfers to emotional situations without biofeedback. First, participants learned to decrease heart rate using biofeedback. Then, inter-individual differences in the acquired skill predicted how well they could decrease heart rate reactivity when later exposed to negative arousing pictures without biofeedback. These findings suggest that (i short lasting biofeedback training improves heart rate regulation and (ii the learned ability transfers to emotion challenging situations without biofeedback. Thus, heart rate biofeedback training may enable regulation of bodily aspects of emotion also when feedback is not available.

  20. Learned cardiac control with heart rate biofeedback transfers to emotional reactions.

    Science.gov (United States)

    Peira, Nathalie; Pourtois, Gilles; Fredrikson, Mats

    2013-01-01

    Emotions involve subjective feelings, action tendencies and physiological reactions. Earlier findings suggest that biofeedback might provide a way to regulate the physiological components of emotions. The present study investigates if learned heart rate regulation with biofeedback transfers to emotional situations without biofeedback. First, participants learned to decrease heart rate using biofeedback. Then, inter-individual differences in the acquired skill predicted how well they could decrease heart rate reactivity when later exposed to negative arousing pictures without biofeedback. These findings suggest that (i) short lasting biofeedback training improves heart rate regulation and (ii) the learned ability transfers to emotion challenging situations without biofeedback. Thus, heart rate biofeedback training may enable regulation of bodily aspects of emotion also when feedback is not available.

  1. High rates of unsuccessful transfer to adult care among young adults with juvenile idiopathic arthritis

    Directory of Open Access Journals (Sweden)

    Duffy Ciarán M

    2010-01-01

    Full Text Available Abstract Background This study aimed to describe the proportion of patients with juvenile idiopathic arthritis (JIA who had experienced an unsuccessful transfer from a pediatric rheumatology team to an adult rheumatologist and to compare the characteristics of those who achieved successful transfer to those who did not. Methods We conducted a systematic chart review of all patients with JIA who attended their final Montreal Children's Hospital JIA clinic appointment between 1992 and 2005. We tracked these patients for the two years after transfer to an adult rheumatologist. We then compared characteristics of patients with successful and unsuccessful transfers of care. Variables pertaining to disease characteristics, disease severity and psychosocial factors were examined. Univariate analyses were performed to determine if any single factor was associated with the outcome of unsuccessful transfer of care. Results 52% of patients fulfilled our criteria for unsuccessful transfer. Of the variables tested, an active joint count (AJC of zero at last visit was associated with the outcome of unsuccessful transfer (OR = 2.67 (CI 1.16-6.16; p = 0.0199. Conclusions Despite the presence of a coordinated process of transfer from pediatric to adult health care for the majority of the patients in this study, there was a high rate of unsuccessful transfer and/or sustained follow up which is disheartening. We found that patients with less active disease at the time of transfer, as indicated by a lower AJC, were more likely to be lost to follow up. Recent literature suggests that even in the least severe categories of JIA, 50% of patients persist with active disease into adulthood. Thus educating all JIA patients about the possibility of disease flare in adulthood may improve their adherence to recommendations for sustained follow-up in the adult milieu. This may lead to improvement of longitudinal outcomes for all JIA patients.

  2. 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.

  3. Rapid policy change to single-embryo transfer while maintaining pregnancy rates per initiated cycle.

    Science.gov (United States)

    Vélez, M P; Kadoch, I-J; Phillips, S J; Bissonnette, F

    2013-05-01

    Public financing of IVF aims at increasing access to treatment while decreasing the expenses associated with multiple pregnancies. Critics argue that it is associated with lower pregnancy rates. This study compared cycles performed during 2009 (before implementation of Quebec's public IVF programme; period I) to those performed in the year following implementation (period II) in a single IVF centre. First fresh cycles in period I (499 women) and first fresh cycles (815 women) along with their corresponding first vitrified-warmed transfer (271 women) in period II were evaluated. From period I to period II, single-embryo transfer increased from 17.3% to 85.0% (Ppregnancy rate decreased from 25.8% to 1.6% (Ppregnancy rate decreased from 31.9% to 23.3% (P=0.001). During period II, the ongoing pregnancy rate per vitrified-warmed embryo transfer was 19.2%, leading to a cumulative ongoing pregnancy rate per initiated cycle of 29.7%, which was not different to the pregnancy rate per fresh cycle during period I (31.9%). To conclude, Quebec's public IVF programme decreased multiple pregnancy rates while maintaining an acceptable cumulative ongoing pregnancy rate, a more precise outcome to evaluate the impact of public IVF programmes.

  4. Tuning of Heat Transfer Rate of Cobalt Manganese Ferrite Based Magnetic Fluids in Varying Magnetic Field

    Directory of Open Access Journals (Sweden)

    Margabandhu MARIMUTHU

    2017-08-01

    Full Text Available Magnetic fluids are the colloidal solutions containing suspended magnetic nanoparticles in carrier fluids. The present work analyzed the heat transfer characteristics of de-ionized water and transformer oil (base fluids based cobalt manganese ferrite (Co1-xMnxFe2O4 coated with oleic acid synthesized via co-precipitation technique magnetic fluids in  varying magnetic field. Experimental investigations were carried out to analyze the heat transfer property of synthesized magnetic fluids (MNF in varying magnetic field applied in perpendicular direction to the thermal gradient of magnetic fluids. The experimental results indicate that the magnetic fluids show enhancement in heat transfer rate than carrier fluids in absence of magnetic field and it shows decrement in heat transfer rate in presence of varying magnetic field. Thus, the results reveal that the heat transfer characteristics of cobalt manganese ferrite based magnetic fluids was tunable by controlling the direction and influence of magnetic field strength. This tunable heat transfer property of cobalt manganese ferrite based magnetic fluids could be applicable in heat transport phenomena of transformers and in microelectronic devices.DOI: http://dx.doi.org/10.5755/j01.ms.23.3.16662

  5. 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.

  6. Lessons Learned From Root Cause Analyses of Transfers of Skilled Nursing Facility (SNF) Patients to Acute Hospitals: Transfers Rated as Preventable Versus Nonpreventable by SNF Staff.

    Science.gov (United States)

    Ouslander, Joseph G; Naharci, Ilkin; Engstrom, Gabriella; Shutes, Jill; Wolf, David G; Alpert, Graig; Rojido, Carolina; Tappen, Ruth; Newman, David

    2016-07-01

    Determining if a transfer of a skilled nursing facility (SNF) patient/resident to an acute hospital is potentially avoidable or preventable is challenging. Most previous research on potentially avoidable or preventable hospitalizations is based on diagnoses without in-depth root cause analysis (RCA), and few studies have examined SNF staff perspective on preventability of transfers. To examine factors associated with hospital transfers rated as potentially preventable versus nonpreventable by SNF staff. Trained staff from SNFs enrolled in a randomized controlled clinical trial of the INTERACT (Interventions to Reduce Acute Care Transfers) quality improvement program performed retrospective RCAs on hospital transfers during a 12-month implementation period. SNFs from across the United States. Sixty-four of 88 SNFs randomized to the intervention group submitted RCAs with a rating of whether the transfer was determined to be potentially preventable or nonpreventable. SNFs were implementing the INTERACT Quality Improvement (QI) program. Data were abstracted from the INTERACT QI tool, a structured, retrospective RCA on hospital transfers. A total of 4527 RCAs with a rating of preventability were submitted during the 12-month implementation period, of which 1044 (23%) were rated as potentially preventable by SNF staff. In unadjusted univariate analyses, factors associated with ratings of potentially preventable included acute changes in condition of fever, decreased food or fluid intake, functional decline, shortness of breath, and new urinary incontinence; other factors included the clinician, resident, and/or family insisting on the transfer, transfers that occurred fewer than 30 days from SNF admission and that occurred on weekends, transfers ordered by a covering physician (as opposed to the primary physician), and transfers that resulted in an emergency department (ED) visit with return to the SNF. Factors associated with ratings of nonpreventable included on

  7. Electron transfer by excited benzoquinone anions: slow rates for two-electron transitions.

    Science.gov (United States)

    Zamadar, Matibur; Cook, Andrew R; Lewandowska-Andralojc, Anna; Holroyd, Richard; Jiang, Yan; Bikalis, Jin; Miller, John R

    2013-09-05

    Electron transfer (ET) rate constants from the lowest excited state of the radical anion of benzoquinone, BQ(-•)*, were measured in THF solution. Rate constants for bimolecular electron transfer reactions typically reach the diffusion-controlled limit when the free-energy change, ΔG°, reaches -0.3 eV. The rate constants for ET from BQ(-•)* are one-to-two decades smaller at this energy and do not reach the diffusion-controlled limit until -ΔG° is 1.5-2.0 eV. The rates are so slow probably because a second electron must also undergo a transition to make use of the energy of the excited state. Similarly, ET, from solvated electrons to neutral BQ to form the lowest excited state, is slow, while fast ET is observed at a higher excited state, which can be populated in a transition involving only one electron. A simple picture based on perturbation theory can roughly account for the control of electron transfer by the need for transition of a second electron. The picture also explains how extra driving force (-ΔG°) can restore fast rates of electron transfer.

  8. Distance dependence of the energy transfer rate from a single semiconductor nanostructure to graphene.

    Science.gov (United States)

    Federspiel, François; Froehlicher, Guillaume; Nasilowski, Michel; Pedetti, Silvia; Mahmood, Ather; Doudin, Bernard; Park, Serin; Lee, Jeong-O; Halley, David; Dubertret, Benoît; Gilliot, Pierre; Berciaud, Stéphane

    2015-02-11

    The near-field Coulomb interaction between a nanoemitter and a graphene monolayer results in strong Förster-type resonant energy transfer and subsequent fluorescence quenching. Here, we investigate the distance dependence of the energy transfer rate from individual, (i) zero-dimensional CdSe/CdS nanocrystals and (ii) two-dimensional CdSe/CdS/ZnS nanoplatelets to a graphene monolayer. For increasing distances d, the energy transfer rate from individual nanocrystals to graphene decays as 1/d(4). In contrast, the distance dependence of the energy transfer rate from a two-dimensional nanoplatelet to graphene deviates from a simple power law but is well described by a theoretical model, which considers a thermal distribution of free excitons in a two-dimensional quantum well. Our results show that accurate distance measurements can be performed at the single particle level using graphene-based molecular rulers and that energy transfer allows probing dimensionality effects at the nanoscale.

  9. Three-dimensional transient mathematical model to predict the heat transfer rate of a heat pipe

    Directory of Open Access Journals (Sweden)

    S Boothaisong

    2015-02-01

    Full Text Available A three-dimensional model was developed to simulate the heat transfer rate on a heat pipe in a transient condition. This article presents the details of a calculation domain consisting of a wall, a wick, and a vapor core. The governing equation based on the shape of the pipe was numerically simulated using the finite element method. The developed three-dimensional model attempted to predict the transient temperature, the velocity, and the heat transfer rate profiles at any domain. The values obtained from the model calculation were then compared with the actual results from the experiments. The experiment showed that the time required to attain a steady state (where transient temperature is constant was reasonably consistent with the model. The working fluid r134a (tetrafluoroethane was the quickest to reach the steady state and transferred the greatest amount of heat.

  10. Theory and experiment on the cuprous-cupric electron transfer rate at a copper electrode.

    Energy Technology Data Exchange (ETDEWEB)

    Halley, J. W.; Smith, B. B.; Walbran, S.; Curtiss, L. A.; Rigney, R. O.; Sutjianto, A.; Hung, N. C.; Yonco, R. M.; Nagy, Z.; Univ. of Minnesota; NREL

    1999-04-01

    We describe results of experiment and theory of the cuprous-cupric electron transfer rate in an aqueous solution at a copper electrode. The methods are similar to those we reported earlier for the ferrous-ferric rate. The comparison strongly suggests that, in marked distinction to the ferrous-ferric case, the electron transfer reaction is adiabatic. The model shows that the activation barrier is dominated by the energy required for the ion to approach the electrode, rather than by the energy required for rearrangement of the solvation shell, also in sharp distinction to the case of the ferric-ferrous electron transfer at a gold electrode. Calculated activation barriers based on this image agree with the experimental results reported here.

  11. Theory and experiment on the cuprous{endash}cupric electron transfer rate at a copper electrode

    Energy Technology Data Exchange (ETDEWEB)

    Halley, J.W. [School of Physics and Astronomy, University of Minnesota, Minneapolis, Minnesota 55455 (United States); Smith, B.B. [National Renewable Energy Laboratory, Golden, Colorado (United States); Walbran, S. [School of Physics and Astronomy, University of Minnesota, Minneapolis, Minnesota 55455 (United States); Curtiss, L.A.; Rigney, R.O.; Sutjianto, A.; Hung, N.C.; Yonco, R.M.; Nagy, Z. [Argonne National Laboratory, Divisions of Materials Science, Chemistry and Chemical Technology, Argonne, Illinois 60439-4837 (United States)

    1999-04-01

    We describe results of experiment and theory of the cuprous{endash}cupric electron transfer rate in an aqueous solution at a copper electrode. The methods are similar to those we reported earlier for the ferrous{endash}ferric rate. The comparison strongly suggests that, in marked distinction to the ferrous{endash}ferric case, the electron transfer reaction is adiabatic. The model shows that the activation barrier is dominated by the energy required for the ion to approach the electrode, rather than by the energy required for rearrangement of the solvation shell, also in sharp distinction to the case of the ferric{endash}ferrous electron transfer at a gold electrode. Calculated activation barriers based on this image agree with the experimental results reported here. {copyright} {ital 1999 American Institute of Physics.}

  12. Nanophotonic enhancement of the F\\"orster resonance energy transfer rate on single DNA molecules

    CERN Document Server

    Ghenuche, Petru; Moparthi, Satish Babu; Grigoriev, Victor; Wenger, Jérôme

    2014-01-01

    Nanophotonics achieves accurate control over the luminescence properties of a single quantum emitter by tailoring the light-matter interaction at the nanoscale and modifying the local density of optical states (LDOS). This paradigm could also benefit to F\\"orster resonance energy transfer (FRET) by enhancing the near-field electromagnetic interaction between two fluorescent emitters. Despite the wide applications of FRET in nanosciences, using nanophotonics to enhance FRET remains a debated and complex challenge. Here, we demonstrate enhanced energy transfer within single donor-acceptor fluorophore pairs confined in gold nanoapertures. Experiments monitoring both the donor and the acceptor emission photodynamics at the single molecule level clearly establish a linear dependence of the FRET rate on the LDOS in nanoapertures. These findings are applied to enhance the FRET rate in nanoapertures up to six times, demonstrating that nanophotonics can be used to intensify the near-field energy transfer and improve t...

  13. Heat and mass transfer rates during flow of dissociated hydrogen gas over graphite surface

    Science.gov (United States)

    Nema, V. K.; Sharma, O. P.

    1986-01-01

    To improve upon the performance of chemical rockets, the nuclear reactor has been applied to a rocket propulsion system using hydrogen gas as working fluid and a graphite-composite forming a part of the structure. Under the boundary layer approximation, theoretical predictions of skin friction coefficient, surface heat transfer rate and surface regression rate have been made for laminar/turbulent dissociated hydrogen gas flowing over a flat graphite surface. The external stream is assumed to be frozen. The analysis is restricted to Mach numbers low enough to deal with the situation of only surface-reaction between hydrogen and graphite. Empirical correlations of displacement thickness, local skin friction coefficient, local Nusselt number and local non-dimensional heat transfer rate have been obtained. The magnitude of the surface regression rate is found low enough to ensure the use of graphite as a linear or a component of the system over an extended period without loss of performance.

  14. Large electron transfer rate effects from the Duschinsky mixing of vibrations

    DEFF Research Database (Denmark)

    Sando, Gerald M.; Spears, Kenneth G; Hupp, Joseph T

    2001-01-01

    We systematically test how the Duschinsky mixing of normal coordinate vibrations affects transition rates for electron transfer (ET). We find that ET rates in the inverted region can increase many orders of magnitude from Duschinsky mixing, and both totally symmetric and nontotally symmetric...... of magnitude for inverted electron transfer reactions and modest mixing. The peak location in a rate vs energy gap plot can depend on the degree of Duschinsky mixing, and therefore it corresponds to a sum of solvent and an effective vibrational reorganization energy that is not predictable by simple models...... vibrations are very important. The Duschinsky effect arises when two electronic states have vibrational normal mode coordinate systems that are rotated and translated relative to each other. We use a conventional quantum rate model for ET, and the examples include 6-8 vibrations, where two vibrational modes...

  15. Electron-impact vibrational excitation rates in the flow field of aeroassisted orbital transfer vehicles

    Science.gov (United States)

    Lee, J.-H.

    1985-01-01

    This paper examines the vibrational excitation rate processes expected in the flow field of aeroassisted orbital transfer vehicles (AOTVs). An analysis of the multiple-quantum vibrational excitation processes by electron impact is made to predict the vibrational excitation cross sections, rate coefficients, and relaxation times which control vibrational temperature. The expression for the rate of electron-vibration energy transfer is derived by solving the system of master equations which account for the multiple-level transitions. The vibrational excitation coefficients, which are the prerequisite physical quantities in solving the obtained vibrational equation, are calculated based on the theoretically predicted cross sections. These cross sections are obtained from quantum mechanical calculations, based on the concept that vibrational excitation of molecules by electron impact occurs through formation of an intermediate negative ion state. Finally, the modified Landau-Teller-type rate equation, which is suitable for the numerical calculations for the AOTV flow fields, is suggested.

  16. Large electron transfer rate effects from the Duschinsky mixing of vibrations

    DEFF Research Database (Denmark)

    Sando, Gerald M.; Spears, Kenneth G; Hupp, Joseph T

    2001-01-01

    We systematically test how the Duschinsky mixing of normal coordinate vibrations affects transition rates for electron transfer (ET). We find that ET rates in the inverted region can increase many orders of magnitude from Duschinsky mixing, and both totally symmetric and nontotally symmetric...... are mixed with different amounts of coordinate rotation. The multidimensional Franck-Condon factors (FCF) are computed with standard algorithms and recently developed recursion relations. When displaced, totally symmetric modes are involved, rates with Duschinsky mixing can increase several orders...... of magnitude for inverted electron transfer reactions and modest mixing. The peak location in a rate vs energy gap plot can depend on the degree of Duschinsky mixing, and therefore it corresponds to a sum of solvent and an effective vibrational reorganization energy that is not predictable by simple models...

  17. Cross-contamination in the kitchen: estimation of transfer rates for cutting boards, hands and knives

    NARCIS (Netherlands)

    Asselt, van E.D.; Jong, de A.E.I.; Jonge, de R.; Nauta, M.J.

    2008-01-01

    Aims: To quantify cross-contamination in the home from chicken to readyto-eat salad. Methods and Results: Based on laboratory scenarios performed by de Jong et al. (2008), transfer rates were estimated for Campylobacter jejuni and Lactobacillus casei as a tracer organism. This study showed that tran

  18. Real Time Quantification of Ultrafast Photoinduced Bimolecular Electron Transfer Rate: Direct Probing of the Transient Intermediate.

    Science.gov (United States)

    Mukherjee, Puspal; Biswas, Somnath; Sen, Pratik

    2015-08-27

    Fluorescence quenching studies through steady-state and time-resolved measurements are inadequate to quantify the bimolecular electron transfer rate in bulk homogeneous solution due to constraints from diffusion. To nullify the effect of diffusion, direct evaluation of the rate of formation of a transient intermediate produced upon the electron transfer is essential. Methyl viologen, a well-known electron acceptor, produces a radical cation after accepting an electron, which has a characteristic strong and broad absorption band centered at 600 nm. Hence it is a good choice to evaluate the rate of photoinduced electron transfer reaction employing femtosecond broadband transient absorption spectroscopy. The time constant of the aforementioned process between pyrene and methyl viologen in methanol has been estimated to be 2.5 ± 0.4 ps using the same technique. The time constant for the backward reaction was found to be 14 ± 1 ps. These values did not change with variation of concentration of quencher, i.e., methyl viologen. Hence, we can infer that diffusion has no contribution in the estimation of rate constants. However, on changing the solvent from methanol to ethanol, the time constant of the electron transfer reaction has been found to increase and has accounted for the change in solvent reorganization energy.

  19. Scaling Relations and Optimization of Excitonic Energy Transfer Rates between One-Dimensional Molecular Aggregates

    NARCIS (Netherlands)

    Chuang, Chern; Knoester, Jasper; Cao, Jianshu

    2014-01-01

    We theoretically study the distance, chain length, and temperature dependence of the electronic couplings as well as the excitonic energy transfer rates between one-dimensional (1D) chromophore aggregates. In addition to the well-known geometry dependent factor that leads to the deviation from Forst

  20. Cross-contamination in the kitchen: estimation of transfer rates for cutting boards, hands and knives

    NARCIS (Netherlands)

    Asselt, van E.D.; Jong, de A.E.I.; Jonge, de R.; Nauta, M.J.

    2008-01-01

    Aims: To quantify cross-contamination in the home from chicken to readyto-eat salad. Methods and Results: Based on laboratory scenarios performed by de Jong et al. (2008), transfer rates were estimated for Campylobacter jejuni and Lactobacillus casei as a tracer organism. This study showed that

  1. Experimental Investigation of Heat transfer rate of Nano fluids using a Shell and Tube Heat exchanger

    Science.gov (United States)

    SIVA ESWARA RAO, M.; SREERAMULU, DOWLURU; ASIRI NAIDU, D.

    2016-09-01

    Nano fluids are used for increasing thermal properties in heat transfer equipment like heat exchangers, radiators etc. This paper investigates the heat transfer rate of Nano fluids using a shell and tube heat exchanger in single and multi tubes under turbulent flow condition by a forced convection mode. Alumina Nanoparticles are prepared by using Sol-Gel method. Heat transfer rate increases with decreasing particle size. In this experiment Alumina Nano particles of about 22 nm diameter used. Alumina Nano fluids are prepared with different concentrations of Alumina particles (0.13%, 0.27%, 0.4%, and 0.53%) with water as a base fluid using ultra-sonicator. Experiment have been conducted on shell and tube heat exchanger for the above concentrations on parallel and counter flow conditions by keeping constant inlet temperatures and mass flow rate. The result shows that the heat transfer rate is good compared to conventional fluids. The properties of Nano fluids and non-dimensional numbers have been calculated.

  2. Quantum chemical calculations of tryptophan → heme electron and excitation energy transfer rates in myoglobin.

    Science.gov (United States)

    Suess, Christian J; Hirst, Jonathan D; Besley, Nicholas A

    2017-04-01

    The development of optical multidimensional spectroscopic techniques has opened up new possibilities for the study of biological processes. Recently, ultrafast two-dimensional ultraviolet spectroscopy experiments have determined the rates of tryptophan → heme electron transfer and excitation energy transfer for the two tryptophan residues in myoglobin (Consani et al., Science, 2013, 339, 1586). Here, we show that accurate prediction of these rates can be achieved using Marcus theory in conjunction with time-dependent density functional theory. Key intermediate residues between the donor and acceptor are identified, and in particular the residues Val68 and Ile75 play a critical role in calculations of the electron coupling matrix elements. Our calculations demonstrate how small changes in structure can have a large effect on the rates, and show that the different rates of electron transfer are dictated by the distance between the heme and tryptophan residues, while for excitation energy transfer the orientation of the tryptophan residues relative to the heme is important. © 2017 The Authors Journal of Computational Chemistry Published by Wiley Periodicals, Inc.

  3. The effect of medical clowning on pregnancy rates after in vitro fertilization and embryo transfer.

    Science.gov (United States)

    Friedler, Shevach; Glasser, Saralee; Azani, Liat; Freedman, Laurence S; Raziel, Arie; Strassburger, Dvora; Ron-El, Raphael; Lerner-Geva, Liat

    2011-05-01

    This experimental prospective quasi-randomized study examining the impact of a medical clowning encounter after embryo transfer (ET) after in vitro fertilization (IVF) found that the pregnancy rate in the intervention group was 36.4%, compared with 20.2% in the control group (adjusted odds ratio, 2.67; 95% confidence interval, 1.36-5.24). Medical clowning as an adjunct to IVF-ET may have a beneficial effect on pregnancy rates and deserves further investigation.

  4. Horizontal Transfer of the Salmonella enterica Serovar Infantis Resistance and Virulence Plasmid pESI to the Gut Microbiota of Warm-Blooded Hosts

    Directory of Open Access Journals (Sweden)

    Gili Aviv

    2016-09-01

    Full Text Available Salmonella enterica serovar Infantis is one of the prevalent salmonellae worldwide. Recently, we showed that the emergence of S. Infantis in Israel was facilitated by the acquisition of a unique megaplasmid (pESI conferring multidrug resistance and increased virulence phenotypes. Here we elucidate the ecology, transmission properties, and regulation of pESI. We show that despite its large size (~280 kb, pESI does not impose a significant metabolic burden in vitro and that it has been recently fixed in the domestic S. Infantis population. pESI conjugation and the transcription of its pilus (pil genes are inhibited at the ambient temperature (27°C and by ≥1% bile but increased under temperatures of 37 to 41°C, oxidative stress, moderate osmolarity, and the microaerobic conditions characterizing the intestinal environment of warm-blooded animals. The pESI-encoded protein TraB and the oxygen homeostasis regulator Fnr were identified as transcriptional regulators of pESI conjugation. Using the mouse model, we show that following S. Infantis infection, pESI can be horizontally transferred to the gut microbiota, including to commensal Escherichia coli strains. Possible transfer, but not persistence, of pESI was also observed into Gram-positive mouse microbiota species, especially Lactobacillus reuteri. Moreover, pESI was demonstrated to further disseminate from gut microbiota to S. enterica serovar Typhimurium, in the context of gastrointestinal infection. These findings exhibit the ability of a selfish clinically relevant megaplasmid to distribute to and from the microbiota and suggest an overlooked role of the microbiota as a reservoir of mobile genetic elements and intermediator in the spread of resistance and virulence genes between commensals and pathogenic bacteria.

  5. Confirmed dioestrus in pseudopregnant mice using vaginal exfoliative cytology improves embryo transfer implantation rate.

    Science.gov (United States)

    Mamrot, Jared; Pangestu, Mulyoto; Walker, David; Gardner, David K; Dickinson, Hayley

    2015-10-01

    Embryo transfer is a commonly performed surgical technique. In mice, protocols typically specify pairing recipient females with vasectomized males to induce a receptive uterine environment for embryo implantation. However, this induced receptive state is not always maintained until implantation occurs. The use of a well-characterized correlation between oestrous state and exfoliative vaginal cytology was therefore evaluated to assess uterine receptivity immediately before embryo transfer. Eight- to 12-week-old virgin female CD1 mice (n = 22) were paired overnight with vasectomized males and successfully mated, indicated by the presence of a vaginal plug. These dams underwent embryo transfer 3 days later with embryos obtained from superovulated 4-week-old F1 (C57BL/6 × CBA) females. Non-invasive vaginal lavage was conducted immediately before transfer. Dams were killed 6 days after transfer and the uterus collected for histological analysis. Embryo implantation rate in mice was 96% when cytological analysis of the lavage samples signified dioestrus (n = 6), whereas the implantation rate was cytology signified other stages of oestrous. This simple, quick, non-invasive measure of receptivity was accurate and easily adopted and, when applied prospectively, will avoid unnecessary surgery and subsequent culling of non-suitable recipients, while maximizing the implantation potential of each recipient female. Copyright © 2015 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

  6. What Controls the Rate of Ultrafast Charge Transfer and Charge Separation Efficiency in Organic Photovoltaic Blends.

    Science.gov (United States)

    Jakowetz, Andreas C; Böhm, Marcus L; Zhang, Jiangbin; Sadhanala, Aditya; Huettner, Sven; Bakulin, Artem A; Rao, Akshay; Friend, Richard H

    2016-09-14

    In solar energy harvesting devices based on molecular semiconductors, such as organic photovoltaics (OPVs) and artificial photosynthetic systems, Frenkel excitons must be dissociated via charge transfer at heterojunctions to yield free charges. What controls the rate and efficiency of charge transfer and charge separation is an important question, as it determines the overall power conversion efficiency (PCE) of these systems. In bulk heterojunctions between polymer donor and fullerene acceptors, which provide a model system to understand the fundamental dynamics of electron transfer in molecular systems, it has been established that the first step of photoinduced electron transfer can be fast, of order 100 fs. But here we report the first study which correlates differences in the electron transfer rate with electronic structure and morphology, achieved with sub-20 fs time resolution pump-probe spectroscopy. We vary both the fullerene substitution and donor/fullerene ratio which allow us to control both aggregate size and the energetic driving force for charge transfer. We observe a range of electron transfer times from polymer to fullerene, from 240 fs to as short as 37 fs. Using ultrafast electro-optical pump-push-photocurrent spectroscopy, we find the yield of free versus bound charges to be weakly dependent on the energetic driving force, but to be very strongly dependent on fullerene aggregate size and packing. Our results point toward the importance of state accessibility and charge delocalization and suggest that energetic offsets between donor and acceptor levels are not an important criterion for efficient charge generation. This provides design rules for next-generation materials to minimize losses related to driving energy and boost PCE.

  7. Blastocyst transfer ameliorates live birth rate compared with cleavage-stage embryos transfer in fresh in vitro fertilization or intracytoplasmic sperm injection cycles: reviews and meta-analysis.

    Science.gov (United States)

    Wang, Shan-Shan; Sun, Hai-Xiang

    2014-05-01

    Blastocyst transfer has been recommended to raise the implantation rate without affecting the pregnancy rate. The objective of this meta-analysis is to systematically evaluate whether the live birth rate and other pregnancy outcomes can be improved by blastocyst transfer compared with cleavage-stage embryos transfer. EMBASE and MEDLINE databases were searched for papers published between March 2004 and March 2013. An extensive range of the electronic databases yielded initially 317 studies from which seven trials met the inclusion criteria for further analysis. Our outcome measures were the live birth rate, clinical pregnancy rate, implantation rate, ongoing pregnancy rate, multiple pregnancy rate, first trimester miscarriage rate and ectopic pregnancy rate. Fixed effects models were chosen to calculate the odds ratio (OR). Seven trials (n=1446 cases) were finally analyzed. Compared with cleavage-stage embryos transfer, the blastocyst transfer was statistically significantly associated with an increase in clinical pregnancy rate [OR 1.43; 95% confidence interval (CI), 1.15-1.78], implantation rate (OR 1.38; 95% CI, 1.09-1.74) and ongoing pregnancy rate (OR 2.15; 95% CI, 1.57-2.94), and also a reduction in the probability of first trimester miscarriage rate (OR 0.51; 95% CI, 0.30-0.87). The improvement in the live birth rate was also observed (OR 1.77; 95% CI, 1.32-2.37). Moreover, there was no evidence of difference in multiple pregnancy and ectopic pregnancy rates. The available evidences suggest that live birth and other pregnancy outcomes after fresh in vitro fertilization or intracytoplasmic sperm injection (IVF/ICSI) are significantly improved following blastocyst transfer as compared to cleavage-stage embryo transfer.

  8. Correlation Between Energy Transfer Rate and Atomization Energy of Some Trinitro Aromatic Explosive Molecules

    Institute of Scientific and Technical Information of China (English)

    Su-hong Ge; Xin-lu Cheng; Zheng-lai Liu; Xiang-dong Yang; Fang-fang Dong

    2008-01-01

    An assumptive theoretical relationship is suggested to describe the property of molecular atomization energy and energy transfer rate in the initiation of explosions. To investigate the relationship between atomization energy and energy transfer rate, the number of doorway modes of explosives is estimated by the theory of Dlott and Fayer in which the rate is proportional to the number of normal mode vibrations. It was evaluated frequencies of normal mode vibrations of eight molecules by means of density functional theory (DFT) at the b3p86/6-31G(d,p) level. It is found that the number of doorway modes shows a linear correlation to the atomization energies of the molecules, which were also calculated by means of the same method. A mechanism of this correlation is discussed. It is also noted that in those explosives with similar molecular structure and molecular weight, the correlation between the atomization energy and the number of doorway modes is higher.

  9. Excitation energy transfer rates: comparison of approximate methods to the exact solution

    CERN Document Server

    Jesenko, Simon

    2014-01-01

    We have evaluated excitation energy transfer rates in photosyntetic complexes using the exact HEOM method and various approximate methods frequently used in the literature, namely, the F\\"orster method, the Redfield method, the modified Redfield method and the variational master equation. The rates are evaluated for the case of a simple dimer and a trimer photosynthetic complex, with vibrational environment characterized by the Drude-Lorentz spectral density. Comparing approximate rates to the exact ones, we have confirmed the validity of approximate methods within appropriate limits, however these limits are often well outside parameter ranges that are relevant for the dynamics in real photosynthetic complexes.

  10. 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.

  11. 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.

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

    Directory of Open Access Journals (Sweden)

    Hain Torsten

    2009-06-01

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

  13. Enhanced rate of intramolecular electron transfer in an engineered purple CuA azurin

    DEFF Research Database (Denmark)

    Farver, O; Lu, Y; Ang, M C

    1999-01-01

    V for blue copper azurin). The reorganization energy of the CuA center is calculated to be 0.4 eV, which is only 50% of that found for the wild-type azurin. These results represent a direct comparison of electron transfer properties of the blue and purple CuA sites in the same protein framework and provide...... and CuA centers are placed in the same location in the protein while all other structural elements remain the same. Long-range electron transfer is induced between the disulfide radical anion, produced pulse radiolytically, and the oxidized binuclear CuA center in the purple azurin mutant. The rate...... support for the notion that the binuclear purple CuA center is a more efficient electron transfer agent than the blue single copper center because reactivity of the former involves a lower reorganization energy....

  14. Thermophoretically augmented mass transfer rates to solid walls across laminar boundary layers

    Science.gov (United States)

    Gokoglu, S. A.; Rosner, D. E.

    1986-01-01

    Predictions of mass transfer (heavy vapor and small particle deposition) rates to solid walls, including the effects of thermal (Soret) diffusion ('thermophoresis' for small particles), are made by numerically solving the two-dimensional self-similar forced convection laminar boundary-layer equations with variable properties, covering the particle size range from vapor molecules up to the size threshold for inertial (dynamical nonequilibrium) effects. The effect of thermophoresis is predicted to be particularly important for submicron particle deposition on highly cooled solid surfaces, with corresponding enhancement factors at atmospheric conditions being over a thousand-fold at T(w)/T(e) equal to about 0.6. As a consequence of this mass transfer mechanism, the particle size dependence of the mass transfer coefficient to a cooled wall will be much weaker than for the corresponding case of isothermal capture by Brownian-convective diffusion.

  15. The effect of mare's age on multiple ovulation rate, embryo recovery, post-transfer pregnancy rate, and interovulatory interval in a commercial embryo transfer program in Argentina.

    Science.gov (United States)

    Marinone, A I; Losinno, L; Fumuso, E; Rodríguez, E M; Redolatti, C; Cantatore, S; Cuervo-Arango, J

    2015-07-01

    Advanced maternal age is an important predisposing factor on the reduction of reproductive efficiency. The aim of this study was to evaluate the effect of donor's age on several reproductive parameters in a commercial equine embryo transfer program. Donors were classified into 3 age groups: Group 1=fillies (3 and 4 years old), Group 2=middle age mares (aged 5-10) and Group 3=old mares (aged 13-25). Embryo recovery, multiple ovulation and pregnancy rates and interovulatory intervals were compared amongst age groups. Group 1 (171/244, 70.1%) and Group 2 (774/1081, 71.6%) had a higher (Precovery rate than Group 3 (385/701, 54.9%). Groups 2 and 3 were 2.5 and 3.4 times more likely to have multiple ovulations than Group 1 (Prate was not significant (P>0.05). The interovulatory intervals length was influenced by individual mare (P0.05), but was shorter than the one of Group 3 (17.4±0.15 days; Precovery rate of flushings from Groups 1 and 2 was influenced by the length of the previous interovulatory interval (P=0.03).

  16. The mass transfer rate in X1916-053 - It is driven by gravitational radiation?

    Science.gov (United States)

    Swank, J. H.; Taam, R. E.; White, N. E.

    1985-01-01

    A 50-minute period for a binary system harboring an X-ray burster would allow several alternatives for the mass-giving secondary, including an H-shell burning-plus-He degenerate core composite model. The burst properties of X1916-053 are presently used to argue against the He degenerate as well as the He main sequence solutions and to estimate whether, for any of the other solutions, the mass transfer rate could be consistent with that expected from gravitational radiation (GR). Within an uncertainty of a factor of 2, the transfer rate for the composite model solution is consistent with gravitational radiation, but enhancement by other mechanisms should be investigated.

  17. Three-fluid plasmas in star formation II. Momentum transfer rate coefficients

    CERN Document Server

    Pinto, Cecilia

    2008-01-01

    The charged component of the insterstellar medium consists of atomic and molecular ions, electrons, and charged dust grains, coupled to the local Galactic magnetic field. Collisions between neutral particles (mostly atomic or molecular hydrogen) and charged species, and between the charged species themselves, affect the magnetohydrodynamical behaviour of the medium and the dissipation of electric currents. The friction force due to elastic collisions between particles of different species in the multi-component interstellar plasma is a nonlinear function of the temperature of each species and the Mach number of the relative drift velocity. The aim of this paper is to provide an accurate and, as far as possible, complete set of momentum transfer rate coefficients for magnetohydrodynamical studies of the interstellar medium. Momentum transfer rates are derived from available experimental data and theoretical calculations of cross sections within the classic approach developed by Boltzmann and Langevin for a wid...

  18. On the stream-accretion disk interaction - Response to increased mass transfer rate

    Science.gov (United States)

    Dgani, Ruth; Livio, Mario; Soker, Noam

    1989-01-01

    The time-dependent interaction between the stream of mass from the inner Lagrangian point and the accretion disk, resulting from an increasing mass transfer rate is calculated. The calculation is fully three-dimensional, using a pseudoparticle description of the hydrodynamics. It is demonstrated that the results of such calculations, when combined with specific observations, have the potential of both determining essential parameters, such as the viscosity parameter alpha, and can distinguish between different models of dwarf nova eruptions.

  19. High-rate nanoscale offset printing process using directed assembly and transfer of nanomaterials.

    Science.gov (United States)

    Cho, Hanchul; Somu, Sivasubramanian; Lee, Jin Young; Jeong, Hobin; Busnaina, Ahmed

    2015-03-11

    High-rate nanoscale offset printing using a newly developed reusable template enables the assembly of nanomaterials into nanostructures followed by their transfer onto a flexible substrate in a few minutes. The developed template can potentially be reused more than 100 times in the offset printing process without any additional functionalization. This approach provides a new way for the printing of flexible devices with nanoscale patterns.

  20. Dependence of dose coefficients for {sup 239}Pu on transfer rates and absorption parameters

    Energy Technology Data Exchange (ETDEWEB)

    Suzuki, K.; Sekimoto, H. [Research Laboratory for Nuclear Reactors, Tokyo Institute of Technology, Tokyo (Japan); Ishigure, N. [Division of Radiotoxicology and Protection, National Institute of Radiological Sciences, Chiba (Japan)

    2000-05-01

    As it is reported of the biokinetic models and parameter values of the International Commission on Radiological Protection (ICRP) for dose estimation have uncertainties owing to insufficiency of human data. For most radionuclides, the data underlying such models and parameters of ICRP usually depend on animal experiments. Moreover, these values or model parameter are also greatly different between mammalian species. Recently, various radiation protection organizations are considering the biokinetic uncertainties from standpoints of data's sources, quality and completeness. In practice, a sensitivity analysis of doses to parameters is significant for the purpose of risk assessment. In general, movement or material in the body is depicted as a system of first-order processes, and parameter values are expressed as transfer rates between compartments. In this study, we made a code to reproduce the ICRP's dose coefficients for {sup 239}Pu, which is one of the most important elements for occupational exposure and its effective dose is much concerned with its own distribution in the body for dominance of alpha-decay. By using this code, we modified each transfer rate in a factor of 2, 3 and 4 in order to evaluate the effects, and calculated the sensitivities of effective doses due to these changes. Additionally, we examined the effects of modification of absorption parameters f{sub r}, S{sub r} and S{sub s}, which represent the absorption of particles from respiratory tract into blood. Consequently, the transfer rates that give a large sensitivity were specified, and it was shown that changes of transfer rates and absorption parameters are not so influential on effective doses for {sup 239}Pu in many cases. (author)

  1. 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

  2. Standard Test Method for Measuring Heat Transfer Rate Using a Thin-Skin Calorimeter

    CERN Document Server

    American Society for Testing and Materials. Philadelphia

    2005-01-01

    1.1 This test method covers the design and use of a thin metallic calorimeter for measuring heat transfer rate (also called heat flux). Thermocouples are attached to the unexposed surface of the calorimeter. A one-dimensional heat flow analysis is used for calculating the heat transfer rate from the temperature measurements. Applications include aerodynamic heating, laser and radiation power measurements, and fire safety testing. 1.2 Advantages 1.2.1 Simplicity of ConstructionThe calorimeter may be constructed from a number of materials. The size and shape can often be made to match the actual application. Thermocouples may be attached to the metal by spot, electron beam, or laser welding. 1.2.2 Heat transfer rate distributions may be obtained if metals with low thermal conductivity, such as some stainless steels, are used. 1.2.3 The calorimeters can be fabricated with smooth surfaces, without insulators or plugs and the attendant temperature discontinuities, to provide more realistic flow conditions for ...

  3. A limit on the energy transfer rate from the human fat store in hypophagia.

    Science.gov (United States)

    Alpert, Seymour S

    2005-03-07

    A limit on the maximum energy transfer rate from the human fat store in hypophagia is deduced from experimental data of underfed subjects maintaining moderate activity levels and is found to have a value of (290+/-25) kJ/kgd. A dietary restriction which exceeds the limited capability of the fat store to compensate for the energy deficiency results in an immediate decrease in the fat free mass (FFM). In cases of a less severe dietary deficiency, the FFM will not be depleted. The transition between these two dietary regions is developed and a criterion to distinguish the regions is defined. An exact mathematical solution for the decrease of the FFM is derived for the case where the fat mass (FM) is in its limited energy transfer mode. The solution shows a steady-state term which is in agreement with conventional ideas, a term indicating a slow decrease of much of the FFM moderated by the limited energy transferred from the fat store, and a final term showing an unprotected rapid decrease of the remaining part of the FFM. The average resting metabolic rate of subjects undergoing hypophagia is shown to decrease linearly as a function of the FFM with a slope of (249+/-25) kJ/kgd. This value disagrees with the results of other observers who have measured metabolic rates of diverse groups. The disagreement is explained in terms of individual metabolic properties as opposed to those of the larger population.

  4. The influence of the magnetic field on the heat transfer rate in rotating spherical shells

    Science.gov (United States)

    Cabello, Ares; Avila, Ruben

    2016-11-01

    Studies of the relationship between natural convection and magnetic field generation in spherical annular geometries with rotation are essential to understand the internal dynamics of the terrestrial planets. In such studies it is important to calculate and analyze the heat transfer rate at the inner and the outer spheres that confine the spherical gap. Previous investigations indicate that the magnetic field has a stabilizing effect on the onset of the natural convection, reduces the intensity of convection and modifies the flow patterns. However so far it is still unclear how the magnetic field change the heat transfer rate behaviour. We investigate the heat transfer rate (Nu) in a rotating spherical gap with a self gravity field varying linearly with radius, and its relation with the intensity of the magnetic field induced by the geodynamo effect. The Boussinesq fluid equations are solved by using a spectral element method (SEM). To avoid the singularity at the poles, the cubed-sphere algorithm is used to generate the spherical mesh. Several cases are simulated in which the Rayleigh number, the magnetic Reynolds number and the Taylor number are the variable parameters. The flow patterns, the temperature distribution and the Nusselt numbers at both spheres are calculated. Special thanks to DGAPA-UNAM Project PAPIIT IN11731, sponsor of this investigation.

  5. Quantifying the Interactions between Maternal and Fetal Heart Rates by Transfer Entropy.

    Science.gov (United States)

    Marzbanrad, Faezeh; Kimura, Yoshitaka; Palaniswami, Marimuthu; Khandoker, Ahsan H

    2015-01-01

    Evidence of the short term relationship between maternal and fetal heart rates has been found in previous studies. However there is still limited knowledge about underlying mechanisms and patterns of the coupling throughout gestation. In this study, Transfer Entropy (TE) was used to quantify directed interactions between maternal and fetal heart rates at various time delays and gestational ages. Experimental results using maternal and fetal electrocardiograms showed significant coupling for 63 out of 65 fetuses, by statistically validating against surrogate pairs. Analysis of TE showed a decrease in transfer of information from fetus to the mother with gestational age, alongside the maturation of the fetus. On the other hand, maternal to fetal TE was significantly greater in mid (26-31 weeks) and late (32-41 weeks) gestation compared to early (16-25 weeks) gestation (Mann Whitney Wilcoxon (MWW) pheart rate being larger than 4 msec in the late gestation. This difference was not observed for the fetuses with smaller RMSSD, which could be associated with the quiet sleep state. Delay in the information transfer from mother to fetus significantly decreased (p = 0.03) from mid to late gestation, implying a decrease in fetal response time. These changes occur concomitant with the maturation of the fetal sensory and autonomic nervous systems with advancing gestational age. The effect of maternal respiratory rate derived from maternal ECG was also investigated and no significant relationship was found between breathing rate and TE at any lag. In conclusion, the application of TE with delays revealed detailed information on the fetal-maternal heart rate coupling strength and latency throughout gestation, which could provide novel clinical markers of fetal development and well-being.

  6. 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.

  7. Information transfer with rate-modulated Poisson processes: a simple model for nonstationary stochastic resonance.

    Science.gov (United States)

    Goychuk, I

    2001-08-01

    Stochastic resonance in a simple model of information transfer is studied for sensory neurons and ensembles of ion channels. An exact expression for the information gain is obtained for the Poisson process with the signal-modulated spiking rate. This result allows one to generalize the conventional stochastic resonance (SR) problem (with periodic input signal) to the arbitrary signals of finite duration (nonstationary SR). Moreover, in the case of a periodic signal, the rate of information gain is compared with the conventional signal-to-noise ratio. The paper establishes the general nonequivalence between both measures notwithstanding their apparent similarity in the limit of weak signals.

  8. Information transfer with rate-modulated Poisson processes: A simple model for nonstationary stochastic resonance

    Science.gov (United States)

    Goychuk, Igor

    2001-08-01

    Stochastic resonance in a simple model of information transfer is studied for sensory neurons and ensembles of ion channels. An exact expression for the information gain is obtained for the Poisson process with the signal-modulated spiking rate. This result allows one to generalize the conventional stochastic resonance (SR) problem (with periodic input signal) to the arbitrary signals of finite duration (nonstationary SR). Moreover, in the case of a periodic signal, the rate of information gain is compared with the conventional signal-to-noise ratio. The paper establishes the general nonequivalence between both measures notwithstanding their apparent similarity in the limit of weak signals.

  9. Influence of medium composition on oxygen transfer rate in animal cell culture

    OpenAIRE

    Toye, Dominique; Galifi, Alain; Salmon, Thierry; Verdin, Emeline; Crine, Michel

    2009-01-01

    Experiments were conducted in a 0.25 m diameter bubble column to investigate the effect of animal cell culture medium composition on oxygen transfer rate. Air is used as the dispersed phase. The gas superficial velocity is varied between 0.4 and 2 cm/s (aeration rate ranging between 0.05 and 0.25 vvm) and the bubble column is thus operated in the homogenous regime. Aqueous solutions the composition (electrolyte, protein concentrations) of which mimics a mammalian cell culture medium) are us...

  10. Physical structure and genetic expression of the sulfonamide-resistance plasmid pLS80 and its derivatives in Streptococcus pneumoniae and Bacillus subtilis

    Energy Technology Data Exchange (ETDEWEB)

    Lopez, P.; Espinosa, M.; Lacks, S.A.

    1984-01-01

    The 10-kb chromosomal fragment of Streptococcus pneumoniae cloned in pLS80 contains the sul-d allele of the pneumococcal gene for dihydropteroate synthase. As a single copy in the chromosome this allele confers resistance to sulfanilamide at 0.2 mg/ml; in the multicopy plasmid it confers resistance to 2.0 mg/ml. The sul-d mutation was mapped by restriction analysis to a 0.4-kb region. A spontaneous deletion beginning approx. 1.5 kb to the right of the sul-d mutation prevented gene function, possibly by removing a promoter. This region could be restored by chromosomal facilitation and be demonstrated in the plasmid by selection for sulfonamide resistance. Under selection for a vector marker, tetracycline resistance, only the deleted plasmid was detectable, apparently as a result of plasmid segregation and the advantageous growth rates of cells with smaller plasmids. When such cells were selected for sulfonamide resistance, the deleted region returned to the plasmid, presumably by equilibration between the chromosome and the plasmid pool, to give a low frequency (approx. 10/sup -3/) of cells resistant to sulfanilamide at 2.0 mg/ml. Models for the mechanisms of chromosomal facilitation and equilibration are proposed. Several derivatives of pLS80 could be transferred to Bacillus subtilis, where they conferred resistance to sulfanilamide at 2 mg/ml, thereby demonstrating cross-species expression of the pneumococcal gene. Transfer of the plasmids to B. subtilis gave rise to large deletions to the left of the sul-d marker, but these deletions did not interfere with the sul-d gene function. Restriction maps of pLS80 and its variously deleted derivatives are presented.

  11. Effect of oxygen transfer rate on cellulases production in stirred tank and internal-loop airlift bioreactors

    OpenAIRE

    Michelin, Michele; Mota,André; Silva,Daniel Pereira da; A.A. Vicente; Teixeira, J. A; Polizeli, Maria de Lourdes T. M.

    2013-01-01

    In an aerobic process, such as enzymes production by fungi, the oxygen supply into fermentation medium is an important factor in order to achieve good productivities. Oxygen has an important role in metabolism and microorganism growth, being of extreme importance the control of both the dissolved oxygen transfer rate into the bioreactor and the oxygen consumption by the microorganism [1,2]. Dissolved oxygen transfer rate can be analyzed and described by means of the mass transfer coefficient,...

  12. [Investigation of plasmid-mediated quinolone resistance in Escherichia coli strains].

    Science.gov (United States)

    Aktepe, Orhan Cem; Aşık, Gülşah; Cetinkol, Yeliz; Biçmen, Meral; Gülay, Zeynep

    2012-01-01

    Quinolones are widely used antimicrobial agents, particularly for the treatment of infections caused by gram-negative bacilli such as E.coli. As a consequence, quinolone resistance has been increasing among this species in recent years. Bacterial resistance to quinolones usually results from mutations in the chromosomal genes which encode topoisomerases and also the expression of efflux pumps and loss of porines contributed to development of quinolone resistance. However, recent studies have shown that the spread and increase of quinolone resistance may be due to the transfer of plasmid-mediated genes. To date, three groups of plasmid-mediated quinolone resistance genes, namely qnr, aac(6')-Ib-cr, and qepA, have been described. The aim of this study was to investigate the presence of plasmid-mediated quinolone resistance genes in E.coli clinical isolates. A total of 112 quinolone-resistant E.coli strains isolated from different clinical specimens (84 urine, 16 blood, 10 wound, 2 bronchoalveolar lavage) of which 78 (69.6%) were extended-spectrum beta-lactamase (ESBL) positive, in Afyon Kocatepe University Hospital, Microbiology Laboratory were included in the study. In the isolates, qnrA, qnrB, qnrS, qnrC, qepA, and aac(6')-1b-cr plasmid genes were analysed by polymerase chain reaction (PCR). After aac(6')- 1b determinant was amplified by PCR, all aac(6')-1b positive amplicons were analyzed by digestion with BseGI restriction enzyme to identify aac(6')-1b-cr variant. It was found that, none of the strains horboured qnrA, qnrB, qnrS, qnrC and qepA genes, however, plasmid-mediated quinolone resistance gene aac(6')-1b-cr was found positive in 59.8% (67/112) of the strains. It was notable that 86.6% (58/67) of those isolates were ESBL producers. The rates of quinolone resistance among E.coli isolates infections were high in our region and an increasing trend has been observed in recent years. Our data indicated that the presence of plasmid- mediated resistance genes

  13. Effect of Radiocesium Transfer on Ambient Dose Rate in Forest Environment

    Science.gov (United States)

    Kato, Hiroaki; Onda, Yuichi; Loffredo, Nicolas; Hisadome, Keigo; Kawamori, Ayumi

    2014-05-01

    We investigated the transfer of canopy-intercepted radiocesium to the forest floor following the Fukushima Daiichi nuclear power plant accident. The cesium-137 (Cs-137) contents of throughfall, stemflow, and litterfall were monitored in two coniferous stands (plantation of Japanese cedar) and a deciduous broad-leaved forest stand (beech with red pine). We also measured an ambient dose rate at different height in the forest by using a survey meter (TCS-172B, Hitachi-Aloka Medical, LTD.) and a portable Ge gamma-ray detector (Detective-DX-100T, Ortec, Ametek, Inc.). In decreasing order of total Cs-137 deposition from the canopy to forest floor were the mature cedar stand, the young cedar stand, and the broad-leaved forest. The ambient dose rate in forest exhibited height dependency and its vertical distribution varied by forest type and stand age. The ambient dose rate showed an exponential decrease with time for all the forest sites, however the decreasing trend differed depending on the height of dose measurement and forest type. The ambient dose rates at the canopy (approx. 10 m-) decreased earlier than physical attenuation of radiocesium, whereas those at the forest floor varied among three forest stands. These data suggested that an ambient dose rate in forest environment can be variable in spatially and temporally reflecting the transfer of radiocesium from canopy to forest floor.

  14. Temperature and detection-wavelength dependence of the electron transfer rates in initial stages of photosynthesis.

    Science.gov (United States)

    Kurzynski, Michal; Chelminiak, Przemyslaw

    2013-10-17

    Unusual temperature behavior, observed in the initial electron transfer stages in the photosynthetic reaction centers of the purple bacteria, and a strong probing pulse wavelength dependence of transfer rates, determined in transient absorption spectroscopy, can easily be explained on assuming that the transfer takes place from dynamically unrelaxed states of protein environment. The transitions from the primary special pair (P) to a single bacteriochlorophyll (B) and next to a bacteriopheophytin (H) are controlled by diffusion down the energy value of underdamped vibrational modes of frequency 200 K, probably determining distances between the succeeding cofactors. The subsequent transition to the quinone A (Q) is controlled by diffusion in the position value of an overdamped conformational mode, probably corresponding to the local polarization. From the fit of available experimental data to simple theoretical formulas, the important physical conclusion arises that the very electronic transitions are fast as compared to the relaxation processes and, in the first approximation, only the latter contribute to the overall times of the initial electron transfer stages in photosynthesis.

  15. ECTOPIC PREGNANCY RATES WITH CLEAVAGE STAGE EMBRYO TRANSFER AT DAY 3 VERSUS BLASTOCYST STAGE TRANSFER AT DAY 5: A PROSPECTIVE ANALYSIS

    Directory of Open Access Journals (Sweden)

    Prabhleen

    2014-10-01

    Full Text Available OBJECTIVE: The purpose of our study was to compare the tubal pregnancy rates between day 3 and day 5 transfers. As theoretically blastocyst transfer is said to decrease the incidence of ectopic pregnancy following IVF-ET due to the decreased uterine contractility reported on day 5. METHODS: A prospective analysis of all clinical pregnancies conceived in our IVF program between May 2010 to April 2011 was performed. The ectopic pregnancy rates were compared for day 3 and day 5 transfers. RESULTS: There were 44 pregnancies resulting from day 3 transfers of which one was ectopic (2.27%. In day 5 transfers, there was also one ectopic pregnancies out of 66 clinical pregnancies (1.52%, difference between these rates was not statistically significant (P>0.05 CONCLUSION: This suggests that the ectopic pregnancy rate is not reduced following blastocyst transfer on day 5 compared to cleavage stage embryo on day 3. While there may be several benefits to extended culture in IVF, the decision to offer blastocyst transfer should be made independently from the issue of ectopic pregnancy risk.

  16. Rate limiting activity of charge transfer during lithiation from ionic liquids

    Science.gov (United States)

    Rodrigues, Marco-Tulio F.; Lin, Xinrong; Gullapalli, Hemtej; Grinstaff, Mark W.; Ajayan, Pulickel M.

    2016-10-01

    Given the increased use of room temperature ionic liquid electrolytes in Li-ion batteries, due to their non-flammability and negligible volatility, this study evaluates the lithiation kinetics to understand and improve the rate performance of Li-ion batteries. Lithium titanate spinel is used as a model electrode and the electrolyte is composed of LiTFSI and TFSI-coordinated alkoxy-modified phosphonium ionic liquid. Based on the analysis of activation energies for each process, we report that the charge-transfer reaction at the electrode/electrolyte interface is the rate-limiting step for cell operation. This finding is further supported by the observation that a 50-fold decrease in charge-transfer resistance at higher temperatures leads to a significant performance improvement over that of a traditional organic electrolyte at room temperature. Charge-transfer resistance and electrolyte wetting on the electrode surface are critical processes for optimal battery performance, and such processes need to be included when designing new ionic liquids in order to exceed the power density obtained with the use of current carbonate-based electrolytes.

  17. Estimation of snag carbon transfer rates by ecozone and lead species for forests in Canada.

    Science.gov (United States)

    Hilger, A B; Shaw, C H; Metsaranta, J M; Kurz, W A

    2012-12-01

    Standing dead trees (snags) and downed woody debris contribute substantially to the carbon (C) budget of Canada's forest. Accurate parameterization of the C transfer rates (CTRs) from snags to downed woody debris is important for forest C dynamics models such as the Carbon Budget Model of the Canadian Forest Sector (CBM-CFS3), but CTRs are rarely measured or reported in the literature. Therefore, forest C models generally use snag fall rates (FRs) available in the literature, as a proxy for CTRs. However, FRs are based on stem counts while CTRs refer to mass transfers. Stem mass and stem number are not linearly related, with small diameter trees representing disproportionately lower C mass transfers. Therefore this proxy, while convenient, may bias C transfer from standing dead to downed woody material. Here, we combined tree data from 10802 sample plots and previously published species-specific individual-tree relationships between tree diameter (diameter at breast height, dbh) and fall rate to derive stand-level estimates of CTRs for the CBM-CFS3. We estimated CTRs and FRs and used the FR values to validate this approach by comparing them with standardized FR values compiled from the literature. FRs generally differed from CTRs. The overall CTR (4.78% +/- 0.02% per year, mean +/- SE) was significantly smaller than the overall FR (5.40% +/- 0.02% per year; mean +/- SE). Both the difference between FR and CTR (FR - CTR) and the CTR itself varied by ecozone, with ecozone means for CTR ranging from 3.94% per year to 10.02% per year. This variation was explained, in part, by heterogeneity in species composition, size (dbh distribution), structure, and age of the stands. The overall mean CTR estimated for the Snag_Stemwood (4.78% per year) and the Snag_Branches (11.95% per year) pools of the CBM-CFS3 were approximately 50% and 20% higher than the current default rates used in the CBM-CFS3 of 3.2% and 10.0%, respectively. Our results demonstrate that using CTRs to

  18. Semiclassical Green's functions and an instanton formulation of electron-transfer rates in the nonadiabatic limit

    CERN Document Server

    Richardson, Jeremy O; Thoss, Michael

    2015-01-01

    We present semiclassical approximations to Green's functions of multidimensional systems, extending Gutzwiller's work to the classically forbidden region. Based on steepest-descent integrals over these functions, we derive an instanton method for computing the rate of nonadiabatic reactions, such as electron transfer, in the weak-coupling limit, where Fermi's golden-rule can be employed. This generalizes Marcus theory to systems for which the environment free-energy curves are not harmonic and where nuclear tunnelling plays a role. The derivation avoids using the Im F method or short-time approximations to real-time correlation functions. A clear physical interpretation of the nuclear tunnelling processes involved in an electron-transfer reaction is thus provided. In the following paper, we discuss numerical evaluation of the formulae.

  19. A general real-time formulation for multi-rate mass transfer problems

    Directory of Open Access Journals (Sweden)

    O. Silva

    2009-08-01

    Full Text Available Many flow and transport phenomena, ranging from delayed storage in pumping tests to tailing in river or aquifer tracer breakthrough curves or slow kinetics in reactive transport, display non-equilibrium (NE behavior. These phenomena are usually modeled by non-local in time formulations, such as multi-porosity, multiple processes non equilibrium, continuous time random walk, memory functions, integro-differential equations, fractional derivatives or multi-rate mass transfer (MRMT, among others. We present a MRMT formulation that can be used to represent all these models of non equilibrium. The formulation can be extended to non-linear phenomena. Here, we develop an algorithm for linear mass transfer, which is accurate, computationally inexpensive and easy to implement in existing groundwater or river flow and transport codes. We illustrate this approach by application to published data involving NE groundwater flow and solute transport in rivers and aquifers.

  20. Semiclassical Green's functions and an instanton formulation of electron-transfer rates in the nonadiabatic limit

    Science.gov (United States)

    Richardson, Jeremy O.; Bauer, Rainer; Thoss, Michael

    2015-10-01

    We present semiclassical approximations to Green's functions of multidimensional systems, extending Gutzwiller's work to the classically forbidden region. Based on steepest-descent integrals over these functions, we derive an instanton method for computing the rate of nonadiabatic reactions, such as electron transfer, in the weak-coupling limit, where Fermi's golden-rule can be employed. This generalizes Marcus theory to systems for which the environment free-energy curves are not harmonic and where nuclear tunnelling plays a role. The derivation avoids using the Im F method or short-time approximations to real-time correlation functions. A clear physical interpretation of the nuclear tunnelling processes involved in an electron-transfer reaction is thus provided. In Paper II [J. O. Richardson, J. Chem. Phys. 143, 134116 (2015)], we discuss numerical evaluation of the formulae.

  1. Determination of Erosion/Corrosion Rates in Hanford Tank Farms Radioactive Waste Transfer System Pipelines

    Energy Technology Data Exchange (ETDEWEB)

    Washenfelder, D. J.; Girardot, C. L.; Wilson, E. R.; Page, J. A.; Engeman, J. K.; Gunter, J. R.; Johnson, J. M.; Baide, D. G.; Cooke, G. A.; Larson, J. D.; Castleberry, J. L.; Boomer, K. D.

    2015-11-05

    The twenty-eight double-shell underground radioactive waste storage tanks at the U. S. Department of Energy’s Hanford Site near Richland, WA are interconnected by the Waste Transfer System network of buried steel encased pipelines and pipe jumpers in below-grade pits. The pipeline material is stainless steel or carbon steel in 51 mm to 152 mm (2 in. to 6 in.) sizes. The pipelines carry slurries ranging up to 20 volume percent solids and supernatants with less than one volume percent solids at velocities necessary to prevent settling. The pipelines, installed between 1976 and 2011, were originally intended to last until the 2028 completion of the double-shell tank storage mission. The mission has been subsequently extended. In 2010 the Tank Operating Contractor began a systematic evaluation of the Waste Transfer System pipeline conditions applying guidelines from API 579-1/ASME FFS-1 (2007), Fitness-For-Service. Between 2010 and 2014 Fitness-for-Service examinations of the Waste Transfer System pipeline materials, sizes, and components were completed. In parallel, waste throughput histories were prepared allowing side-by-side pipeline wall thinning rate comparisons between carbon and stainless steel, slurries and supernatants and throughput volumes. The work showed that for transfer volumes up to 6.1E+05 m3 (161 million gallons), the highest throughput of any pipeline segment examined, there has been no detectable wall thinning in either stainless or carbon steel pipeline material regardless of waste fluid characteristics or throughput. The paper describes the field and laboratory evaluation methods used for the Fitness-for-Service examinations, the results of the examinations, and the data reduction methodologies used to support Hanford Waste Transfer System pipeline wall thinning conclusions.

  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. Risk Factors Influencing Conception Rate in Holstein Heifers before Artificial Insemination or Embryo Transfer

    Directory of Open Access Journals (Sweden)

    M. Yusuf

    2016-12-01

    Full Text Available The objective of this study was to show the risk factors affecting the conception rate in Holstein heifers after synchronization of estrus. A total of 275 Holstein heifers housed in a free barn were used for the experiment. The herd was visited regularly at four week intervals for synchronization of estrus using Heatsynch and CIDR-Heatsynch protocols. A group of four to 14 animals, depending on the availability, were referred to the experiment at each visit. Estrus induction rates in the two protocols were 93.9% and 94.9%, respectively. There was no difference in the conception rate between the two protocols. Conception rate after artificial insemination (AI or embryo transfer (ET were 46.3% and 51.4%, respectively. The risk factors affecting conception rate in heifers were daily weight gain (odds ratio [OR]= 4.673; P= 0.036 and body condition score (BCS (OR= 3.642; P= 0.018. Furthermore, estrus synchronization protocol (OR= 1.774; P= 0.083 and the absence of corpus luteum (CL at the initiation of treatment (OR= 0.512; P= 0.061 had a tendency to affect the conception rate, while age (OR= 0.715; P= 0.008 was a protective factor to conception rate.  In conclusion, positive daily weight gain before AI or ET, higher BCS, younger age, and the presence of CL at the initiation of estrus synchronization in dairy heifers increased the likelihood to conceive.

  4. Cross-language differences of articulation rate and its transfer into Japanese as a second language.

    Science.gov (United States)

    Amino, Kanae; Osanai, Takashi

    2015-04-01

    Recently, the articulation rate has been attracting attention in forensic speech investigation as an acoustic feature that varies across speakers, dialects, and languages. The present study investigates how cross-language differences in the articulation rate are transferred into Japanese as a second language. Participants were speakers of Japanese, Chinese, Korean, and Thai. They were recorded while they read a passage in their native language and in Japanese. Local and global articulation rates were calculated based on the number of syllables as well as the number of morae for Japanese speech. When we compared the articulation rate of the native languages, Japanese was the fastest, then Korean, Chinese, and Thai in that order. Also, a significant positive correlation was observed between the articulation rate of the native language and that of the second language. A gender difference was found in the articulation rate of some languages, with males speaking faster than females. The effect of age was limited to Thai speakers only. Accent discrimination and identification experiments were conducted and the results revealed that native and non-native accents could be correctly discriminated just by the articulation rate.

  5. Predicted Modulated Differential Rates for Direct WIMP Searches at Low Energy Transfers

    Institute of Scientific and Technical Information of China (English)

    J.D. Vergados

    2012-01-01

    The differential event rate for direct detection of dark matter, both the time averaged and the modulated one due to the motion of the Earth, are discussed. The calculations focus on relatively light cold dark matter candidates (WIMP) and low energy transfers. It is shown that for sufficiently light WIMPs the extraction of relatively large nucleon cross sections is possible. Furthermore for some WIMP masses the modulation amplitude may change sign, meaning that, in such a case, the maximum rate may occur six months later than naively expected. This effect can be exploited to yield information about the mass of the dark matter candidate, if and when the observation of the modulation of the event rate is established.

  6. Negative transfer of heart rate control following biofeedback training: a partial replication.

    Science.gov (United States)

    Steptoe, A; Macready, D

    1985-09-01

    Ability to raise and lower heart rate (HR) on instruction was tested before and after unidirectional biofeedback training in two groups of 10 male volunteers. Instructional control was assessed in 2-min trials before training, and after 5 and 10 biofeedback trials of increasing (Group I) and decreasing (Group D) HR. The magnitude of HR elevations produced by Group D diminished following training, while modifications in Group I were unchanged. This negative transfer effect is discussed in relation to whether voluntary speeding and slowing HR reflect distinct capacities.

  7. 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...

  8. Klebsiella pneumoniae multiresistance plasmid pMET1: similarity with the Yersinia pestis plasmid pCRY and integrative conjugative elements.

    Directory of Open Access Journals (Sweden)

    Alfonso J C Soler Bistué

    Full Text Available BACKGROUND: Dissemination of antimicrobial resistance genes has become an important public health and biodefense threat. Plasmids are important contributors to the rapid acquisition of antibiotic resistance by pathogenic bacteria. PRINCIPAL FINDINGS: The nucleotide sequence of the Klebsiella pneumoniae multiresistance plasmid pMET1 comprises 41,723 bp and includes Tn1331.2, a transposon that carries the bla(TEM-1 gene and a perfect duplication of a 3-kbp region including the aac(6'-Ib, aadA1, and bla(OXA-9 genes. The replication region of pMET1 has been identified. Replication is independent of DNA polymerase I, and the replication region is highly related to that of the cryptic Yersinia pestis 91001 plasmid pCRY. The potential partition region has the general organization known as the parFG locus. The self-transmissible pMET1 plasmid includes a type IV secretion system consisting of proteins that make up the mating pair formation complex (Mpf and the DNA transfer (Dtr system. The Mpf is highly related to those in the plasmid pCRY, the mobilizable high-pathogenicity island from E. coli ECOR31 (HPI(ECOR31, which has been proposed to be an integrative conjugative element (ICE progenitor of high-pathogenicity islands in other Enterobacteriaceae including Yersinia species, and ICE(Kp1, an ICE found in a K. pneumoniae strain causing primary liver abscess. The Dtr MobB and MobC proteins are highly related to those of pCRY, but the endonuclease is related to that of plasmid pK245 and has no significant homology with the protein of similar function in pCRY. The region upstream of mobB includes the putative oriT and shares 90% identity with the same region in the HPI(ECOR31. CONCLUSIONS: The comparative analyses of pMET1 with pCRY, HPI(ECOR31, and ICE(Kp1 show a very active rate of genetic exchanges between Enterobacteriaceae including Yersinia species, which represents a high public health and biodefense threat due to transfer of multiple resistance

  9. 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.

  10. 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.

  11. 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.

  12. 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.

  13. Effect of meloxicam on pregnancy rate of recipient heifers following transfer of in vitro produced embryos.

    Science.gov (United States)

    Aguiar, T S; Araújo, C V; Tirloni, R R; Martins, L R

    2013-12-01

    The main objective of this study was to determine if administration of meloxicam, a cyclooxygenase (COX) two inhibitor, to heifers in which embryo transfer (ET) is more difficult and requires a greater manipulation of the tract, would be beneficial. Nulliparous recipient heifers were divided in two groups: CON (n = 102), in which animals received 10 ml of saline IM (the same volume of meloxicam) and MEL (n = 105) animals that were treated with meloxicam. According to the degree in passing the catheter, recipients from both groups were classified as Grade I, easy (meloxicam (10 ml).There was no difference in the pregnancy rates on Day 35 considering animals which presented Grade I cervix independently whether the treatment was performed or not (p = 0.22). There was a statistical difference in the pregnancy rates (p meloxicam had a positive influence on general pregnancy rate of treated heifers in comparison to non-treated heifers. It was also observed that pregnancy rate was not influenced by meloxicam administration in Grade I heifers. Treatment increased the pregnancy rate of Grade II heifers.

  14. Investigating rate-limiting barriers to nanoscale nonviral gene transfer with nanobiophotonics

    Science.gov (United States)

    Chen, Hunter H.

    Nucleic acids are a novel class of therapeutics poised to address many unmet clinical needs. Safe and efficient delivery remains a significant challenge that has delayed the realization of the full therapeutic potential of nucleic acids. Nanoscale nonviral vectors offer an attractive alternative to viral vectors as natural and synthetic polymers or polypeptides may be rationally designed to meet the unique demands of individual applications. A mechanistic understanding of cellular barriers is necessary to develop guidelines for designing custom gene carriers which are expected to greatly impact this delivery challenge. The work herein focused on the relationships among nanocomplex stability, intracellular trafficking and unpacking kinetics, and DNA degradation. Ultrasensitive nanosensors based on QD-FRET were developed to characterize the biophysical properties of nanocomplexes and study these rate-limiting steps. Quantitative image analysis enabled the distributions of the subpopulation of condensed or released DNA to be determined within the major cellular compartments encountered during gene transfer. The steady state stability and unpacking kinetics within these compartments were found to impact transgene expression, elucidating multiple design strategies to achieve efficient gene transfer. To address enzymatic barriers, a novel two-step QD-FRET nanosensor was developed to analyze unpacking and DNA degradation simultaneously, which has not been accomplished previously. Bioresponsive strategies such as disulfide crosslinking and thermosensitivity were evaluated by QD-FRET and quantitative compartmental analysis as case studies to determine appropriate design specifications for thiolated polymers and thermoresponsive polypeptides. Relevant nanobiophotonic tools were developed as a platform to study major rate-limiting barriers to nanomedicine and demonstrated the feasibility of using mechanistic information gained from these tools to guide the rational design of

  15. A novel estimator for the rate of information transfer by continuous signals.

    Directory of Open Access Journals (Sweden)

    Jouni Takalo

    Full Text Available The information transfer rate provides an objective and rigorous way to quantify how much information is being transmitted through a communications channel whose input and output consist of time-varying signals. However, current estimators of information content in continuous signals are typically based on assumptions about the system's linearity and signal statistics, or they require prohibitive amounts of data. Here we present a novel information rate estimator without these limitations that is also optimized for computational efficiency. We validate the method with a simulated Gaussian information channel and demonstrate its performance with two example applications. Information transfer between the input and output signals of a nonlinear system is analyzed using a sensory receptor neuron as the model system. Then, a climate data set is analyzed to demonstrate that the method can be applied to a system based on two outputs generated by interrelated random processes. These analyses also demonstrate that the new method offers consistent performance in situations where classical methods fail. In addition to these examples, the method is applicable to a wide range of continuous time series commonly observed in the natural sciences, economics and engineering.

  16. Experimental investigation on heat transfer rate of Co–Mn ferrofluids in external magnetic field

    Directory of Open Access Journals (Sweden)

    Margabandhu M.

    2016-06-01

    Full Text Available Manganese substituted cobalt ferrite (Co1–xMnxFe2O4 with x = 0, 0.3, 0.5, 0.7 and 1 nanopowders were synthesized by chemical coprecipitation method. The synthesized magnetic nanoparticles were investigated by various characterization techniques, such as X-ray diffraction (XRD, vibrating sample magnetometry (VSM, scanning electron microscopy (SEM and thermogravimetric and differential thermal analysis (TG/DTA. The XRD results confirmed the presence of cubic spinel structure of the prepared powders and the average crystallite size of magnetic particles ranging from 23 to 45 nm. The VSM results showed that the magnetic properties varied with an increase in substituted manganese while SEM analysis showed the change in the morphology of obtained magnetic nanoparticles. The TG/DTA analysis indicated the formation of crystalline structure of the synthesized samples. The heat transfer rate was measured in specially prepared magnetic nanofluids (nanoparticles dispersed in carrier fluid transformer oil as a function of time and temperature in presence of external magnetic fields. The experimental analysis indicated enhanced heat transfer rate of the magnetic nanofluids which depended upon the strength of external magnetic field and chemical composition.

  17. 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.

  18. Effect of injection rate on contrast-enhanced MR angiography image quality: Modulation transfer function analysis.

    Science.gov (United States)

    Clark, Toshimasa J; Wilson, Gregory J; Maki, Jeffrey H

    2017-07-01

    Contrast-enhanced (CE)-MRA optimization involves interactions of sequence duration, bolus timing, contrast recirculation, and both R1 relaxivity and R2*-related reduction of signal. Prior data suggest superior image quality with slower gadolinium injection rates than typically used. A computer-based model of CE-MRA was developed, with contrast injection, physiologic, and image acquisition parameters varied over a wide gamut. Gadolinium concentration was derived using Verhoeven's model with recirculation, R1 and R2* calculated at each time point, and modulation transfer curves used to determine injection rates, resulting in optimal resolution and image contrast for renal and carotid artery CE-MRA. Validation was via a vessel stenosis phantom and example patients who underwent carotid CE-MRA with low effective injection rates. Optimal resolution for renal and carotid CE-MRA is achieved with injection rates between 0.5 to 0.9 mL/s and 0.2 to 0.3 mL/s, respectively, dependent on contrast volume. Optimal image contrast requires slightly faster injection rates. Expected signal-to-noise ratio varies with both contrast volume and cardiac output. Simulated vessel phantom and clinical carotid CE-MRA exams at an effective contrast injection rate of 0.4 to 0.5 mL/s demonstrate increased resolution. Optimal image resolution is achieved at intuitively low, effective injection rates (0.2-0.9 mL/s, dependent on imaging parameters and contrast injection volume). Magn Reson Med 78:357-369, 2017. © 2016 International Society for Magnetic Resonance in Medicine. © 2016 International Society for Magnetic Resonance in Medicine.

  19. Solvent effect on intramolecular electron transfer rates of mixed-valence biferrocene monocation derivatives.

    Science.gov (United States)

    Masuda, Y; Shimizu, C

    2006-06-01

    Intramolecular electron transfer (ET) rates in various solvents of mixed-valence biferrocene monocation (Fe(II), Fe(III)) and the 1',1' ''-diiodo and 1',1' ''-diethyl derivatives (respectively abbreviated as BFC(+), I(2)BFC(+), and Et(2)BFC(+)) were determined by means of the spin-lattice relaxation times of the protons, taking into account the local magnetic field fluctuation caused by the electron hopping between the two ferrocene units. We also determined the ET rates of a mixed-valence diferrocenylacetylene monocation (DFA(+)) in order to examine the effect of the insertion of an acetylene bridge between the two ferrocene units. The insertion of the bridge decreased the ET rate, while the effect of substitution on the cyclopentadienyl rings on the rate was minor. The observed rates for each mixed-valence monocation in various solvents did not correlate with the reorganization energies, but we did find a significant contribution of the solvent dynamics. The observed rates were considerably higher than those expected on the basis of the Sumi-Marcus-Nalder model in which the solvents were regarded as dielectric continua. The slope of the logarithm plot of the pre-exponential factors in various solvents for each mixed-valence monocation versus the inverse of the longitudinal dielectric relaxation times of the solvents was significantly smaller than unity, and the slope for DFA(+) was larger than those for BFC(+), I(2)BFC(+), and Et(2)BFC(+). These results were ascribed to a partial contribution of the dielectric friction to the dynamics along the solvent coordinate; the extent of the contribution decreased with a reduction in the ET distance. For the dynamics along the solvent coordinate of the ET reactions in methanol, the observed rates indicated an important contribution by the minor dielectric relaxation components with faster relaxation times, rather than the major component with an extraordinarily long relaxation time.

  20. Low vertical transfer rates of carbon inferred from radiocarbon analysis in an Amazon Podzol

    Directory of Open Access Journals (Sweden)

    C. A. Sierra

    2013-06-01

    Full Text Available Hydromorphic Podzol soils in the Amazon Basin generally support low-stature forests with some of the lowest amounts of aboveground net primary production (NPP in the region. However, they can also exhibit large values of belowground NPP that can contribute significantly to the total annual inputs of organic matter into the soil. These hydromorphic Podzol soils also exhibit a horizon rich in organic matter at around 1–2 m depth, presumably as a result of eluviation of dissolved organic matter and sesquioxides of Fe and Al. Therefore, it is likely that these ecosystems store large quantities of carbon by (1 large amounts of C inputs to soils dominated by their high levels of fine-root production, (2 stabilization of organic matter in an illuviation horizon due to significant vertical transfers of C. To assess these ideas we studied soil carbon dynamics using radiocarbon in two adjacent Amazon forests growing on contrasting soils: a hydromorphic Podzol and a well-drained Alisol supporting a high-stature terra firme forest. Our measurements showed similar concentrations of C and radiocarbon in the litter layer and the first 5 cm of the mineral soil for both sites. This result is consistent with the idea that the hydromorphic Podzol soil has similar soil C storage and cycling rates compared to the well-drained Alisol that supports a more opulent vegetation. However, we found important differences in carbon dynamics and transfers along the vertical profile. At both soils, we found similar radiocarbon concentrations in the subsoil, but the carbon released after incubating soil samples presented radiocarbon concentrations of recent origin in the Alisol, but not in the Podzol. There were no indications of incorporation of C fixed after 1950 in the illuvial horizon of the Podzol. With the aid of a simulation model, we predicted that only a minor fraction (1.7% of the labile carbon decomposed in the topsoil is transferred to the subsoil of the Podzol

  1. Low vertical transfer rates of carbon inferred from radiocarbon analysis in an Amazon podzol

    Directory of Open Access Journals (Sweden)

    C. A. Sierra

    2013-02-01

    Full Text Available Hydromorphic podzol soils in the Amazon Basin generally support low-stature forests with some of the lowest amounts of aboveground net primary production (NPP in the region. However, they can also exhibit large values of belowground NPP that can contribute significantly to the total annual inputs of organic matter into the soil. These hydromorphic podzol soils also exhibit a horizon rich in organic matter at around 1 m depth, presumably as a result of elluviation of dissolved organic matter and sesquioxides of Fe and Al. Therefore, it is likely that these ecosystems store large quantities of carbon by (1 large amounts of C inputs to soils dominated by their high levels of fine-root production, (2 stabilization of organic matter in an illuviation horizon due to significant vertical transfers of C. To assess these ideas we studied soil carbon dynamics using radiocarbon in two adjacent Amazon forests growing on contrasting soils, a hydromorphic podzol and a well-drained alisol supporting a high-stature terra firme forest. Our measurements showed similar concentrations of C and radiocarbon in the litter layer and the first 5 cm of the mineral soil for both sites. This result is consistent with the idea that the hydromorphic podzol soil has similar soil C storage and cycling rates compared to the well-drained alisol that supports a more opulent vegetation. However, we found important differences in carbon dynamics and transfers along the vertical profile. At both soils, we found similar radiocarbon concentrations in the subsoil, but the carbon released after incubating soil samples presented radiocarbon concentrations of recent origin in the alisol, but not in the podzol. There were no indications of incorporation of C fixed after 1950 in the illuvial horizon of the podzol. With the aid of a simulation model, we predicted that only a minor fraction (1.7% of the labile carbon decomposed in the topsoil is transferred to the subsoil of the podzol, while

  2. Characterization of plasmids that encode streptomycin-resistance in bacterial epiphytes of apple.

    Science.gov (United States)

    Huang, T C; Burr, T J

    1999-05-01

    Streptomycin resistance in strains of Pseudomonas syringae pv. papulans, Pantoea agglomerans and a yellow-pigmented, non-fluorescent Pseudomonas sp. (Py), isolated from apple orchards in New York and Washington states, is predominantly associated with strA-strB genes carried on conjugal plasmids (R plasmids). None of 128 resistant Erwinia amylovora strains from the eastern and western USA hybridized with a strA-strB probe, SMP3. Resistant Py strains transfered R plasmids to Ps. syringae pv. papulans and to Py in vitro at frequencies of 10(-1)-10(-2) per recipient cell whereas Ps. syringae pv. papulans transferred its plasmids at frequencies of 10(-2) to below detectable levels. Transfer of R plasmids to P. agglomerans was not detected and resistant P. agglomerans did not transfer their R plasmids to any recipients. R plasmids were found to be highly diverse as measured by DNA fingerprint analysis. Transfer-deficient transposon mutants of R plasmid pCPP519 were generated, and 3.9 kb EcoRI and 3.0 kb SmaI fragments that hybridized with a Tn5 probe were cloned and sequenced. The deduced amino acid sequences of the 3.9 kb fragment were similar to proteins involved in replication, nicking at oriT, and piliation in other bacteria.

  3. 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

  4. Enhanced rate of intramolecular electron transfer in an engineered purple CuA azurin.

    Science.gov (United States)

    Farver, O; Lu, Y; Ang, M C; Pecht, I

    1999-02-02

    The recent expression of an azurin mutant where the blue type 1 copper site is replaced by the purple CuA site of Paracoccus denitrificans cytochrome c oxidase has yielded an optimal system for examining the unique electron mediation properties of the binuclear CuA center, because both type 1 and CuA centers are placed in the same location in the protein while all other structural elements remain the same. Long-range electron transfer is induced between the disulfide radical anion, produced pulse radiolytically, and the oxidized binuclear CuA center in the purple azurin mutant. The rate constant of this intramolecular process, kET = 650 +/- 60 s-1 at 298 K and pH 5.1, is almost 3-fold faster than for the same process in the wild-type single blue copper azurin from Pseudomonas aeruginosa (250 +/- 20 s-1), in spite of a smaller driving force (0.69 eV for purple CuA azurin vs. 0.76 eV for blue copper azurin). The reorganization energy of the CuA center is calculated to be 0.4 eV, which is only 50% of that found for the wild-type azurin. These results represent a direct comparison of electron transfer properties of the blue and purple CuA sites in the same protein framework and provide support for the notion that the binuclear purple CuA center is a more efficient electron transfer agent than the blue single copper center because reactivity of the former involves a lower reorganization energy.

  5. 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.

  6. 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.

  7. Spatial statistical point prediction guidance for heating-rate-limited aeroassisted orbital transfer

    Science.gov (United States)

    Ghosh, Pradipto; Conway, Bruce A.

    2015-06-01

    Feedback control of constrained non-linear dynamical systems satisfying a certain optimality criterion and meeting a specified transfer objective in the state space is recognized as one of the most challenging problems in control theory. One approach to computing optimal feedback policies is the dynamic programming route of numerically solving the Hamilton-Jacobi-Bellman (HJB) partial differential equation directly. In this paper an alternate and more tractable dynamic programming approach, the optimal feedback synthesis method, is utilized. The effectiveness of this method is demonstrated through an explicit guidance scheme for the heating-rate-constrained maneuver of an Aeroassisted Transfer Vehicle (AOTV). In optimal feedback synthesis, a feedback chart is constructed from a family of open-loop extremals, thus ensuring optimality with respect to any initial condition in the family. This paper presents a solution to the AOTV optimal feedback synthesis problem using the Gaussian process spatial prediction method of universal kriging. A closed-form expression for a near-optimal guidance law is derived. Its performance is found to be very promising; initial atmospheric entry errors due to simulated thruster misfiring are seen to be accurately corrected while the algebraic state-inequality constraint is closely respected.

  8. Increasing The Number of Embryos Transferred from Two to Three, Does not Increase Pregnancy Rates in Good Prognosis Patients

    Directory of Open Access Journals (Sweden)

    Mahnaz Ashrafi

    2015-10-01

    Full Text Available Background: To compare the pregnancy outcomes after two embryos versus three embryos transfers (ETs in women undergoing in vitro fertilization (IVF/intracytoplasmic sperm injection (ICSI cycles. Materials and Methods: This retrospective study was performed on three hundred eighty seven women with primary infertility and with at least one fresh embryo in good quality in order to transfer at each IVF/ICSI cycle, from September 2006 to June 2010. Patients were categorized into two groups according to the number of ET as follows: ET2 and ET3 groups, indicating two and three embryos were respectively transferred. Pregnancy outcomes were compared between ET2 and ET3 groups. Chi square and student t tests were used for data analysis. Results: Clinical pregnancy and live birth rates were similar between two groups. The rates of multiple pregnancies were 27 and 45.2% in ET2 and ET3 groups, respectively. The rate of multiple pregnancies in young women was significantly increased when triple instead of double embryos were transferred. Logistic regression analysis indicated two significant prognostic variables for live birth that included number and quality of transferred embryos; it means that the chance of live birth following ICSI treatment increased 3.2-fold when the embryo with top quality (grade A was transferred, but the number of ET had an inverse relationship with live birth rate; it means that probability of live birth in women with transfer of two embryos was three times greater than those who had three ET. Conclusion: Due to the difficulty of implementation of the elective single-ET technique in some infertility centers in the world, we suggest transfer of double instead of triple embryos when at least one good quality embryo is available for transfer in women aged 39 years or younger. However, to reduce the rate of multiple pregnancies, it is recommended to consider the elective single ET strategy.

  9. Identification of a Novel Conjugative Plasmid in Mycobacteria That Requires Both Type IV and Type VII Secretion

    KAUST Repository

    Ummels, R.

    2014-09-23

    Conjugative plasmids have been identified in a wide variety of different bacteria, ranging from proteobacteria to firmicutes, and conjugation is one of the most efficient routes for horizontal gene transfer. The most widespread mechanism of plasmid conjugation relies on different variants of the type IV secretion pathway. Here, we describe the identification of a novel type of conjugative plasmid that seems to be unique for mycobacteria. Interestingly, while this plasmid is efficiently exchanged between different species of slow-growing mycobacteria, including Mycobacterium tuberculosis, it could not be transferred to any of the fast-growing mycobacteria tested. Genetic analysis of the conjugative plasmid showed the presence of a locus containing homologues of three type IV secretion system components and a relaxase. In addition, a new type VII secretion locus was present. Using transposon insertion mutagenesis, we show that in fact both these secretion systems are essential for conjugation, indicating that this plasmid represents a new class of conjugative plasmids requiring two secretion machineries. This plasmid could form a useful new tool to exchange or introduce DNA in slow-growing mycobacteria. IMPORTANCE: Conjugative plasmids play an important role in horizontal gene transfer between different bacteria and, as such, in their adaptation and evolution. This effect is most obvious in the spread of antibiotic resistance genes. Thus far, conjugation of natural plasmids has been described only rarely for mycobacterial species. In fact, it is generally accepted that M. tuberculosis does not show any recent sign of horizontal gene transfer. In this study, we describe the identification of a new widespread conjugative plasmid that can also be efficiently transferred to M. tuberculosis. This plasmid therefore poses both a threat and an opportunity. The threat is that, through the acquisition of antibiotic resistance markers, this plasmid could start a rapid spread of

  10. A general real-time formulation for multi-rate mass transfer problems

    Directory of Open Access Journals (Sweden)

    O. Silva

    2009-03-01

    Full Text Available Many flow and transport phenomena, ranging from delayed storage observed in pumping tests to tailing in river or aquifer tracer breakthrough curves, display non-equilibrium behavior. Usually, they are modeled by non-local in time formulations, such as multi-porosity, multiple processes non equilibrium, continuous time random walk, memory functions, integro-differential equations, fractional derivatives or multi-rate mass transfer (MRMT, among others. We develop a MRMT algorithm that can be used to represent all these formulations. The method is accurate, computationally inexpensive and easy to implement in groundwater or river flow and transport codes. In fact, we present a module that can be linked to existing programs with minimal programming effort. Its accuracy is verified by comparison with existing solutions.

  11. Wireless Information and Power Transfer: Architecture Design and Rate-Energy Tradeoff

    CERN Document Server

    Zhou, Xun; Ho, Chin Keong

    2012-01-01

    Simultaneous information and power transfer over the wireless channels potentially offers great convenience to mobile users. Yet practical receiver designs impose technical constraints on its hardware realization, as practical circuits for harvesting energy from radio signals are not yet able to decode the carried information directly. To make theoretical progress, we propose a general receiver operation, namely, dynamic power splitting (DPS), which splits the received signal with adjustable power for energy harvesting and for information decoding. Moreover, we propose two types of practical receiver architectures, namely, separated versus integrated information and energy receivers. The integrated receiver integrates the front-end components of the separated receiver, thus achieving a smaller form factor. The rateenergy tradeoff for these two architectures are characterized by a so-called rate-energy (R-E) region. Numerical results show that the R-E region of the integrated receiver is superior to that of th...

  12. Heat transfer and heating rate of food stuffs in commercial shop ovens

    Indian Academy of Sciences (India)

    P Navaneethakrishnan; P S S Srinivasan; S Dhandapani

    2007-10-01

    The CFD analysis of flow and temperature distribution in heating ovens used in bakery shop, to keep the foodstuffs warm, is attempted using finite element technique. The oven is modelled as a two-dimensional steady state natural convection heat transfer problem. Effects of heater location and total heat input on temperature uniformity of foodstuffs are studied. Placing the heater at the bottom of the oven improves the air circulation rate by 17 times and 10 times than that at the top and side of the oven. But the top location provides better uniformity in foodstuff temperature than the other cases. Side location is not preferable. In the present ovens, the heating elements are located at the top. The analysis shows that if heaters are located at the bottom along with additional flow guidance arrangements, energy efficient oven configuration can be obtained.

  13. 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.

  14. Vibrational energy transfer in selectively excited diatomic molecules. [Relaxation rates, self-relaxation, upper limits

    Energy Technology Data Exchange (ETDEWEB)

    Dasch, C.J.

    1978-09-01

    Single rovibrational states of HCl(v=2), HBr(v=2), DCl(v=2), and CO(v=2) were excited with a pulsed optical parametric oscillator (OPO). Total vibrational relaxation rates near - resonance quenchers were measured at 295/sup 0/K using time resolved infrared fluorescence. These rates are attributed primarily to V - V energy transfer, and they generally conform to a simple energy gap law. A small deviation was found for the CO(v) + DCl(v') relaxation rates. Upper limits for the self relaxation by V - R,T of HCl(v=2) and HBr(v=2) and for the two quantum exchange between HCl and HBr were determined. The HF dimer was detected at 295/sup 0/K and 30 torr HF pressure with an optoacoustic spectrometer using the OPO. Pulsed and chopped, resonant and non-resonant spectrophones are analyzed in detail. From experiments and first order perturbation theory, these V - V exchange rates appear to behave as a first order perturbation in the vibrational coordinates. The rotational dynamics are known to be complicated however, and the coupled rotational - vibrational dynamics were investigated theoreticaly in infinite order by the Dillon and Stephenson and the first Magnus approximations. Large ..delta..J transitions appear to be important, but these calculations differ by orders of magnitude on specific rovibrational transition rates. Integration of the time dependent semiclassical equations by a modified Gordon method and a rotationally distorted wave approximation are discussed as methods which would treat the rotational motion more accurately. 225 references.

  15. Prediction and improvement of the solid particles transfer rate for the bulk handing system design of offshore drilling vessels

    Directory of Open Access Journals (Sweden)

    Mincheol Ryu

    2015-11-01

    Full Text Available Numerous experiments with a scaled pilot facility were carried out to compare the relative bulk transfer performance of three special devices for applications to drilling systems. The pipe diameter for bulk transportation was 3 in., which corresponds to around half of the actual system dimensions. Two different pressures, 3 and 4 bar, were considered to check the relative performance under different pressure conditions at a bulk storage tank. And to make a practical estimation method of the bulk transfer rate at the early design stages of the bulk handling system, a series of experiments were conducted for real scaled bulk handing systems of two drilling vessels. The pressure drops at each pipe element as well as the bulk transfer rates were measured under different operating conditions. Using the measured results, the friction factor for each pipe element was calculated and a procedure for transfer rate estimation was developed. Compared to the measured transfer rate results for other drilling vessels, the estimated transfer rates were within a maximum 15% error bound.

  16. 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。

  17. A comparison of implantation, miscarriage and pregnancy rates of single and double day 3 embryo transfer between fresh and frozen thawed transfer cycles: a retrospective study

    Institute of Scientific and Technical Information of China (English)

    Liu Liu; Tong Xiaomei; Jiang Lingying; Tinchiu Li; Zhou Feng; Zhang Songying

    2014-01-01

    Background Reduced endometrial receptivity in hyperstimulated cycles may lead to a lower implantation rate and a lower clinical pregnancy rate,but it is unclear if it is also associated with an increase in pregnancy loss rate.The aim of this study was to compare the implantation,miscarriage,and pregnancy rates between fresh and frozen thawed transfer of one or two day-3 embryos,with a view to understanding whether or not reduced endometrial receptivity encountered in hyperstimulated cycles is associated with an increase in miscarriage rate.Methods This study involved a consecutive series of 1 551 single day-3 embryo transfer cycles and consecutive 5 919 double day-3 embryo transfer cycles in the Assisted Reproductive Unit of the Sir Run Run Shaw Hospital,Hangzhou,China,between January 2010 and December 2012.Results The implantation and clinical pregnancy rates (single embryo 30.7% and double embryos 33.4% and 51.4%)using fresh cycle were both significantly lower than that of frozen-thawed cycles (single embryo 35.8% and double embryos 38.1% and 57.8%).There was no difference in biochemical loss or clinical miscarriage rates between the two groups.Conclusions Impairment of endometrial receptivity associated with ovarian hyperstimulation leads to implantation failure at a very early stage,resulting in an increased number of non-pregnancy.It does not lead to increase in biochemical or clinical losses.The significantly reduced ongoing pregnancy rates in both fresh single and double embryo transfer are therefore due to failure to achieve a pregnancy,rather than pregnancy loss after conception.

  18. Plasmid Classification in an Era of Whole-Genome Sequencing: Application in Studies of Antibiotic Resistance Epidemiology

    Science.gov (United States)

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

    2017-01-01

    Plasmids are extra-chromosomal genetic elements ubiquitous in bacteria, and commonly transmissible between host cells. Their genomes include variable repertoires of ‘accessory genes,’ such as antibiotic resistance genes, as well as ‘backbone’ loci which are largely conserved within plasmid families, and often involved in key plasmid-specific functions (e.g., replication, stable inheritance, mobility). Classifying plasmids into different types according to their phylogenetic relatedness provides insight into the epidemiology of plasmid-mediated antibiotic resistance. Current typing schemes exploit backbone loci associated with replication (replicon typing), or plasmid mobility (MOB typing). Conventional PCR-based methods for plasmid typing remain widely used. With the emergence of whole-genome sequencing (WGS), large datasets can be analyzed using in silico plasmid typing methods. However, short reads from popular high-throughput sequencers can be challenging to assemble, so complete plasmid sequences may not be accurately reconstructed. Therefore, localizing resistance genes to specific plasmids may be difficult, limiting epidemiological insight. Long-read sequencing will become increasingly popular as costs decline, especially when resolving accurate plasmid structures is the primary goal. This review discusses the application of plasmid classification in WGS-based studies of antibiotic resistance epidemiology; novel in silico plasmid analysis tools are highlighted. Due to the diverse and plastic nature of plasmid genomes, current typing schemes do not classify all plasmids, and identifying conserved, phylogenetically concordant genes for subtyping and phylogenetics is challenging. Analyzing plasmids as nodes in a network that represents gene-sharing relationships between plasmids provides a complementary way to assess plasmid diversity, and allows inferences about horizontal gene transfer to be made. PMID:28232822

  19. Application of a plasmid classification system to determine prevalence of replicon families among multidrug resistant enterococci

    Science.gov (United States)

    The presence and transfer of plasmids from commensal bacteria to more pathogenic bacteria may contribute to dissemination of antimicrobial resistance. However, prevalence of plasmids from commensal bacteria in food animals such as the enterococci remains largely unknown. In this study, the prevale...

  20. Single-molecule sequencing to track plasmid diversity of hospital-associated carbapenemase-producing Enterobacteriaceae.

    Science.gov (United States)

    Conlan, Sean; Thomas, Pamela J; Deming, Clayton; Park, Morgan; Lau, Anna F; Dekker, John P; Snitkin, Evan S; Clark, Tyson A; Luong, Khai; Song, Yi; Tsai, Yu-Chih; Boitano, Matthew; Dayal, Jyoti; Brooks, Shelise Y; Schmidt, Brian; Young, Alice C; Thomas, James W; Bouffard, Gerard G; Blakesley, Robert W; Mullikin, James C; Korlach, Jonas; Henderson, David K; Frank, Karen M; Palmore, Tara N; Segre, Julia A

    2014-09-17

    Public health officials have raised concerns that plasmid transfer between Enterobacteriaceae species may spread resistance to carbapenems, an antibiotic class of last resort, thereby rendering common health care-associated infections nearly impossible to treat. To determine the diversity of carbapenemase-encoding plasmids and assess their mobility among bacterial species, we performed comprehensive surveillance and genomic sequencing of carbapenem-resistant Enterobacteriaceae in the National Institutes of Health (NIH) Clinical Center patient population and hospital environment. We isolated a repertoire of carbapenemase-encoding Enterobacteriaceae, including multiple strains of Klebsiella pneumoniae, Klebsiella oxytoca, Escherichia coli, Enterobacter cloacae, Citrobacter freundii, and Pantoea species. Long-read genome sequencing with full end-to-end assembly revealed that these organisms carry the carbapenem resistance genes on a wide array of plasmids. K. pneumoniae and E. cloacae isolated simultaneously from a single patient harbored two different carbapenemase-encoding plasmids, indicating that plasmid transfer between organisms was unlikely within this patient. We did, however, find evidence of horizontal transfer of carbapenemase-encoding plasmids between K. pneumoniae, E. cloacae, and C. freundii in the hospital environment. Our data, including full plasmid identification, challenge assumptions about horizontal gene transfer events within patients and identify possible connections between patients and the hospital environment. In addition, we identified a new carbapenemase-encoding plasmid of potentially high clinical impact carried by K. pneumoniae, E. coli, E. cloacae, and Pantoea species, in unrelated patients and in the hospital environment. Copyright © 2014, American Association for the Advancement of Science.

  1. An individual-based approach to explain plasmid invasion in bacterial populations

    DEFF Research Database (Denmark)

    Seoane, Jose Miguel; Yankelevich, Tatiana; Dechesne, Arnaud

    2011-01-01

    observe, however, that transient periods of elevated plasmid transfer in newly formed transconjugant cells are offset by unfavorable cell-to-cell contact mechanics, which ultimately precludes the pWWO TOL plasmid from fully invading tightly packed multicellular P. putida populations such as microcolonies...

  2. Electrocardiograms and heart rate signals a short distance transfer to the Android type of mobile devices, saving energy and bandwidth expenditure

    OpenAIRE

    Dalius Miežinis; Laimutis Telksnys

    2013-01-01

    Electrocardiograms and heart rate signals short distance transfer to mobile Android devices problem is examined. Energy saving and efficient use of bandwidth issues for signals transfer is discussed. Electrocardiograms and heart rate signaling proximity to mobile Android devices implementation possibilities is analyzed. Electrocardiograms and heart rate signals short distance transfer to mobile Android devices is proposed.

  3. Characterization and restriction analysis of the P sex factor and the cryptic plasmid of Vibrio cholerae strain V58.

    Science.gov (United States)

    Bartowsky, E J; Morelli, G; Kamke, M; Manning, P A

    1987-07-01

    The P plasmid of Vibrio cholerae is a derepressed sex factor restricted to V. cholerae and has been shown to express surface exclusion. We have isolated the plasmids of strain V58 and have found that in addition to P, two further cryptic plasmids are also present. P has a size of 68 kb as determined by both electron microscopy and restriction endonuclease analysis. These other plasmids are 34 and 4.7 kb in size. Restriction maps of P and the larger cryptic plasmid have been determined. It has been demonstrated that P differs from the standard Inc group test plasmids and also expresses a surface exclusion system. The ability of the type Inc plasmids to be transferred to V. cholerae by either liquid or filter matings and the stability of these plasmids in V. cholerae have also been examined.

  4. 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...

  5. TRANSFER

    African Journals Online (AJOL)

    “Chemistry Department, Kenyatta University, P. 0. Box 43844 ... harvester (X) [L 2] in a manner consistent with the following Forster equation for long range energy transfer [3-7]. .... sensitive foods, chemical reactors and essences. Recently we ...

  6. The effect of microbubbles on gas-liquid mass transfer coefficient and degradation rate of COD in wastewater treatment.

    Science.gov (United States)

    Yao, Kangning; Chi, Yong; Wang, Fei; Yan, Jianhua; Ni, Mingjiang; Cen, Kefa

    2016-01-01

    A commonly used aeration device at present has the disadvantages of low mass transfer rate because the generated bubbles are several millimeters in diameter which are much bigger than microbubbles. Therefore, the effect of a microbubble on gas-liquid mass transfer and wastewater treatment process was investigated. To evaluate the effect of each bubble type, the volumetric mass transfer coefficients for microbubbles and conventional bubbles were determined. The volumetric mass transfer coefficient was 0.02905 s(-1) and 0.02191 s(-1) at a gas flow rate of 0.67 L min(-1) in tap water for microbubbles and conventional bubbles, respectively. The degradation rate of simulated municipal wastewater was also investigated, using aerobic activated sludge and ozone. Compared with the conventional bubble generator, the chemical oxygen demand (COD) removal rate was 2.04, 5.9, 3.26 times higher than those of the conventional bubble contactor at the same initial COD concentration of COD 200 mg L(-1), 400 mg L(-1), and 600 mg L(-1), while aerobic activated sludge was used. For the ozonation process, the rate of COD removal using microbubble generator was 2.38, 2.51, 2.89 times of those of the conventional bubble generator. Based on the results, the effect of initial COD concentration on the specific COD degradation rate were discussed in different systems. Thus, the results revealed that microbubbles could enhance mass transfer in wastewater treatment and be an effective method to improve the degradation of wastewater.

  7. Rate of Iron Transfer Through the Horse Spleen Ferritin Shell Determined by the Rate of Formation of Prussian Blue and Fe-desferrioxamine Within the Ferritin Cavity

    Science.gov (United States)

    Zhang, Bo; Watt, Richard K.; Galvez, Natividad; Dominquez-Vera, Jose M.; Watt, Gerald D.

    2005-01-01

    Iron (2+ and 3+) is believed to transfer through the three-fold channels in the ferritin shell during iron deposition and release in animal ferritins. However, the rate of iron transit in and out through these channels has not been reported. The recent synthesis of [Fe(CN)(sub 6)](3-), Prussian Blue (PB) and desferrioxamine (DES) all trapped within the horse spleen ferritin (HoSF) interior makes these measurements feasible. We report the rate of Fe(2+) penetrating into the ferritin interior by adding external Fe(2+) to [Fe(CN)(sub 6)](3-) encapsulated in the HoSF interior and measuring the rate of formation of the resulting encapsulated PB. The rate at which Fe(2+) reacts with [Fe(CN)(sub 6)](3-) in the HoSF interior is much slower than the formation of free PB in solution and is proceeded by a lag period. We assume this lag period and the difference in rate represent the transfer of Fe(2+) through the HoSF protein shell. The calculated diffusion coefficient, D approx. 5.8 x 10(exp -20) square meters per second corresponds to the measured lag time of 10-20 s before PB forms within the HoSF interior. The activation energy for Fe(2+) transfer from the outside solution through the protein shell was determined to be 52.9 kJ/mol by conducting the reactions at 10 to approximately 40 C. The reaction of Fe(3+) with encapsulated [Fe(CN)6](4-) also readily forms PB in the HoSF interior, but the rate is faster than the corresponding Fe(2+) reaction. The rate for Fe(3+) transfer through the ferritin shell was confirmed by measuring the rate of the formation of Fe-DES inside HoSF and an activation energy of 58.4 kJ/mol was determined. An attempt was made to determine the rate of iron (2+ and 3+) transit out from the ferritin interior by adding excess bipyridine or DES to PB trapped within the HoSF interior. However, the reactions are slow and occur at almost identical rates for free and HoSF-encapsulated PB, indicating that the transfer of iron from the interior through the

  8. The interconnection between biofilm formation and horizontal gene transfer.

    Science.gov (United States)

    Madsen, Jonas Stenløkke; Burmølle, Mette; Hansen, Lars Hestbjerg; Sørensen, Søren Johannes

    2012-07-01

    Recent research has revealed that horizontal gene transfer and biofilm formation are connected processes. Although published research investigating this interconnectedness is still limited, we will review this subject in order to highlight the potential of these observations because of their believed importance in the understanding of the adaptation and subsequent evolution of social traits in bacteria. Here, we discuss current evidence for such interconnectedness centred on plasmids. Horizontal transfer rates are typically higher in biofilm communities compared with those in planktonic states. Biofilms, furthermore, promote plasmid stability and may enhance the host range of mobile genetic elements that are transferred horizontally. Plasmids, on the other hand, are very well suited to promote the evolution of social traits such as biofilm formation. This, essentially, transpires because plasmids are independent replicons that enhance their own success by promoting inter-bacterial interactions. They typically also carry genes that heighten their hosts' direct fitness. Furthermore, current research shows that the so-called mafia traits encoded on mobile genetic elements can enforce bacteria to maintain stable social interactions. It also indicates that horizontal gene transfer ultimately enhances the relatedness of bacteria carrying the mobile genetic elements of the same origin. The perspective of this review extends to an overall interconnectedness between horizontal gene transfer, mobile genetic elements and social evolution of bacteria.

  9. Plasmids of Carotenoid-Producing Paracoccus spp. (Alphaproteobacteria) - Structure, Diversity and Evolution

    Science.gov (United States)

    Maj, Anna; Dziewit, Lukasz; Czarnecki, Jakub; Wlodarczyk, Miroslawa; Baj, Jadwiga; Skrzypczyk, Grazyna; Giersz, Dorota; Bartosik, Dariusz

    2013-01-01

    Plasmids are components of many bacterial genomes. They enable the spread of a large pool of genetic information via lateral gene transfer. Many bacterial strains contain mega-sized replicons and these are particularly common in Alphaproteobacteria. Considerably less is known about smaller alphaproteobacterial plasmids. We analyzed the genomes of 14 such plasmids residing in 4 multireplicon carotenoid-producing strains of the genus Paracoccus (Alphaproteobacteria): P. aestuarii DSM 19484, P. haeundaensis LG P-21903, P. marcusii DSM 11574 and P. marcusii OS22. Comparative analyses revealed mosaic structures of the plasmids and recombinational shuffling of diverse genetic modules involved in (i) plasmid replication, (ii) stabilization (including toxin-antitoxin systems of the relBE/parDE, tad-ata, higBA, mazEF and toxBA families) and (iii) mobilization for conjugal transfer (encoding relaxases of the MobQ, MobP or MobV families). A common feature of the majority of the plasmids is the presence of AT-rich sequence islets (located downstream of exc1-like genes) containing genes, whose homologs are conserved in the chromosomes of many bacteria (encoding e.g. RelA/SpoT, SMC-like proteins and a retron-type reverse transcriptase). The results of this study have provided insight into the diversity and plasticity of plasmids of Paracoccus spp., and of the entire Alphaproteobacteria. Some of the identified plasmids contain replication systems not described previously in this class of bacteria. The composition of the plasmid genomes revealed frequent transfer of chromosomal genes into plasmids, which significantly enriches the pool of mobile DNA that can participate in lateral transfer. Many strains of Paracoccus spp. have great biotechnological potential, and the plasmid vectors constructed in this study will facilitate genetic studies of these bacteria. PMID:24260361

  10. Effect of surface-plasmon polaritons on spontaneous emission and intermolecular energy-transfer rates in multilayered geometries

    NARCIS (Netherlands)

    Marocico, C. A.; Knoester, J.

    2011-01-01

    We use a Green's tensor method to investigate the spontaneous emission rate of a molecule and the energy-transfer rate between molecules placed in two types of layered geometries: a slab geometry and a planar waveguide. We focus especially on the role played by surface-plasmon polaritons in

  11. 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...

  12. War wound treatment complications due to transfer of an IncN plasmid harboring bla(OXA-181) from Morganella morganii to CTX-M-27-producing sequence type 131 Escherichia coli.

    Science.gov (United States)

    McGann, Patrick; Snesrud, Erik; Ong, Ana C; Appalla, Lakshmi; Koren, Michael; Kwak, Yoon I; Waterman, Paige E; Lesho, Emil P

    2015-01-01

    A 22-year-old male developed a recurrent sacral abscess associated with embedded shrapnel following a blast injury. Cultures grew extended-spectrum β-lactamase (ESBL)-producing, carbapenem-susceptible Escherichia coli. Ertapenem was administered, but the infection recurred after each course of antibiotics. Initial surgical interventions were unsuccessful, and subsequent cultures yielded E. coli and Morganella morganii, both nonsusceptible to carbapenems. The isolates were Carba NP test negative, gave ambiguous results with the modified Hodge test, and amplified the bla(OXA48)-like gene by real-time PCR. All E. coli isolates were sequence type 131 (ST131), carried nine resistance genes (including bla(CTX-M-27)) on an IncF plasmid, and were identical by genome sequencing, except for 150 kb of plasmid DNA in carbapenem-nonsusceptible isolates only. Sixty kilobases of this was shared by M. morganii and represented an IncN plasmid harboring bla(OXA-181). In M. morganii, the gene was flanked by IS3000 and ISKpn19, but in all but one of the E. coli isolates containing bla(OXA-181), a second copy of ISKpn19 had inserted adjacent to IS3000. To the best of our knowledge, this is the first report of bla(OXA-181) in the virulent ST131 clonal group and carried by the promiscuous IncN family of plasmids. The tendency of M. morganii to have high MICs of imipenem, a bla(OXA-181) substrate profile that includes penicillins but not extended-spectrum cephalosporins, and weak carbapenemase activity almost resulted in the presence of bla(OXA-181) being overlooked. We highlight the importance of surveillance for carbapenem resistance in all species, even those with intrinsic resistances, and the value of advanced molecular techniques in detecting subtle genetic changes.

  13. The Optimum Plate to Plate Spacing for Maximum Heat Transfer Rate from a Flat Plate Type Heat Exchanger

    Science.gov (United States)

    Ambarita, Himsar; Kishinami, Koki; Daimaruya, Mashashi; Tokura, Ikuo; Kawai, Hideki; Suzuki, Jun; Kobiyama, Mashayosi; Ginting, Armansyah

    The present paper is a study on the optimum plate to plate spacing for maximum heat transfer rate from a flat plate type heat exchanger. The heat exchanger consists of a number of parallel flat plates. The working fluids are flowed at the same operational conditions, either fixed pressure head or fixed fan power input. Parallel and counter flow directions of the working fluids were considered. While the volume of the heat exchanger is kept constant, plate number was varied. Hence, the spacing between plates as well as heat transfer rate will vary and there exists a maximum heat transfer rate. The objective of this paper is to seek the optimum plate to plate spacing for maximum heat transfer rate. In order to solve the problem, analytical and numerical solutions have been carried out. In the analytical solution, the correlations of the optimum plate to plate spacing as a function of the non-dimensional parameters were developed. Furthermore, the numerical simulation is carried out to evaluate the correlations. The results show that the optimum plate to plate spacing for a counter flow heat exchanger is smaller than parallel flow ones. On the other hand, the maximum heat transfer rate for a counter flow heat exchanger is bigger than parallel flow ones.

  14. Origin and Evolution of Nitrogen Fixation Genes on Symbiosis Islands and Plasmid in Bradyrhizobium

    Science.gov (United States)

    Okubo, Takashi; Piromyou, Pongdet; Tittabutr, Panlada; Teaumroong, Neung; Minamisawa, Kiwamu

    2016-01-01

    The nitrogen fixation (nif) genes of nodule-forming Bradyrhizobium strains are generally located on symbiosis islands or symbiosis plasmids, suggesting that these genes have been transferred laterally. The nif genes of rhizobial and non-rhizobial Bradyrhizobium strains were compared in order to infer the evolutionary histories of nif genes. Based on all codon positions, the phylogenetic tree of concatenated nifD and nifK sequences showed that nifDK on symbiosis islands formed a different clade from nifDK on non-symbiotic loci (located outside of symbiosis islands and plasmids) with elongated branches; however, these genes were located in close proximity, when only the 1st and 2nd codon positions were analyzed. The guanine (G) and cytosine (C) content of the 3rd codon position of nifDK on symbiosis islands was lower than that on non-symbiotic loci. These results suggest that nif genes on symbiosis islands were derived from the non-symbiotic loci of Bradyrhizobium or closely related strains and have evolved toward a lower GC content with a higher substitution rate than the ancestral state. Meanwhile, nifDK on symbiosis plasmids clustered with nifDK on non-symbiotic loci in the tree representing all codon positions, and the GC content of symbiotic and non-symbiotic loci were similar. These results suggest that nif genes on symbiosis plasmids were derived from the non-symbiotic loci of Bradyrhizobium and have evolved with a similar evolutionary pattern and rate as the ancestral state. PMID:27431195

  15. Origin and Evolution of Nitrogen Fixation Genes on Symbiosis Islands and Plasmid in Bradyrhizobium.

    Science.gov (United States)

    Okubo, Takashi; Piromyou, Pongdet; Tittabutr, Panlada; Teaumroong, Neung; Minamisawa, Kiwamu

    2016-09-29

    The nitrogen fixation (nif) genes of nodule-forming Bradyrhizobium strains are generally located on symbiosis islands or symbiosis plasmids, suggesting that these genes have been transferred laterally. The nif genes of rhizobial and non-rhizobial Bradyrhizobium strains were compared in order to infer the evolutionary histories of nif genes. Based on all codon positions, the phylogenetic tree of concatenated nifD and nifK sequences showed that nifDK on symbiosis islands formed a different clade from nifDK on non-symbiotic loci (located outside of symbiosis islands and plasmids) with elongated branches; however, these genes were located in close proximity, when only the 1st and 2nd codon positions were analyzed. The guanine (G) and cytosine (C) content of the 3rd codon position of nifDK on symbiosis islands was lower than that on non-symbiotic loci. These results suggest that nif genes on symbiosis islands were derived from the non-symbiotic loci of Bradyrhizobium or closely related strains and have evolved toward a lower GC content with a higher substitution rate than the ancestral state. Meanwhile, nifDK on symbiosis plasmids clustered with nifDK on non-symbiotic loci in the tree representing all codon positions, and the GC content of symbiotic and non-symbiotic loci were similar. These results suggest that nif genes on symbiosis plasmids were derived from the non-symbiotic loci of Bradyrhizobium and have evolved with a similar evolutionary pattern and rate as the ancestral state.

  16. A novel plasmid for delivering genes into mammalian cells with noninvasive food and commensal lactic acid bacteria.

    Science.gov (United States)

    Tao, Lin; Pavlova, Sylvia I; Ji, Xin; Jin, Ling; Spear, Gregory

    2011-01-01

    Using food and commensal lactic acid bacteria (LAB) as vehicles for DNA delivery into epithelial cells is a new strategy for vaccine delivery or gene therapy. However, present methods for DNA delivery with LAB have suffered low efficiency. Our goal was to develop a new system to deliver DNA into epithelial cells with high efficiency using food and commensal LAB. An Escherichia coli-LAB shuttle plasmid, pLKV1, for DNA delivery into eukaryotic cells was constructed. Two reporter plasmids with green and red fluorescent protein genes were also constructed to monitor the uptake of protein and DNA, respectively. Bacteria delivering these reporter plasmids into Caco-2 cells were monitored by fluorescence microscopy. Several methods that weaken the bacterial cell wall prior to co-culture with Caco-2 cells were evaluated for their role in the improvement of gene transfer efficiency. Treating Streptococcus gordonii with penicillin and lysozyme greatly increased its rate of gene delivery to mammalian cells compared to untreated control bacteria, while glycine pretreatment promoted the highest gene transfer rate for Lactococcus lactis. Uptake of green fluorescent bacteria by Caco-2 cells showed that the cell wall-weakening treatment promoted the internalization of the noninvasive bacteria into Caco-2 cells. In conclusion, we have developed a noninvasive system using LAB as a vehicle for vaccine delivery or gene therapy, and tested this system in vitro with Caco-2 cells.

  17. 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...

  18. 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.

  19. 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.

  20. 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...

  1. Measurement of the energy dependence of the muon transfer rate from hydrogen to higher-Z gases

    CERN Document Server

    Bakalov, Dimitar; Stoilov, Mihail; Vacchi, Andrea

    2014-01-01

    The recent Lamb shift experiment at PSI and the apparent incompatibility of the proton radii extracted using different methods revived the interest in the measurement of the hyperfine splitting in the ground state of muonic hydrogen as an alternative possibility for the experimental comparison of ordinary and muonic hydrogen spectroscopy data about the proton electromagnetic structure. The efficiency of the method developed for this measurement has been shown to critically depend on the energy dependence of the rate of muon transfer from hydrogen to heavier gases in the epithermal range. The available experimental data provide only qualitative information on the energy dependence, and the detailed theoretical predictions have not yet been tested. The present paper outlines an experimental method for the quantitative measurement of the muon transfer rate based on a series of repeated measurements of the muon transfer rate in a mixture of hydrogen and the gas of interest with appropriate concentration and densi...

  2. Host range of enterococcal vanA plasmids among Gram-positive intestinal bacteria

    DEFF Research Database (Denmark)

    Werner, Guido; Freitas, Ana R.; Coque, Teresa M.

    2010-01-01

    OBJECTIVES: The most prevalent type of acquired glycopeptide resistance is encoded by the vanA transposon Tn1546 located mainly on transferable plasmids in Enterococcus faecium. The limited occurrence in other species could be due to the lack of inter-species transferability and/or stability of Tn......-genus matings were performed and evaluated. In summary, intra-species transfer was far more frequent than inter-species transfer, if that was detectable at all. All recipients of the same species behaved similarly. Inter-genus transfer was shown for broad host range control plasmids (pIP501/pAMβ1) only...

  3. Foerster resonance energy transfer rate and local density of optical states are uncorrelated in any dielectric nanophotonic medium

    CERN Document Server

    Wubs, Martijn

    2015-01-01

    Motivated by the ongoing debate about nanophotonic control of Foerster resonance energy transfer (FRET), notably by the local density of optical states (LDOS), we study an analytic model system wherein a pair of ideal dipole emitters - donor and acceptor - exhibit energy transfer in the vicinity of an ideal mirror. The FRET rate is controlled by the mirror up to distances comparable to the donor-acceptor distance, that is, the few-nanometer range. For vanishing distance, we find a complete inhibition or a four-fold enhancement, depending on dipole orientation. For mirror distances on the wavelength scale, where the well-known `Drexhage' modification of the spontaneous-emission rate occurs, the FRET rate is constant. Hence there is no correlation between the Foerster (or total) energy transfer rate and the LDOS. At any distance to the mirror, the total energy transfer between a closely-spaced donor and acceptor is dominated by Foerster transfer, i.e., by the static dipole-dipole interaction that yields the cha...

  4. Effect of Gutai Decoction (固胎汤) on the Abortion Rate of in vitro Fertilization and Embryo Transfer

    Institute of Scientific and Technical Information of China (English)

    LIU Ying; WU Jing-zhi

    2006-01-01

    Objective: To study the effect of Chinese herbal medicine Gutai Decoction (固胎汤, GTD) on the abortion rate of in vitro fertilization and embryo transfer (IVF-ET). Methods: Observed were two hundred and forty-seven women having received IVF-ET and with β-human chorionic gonadotropin (β-HCG) > 25 IU/L on the 14th day after transferring. All were treated conventionally with progesterone 20-80 mg per day after transferring and if necessary the treatment was supplemented with Progynova 2 - 4 mg per day, with the medication withdrawn gradually from the 9th week of pregnancy till stopped completely. Among them 131 cases received GTD medication additionally, for 109 cases of whom the medication started from the 2nd day of transferring (taken as Group A) and for the other 22 cases from the 14th day after transferring (taken as Group B), the other 116 cases with no additional GTD treatment given were taken as the control group, with the medication lasting to the 12th week. The abortion rate in them was observed. Results: The abortion rate in Group A, Group B and the control group was 12.84%, 13.64% and 23.28%, respectively, the difference between the GTD treated groups and the control group was significant (P<0.05). Conclusion: Chinese medicine GTD could reduce abortion rate in women receiving IVF-ET.

  5. Factors affecting conception rates following artificial insemination or embryo transfer in lactating Holstein cows.

    Science.gov (United States)

    Demetrio, D G B; Santos, R M; Demetrio, C G B; Vasconcelos, J L M

    2007-11-01

    The objective of this study was to evaluate the factors that may affect conception rates (CR) following artificial insemination (AI) or embryo transfer (ET) in lactating Holstein cows. Estrous cycling cows producing 33.1 +/- 7.2 kg of milk/d received PGF2alpha injections and were assigned randomly to 1 of 2 groups (AI or ET). Cows detected in estrus (n = 387) between 48 and 96 h after the PGF2alpha injection received AI (n = 227) 12 h after detection of estrus or ET (n = 160) 6 to 8 d later (1 fresh embryo, grade 1 or 2, produced from nonlactating cows). Pregnancy was diagnosed at 28 and 42 d after estrus, and embryonic loss occurred when a cow was pregnant on d 28 but not pregnant on d 42. Ovulation, conception, and embryonic loss were analyzed by a logistic model to evaluate the effects of covariates [days in milk (DIM), milk yield, body temperature (BT) at d 7 and 14 post-AI, and serum concentration of progesterone (P4) at d 7 and 14 post-AI] on the probability of success. The first analysis included all cows that were detected in estrus. The CR of AI and ET were different on d 28 (AI, 32.6% vs. ET, 49.4%) and 42 (AI, 29.1% vs. ET, 38.8%) and were negatively influenced by high BT (d 7) and DIM. The second analysis included only cows with a corpus luteum on d 7. Ovulation rate was 84.8% and was only negatively affected by DIM. Conception rates of AI and ET were different on d 28 (AI, 37.9% vs. ET, 59.4%) and 42 (AI, 33.8% vs. ET, 46.6%) and were negatively influenced by high BT (d 7). The third analysis included only ovulating cows that were 7 d postestrus. Conception rates of AI and ET were different on d 28 (AI, 37.5% vs. ET, 63.2%) and 42 (AI, 31.7% vs. ET, 51.7%) and were negatively influenced by high BT (d 7). There was a positive effect of serum concentration of P4 and a negative effect of milk production on the probability of conception for the AI group but not for the ET group. The fourth analysis was embryonic loss (AI, 10.8% vs. ET, 21.5%). The transfer

  6. 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.

  7. Application of H∞ Filter on the Angular Rate Matching in the Transfer Alignment

    Directory of Open Access Journals (Sweden)

    Lijun Song

    2016-01-01

    Full Text Available The transfer alignment (TA scheme is used for the initial alignment of Inertial Navigation System (INS on dynamical base. The Kalman filter is often used in TA to improve the precision of TA. And the statistical characteristics of interference signal which is difficult to get must be known before the Kalman filter is used in the TA, because the interference signal is a random signal and there are some changes on the dynamic model of system. In this paper, the H∞ filter is adopted in the TA scheme of the angular rate matching when the various stages of disturbance in measurement are unknown. And it is compared with the Kalman filter in the same environment of simulation and evaluation. The result of simulation shows that the H∞ filter and the Kalman filter are both effective. The Kalman filter is more accurate than the H∞ filter when system noise and measurement noise are white noise, but the H∞ filter is more accurate and quicker than the Kalman filter when system noise and measurement noise are color noise. In the engineering practice, system noise and measurement noise are always color noise, so the H∞ filter is more suitable for engineering practice than the Kalman filter.

  8. Implications of rate-limited mass transfer for aquifer storage and recovery.

    Science.gov (United States)

    Culkin, Sean L; Singha, Kamini; Day-Lewis, Frederick D

    2008-01-01

    Pressure to decrease reliance on surface water storage has led to increased interest in aquifer storage and recovery (ASR) systems. Recovery efficiency, which is the ratio of the volume of recovered water that meets a predefined standard to total volume of injected fluid, is a common criterion of ASR viability. Recovery efficiency can be degraded by a number of physical and geochemical processes, including rate-limited mass transfer (RLMT), which describes the exchange of solutes between mobile and immobile pore fluids. RLMT may control transport behavior that cannot be explained by advection and dispersion. We present data from a pilot-scale ASR study in Charleston, South Carolina, and develop a three-dimensional finite-difference model to evaluate the impact of RLMT processes on ASR efficiency. The modeling shows that RLMT can explain a rebound in salinity during fresh water storage in a brackish aquifer. Multicycle model results show low efficiencies over one to three ASR cycles due to RLMT degrading water quality during storage; efficiencies can evolve and improve markedly, however, over multiple cycles, even exceeding efficiencies generated by advection-dispersion only models. For an idealized ASR model where RLMT is active, our simulations show a discrete range of diffusive length scales over which the viability of ASR schemes in brackish aquifers would be hindered.

  9. Thermal conductance and basal metabolic rate are part of a coordinated system for heat transfer regulation.

    Science.gov (United States)

    Naya, Daniel E; Spangenberg, Lucía; Naya, Hugo; Bozinovic, Francisco

    2013-09-22

    Thermal conductance measures the ease with which heat leaves or enters an organism's body. Although the analysis of this physiological variable in relation to climatic and ecological factors can be traced to studies by Scholander and colleagues, only small advances have occurred ever since. Here, we analyse the relationship between minimal thermal conductance estimated during summer (Cmin) and several ecological, climatic and geographical factors for 127 rodent species, in order to identify the exogenous factors that have potentially affected the evolution of thermal conductance. In addition, we evaluate whether there is compensation between Cmin and basal metabolic rate (BMR)-in such a way that a scale-invariant ratio between both variables is equal to one-as could be expected from the Scholander-Irving model of heat transfer. Our major findings are (i) annual mean temperature is the best single predictor of mass-independent Cmin. (ii) After controlling for the effect of body mass, there is a strong positive correlation between log10 (Cmin) and log10 (BMR). Further, the slope of this correlation is close to one, indicating an almost perfect compensation between both physiological variables. (iii) Structural equation modelling indicated that Cmin values are adjusted to BMR values and not the other way around. Thus, our results strongly suggest that BMR and thermal conductance integrate a coordinated system for heat regulation in endothermic animals and that summer conductance values are adjusted (in an evolutionary sense) to track changes in BMRs.

  10. Efficient Data Transfer Rate and Speed of Secured Ethernet Interface System.

    Science.gov (United States)

    Ghanti, Shaila; Naik, G M

    2016-01-01

    Embedded systems are extensively used in home automation systems, small office systems, vehicle communication systems, and health service systems. The services provided by these systems are available on the Internet and these services need to be protected. Security features like IP filtering, UDP protection, or TCP protection need to be implemented depending on the specific application used by the device. Every device on the Internet must have network interface. This paper proposes the design of the embedded Secured Ethernet Interface System to protect the service available on the Internet against the SYN flood attack. In this experimental study, Secured Ethernet Interface System is customized to protect the web service against the SYN flood attack. Secured Ethernet Interface System is implemented on ALTERA Stratix IV FPGA as a system on chip and uses the modified SYN flood attack protection method. The experimental results using Secured Ethernet Interface System indicate increase in number of genuine clients getting service from the server, considerable improvement in the data transfer rate, and better response time during the SYN flood attack.

  11. Structural and dynamical control of the reaction rate in protein electron transfer

    Science.gov (United States)

    Balabin, Ilya A.

    Electron transfer (ET) reactions in proteins are key steps in many vital bioenergetic processes, and the reaction rate is known to be highly sensitive to the protein structure in some cases. For most bioenergetic reactions, as described by the Fermi Golden rule, the rate is proportional to a product of the average square of the effective electronic donor to acceptor coupling and a Franck-Condon factor, which accounts for the nuclear control of the energy gap. The nuclear factor is reasonably well described in Marcus theory and its modifications, and this work is focused on the mechanisms that control the effective coupling. About ten years ago, the Pathways model described for the first time how protein environment may control the effective coupling. In this work, a novel theoretical approach is developed to explore the mechanisms of structural and dynamical control beyond the qualitative level of the Pathways model. In Chapter 1, the assumptions of the Pathways model, its limitations and effects of the structure and the electronic Hamiltonian are investigated for model chain-like bridges using the Dyson's equations. In Chapter II, the framework to explore the sensitivity of the effective coupling to quality of the electronic Hamiltonian, the interference among the dominant pathways and the bridge dynamics is presented. This analysis employs the Green's function technique and includes combined molecular dynamics and electronic structure calculations. Finally, in Chapter III, this framework is tested on the bacterial photosynthetic reaction center, and the mechanisms of the structural and dynamical control for different ET steps are discussed.

  12. 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...

  13. 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.

  14. 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

  15. 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, ...

  16. A Theoretical Model of Non-conservative Mass Transfer with Non-uniform Mass Accretion Rate in Close Binary Stars

    CERN Document Server

    Gharami, Prabir; Rahaman, Farook

    2014-01-01

    Mass transfer in close binaries is often non-conservative and the modeling of this kind of mass transfer is mathematically challenging as in this case due to the loss of mass as well as angular momentum the governing system gets complicated and uncertain. In the present work a new mathematical model has been prescribed for the non-conservative mass transfer in a close binary system taking in to account the gradually decreasing profile of the mass accretion rate by the accreting star with respect to time as well as with respect to the increase in mass of the accreting star. The process of mass transfer is understood to occur up to a critical mass limit of the accreting star beyond which this process may cease to work.

  17. Theoretical Model of Non-Conservative Mass Transfer with Uniform Mass Accretion Rate in Contact Binary Stars

    Science.gov (United States)

    Gharami, Prabir; Ghosh, Koushik; Rahaman, Farook

    2016-01-01

    In contact binaries mass transfer is usually non-conservative which ends into loss of mass as well as angular momentum in the system. In the present work we have presented a new mathematical model of the non-conservative mass transfer with a uniform mass accretion rate in a contact binary system with lower angular momentum. The model has been developed under the consideration of reverse mass transfer which may occur simultaneously with the original mass transfer as a result of the large scale circulations encircling the entire donor and a significant portion of the gainer. These circulations in contact binaries with lower angular momentum are caused by the overflow of the critical equipotential surface by both the components of the binary system making the governing system more intricate and uncertain.

  18. Absorption and desorption mass transfer rates in non-reactive systems

    NARCIS (Netherlands)

    Hamborg, Espen S.; Kersten, Sascha R. A.; Versteeg, Geert F.

    2010-01-01

    Liquid phase mass transfer coefficients have been measured in a controlled environment during gas absorption into a liquid and gas desorption from a liquid in a batch operated stirred tank reactor over a wide range of operating conditions. At identical operating conditions, the mass transfer

  19. ENHANCEMENT OF GAS TO WATER MASS-TRANSFER RATES BY A DISPERSED ORGANIC-PHASE

    NARCIS (Netherlands)

    VANEDE, CJ; VANHOUTEN, R; BEENACKERS, AACM

    1995-01-01

    Experimental data are presented on the enhancement of oxygen mass transfer into an aqueous sulphite solution in a stirred cell, due to the presence of a dispersed liquid octene phase. The observed enhancement factors can be described with a new mass transfer theory, called the Film Variable Hold-up

  20. The interconnection between biofilm formation and horizontal gene transfer

    DEFF Research Database (Denmark)

    Madsen, Jonas Stenløkke; Burmølle, Mette; Hansen, Lars H.

    2012-01-01

    Recent research has revealed that horizontal gene transfer and biofilm formation are connected processes. Although published research investigating this interconnectedness is still limited, we will review this subject in order to highlight the potential of these observations because of their beli......Recent research has revealed that horizontal gene transfer and biofilm formation are connected processes. Although published research investigating this interconnectedness is still limited, we will review this subject in order to highlight the potential of these observations because...... of their believed importance in the understanding of the adaptation and subsequent evolution of social traits in bacteria. Here, we discuss current evidence for such interconnectedness centred on plasmids. Horizontal transfer rates are typically higher in biofilm communities compared with those in planktonic states....... Biofilms, furthermore, promote plasmid stability and may enhance the host range of mobile genetic elements that are transferred horizontally. Plasmids, on the other hand, are very well suited to promote the evolution of social traits such as biofilm formation. This, essentially, transpires because plasmids...

  1. 阳离子脂质体介导增强型绿色荧光蛋白质粒转染骨骼肌卫星细胞%Cationic liposome-mediated enhanced green fluorescent protein plasmid transferred into skeletal muscle satellite cells

    Institute of Scientific and Technical Information of China (English)

    许志锋; 李敬来; 韩振; 冯钢; 任明明

    2013-01-01

    BACKGROUND:Skeletal muscle satel ite cells are totipotential stem cells with multi-directional differentiation potential, locate in skeletal muscle interstitium, have a certain tolerance to ischemia and hypoxia, and are important cells in stem cellengineering. OBJECTIVE:To establish a thrifty, convenient culture procedure and create a simple, efficient method to transfect skeletal muscle satel ite cells, and investigate genetic expression after the transfection for cellular cardiomyoplasty. METHODS:Skeletal muscle satel ite cells were isolated from rabbit thigh and cultured. Their growth curves were determined by CKK-8 method. Grouped by different proportions of the plasmid and liposome, skeletal muscle satel ite cells were transfered by the enhanced green fluorescent protein plasmid based on liposome. After transfection, the efficiency and character of target genetic expression was determined. RESULTS AND CONCLUSION:Satel ite cells were isolated, cultured and transfected successful y. In suitable ratio of plasmid and liposomes, the transfection efficiency reached up to above 35%. The target protein was expressed within 12 hours after transfection, reached maximum in 48-72 hours and decreased gradual y after one week. The expression stil could be observed two weeks latter. The enhanced green fluorescent protein plasmid conducted by cationic liposome could be transfered into skeletal muscle satel ite cells efficiently. The transfection efficiency was correlated closely to the ratio of plasmid and lipofectamine. The change of target gene expression depended on time.%背景:骨骼肌卫星细胞是一种具有多向分化能力的全能干细胞,存在于骨骼肌间质中,对缺血、缺氧有一定的耐受力,是干细胞工程中重要来源细胞。  目的:为联合基因工程细胞心肌成形治疗初步探讨较为简便、经济的骨骼肌卫星细胞体外培养方法,建立一种简单、高效的转染骨骼肌卫星细胞的方法及

  2. Does sequential embryo transfer improve pregnancy rate in patients with repeated implantation failure? A randomized control study

    Directory of Open Access Journals (Sweden)

    Wael A. Ismail Madkour

    2015-12-01

    Full Text Available Background: Repeated failure of in vitro fertilization treatment is frustrating to the patients and their clinicians. Various treatment plans and a change of protocol have been suggested for “low responders”; however, patients who fail treatment repeatedly inspite of good quality embryos pose a special therapeutic challenge. Additional challenge would be imposed on that particular group when the local IVF regulating low does not permit surplus embryo freezing. Objective: To examine whether sequential transfer of embryos on day 3 and on day 5 after ovum pick-up improves IVF/ET success rates in patients with repeated consecutive IVF failures (⩾ 3 trials compared to day 3 alone, with the background that local regulation prohibits embryo freezing. Study design: Randomized controlled study. Women scheduled for IVF/ET with repeated consecutive IVF failures (⩾ 3 trials were randomized to either sequential transfer of embryos on day 3 and on day 5 after ovum pick-up (Group I = 74 or conventional day 3 transfer (Group II = 73 as a control. The primary outcome measures were clinical pregnancy rate and implantation rate. The secondary outcome measures were ongoing pregnancy rate and early pregnancy loss. Results: Baseline and cycle characteristics were comparable in both groups. Clinical pregnancy rate (per embryo transfer was significantly higher in sequential ET group (37. 8% compared to that in day 3 group (21.9% (P value <0.05. Also, implantation rate (per embryos transferred was significantly higher in sequential ET group (17.1% compared to that in control group (10.5% (P value <0.01. Similarly, ongoing pregnancy (per embryo transfer was significantly higher in sequential ET group (33. 8% compared to that in day 3 group (19.2% (P value <0.05. Conclusions: Patients with repeated implantation failures, treatment with the sequential embryo transfer approach had significantly improved pregnancy outcomes compared to regular day 3 transfers

  3. Obstacle avoidance during human walking: learning rate and cross-modal transfer.

    Science.gov (United States)

    Erni, T; Dietz, V

    2001-07-01

    1. The aim of this study was to investigate the significance of specific afferent information during motor learning. Blindfolded subjects stepped over an obstacle on a treadmill while different stimuli (acoustic (ACU), somatosensory (SOM) and light flash (LED)) signalled the approaching obstacle. The effect of the above stimuli was then evaluated and compared to full vision (VIS) locomotion. In the non-visual conditions feedback information about the performance was provided by an acoustic signal. 2. Using each of the different stimuli for information the level of subject performance was assessed by noting foot clearance and analysing both leg muscle electromyographic activity and movement trajectories during three successive runs. Each of these runs consisted of 100 steps over the obstacle. 3. The best performance at the onset of the first run was achieved during the VIS condition. When the VIS condition (run 1 + 2) was followed by ACU or SOM information or when the ACU condition (run 1 + 2) was followed by LED, little cross-modal transfer (CMT) occurred, i.e. adaptation in run 3 started again at a low level of performance. In contrast, if adaptation started with ACU stimuli followed by SOM stimuli, almost full CMT occurred. The absolute level of performance achieved after the second or third runs was similar in the VIS and non-VIS conditions. 4. In conclusion, the course of motor learning depends on specific afferent information, and feedforward control has a special influence on the performance only at the onset of the experiment but not on the rate of learning. The fact that little CMT occurs from visual to non-visual stimuli and from ACU to LED suggests that visual afferent input is processed in a different way to non-visual stimuli.

  4. Impact of Increased Academic Intensity on Transfer Rates: An Application of Matching Estimators to Student-Unit Record Data

    Science.gov (United States)

    Doyle, William R.

    2009-01-01

    The impact of increased academic intensity on transfer rates from community colleges to 4-year institutions has been estimated only from observational data, with the possibility of selection bias. This study uses matching estimators to overcome possible selection bias and estimate the causal impact of increased academic intensity on transfer…

  5. Mid-luteal serum progesterone concentrations govern implantation rates for cryopreserved embryo transfers conducted under hormone replacement.

    Science.gov (United States)

    Yovich, John L; Conceicao, Jason L; Stanger, James D; Hinchliffe, Peter M; Keane, Kevin N

    2015-08-01

    This study explores the relevance of mid-luteal serum hormonal concentrations in cryopreserved embryo transfer cycles conducted under hormone replacement therapy (HRT) control and which involved single-embryo transfer (SET) of 529 vitrified blastocysts. Widely ranging mid-luteal oestradiol and progesterone concentrations ensued from the unique HRT regimen. Oestradiol had no influence on clinical pregnancy or live birth rates, but an optimal progesterone range between 70 and 99 nmol/l (P decreased implantation rates. There was no clear interaction between oestradiol and progesterone concentrations but embryo quality grading did show a significant influence on outcomes (P pregnancy and live birth rates, respectively). Multiple comparison analysis showed that the progesterone effect was influential regardless of embryo grading, body mass index or the woman's age, either at vitrification or at cryopreserved embryo transfer. The results support the argument that careful monitoring of serum progesterone concentrations in HRT-cryopreserved embryo transfer is warranted and that further studies should explore pessary adjustments to optimize concentrations for individual women to enhance implantation rates.

  6. Effect of salinity and pressure on the rate of mass transfer in aquifer storage of CO2

    NARCIS (Netherlands)

    Khosrokhavar, R.; Eftekhari, A.A.; Farajzadeh, R.; Wolf, K.H.A.A.; Bruining, J.

    2015-01-01

    The growing concern about global warming has increased interest in improving the technology for the geological storage of CO2 in aquifers. One important aspect for aquifer storage is the rate of transfer between the overlying gas layer and the aquifer below. It is generally accepted that density dri

  7. The effect of temperature and transmembrane potentials on the rates of electron transfer between membrane-bound biological redox components.

    Science.gov (United States)

    Kuznetsov, A M; Ulstrup, J

    1981-06-12

    We have investigated rate data for the temperature and free energy dependence of the primary electron-transfer processes in bacterial photosynthesis. Rather than representing the whole electronic-nuclear coupling by a frequently applied discrete single-mode model, we have incorporated a continuum of modes characterized by a certain distribution function. In this way, we can illuminate the role of both a broad distribution of low-frequency modes representing the medium and a narrow distribution representing local nuclear modes. Furthermore, it emerges from the calculations that both sets are important in the overall scheme of primary photosynthetic electron-transfer processes. By means of this model and quantum-mechanical rate theory, we can reproduce a number of important features of the primary photosynthetic processes concerning in particular the temperature (tunnelling or thermally activated nuclear motion) and free energy dependence ('normal', 'activation-less', or 'inverted' regions) of the rate constants and estimate such parameters as nuclear-reorganization energy electron-exchange integrals and electron-transfer distances. We have finally considered some of the important factors which determine the potential drop across the membrane and estimated the extent to which variations in the potential drop affect the rate constants of the electron-transfer processes.

  8. Background for a new standard on pass-by measurement of combined roughness, track decay rate and vibroacoustic transfer functions

    NARCIS (Netherlands)

    Dittrich, M.G.; Létourneaux, F.; Dupuis, H.

    2013-01-01

    A measurement method for combined roughness, track decay rates and transfer functions derived from rail vibration during a train pass-by was initially developed in the late nineties [1]. This method has been then later implemented in software tools [2] and applied in several countries for various pu

  9. 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

  10. 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.

  11. Forster resonance energy transfer rate in any dielectric nanophotonic medium with weak dispersion

    DEFF Research Database (Denmark)

    Wubs, Martijn; Vos, Willem L.

    2016-01-01

    Motivated by the ongoing debate about nanophotonic control of Forster resonance energy transfer (FRET), notably by the local density of optical states (LDOS), we study FRET and spontaneous emission in arbitrary nanophotonic media with weak dispersion and weak absorption in the frequency overlap...... to the mirror, typically a few nm. Finally, we discuss the consequences of our results to applications of Forster resonance energy transfer, for instance in quantum information processing....

  12. Characterization of plasmids in extensively drug-resistant acinetobacter strains isolated in India and Pakistan.

    Science.gov (United States)

    Jones, Lim S; Carvalho, Maria J; Toleman, Mark A; White, P Lewis; Connor, Thomas R; Mushtaq, Ammara; Weeks, Janis L; Kumarasamy, Karthikeyan K; Raven, Katherine E; Török, M Estée; Peacock, Sharon J; Howe, Robin A; Walsh, Timothy R

    2015-02-01

    The blaNDM-1 gene is associated with extensive drug resistance in Gram-negative bacteria. This probably spread to Enterobacteriaceae from Acinetobacter spp., and we characterized plasmids associated with blaNDM-1 in Acinetobacter spp. to gain insight into their role in this dissemination. Four clinical NDM-1-producing Acinetobacter species strains from India and Pakistan were investigated. A plasmid harboring blaNDM-1, pNDM-40-1, was characterized by whole-genome sequencing of Acinetobacter bereziniae CHI-40-1 and comparison with related plasmids. The presence of similar plasmids in strains from Pakistan was sought by PCR and sequencing of amplicons. Conjugation frequency was tested and stability of pNDM-40-1 investigated by real-time PCR of isolates passaged with and without antimicrobial selection pressure. A. bereziniae and Acinetobacter haemolyticus strains contained plasmids similar to the pNDM-BJ01-like plasmids identified in Acinetobacter spp. in China. The backbone of pNDM-40-1 was almost identical to that of pNDM-BJ01-like plasmids, but the transposon harboring blaNDM-1, Tn125, contained two short deletions. Escherichia coli and Acinetobacter pittii transconjugants were readily obtained. Transconjugants retained pNDM-40-1 after a 14-day passage experiment, although stability was greater with meropenem selection. Fragments of pNDM-BJ01-like plasmid backbones are found near blaNDM-1 in some genetic contexts from Enterobacteriaceae, suggesting that cross-genus transfer has occurred. pNDM-BJ01-like plasmids have been described in isolates originating from a wide geographical region in southern Asia. In vitro data on plasmid transfer and stability suggest that these plasmids could have contributed to the spread of blaNDM-1 into Enterobacteriaceae.

  13. Effect of intrauterine injection of human chorionic gonadotropin before embryo transfer on pregnancy rate: A prospective randomized study.

    Science.gov (United States)

    Mostajeran, Fatemeh; Godazandeh, Farzaneh; Ahmadi, Sayed Mehdi; Movahedi, Minoo; Jabalamelian, Seyed Abolfazl

    2017-01-01

    Human chorionic gonadotropin (hCG) as the most important factor to controlled implantation is one of the early embryonic signals in primates that is secreted by the embryo before its implantation. This study was designed to assess the effects of intrauterine injection of hCG before the embryo transfer in an in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) cycle on pregnancy rate in infertile patients. This randomized study was done on 100 infertile patients in two groups: intervention group received injection of 700 IU of intrauterine hCG 10 min before embryo transfer and control group did not receive hCG. The pregnancy rate was tested 2 weeks after embryo transfer, and if the pregnancy test was positive, a transvaginal ultrasound was performed 3 weeks later to search for signs of pregnancy, such as the presence of a gestational sac, embryo, and fetal heart rate, and confirmed as successful pregnancy. Pregnancy test was positive in 13 (28.6%) of 46 patients in hCG group and in control group was positive in 6 (12.5%) of 48 patients. The pregnancy rate between hCG group and control group was not significantly different (P = 0.54). The pregnancy rate in hCG group with IVF fertilization was 20.8% and in their controls was 7.4% (P = 0.51). The pregnancy rate in hCG group with ICSI fertilization was 36.4% and in their controls was 19% (P = 0.16). The intrauterine injection of 700 IU of hCG before embryo transfer improved pregnancy rate compared to control group but was not significantly different.

  14. 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

  15. Embryo quality is the main factor affecting cumulative live birth rate after elective single embryo transfer in fresh stimulation cycles.

    Science.gov (United States)

    Niinimäki, Maarit; Veleva, Zdravka; Martikainen, Hannu

    2015-11-01

    The study was aimed to evaluate which factors affect the cumulative live birth rate after elective single embryo transfer in women younger than 36 years. Additionally, number of children in women with more than one delivery per ovum pick-up after fresh elective single embryo transfer and subsequent frozen embryo transfers was assessed. Retrospective cohort study analysing data of a university hospital's infertility clinic in 2001-2010. A total of 739 IVF/ICSI cycles with elective single embryo transfer were included. Analyses were made per ovum pick-up including fresh and subsequent frozen embryo transfers. Factors affecting cumulative live birth rates were examined in uni- and multivariate analyses. A secondary endpoint was the number of children born after all treatments. In the fresh cycles, the live birth rate was 29.2% and the cumulative live birth rate was 51.3%, with a twin rate of 3.4%. In the multivariate analysis, having two (odds ratio (OR) 1.73; 95% confidence interval (CI) 1.12-2.67) or ≥3 top embryos (OR 2.66; 95% CI 1.79-3.95) was associated with higher odds for live birth after fresh and frozen embryo cycles. Age, body mass index, duration of infertility, diagnosis or total gonadotropin dose were not associated with the cumulative live birth rate. In cycles with one top embryo, the cumulative live birth rate was 40.2%, whereas it was 64.1% in those with at least three top embryos. Of women who had a live birth in the fresh cycle, 20.4% had more than one child after all frozen embryo transfers. Among women with three or more top embryos after ovum pick-up, 16.1% gave birth to more than one child. The cumulative live birth rate in this age group varies from 40% to 64% and is dependent on the quality of embryos. Women with three or more top embryos have good chance of having more than one child per ovum pick-up without elevated risk of multiple pregnancies. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  16. Prediction of Heat Transfer Rates for Shell-and-Tube Heat Exchangers by Artificial Neural Networks Approach

    Institute of Scientific and Technical Information of China (English)

    Qiuwang WANG; Gongnan XIE; Ming ZENG; Laiqin LUO

    2006-01-01

    This work used artificial neural network (ANN) to predict the heat transfer rates of shell-and-tube heat exchangers with segmental baffles or continuous helical baffles, based on limited experimental data. The Back Propagation (BP) algorithm was used in training the networks. Different network configurations were also studied. The deviation between the predicted results and experimental data was less than 2%. Comparison with correlation for prediction shows ANN superiority. It is recommended that ANN can be easily used to predict the performances of thermal systems in engineering applications, especially to model heat exchangers for heat transfer analysis.

  17. 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.

  18. Epsilon-toxin plasmids of Clostridium perfringens type D are conjugative.

    Science.gov (United States)

    Hughes, Meredith L; Poon, Rachael; Adams, Vicki; Sayeed, Sameera; Saputo, Juliann; Uzal, Francisco A; McClane, Bruce A; Rood, Julian I

    2007-11-01

    Isolates of Clostridium perfringens type D produce the potent epsilon-toxin (a CDC/U.S. Department of Agriculture overlap class B select agent) and are responsible for several economically significant enterotoxemias of domestic livestock. It is well established that the epsilon-toxin structural gene, etx, occurs on large plasmids. We show here that at least two of these plasmids are conjugative. The etx gene on these plasmids was insertionally inactivated using a chloramphenicol resistance cassette to phenotypically tag the plasmid. High-frequency conjugative transfer of the tagged plasmids into the C. perfringens type A strain JIR325 was demonstrated, and the resultant transconjugants were shown to act as donors in subsequent mating experiments. We also demonstrated the transfer of "unmarked" native epsilon-toxin plasmids into strain JIR325 by exploiting the high transfer frequency. The transconjugants isolated in these experiments expressed functional epsilon-toxin since their supernatants had cytopathic effects on MDCK cells and were toxic in mice. Using the widely accepted multiplex PCR approach for toxin genotyping, these type A-derived transconjugants were genotypically type D. These findings have significant implications for the C. perfringens typing system since it is based on the toxin profile of each strain. Our study demonstrated the fluid nature of the toxinotypes and their dependence upon the presence or absence of toxin plasmids, some of which have for the first time been shown to be conjugative.

  19. Epsilon-Toxin Plasmids of Clostridium perfringens Type D Are Conjugative▿ †

    Science.gov (United States)

    Hughes, Meredith L.; Poon, Rachael; Adams, Vicki; Sayeed, Sameera; Saputo, Juliann; Uzal, Francisco A.; McClane, Bruce A.; Rood, Julian I.

    2007-01-01

    Isolates of Clostridium perfringens type D produce the potent epsilon-toxin (a CDC/U.S. Department of Agriculture overlap class B select agent) and are responsible for several economically significant enterotoxemias of domestic livestock. It is well established that the epsilon-toxin structural gene, etx, occurs on large plasmids. We show here that at least two of these plasmids are conjugative. The etx gene on these plasmids was insertionally inactivated using a chloramphenicol resistance cassette to phenotypically tag the plasmid. High-frequency conjugative transfer of the tagged plasmids into the C. perfringens type A strain JIR325 was demonstrated, and the resultant transconjugants were shown to act as donors in subsequent mating experiments. We also demonstrated the transfer of “unmarked” native ɛ-toxin plasmids into strain JIR325 by exploiting the high transfer frequency. The transconjugants isolated in these experiments expressed functional ɛ-toxin since their supernatants had cytopathic effects on MDCK cells and were toxic in mice. Using the widely accepted multiplex PCR approach for toxin genotyping, these type A-derived transconjugants were genotypically type D. These findings have significant implications for the C. perfringens typing system since it is based on the toxin profile of each strain. Our study demonstrated the fluid nature of the toxinotypes and their dependence upon the presence or absence of toxin plasmids, some of which have for the first time been shown to be conjugative. PMID:17720791

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

    Science.gov (United States)

    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.

  1. Application of multivariate adaptive regression spine-assisted objective function on optimization of heat transfer rate around a cylinder

    Energy Technology Data Exchange (ETDEWEB)

    Dey, Prasenjit; Dad, Ajoy K. [Mechanical Engineering Department, National Institute of Technology, Agartala (India)

    2016-12-15

    The present study aims to predict the heat transfer characteristics around a square cylinder with different corner radii using multivariate adaptive regression splines (MARS). Further, the MARS-generated objective function is optimized by particle swarm optimization. The data for the prediction are taken from the recently published article by the present authors [P. Dey, A. Sarkar, A.K. Das, Development of GEP and ANN model to predict the unsteady forced convection over a cylinder, Neural Comput. Appl. (2015). Further, the MARS model is compared with artificial neural network and gene expression programming. It has been found that the MARS model is very efficient in predicting the heat transfer characteristics. It has also been found that MARS is more efficient than artificial neural network and gene expression programming in predicting the forced convection data, and also particle swarm optimization can efficiently optimize the heat transfer rate.

  2. Exciton interference revealed by energy dependent exciton transfer rate for ring-structured molecular systems

    Energy Technology Data Exchange (ETDEWEB)

    Yan, Yun-An, E-mail: yunan@gznc.edu.cn [Guizhou Provincial Key Laboratory of Computational Nanomaterial Science, Guizhou Education University, Guiyang, Guizhou 550018 (China)

    2016-01-14

    The quantum interference is an intrinsic phenomenon in quantum physics for photon and massive quantum particles. In principle, the quantum interference may also occur with quasi-particles, such as the exciton. In this study, we show how the exciton quantum interference can be significant in aggregates through theoretical simulations with hierarchical equations of motion. The systems under investigation are generalized donor-bridge-acceptor model aggregates with the donor consisting of six homogeneous sites assuming the nearest neighbor coupling. For the models with single-path bridge, the exciton transfer time only shows a weak excitation energy dependence. But models with double-path bridge have a new short transfer time scale and the excitation energy dependence of the exciton transfer time assumes clear peak structure which is detectable with today’s nonlinear spectroscopy. This abnormality is attributed to the exciton quantum interference and the condition for a clear observation in experiment is also explored.

  3. 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.

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

    Science.gov (United States)

    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

  5. Model system-bath Hamiltonian and nonadiabatic rate constants for proton-coupled electron transfer at electrode-solution interfaces.

    Science.gov (United States)

    Navrotskaya, Irina; Soudackov, Alexander V; Hammes-Schiffer, Sharon

    2008-06-28

    An extension of the Anderson-Newns-Schmickler model for electrochemical proton-coupled electron transfer (PCET) is presented. This model describes reactions in which electron transfer between a solute complex in solution and an electrode is coupled to proton transfer within the solute complex. The model Hamiltonian is derived in a basis of electron-proton vibronic states defined within a double adiabatic approximation for the electrons, transferring proton, and bath modes. The interaction term responsible for electronic transitions between the solute complex and the electrode depends on the proton donor-acceptor vibrational mode within the solute complex. This model Hamiltonian is used to derive the anodic and cathodic rate constants for nonadiabatic electrochemical PCET. The derivation is based on the master equations for the reduced density matrix of the electron-proton subsystem, which includes the electrons of the solute complex and the electrode, as well as the transferring proton. The rate constant expressions differ from analogous expressions for electrochemical electron transfer because of the summation over electron-proton vibronic states and the dependence of the couplings on the proton donor-acceptor vibrational motion. These differences lead to additional contributions to the total reorganization energy, an additional exponential temperature-dependent prefactor, and a temperature-dependent term in the effective activation energy that has different signs for the anodic and cathodic processes. This model can be generalized to describe both nonadiabatic and adiabatic electrochemical PCET reactions and provides the framework for the inclusion of additional effects, such as the breaking and forming of other chemical bonds.

  6. Determination of the Astrophysical S(E) Factors or Rates for Radiative Capture Reaction with One Nucleon Transfer Reaction%Determination of the Astrophysical S(E) Factors or Rates for Radiative Capture Reaction with One Nucleon Transfer Reaction

    Institute of Scientific and Technical Information of China (English)

    李志宏; 郭冰; 李云居; 苏俊; 李二涛; 白希祥; 王友宝; 曾晟; 王宝祥; 颜胜权; 李志常; 刘建成; 连钢; 金孙均; 刘鑫; 柳卫平

    2012-01-01

    The radiative capture reaction plays an important role in nuclear astrophysics. We have indirectly measured the astrophysical S(E) factors for some proton capture reactions and reaction rates for several neutron capture reactions with one nucleon transfer reactions at HI-13 tandem accelerator in recent years. Some of them are compiled into IAEA EXFOR database and JINA REACLIB project, and used in the network calculations of Big Bang nucleosynthesis and type-I X-ray bursts.

  7. Convective Heat Transfer Scaling of Ignition Delay and Burning Rate with Heat Flux and Stretch Rate in the Equivalent Low Stretch Apparatus

    Science.gov (United States)

    Olson, Sandra

    2011-01-01

    To better evaluate the buoyant contributions to the convective cooling (or heating) inherent in normal-gravity material flammability test methods, we derive a convective heat transfer correlation that can be used to account for the forced convective stretch effects on the net radiant heat flux for both ignition delay time and burning rate. The Equivalent Low Stretch Apparatus (ELSA) uses an inverted cone heater to minimize buoyant effects while at the same time providing a forced stagnation flow on the sample, which ignites and burns as a ceiling fire. Ignition delay and burning rate data is correlated with incident heat flux and convective heat transfer and compared to results from other test methods and fuel geometries using similarity to determine the equivalent stretch rates and thus convective cooling (or heating) rates for those geometries. With this correlation methodology, buoyant effects inherent in normal gravity material flammability test methods can be estimated, to better apply the test results to low stretch environments relevant to spacecraft material selection.

  8. Mobilization of Bacillus thuringiensis plasmid pTX14-3.

    Science.gov (United States)

    Andrup, L; Bendixen, H H; Jensen, G B

    1995-05-01

    The Bacillus thuringiensis subsp. israelensis (Bti) plasmid pTX14-3 has been reported to contain a gene, mob14-3, with considerable homology to genes encoding mobilization proteins from other gram-positive bacteria. We have used the aggregation-mediated conjugation system recently discovered in Bti to compare the mobilization kinetics of different derivatives of plasmid pTX14-3. Plasmid pTX14-3 has been found to replicate by the rolling-circle mechanism and to contain a locus suppressing the formation of high-molecular-weight DNA. We found that deleting a DNA fragment containing this locus increased the transfer frequency about twofold. The mobilization frequency of the plasmid containing the intact mob14-3 gene did not indicate a mobilization-enhancing activity of the encoded polypeptide. However, the presence of the mob14-3 gene seemed to increase the stability of the plasmid in exponential growth.

  9. The Impact of Collective Bargaining on Teacher Transfer Rates in Urban High-Poverty Schools

    Science.gov (United States)

    Nelson, F. Howard

    2006-01-01

    Data in this report reveals that collectively bargaining agreements are not the source of the teacher quality problem in urban school districts. The data shows that collective bargaining agreements are associated with reduced teacher transfer activity, especially in high-poverty schools, and less reliance on first-year teachers to staff…

  10. Absorption and desorption mass transfer rates in chemically enhanced reactive systems. Part I : Chemical enhancement factors

    NARCIS (Netherlands)

    Hamborg, Espen S.; Versteeg, Geert F.

    2012-01-01

    The chemical enhancement factors have been measured in a controlled environment for absorption and desorption mass transfer processes in aqueous 2.0 M MDEA solutions at temperatures of 298.15, 313.15, and 333.15 K and the loading of CO2 ranging from 0 to 0.8 in a batch-operated stirred tank reactor.

  11. Gas transfer rates from airlifts used for concurrent aeration, carbon dioxide stripping, and recirculation

    Science.gov (United States)

    Airlifts simplify recirculating aquaculture systems and can potentially reduce capital costs and minimize maintenance issues. Airlifts have the ability to move and aerate water as well as degass the water of any carbon dioxide. This study evaluated the oxygen transfer and carbon dioxide removal abil...

  12. Heat transfer intensification by increasing vapor flow rate in flat heat pipes

    Science.gov (United States)

    Sprinceana, Silviu; Mihai, Ioan; Beniuga, Marius; Suciu, Cornel

    2015-02-01

    Flat heat pipes have various technical applications, one of the most important being the cooling of electronic components[9]. Their continuous development is due to the fact that these devices permit heat transfer without external energetic contribution. The practical exploitation of flat heat pipes however is limited by the fact that dissipated power can only reach a few hundred watts. The present paper aims to advance a new method for the intensification of convective heat transfer. A centrifugal mini impeller, driven by a turntable which incorporates four permanent magnets was designed. These magnets are put in motion by another rotor, which in its turn includes two permanent magnets and is driven by a mini electrical motor. Rotation of the centrifugal blades generates speed and pressure increase of the cooling agent brought to vapor state within the flat micro heat pipe. It's well known that the liquid suffers biphasic transformations during heat transfer inside the heat pipe. Over the hotspot (the heat source being the electronic component) generated at one end of the heat pipe, convective heat transfer occurs, leading to sudden vaporization of the liquid. Pressures generated by newly formed vapors push them towards the opposite end of the flat heat pipe, where a finned mini heat sink is usually placed. The mini-heat exchanger is air-cooled, thus creating a cold spot, where vapors condensate. The proposed method contributes to vapor flow intensification by increasing their transport speed and thus leading to more intense cooling of the heat pipe.

  13. 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...

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

    Science.gov (United States)

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

    2015-05-15

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

  15. Respiration and heart rate complexity: effects of age and gender assessed by band-limited transfer entropy.

    Science.gov (United States)

    Nemati, Shamim; Edwards, Bradley A; Lee, Joon; Pittman-Polletta, Benjamin; Butler, James P; Malhotra, Atul

    2013-10-01

    Aging and disease are accompanied with a reduction of complex variability in the temporal patterns of heart rate. This reduction has been attributed to a break down of the underlying regulatory feedback mechanisms that maintain a homeodynamic state. Previous work has established the utility of entropy as an index of disorder, for quantification of changes in heart rate complexity. However, questions remain regarding the origin of heart rate complexity and the mechanisms involved in its reduction with aging and disease. In this work we use a newly developed technique based on the concept of band-limited transfer entropy to assess the aging-related changes in contribution of respiration and blood pressure to entropy of heart rate at different frequency bands. Noninvasive measurements of heart beat interval, respiration, and systolic blood pressure were recorded from 20 young (21-34 years) and 20 older (68-85 years) healthy adults. Band-limited transfer entropy analysis revealed a reduction in high-frequency contribution of respiration to heart rate complexity (p<0.001) with normal aging, particularly in men. These results have the potential for dissecting the relative contributions of respiration and blood pressure-related reflexes to heart rate complexity and their degeneration with normal aging.

  16. Influence of radiant energy exchange on the determination of convective heat transfer rates to Orbiter leeside surfaces during entry

    Science.gov (United States)

    Throckmorton, D. A.

    1982-01-01

    Temperatures measured at the aerodynamic surface of the Orbiter's thermal protection system (TPS), and calorimeter measurements, are used to determine heating rates to the TPS surface during atmospheric entry. On the Orbiter leeside, where convective heating rates are low, it is possible that a significant portion of the total energy input may result from solar radiation, and for the wing, cross radiation from the hot (relatively) Orbiter fuselage. In order to account for the potential impact of these sources, values of solar- and cross-radiation heat transfer are computed, based upon vehicle trajectory and attitude information and measured surface temperatures. Leeside heat-transfer data from the STS-2 mission are presented, and the significance of solar radiation and fuselage-to-wing cross-radiation contributions to total energy input to Orbiter leeside surfaces is assessed.

  17. Aggregate Download Throughput for TCP-controlled long file transfers in a WLAN with multiple STA-AP association rates

    CERN Document Server

    BK, Pradeepa

    2010-01-01

    We consider several WLAN stations associated at rates r1, r2, ..., rk with an Access Point. Each station is downloading a long file from a local server, located on the LAN to which the AP is attached. We model these simultaneous TCP-controlled transfers using a Markov Chain. Our analytical approach leads to a procedure to compute aggregate download throughput numerically, and the results match simulations very well.

  18. Genome Sequence of Listeria monocytogenes Plasmid pLM-C-273 Carrying Genes Related to Stress Resistance.

    Science.gov (United States)

    Liang, Lindsay; Gnaneshan, Saravanamuttu; Garduño, Rafael A; Mallo, Gustavo V

    2016-10-13

    Mobile genetic elements in bacteria, such as plasmids, act as important vectors for the transfer of antibiotic resistance, virulence, and metal resistance genes. Here, we report the genome sequence of a new plasmid pLM-C-273, identified in a Listeria monocytogenes strain isolated from a clinical sample in Ontario, Canada.

  19. Genome Sequence of Listeria monocytogenes Plasmid pLM-C-273 Carrying Genes Related to Stress Resistance

    Science.gov (United States)

    Liang, Lindsay; Gnaneshan, Saravanamuttu; Garduño, Rafael A.

    2016-01-01

    Mobile genetic elements in bacteria, such as plasmids, act as important vectors for the transfer of antibiotic resistance, virulence, and metal resistance genes. Here, we report the genome sequence of a new plasmid pLM-C-273, identified in a Listeria monocytogenes strain isolated from a clinical sample in Ontario, Canada. PMID:27738039

  20. Enhanced transformation efficiency of recalcitrant Bacillus cereus and Bacillus weihenstephanensis isolates upon in vitro methylation of plasmid DNA

    NARCIS (Netherlands)

    Nierop Groot, M.N.; Nieboer, F.; Abee, T.

    2008-01-01

    Digestion patterns of chromosomal DNAs of Bacillus cereus and Bacillus weihenstephanensis strains suggest that Sau3AI-type restriction modification systems are widely present among the isolates tested. In vitro methylation of plasmid DNA was used to enhance poor plasmid transfer upon electroporation

  1. Effectiveness of a conditional cash transfer programme on TB cure rate: a retrospective cohort study in Brazil.

    Science.gov (United States)

    Torrens, Ana W; Rasella, Davide; Boccia, Delia; Maciel, Ethel L N; Nery, Joilda S; Olson, Zachary D; Barreira, Draurio C N; Sanchez, Mauro N

    2016-03-01

    Despite the efforts of the National Tuberculosis Programme, TB cure rates in Brazil are sub-optimal. The End TB Strategy for post-2015 identifies conditional cash transfer interventions as powerful tools to improve TB control indicators, including TB cure rate. This study aims to inform the new policy by evaluating the role of the Bolsa Familia Programme (BFP), one of the largest conditional cash transfer programmes in the world, on TB cure rates in Brazil. We undertook a retrospective cohort study, based on an unprecedented record linkage of socioeconomic and health data, to compare cases of patients newly diagnosed with TB in 2010 receiving BFP cash benefits (n=5788) with those who did not (n=1467) during TB treatment. We used Poisson regression with robust variance to estimate the relative risks for TB cure adjusted for known confounders. The cure rate among patients exposed to BFP during TB treatment was 82.1% (4752/5788), 5.2% higher than among those not exposed. This was confirmed after controlling for TB type, diabetes mellitus, HIV status and other relevant clinical and socioeconomic covariates (RR=1.07, 95% CI 1.04 to 1.11 for cure rates among BFP beneficiaries). This association seemed higher for patients not under directly observed treatment (RR=1.11; 95% CI 1.05 to 1.16). Although further research is needed, this study suggests that conditional cash transfer programmes can contribute to improve TB cure rate in Brazil. © The Author 2016. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  2. On the use of exchange rates as trading rules in a bilateral system of transferable discharge permits

    Energy Technology Data Exchange (ETDEWEB)

    Rodriguez, F. [Department Economia Aplicada, Facultad de Derecho, Campus ' Miguel de Unamuno' , University of Salamanca, Salamanca (Spain)

    2000-04-01

    The use of a system of transferable discharge permits to control the harmful effects of non-uniformly mixed pollutants requires the application of trading rules in order to prevent permit trading among sources from violating environmental standards. The elements and properties of bilateral trading rules can be analyzed more easily once formulated as exchange rates, which would convert, in a cost-effective way, the emission right potentially given up by the seller into an offsetting emission right acquired by the buyer. In this article, a new expression for such exchange rates is proposed and then analyzed to infer some unexplored properties of the system. 8 refs.

  3. Mass transfer during electrodeposition of metals at a periodically changing rate (REVIEW

    Directory of Open Access Journals (Sweden)

    MIODRAG D. MAKSIMOVIC

    1999-05-01

    Full Text Available 1. Introduction 2. Mass transfer in the steady state periodic condition 2.1. Reversing current 2.2. Pulsating current 2.3. Alternating current superimposed on direct current 3. The influence of the charge and discharge of the electrical double layer 4. The validity of the mathematical model 4.1. Reversing current in the millisecond range 4.2. Reversing current in the second range 4.3. Pulsating current 4.4. Pulsating overpotential 5. Conclusion

  4. Local translational diffusion rates of membranous Na+,K(+)-ATPase measured by saturation transfer ESR spectroscopy.

    OpenAIRE

    Esmann, M.; Marsh, D.

    1992-01-01

    Diffusion-controlled Heisenberg spin exchange between spin-labeled Na+,K(+)-ATPase [ATP phosphohydrolase (Na+/K(+)-transporting), EC 3.6.1.37] proteins has been studied by saturation transfer ESR spectroscopy in reconstituted membranes. Na+,K(+)-ATPase from the salt gland of Squalus acanthias was solubilized in a polyoxyethylene ether detergent, octa(ethylene glycol) dodecyl monoether. Part of the solubilized enzyme was covalently spin-labeled with a nitroxide derivative of indanedione and re...

  5. Heat transfer in a microvascular network: the effect of heart rate on heating and cooling in reptiles (Pogona barbata and Varanus varius).

    Science.gov (United States)

    Seebacher, F

    2000-03-21

    Thermally-induced changes in heart rate and blood flow in reptiles are believed to be of selective advantage by allowing animal to exert some control over rates of heating and cooling. This notion has become one of the principal paradigms in reptilian thermal physiology. However, the functional significance of changes in heart rate is unclear, because the effect of heart rate and blood flow on total animal heat transfer is not known. I used heat transfer theory to determine the importance of heat transfer by blood flow relative to conduction. I validated theoretical predictions by comparing them with field data from two species of lizard, bearded dragons (Pogona barbata) and lace monitors (Varanus varius). Heart rates measured in free-ranging lizards in the field were significantly higher during heating than during cooling, and heart rates decreased with body mass. Convective heat transfer by blood flow increased with heart rate. Rates of heat transfer by both blood flow and conduction decreased with mass, but the mass scaling exponents were different. Hence, rate of conductive heat transfer decreased more rapidly with increasing mass than did heat transfer by blood flow, so that the relative importance of blood flow in total animal heat transfer increased with mass. The functional significance of changes in heart rate and, hence, rates of heat transfer, in response to heating and cooling in lizards was quantified. For example, by increasing heart rate when entering a heating environment in the morning, and decreasing heart rate when the environment cools in the evening a Pogona can spend up to 44 min longer per day with body temperature within its preferred range. It was concluded that changes in heart rate in response to heating and cooling confer a selective advantage at least on reptiles of mass similar to that of the study animals (0. 21-5.6 kg). Copyright 2000 Academic Press.

  6. Influence of position and length of uterus on implantation and clinical pregnancy rates in IVF and embryo transfer treatment cycles.

    Science.gov (United States)

    Egbase, P E; Al-Sharhan, M; Grudzinskas, J G

    2000-09-01

    In a prospective study of 807 consecutive women shown to have an apparently normal uterus after hysterosalpingography, hysteroscopy or pelvic ultrasonography prior to IVF or intracytoplasmic sperm injection (ICSI) and embryo transfer, the position and length of the uterine cavity was measured routinely at a pre-treatment mock transfer procedure. The apparent length of the uterine cavity was 9 cm in 85 women (group 3). The uterus was noted to be retroverted in 38. 2% (308) women. The embryo transfer catheter was advanced to 5 mm from the uterine fundus based on the previously determined cavity length in all the embryo transfer procedures at 48 h after oocyte collection. Implantation and clinical pregnancy rates were not significantly different with respect to position of the uterus, difficulties encountered in passage of the catheter, mean age of the women, aetiology or duration of infertility or embryology events. An apparently greater cavity length was seen in older and/or parous women, but the difference was not statistically significant. Although the highest implantation and clinical pregnancy rates were seen in women with a cavity length of 7-9 cm (group 2) the differences were not statistically significant: group 1, 18.9 and 36. 7%; group 2, 21.0 and 46.5%; and group 3, 17.3 and 32.9% respectively. The incidence of ectopic pregnancy per reported clinical pregnancy was highest in group 1 women, being 14.9% (7/47) in comparison with group 2 (1.8%, 5/276) and group 3 (0%, 0/27) (P: size of the uterus is a critical factor in the aetiology of ectopic pregnancy in IVF/ICSI-embryo transfer.

  7. A comparative study between cleavage stage embryo transfer at day 3 and blastocyst stage transfer at day 5 in in-vitro fertilization/intra-cytoplasmic sperm injection on clinical pregnancy rates.

    Science.gov (United States)

    Kaur, Prabhleen; Swarankar, M L; Maheshwari, Manju; Acharya, Veena

    2014-07-01

    To evaluate the efficacy of blastocyst transfer in comparison with cleavage stage transfer. A randomized, prospective study was conducted in Infertility clinic, Department of Obstetrics and Gynecology, Mahatma Gandhi Hospital, Jaipur on 300 patients aged 25-40 years undergoing in-vitro fertilization (IVF)/intra-cytoplasmic sperm injection (ICSI) cycle from May 2010-April 2011. When three or more Grade-I embryos were observed on day 2 of culture, patients were divided randomly into two study groups, cleavage stage transfer and blastocyst transfer group having 150 patients each. Primary outcomes evaluated were, Clinical pregnancy rate and Implantation rate. The results were analyzed using proportions, standard deviation and Chi-square test. Both the groups were similar for age, indication and number of embryos transferred. Clinical pregnancies after blastocyst transfer were significantly higher 66 (44.0%) compared to cleavage stage embryo transfer 44 (29.33%) (P < 0.01). Implantation rate for blastocyst transfer group was also significantly higher (P < 0.001). Blastocyst transfer having higher implantation rate and clinical pregnancy rate lead to reduction in multiple pregnancies.

  8. A comparative study between cleavage stage embryo transfer at day 3 and blastocyst stage transfer at day 5 in in-vitro fertilization/intra-cytoplasmic sperm injection on clinical pregnancy rates

    Directory of Open Access Journals (Sweden)

    Prabhleen Kaur

    2014-01-01

    Full Text Available Objective: To evaluate the efficacy of blastocyst transfer in comparison with cleavage stage transfer. Study Design: A randomized, prospective study was conducted in Infertility clinic, Department of Obstetrics and Gynecology, Mahatma Gandhi Hospital, Jaipur on 300 patients aged 25-40 years undergoing in-vitro fertilization (IVF/intra-cytoplasmic sperm injection (ICSI cycle from May 2010-April 2011. When three or more Grade-I embryos were observed on day 2 of culture, patients were divided randomly into two study groups, cleavage stage transfer and blastocyst transfer group having 150 patients each. Primary outcomes evaluated were, Clinical pregnancy rate and Implantation rate. The results were analyzed using proportions, standard deviation and Chi-square test. Results: Both the groups were similar for age, indication and number of embryos transferred. Clinical pregnancies after blastocyst transfer were significantly higher 66 (44.0% compared to cleavage stage embryo transfer 44 (29.33% (P < 0.01.Implantation rate for blastocyst transfer group was also significantly higher (P < 0.001. Conclusion: Blastocyst transfer having higher implantation rate and clinical pregnancy rate lead to reduction in multiple pregnancies.

  9. Characterization and comparative analysis of antibiotic resistance plasmids isolated from a wastewater treatment plant.

    Directory of Open Access Journals (Sweden)

    Teddie O Rahube

    2014-10-01

    Full Text Available A wastewater treatment plant (WWTP is an environment high in nutrient concentration with diverse bacterial populations and can provide an ideal environment for the proliferation of mobile elements such as plasmids. WWTPs have also been identified as reservoirs for antibiotic resistance genes that are associated with human pathogens. The objectives of this study were to isolate and characterize self-transmissible or mobilizable resistance plasmids associated with effluent from wastewater treatment plant. An enrichment culture approach designed to capture plasmids conferring resistance to high concentrations of erythromycin was used to capture plasmids from an urban wastewater treatment plant servicing a population of ca. 210,000. DNA sequencing of the plasmids revealed diversity of plasmids represented by incompatibility groups IncU, col-E, IncFII and IncP-1β. Genes coding resistance to clinically relevant antibiotics (macrolide, tetracycline, beta-lactam, trimethoprim, chloramphenicol, sulphonamide, quaternary ammonium compounds and heavy metals were co-located on these plasmids, often within transposable and integrative mobile elements. Several of the plasmids were self-transmissible or mobilizable and could be maintained in the absence of antibiotic selection. The IncFII plasmid pEFC36a showed the highest degree of sequence identity to plasmid R1 which has been isolated in England more than fifty years ago from a patient suffering from a Salmonella infection. Functional conservation of key regulatory features of this F-like conjugation module were demonstrated by the finding that the conjugation frequency of pEFC36a could be stimulated by the positive regulator of plasmid R1 DNA transfer genes, TraJ.

  10. Characterization and comparative analysis of antibiotic resistance plasmids isolated from a wastewater treatment plant.

    Science.gov (United States)

    Rahube, Teddie O; Viana, Laia S; Koraimann, Günther; Yost, Christopher K

    2014-01-01

    A wastewater treatment plant (WWTP) is an environment high in nutrient concentration with diverse bacterial populations and can provide an ideal environment for the proliferation of mobile elements such as plasmids. WWTPs have also been identified as reservoirs for antibiotic resistance genes that are associated with human pathogens. The objectives of this study were to isolate and characterize self-transmissible or mobilizable resistance plasmids associated with effluent from WWTP. An enrichment culture approach designed to capture plasmids conferring resistance to high concentrations of erythromycin was used to capture plasmids from an urban WWTP servicing a population of ca. 210,000. DNA sequencing of the plasmids revealed diversity of plasmids represented by incompatibility groups IncU, col-E, IncFII and IncP-1β. Genes coding resistance to clinically relevant antibiotics (macrolide, tetracycline, beta-lactam, trimethoprim, chloramphenicol, sulphonamide), quaternary ammonium compounds and heavy metals were co-located on these plasmids, often within transposable and integrative mobile elements. Several of the plasmids were self-transmissible or mobilizable and could be maintained in the absence of antibiotic selection. The IncFII plasmid pEFC36a showed the highest degree of sequence identity to plasmid R1 which has been isolated in England more than 50 years ago from a patient suffering from a Salmonella infection. Functional conservation of key regulatory features of this F-like conjugation module were demonstrated by the finding that the conjugation frequency of pEFC36a could be stimulated by the positive regulator of plasmid R1 DNA transfer genes, TraJ.

  11. 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

  12. Analysis of Ligand-Receptor Association and Intermediate Transfer Rates in Multienzyme Nanostructures with All-Atom Brownian Dynamics Simulations.

    Science.gov (United States)

    Roberts, Christopher C; Chang, Chia-En A

    2016-08-25

    We present the second-generation GeomBD Brownian dynamics software for determining interenzyme intermediate transfer rates and substrate association rates in biomolecular complexes. Substrate and intermediate association rates for a series of enzymes or biomolecules can be compared between the freely diffusing disorganized configuration and various colocalized or complexed arrangements for kinetic investigation of enhanced intermediate transfer. In addition, enzyme engineering techniques, such as synthetic protein conjugation, can be computationally modeled and analyzed to better understand changes in substrate association relative to native enzymes. Tools are provided to determine nonspecific ligand-receptor association residence times, and to visualize common sites of nonspecific association of substrates on receptor surfaces. To demonstrate features of the software, interenzyme intermediate substrate transfer rate constants are calculated and compared for all-atom models of DNA origami scaffold-bound bienzyme systems of glucose oxidase and horseradish peroxidase. Also, a DNA conjugated horseradish peroxidase enzyme was analyzed for its propensity to increase substrate association rates and substrate local residence times relative to the unmodified enzyme. We also demonstrate the rapid determination and visualization of common sites of nonspecific ligand-receptor association by using HIV-1 protease and an inhibitor, XK263. GeomBD2 accelerates simulations by precomputing van der Waals potential energy grids and electrostatic potential grid maps, and has a flexible and extensible support for all-atom and coarse-grained force fields. Simulation software is written in C++ and utilizes modern parallelization techniques for potential grid preparation and Brownian dynamics simulation processes. Analysis scripts, written in the Python scripting language, are provided for quantitative simulation analysis. GeomBD2 is applicable to the fields of biophysics, bioengineering

  13. Microbial degradation of pyridine using Pseudomonas sp. and isolation of plasmid responsible for degradation.

    Science.gov (United States)

    Mohan, S Venkata; Sistla, Srinivas; Guru, R Kumar; Prasad, K Krishna; Kumar, C Suresh; Ramakrishna, S V; Sarma, P N

    2003-01-01

    Pseudomonas (PI2) capable of degrading pyridine was isolated from the mixed population of the activated sludge unit which was being used for treating complex effluents, the strain was characterized. Aerobic degradation of pyridine was studied with the isolated strain and the growth parameters were evaluated. Pyridine degradation was further conformed by chromatography (HPLC) analysis. The process parameters like biomass growth and dissolved oxygen consumption were monitored during pyridine degradation. In order to conform with the plasmid capability to degrade pyridine, the requisite plasmid was isolated and transferred to DH 5alpha Escherichia coli. The subsequent biodegradation studies revealed the ability of the transformed plasmid capability to degrade the pyridine.

  14. Effect of radiocesium transfer on ambient dose rate in forest environments affected by the Fukushima Nuclear Power Plant accident

    Science.gov (United States)

    Kato, H.

    2015-12-01

    We investigated the transfer of canopy-intercepted radiocesium to the forest floor during 3 years following the Fukushima Daiichi Nuclear Power Plant accident. The cesium-137 (Cs-137) contents in throughfall, stemflow, and litterfall were monitored in two coniferous stands (plantation of Japanese cedar) and a deciduous broad-leaved forest stand (Japanese oak with red pine). We also measured the ambient dose rate (ADR) at different heights in the forest using a survey meter and a portable Ge gamma-ray detector. Total Cs-137 deposition flux from the canopy to forest floor for the mature cedar, young cedar, and the mixed broad-leaved stands were 166 kBq/m2, 174 kBq/m2, and 60 kBq/m2, respectively. These values correspond to 38%, 40% and 13% of total atmospheric input after the accident. The ambient dose rate in forest exhibited height dependency and its vertical distribution varied with forest type and stand age. The ambient dose rate showed an exponential decrease with time for all the forest sites, however the decreasing trend differed depending on the height of dose measurement and forest type. The ambient dose rate at the canopy (approx. 10 m-height) decreased faster than that expected from physical decay of the two radiocesium isotopes, whereas those at the forest floor varied between the three forest stands. The radiocesium deposition via throughfall seemed to increase ambient dose rate during the first 200 days after the accident, however there was no clear relationship between litterfall and ambient dose rate since 400 days after the accident. These data suggested that the ambient dose rate in forest environment varied both spatially and temporally reflecting the transfer of radiocesium from canopy to forest floor. However, further monitoring investigation and analysis are required to determine the effect of litterfall on long-term trend of ambient dose rate in forest environments.

  15. 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.

  16. 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.

  17. Broad host range plasmids can invade an unexpectedly diverse fraction of a soil bacterial community

    DEFF Research Database (Denmark)

    Klümper, Uli; Riber, Leise; Dechesne, Arnaud

    2014-01-01

    range of IncP- and IncPromA-type broad host range plasmids from three proteobacterial donors to a soil bacterial community. We identified transfer to many different recipients belonging to 11 different bacterial phyla. The prevalence of transconjugants belonging to diverse Gram-positive Firmicutes...... bacteria and can, therefore, directly connect large proportions of the soil bacterial gene pool. This finding reinforces the evolutionary and medical significances of these plasmids....

  18. High speed transfer switch with 50 kA and 50 kV ratings

    Science.gov (United States)

    Reass, W. A.; Kasik, R. J.; Wilds, W. A.

    This paper gives the mechanical design and electrical parameters of a pneumatically operated transfer switch. This design is used to switch 3-second 50-kA current pulses, and is easily capable of 75 kA operation, with water-cooled versions capable of 20 kA continuously. Although the switch is not specifically designed to make or break 50 kA, it is provided with auxiliary Elkonite arcing contacts that have proven their value in protecting the main electrodes even under repetitive (50 kA) fault conditions. Included in this presentation will be the results of extensive life testing and associated criteria.

  19. 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.

  20. 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.

  1. 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.

  2. 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.

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

    Science.gov (United States)

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

    2010-01-01

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

  4. NLP modeling for the optimization of LiBr-H2O absorption refrigeration systems with exergy loss rate, heat transfer area, and cost as single objective functions

    DEFF Research Database (Denmark)

    Mussati, Sergio F.; Gernaey, Krist; Morosuk, Tatiana

    2016-01-01

    exergy loss rate, the total heat transfer area, and the total annual cost of the system. It was found that the optimal solution obtained by minimization of the total exergy loss rate provides “theoretical” upper bounds not only for the total heat transfer area of the system but also for each process unit...... and all stream temperatures, while the optimal solution obtained by minimization of the total heat transfer area provides the lower bounds for these model variables, to solve a cost optimization problem. The minimization of the total exergy loss rate by varying parametrically the available total heat...... and quantitatively with increasing available total heat transfer area. These optimization results allowed to find a “practical” value of the total heat transfer area, i.e. no benefits can be obtained by increasing the available total heat transfer area above this value since the minimal total exergy loss value...

  5. Strongly Nonlinear Dependence of Energy Transfer Rate on sp(2) Carbon Content in Reduced Graphene Oxide-Quantum Dot Hybrid Structures.

    Science.gov (United States)

    Dong, Yitong; Son, Dong Hee

    2015-01-02

    The dependence of the energy transfer rate on the content of sp(2)-hybridized carbon atoms in the hybrid structures of reduced graphene oxide (RGO) and Mn-doped quantum dot (QD(Mn)) was investigated. Taking advantage of the sensitivity of QD(Mn)'s dopant luminescence lifetime only to the energy transfer process without interference from the charge transfer process, the correlation between the sp(2) carbon content in RGO and the rate of energy transfer from QD(Mn) to RGO was obtained. The rate of energy transfer showed a strongly superlinear increase with increasing sp(2) carbon content in RGO, suggesting the possible cooperative behavior of sp(2) carbon domains in the energy transfer process as the sp(2) carbon content increases.

  6. Does premature elevated progesterone on the day of trigger increase spontaneous abortion rates in fresh and subsequent frozen embryo transfers?

    Science.gov (United States)

    Healy, Mae; Patounakis, George; Zanelotti, Austin; Devine, Kate; DeCherney, Alan; Levy, Michael; Hill, Micah J

    2017-06-01

    Recent evidence has shown elevated progesterone (P) advances the endometrium in fresh ART cycles, creating asynchrony with the embryo and thus implantation failure and decreased live birth rates. If the window of implantation is closing as the embryo attempts to implant, there may be difficulty with trophoblastic invasion, leading to failure of early pregnancies. Our objective was to evaluate if P on the day of trigger was associated with spontaneous abortion (SAB) rates in fresh ART transfers. This was a retrospective cohort study involving fresh autologous and FET cycles from 2011 to 2013. The main outcome was spontaneous abortion rates. About 4123 fresh and FET transfer cycles were included which resulted in 1547 fresh and 491 FET pregnancies. The overall SAB rate was 20% among fresh cycles and 19% in FET cycles. P on the day of trigger, as a continuous variable or when > 2 ng/mL, was not associated with SAB in fresh cycles. Similar results were found after adjusting for age, embryo quality, and embryo stage. Despite elevated P likely advancing the window of implantation, once implantation occurs, pregnancies were no longer negatively impacted by progesterone.

  7. [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.

  8. 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.

  9. Quantification of magnetization transfer rate and native T1 relaxation time of the brain: correlation with magnetization transfer ratio measurements in patients with multiple sclerosis.

    Science.gov (United States)

    Karampekios, Spyros; Papanikolaou, Nickolas; Papadaki, Eufrosini; Maris, Thomas; Uffman, Kai; Spilioti, Martha; Plaitakis, Andreas; Gourtsoyiannis, Nicholas

    2005-03-01

    The purpose of this paper is to perform quantitative measurements of the magnetization transfer rate (Kfor) and native T1 relaxation time (T1free) in the brain tissue of normal individuals and patients with multiple sclerosis (MS) by means of multiple gradient echo acquisitions, and to correlate these measurements with the magnetization transfer ratio (MTR). Quantitative magnetization transfer imaging was performed in five normal volunteers and 12 patients with relapsing-remitting MS on a 1.5 T magnetic resonance (MR) scanner. The T1 relaxation time under magnetization transfer irradiation (T1sat) was calculated by means of fitting the signal intensity over the flip angle in several 3D spoiled gradient echo acquisitions (3 degrees , 15 degrees , 30 degrees , and 60 degrees ), while a single acquisition without MT irradiation (flip angle of 3 degrees ) was utilized to calculate the MTR. The Kfor and T1free constants were quantified on a pixel-by-pixel basis and parametric maps were reconstructed. We performed 226 measurements of Kfor, T1free, and the MTR on normal white matter (NWM) of healthy volunteers (n=50), and normal-appearing white matter (NAWM) and pathological brain areas of MS patients (n=120 and 56, respectively). Correlation coefficients between Kfor-MTR, T1free-MTR, and T1free-Kfor were calculated. Lesions were classified, according to their characteristics on T1-weighted images, into isointense (compared to white matter), mildly hypointense (showing signal intensity lower than white matter and higher than gray matter), and severely hypointense (revealing signal intensity lower than gray matter). "Dirty" white matter (DWM) corresponded to areas with diffused high signal, as identified on T2-weighted images. Strong correlation coefficients were obtained between MTR and Kfor for all lesions studied (r2=0.9, pKfor and T1free measurements (r2=0.98, pKfor and T1free were found for the rest of the subgroups, except for the NAWM, in which a moderate

  10. Quantification of magnetization transfer rate and native T1 relaxation time of the brain: correlation with magnetization transfer ratio measurements in patients with multiple sclerosis

    Energy Technology Data Exchange (ETDEWEB)

    Karampekios, Spyros; Papanikolaou, Nickolas; Papadaki, Eufrosini; Gourtsoyiannis, Nicholas [University Hospital of Heraklion, Department of Radiology, Heraklion (Greece); Maris, Thomas [University Hospital of Heraklion, Department of Medical Physics, Heraklion (Greece); Uffman, Kai [University Hospital of Essen, Department of Diagnostic Radiology, Essen (Germany); Spilioti, Martha; Plaitakis, Andreas [University Hospital of Heraklion, Department of Neurology, Heraklion (Greece)

    2005-03-01

    The purpose of this paper is to perform quantitative measurements of the magnetization transfer rate (Kfor) and native T1 relaxation time (T1free) in the brain tissue of normal individuals and patients with multiple sclerosis (MS) by means of multiple gradient echo acquisitions, and to correlate these measurements with the magnetization transfer ratio (MTR). Quantitative magnetization transfer imaging was performed in five normal volunteers and 12 patients with relapsing-remitting MS on a 1.5 T magnetic resonance (MR) scanner. The T1 relaxation time under magnetization transfer irradiation (T1sat) was calculated by means of fitting the signal intensity over the flip angle in several 3D spoiled gradient echo acquisitions (3 , 15 , 30 , and 60 ), while a single acquisition without MT irradiation (flip angle of 3 ) was utilized to calculate the MTR. The Kfor and T1free constants were quantified on a pixel-by-pixel basis and parametric maps were reconstructed. We performed 226 measurements of Kfor, T1free, and the MTR on normal white matter (NWM) of healthy volunteers (n=50), and normal-appearing white matter (NAWM) and pathological brain areas of MS patients (n=120 and 56, respectively). Correlation coefficients between Kfor-MTR, T1free-MTR, and T1free-Kfor were calculated. Lesions were classified, according to their characteristics on T1-weighted images, into isointense (compared to white matter), mildly hypointense (showing signal intensity lower than white matter and higher than gray matter), and severely hypointense (revealing signal intensity lower than gray matter). ''Dirty'' white matter (DWM) corresponded to areas with diffused high signal, as identified on T2-weighted images. Strong correlation coefficients were obtained between MTR and Kfor for all lesions studied (r{sup 2}=0.9, p<0.0001), for mildly hypointense plaques (r{sup 2}=0.82, p<0.0001), and for DWM (r{sup 2}=0.78, p=0.0007). In contrast, comparison between MTR and T1free values

  11. A new approach to the correlation of boundary layer mass transfer rates with thermal diffusion and/or variable properties

    Science.gov (United States)

    Srivastava, R.; Rosner, D. E.

    1979-01-01

    A rational approach to the correlation of boundary layer mass transport rates, applicable to many commonly encountered laminar flow conditions with thermal diffusion and/or variable properties, is outlined. The correlation scheme builds upon already available constant property blowing/suction solutions by introducing appropriate correction factors to account for the additional ('pseudo' blowing and source) effects identified with variable properties and thermal diffusion. Applications of the scheme to the particular laminar boundary layer mass transfer problems considered herein (alkali and transition metal compound vapor transport) indicates satisfactory accuracy up to effective blowing factors equivalent to about one third of the 'blow off' value. As a useful by-product of the variable property correlation, we extend the heat-mass transfer analogy, for a wide range of Lewis numbers, to include variable property effects.

  12. Charge transfer rates in organic semiconductors beyond first-order perturbation: from weak to strong coupling regimes.

    Science.gov (United States)

    Nan, Guangjun; Wang, Linjun; Yang, Xiaodi; Shuai, Zhigang; Zhao, Yi

    2009-01-14

    Semiclassical Marcus electron transfer theory is often employed to investigate the charge transport properties of organic semiconductors. However, quite often the electronic couplings vary several orders of magnitude in organic crystals, which goes beyond the application scope of semiclassical Marcus theory with the first-order perturbative nature. In this work, we employ a generalized nonadiabatic transition state theory (GNTST) [Zhao et al., J. Phys. Chem. A 110, 8204 (2004)], which can evaluate the charge transfer rates from weak to strong couplings, to study charge transport properties in prototypical organic semiconductors: quaterthiophene and sexithiophene single crystals. By comparing with GNTST results, we find that the semiclassical Marcus theory is valid for the case of the coupling semiconductors with general electronic coupling terms. Taking oligothiophenes as examples, we find that our GNTST-calculated hole mobility is about three times as large as that from the semiclassical Marcus theory. The difference arises from the quantum nuclear tunneling and the nonperturbative effects.

  13. Prediction of the heat transfer rate of a single layer wire-on-tube type heat exchanger using ANFIS

    Energy Technology Data Exchange (ETDEWEB)

    Hayati, Mohsen [Electrical Engineering Department, Faculty of Engineering, Razi University, Tagh-E-Bostan, Kermanshah 67149 (Iran); Computational Intelligence Research Center, Razi University, Tagh-E-Bostan, Kermanshah 67149 (Iran); Rezaei, Abbas; Seifi, Majid [Electrical Engineering Department, Faculty of Engineering, Razi University, Tagh-E-Bostan, Kermanshah 67149 (Iran)

    2009-12-15

    In this paper, we applied an Adaptive Neuro-Fuzzy Inference System (ANFIS) model for prediction of the heat transfer rate of the wire-on-tube type heat exchanger. Limited experimental data was used for training and testing ANFIS configuration with the help of hybrid learning algorithm consisting of backpropagation and least-squares estimation. The predicted values are found to be in good agreement with the actual values from the experiments with mean relative error less than 2.55%. Also, we compared the proposed ANFIS model to an ANN approach. Results show that the ANFIS model has more accuracy in comparison to ANN approach. Therefore, we can use ANFIS model to predict the performances of thermal systems in engineering applications, such as modeling heat exchangers for heat transfer analysis. (author)

  14. Semiclassical Green’s functions and an instanton formulation of electron-transfer rates in the nonadiabatic limit

    Energy Technology Data Exchange (ETDEWEB)

    Richardson, Jeremy O., E-mail: jeremy.richardson@fau.de; Bauer, Rainer; Thoss, Michael [Institut für Theoretische Physik und Interdisziplinäres Zentrum für Molekulare Materialien, Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), Staudtstraße 7/B2, 91058 Erlangen (Germany)

    2015-10-07

    We present semiclassical approximations to Green’s functions of multidimensional systems, extending Gutzwiller’s work to the classically forbidden region. Based on steepest-descent integrals over these functions, we derive an instanton method for computing the rate of nonadiabatic reactions, such as electron transfer, in the weak-coupling limit, where Fermi’s golden-rule can be employed. This generalizes Marcus theory to systems for which the environment free-energy curves are not harmonic and where nuclear tunnelling plays a role. The derivation avoids using the Im F method or short-time approximations to real-time correlation functions. A clear physical interpretation of the nuclear tunnelling processes involved in an electron-transfer reaction is thus provided. In Paper II [J. O. Richardson, J. Chem. Phys. 143, 134116 (2015)], we discuss numerical evaluation of the formulae.

  15. Influence of heat transfer rates on pressurization of liquid/slush hydrogen propellant tanks

    Science.gov (United States)

    Sasmal, G. P.; Hochstein, J. I.; Hardy, T. L.

    1993-01-01

    A multi-dimensional computational model of the pressurization process in liquid/slush hydrogen tank is developed and used to study the influence of heat flux rates at the ullage boundaries on the process. The new model computes these rates and performs an energy balance for the tank wall whereas previous multi-dimensional models required a priori specification of the boundary heat flux rates. Analyses of both liquid hydrogen and slush hydrogen pressurization were performed to expose differences between the two processes. Graphical displays are presented to establish the dependence of pressurization time, pressurant mass required, and other parameters of interest on ullage boundary heat flux rates and pressurant mass flow rate. Detailed velocity fields and temperature distributions are presented for selected cases to further illuminate the details of the pressurization process. It is demonstrated that ullage boundary heat flux rates do significantly effect the pressurization process and that minimizing heat loss from the ullage and maximizing pressurant flow rate minimizes the mass of pressurant gas required to pressurize the tank. It is further demonstrated that proper dimensionless scaling of pressure and time permit all the pressure histories examined during this study to be displayed as a single curve.

  16. In situ detection of horizontal transfer of mobile genetic elements

    DEFF Research Database (Denmark)

    Haagensen, Janus Anders Juul; Hansen, Susse Kirkelund; Johansen, Tove

    2002-01-01

    Plasmid transfer was investigated in microbial populations associated with different types of surfaces. The general strategy behind these investigations was to label the transferable plasmid with a gene encoding a fluorescent protein in order to make it a transfer reporter. This was achieved by f...

  17. The effect of ultrasound irradiation on the convective heat transfer rate during immersion cooling of a stationary sphere.

    Science.gov (United States)

    Kiani, Hossein; Sun, Da-Wen; Zhang, Zhihang

    2012-11-01

    It has been proven that ultrasound irradiation can enhance the rate of heat transfer processes. The objective of this work was to study the heat transfer phenomenon, mainly the heat exchange at the surface, as affected by ultrasound irradiation around a stationary copper sphere (k=386W m(-1)K(-1), C(p)=384J kg(-1)K(-1), ρ=8660kg m(-3)) during cooling. The sphere (0.01m in diameter) was immersed in an ethylene glycol-water mixture (-10°C) in an ultrasonic cooling system that included a refrigerated circulator, a flow meter, an ultrasound generator and an ultrasonic bath. The temperature of the sphere was recorded using a data logger equipped with a T-type thermocouple in the center of the sphere. The temperature of the cooling medium was also monitored by four thermocouples situated at different places in the bath. The sphere was located at different positions (0.02, 0.04 and 0.06m) above the transducer surface of the bath calculated considering the center of the sphere as the center of the reference system and was exposed to different intensities of ultrasound (0, 120, 190, 450, 890, 1800, 2800, 3400 and 4100W m(-2)) during cooling. The frequency of the ultrasound was 25kHz. It was demonstrated that ultrasound irradiation can increase the rate of heat transfer significantly, resulting in considerably shorter cooling times. Higher intensities caused higher cooling rates, and Nu values were increased from about 23-27 to 25-108 depending on the intensity of ultrasound and the position of the sphere. However, high intensities of ultrasound led to the generation of heat at the surface of the sphere, thus limiting the lowest final temperature achieved. An analytical solution was developed considering the heat generation and was fitted to the experimental data with R(2) values in the range of 0.910-0.998. Visual observations revealed that both cavitation and acoustic streaming were important for heat transfer phenomenon. Cavitation clouds at the surface of the sphere

  18. Risk Factors Influencing Conception Rate in Holstein Heifers before Artificial Insemination or Embryo Transfer

    OpenAIRE

    2016-01-01

    The objective of this study was to show the risk factors affecting the conception rate in Holstein heifers after synchronization of estrus. A total of 275 Holstein heifers housed in a free barn were used for the experiment. The herd was visited regularly at four week intervals for synchronization of estrus using Heatsynch and CIDR-Heatsynch protocols. A group of four to 14 animals, depending on the availability, were referred to the experiment at each visit. Estrus induction rates in the two ...

  19. Methods to increase the rate of mass transfer during osmotic dehydration of foods

    Directory of Open Access Journals (Sweden)

    Anna Chwastek

    2014-12-01

    Full Text Available Traditional methods of food preservation such as freezing, freeze drying (lyophilization, vacuum drying, convection drying are often supplemented by new technologies that enable obtaining of high quality products. Osmotic dehydration is more and more often used during processing of fruits and vegetables. This method allows maintaining good organoleptic and functional properties in the finished product. Obtaining the desired degree of dehydration or saturation of the material with an osmoactive substance often requires  elongation of time or use of high temperatures. In recent years much attention was devoted to techniques aimed at increasing the mass transfer between the dehydrated material and the hypertonic solution. The work reviews the literature focused on methods of streamlining the process of osmotic dehydration which include the use of: ultrasound, high hydrostatic pressure, vacuum osmotic dehydration and pulsed electric field.

  20. Single-embryo transfer reduces clinical pregnancy rates and live births in fresh IVF and Intracytoplasmic Sperm Injection (ICSI cycles: a meta-analysis

    Directory of Open Access Journals (Sweden)

    Oliveira João

    2009-04-01

    Full Text Available Abstract Background It has become an accepted procedure to transfer more than one embryo to the patient to achieve acceptable ongoing pregnancy rates. However, transfers of more than a single embryo increase the probability of establishing a multiple gestation. Single-embryo transfer can minimize twin pregnancies but may also lower live birth rates. This meta-analysis aimed to compare current data on single-embryo versus double-embryo transfer in fresh IVF/ICSI cycles with respect to implantation, ongoing pregnancy and live birth rates. Methods Search strategies included on-line surveys of databases from 1995 to 2008. Data management and analysis were conducted using the Stats Direct statistical software. The fixed-effect model was used for odds ratio (OR. Fixed-effect effectiveness was evaluated by the Mantel Haenszel method. Seven trials fulfilled the inclusion criteria. Results When pooling results under the fixed-effect model, the implantation rate was not significantly different between double-embryo transfer (34.5% and single-embryo transfer group (34.7% (P = 0.96; OR = 0.99, 95% CI 0.78, 1.25. On the other hand, double-embryo transfer produced a statistically significantly higher ongoing clinical pregnancy rate (44.5% than single-embryo transfer (28.3% (P Conclusion Meta-analysis with 95% confidence showed that, despite similar implantation rates, fresh double-embryo transfer had a 1.64 to 2.60 times greater ongoing pregnancy rate and 1.44 to 2.42 times greater live birth rate than single-embryo transfer in a population suitable for ART treatment.

  1. A 2,4-dichlorophenoxyacetic acid degradation plasmid pM7012 discloses distribution of an unclassified megaplasmid group across bacterial species.

    Science.gov (United States)

    Sakai, Yoriko; Ogawa, Naoto; Shimomura, Yumi; Fujii, Takeshi

    2014-03-01

    Analysis of the complete nucleotide sequence of plasmid pM7012 from 2,4-dichlorophenoxyacetic-acid (2,4-D)-degrading bacterium Burkholderia sp. M701 revealed that the plasmid had 582 142 bp, with 541 putative protein-coding sequences and 39 putative tRNA genes for the transport of the standard 20 aa. pM7012 contains sequences homologous to the regions involved in conjugal transfer and plasmid maintenance found in plasmids byi_2p from Burkholderia sp. YI23 and pBVIE01 from Burkholderia sp. G4. No relaxase gene was found in any of these plasmids, although genes for a type IV secretion system and type IV coupling proteins were identified. Plasmids with no relaxase gene have been classified as non-mobile plasmids. However, nucleotide sequences with a high level of similarity to the genes for plasmid transfer, plasmid maintenance, 2,4-D degradation and arsenic resistance contained on pM7012 were also detected in eight other megaplasmids (~600 or 900 kb) found in seven Burkholderia strains and a strain of Cupriavidus, which were isolated as 2,4-D-degrading bacteria in Japan and the United States. These results suggested that the 2,4-D degradation megaplasmids related to pM7012 are mobile and distributed across various bacterial species worldwide, and that the plasmid group could be distinguished from known mobile plasmid groups.

  2. An extranuclear expression system for analysis of cytoplasmic promoters of yeast linear killer plasmids.

    Science.gov (United States)

    Schründer, J; Meinhardt, F

    1995-03-01

    Based on the cytoplasmically localized killer plasmids pGKL1 and pGKL2 of Kluyveromyces lactis two new linear hybrid plasmids were constructed which consist of pGKL1, into which in addition to the previously developed cytoplasmically expressible LEU2* selectable marker a glucose dehydrogenase-encoding bacterial gene (gdh A) has been integrated. One of the hybrid plasmids carries the bacterial gene preceded by an arbitrarily placed cytoplasmic promoter (upstream conserved sequence) in front of the coding region (pRKL121). The other plasmid was constructed in such a way that the ATG start codon of the gdh A gene was fused in frame to the ATG start codon of the killer plasmid's open reading frame 5 (pRKL122). The structures of both linear hybrid plasmids were confirmed by restriction analysis, Southern hybridization, and sequencing of the junction sites. Yeast strains carrying either of the plasmids expressed the glucose dehydrogenase gene; however, expression of the in phase fused gene was 40-fold higher compared to the arbitrarily placed cytoplasmic promoter. In general, an in phase fusion was not required for expression, but efficiency is dramatically enhanced when the 5' noncoding sequences in front of the heterologous genes are the same as those found on the native killer plasmids. The developed system can serve as a reporter for determining the efficiency of the different cytoplasmic promoters present on both linear plasmids. Hybrid plasmids were stably maintained without selective pressure in K. lactis and they were transferred and expressed also in Saccharomyces cerevisiae.

  3. Differential behavior of plasmids containing chromosomal DNA insertions of various sizes during transformation and conjugation in Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

    Balganesh, M.; Setlow, J.K.

    1985-01-01

    Plasmids with chromosomal insertions were constructed by removal of a 1.1-kilobase-pair piece from the 9.8-kilobase-pair vector plasmid pDM2 by EcoRI digestion and inserting in its place various lengths of chromosomal DNA (1.7, 3.4, and 9.0 kilobase pairs) coding for resistance to novobiocin. A fourth plasmid was constructed by insertion of the largest piece of chromosomal DNA into the SmaI site of pDM2. The plasmids without inserts were taken up poorly by competent cells and thus were considered not to contain specific DNA uptake sites. The presence of even the smallest insert of chromosomal DNA caused a large increase in transformation of Rec/sup +/ and Rec/sup -/ strains. The frequency of plasmid establishment in Rec/sup +/ cells by transformation increased exponentially with increasing insert size, but in Rec/sup -/ cells there was less transformation by the larger plasmids. Conjugal transfer of these plasmids was carried out with the 35-kilobase-pair mobilizing plasmid pHD147. The frequency of establishment of plasmids by this method not only was not markedly affected by the presence of the insertions, but also decreased somewhat with increase in insert size and was independent of rec-1 and rec-2 genes. Recombination between plasmid and chromosome was readily detected after transformation, but could not be detected after transconjugation even when the recipient cells were Rec/sup +/ and made competent. These data suggested that there is a special processing of plasmid DNA that enters the competent cells in transformation that makes possible recombination of homologous regions of the plasmid with the chromosome and pairing with the chromosome that aids plasmid establishment.

  4. Live birth rate and number of blastomeres on day 2 transfer

    DEFF Research Database (Denmark)

    Azzarello, Antonino; Hoest, Thomas; Hay-Schmidt, Anders

    2016-01-01

    Purpose To investigate whether the presence of large fragment (LF) and abnormal cell divisions (ACDs) has influenced the correlation between live birth rate and number of blastomeres detected on day 2 by conventional scoring. Methods This study included 578 embryos cultured in time lapse and sele......Purpose To investigate whether the presence of large fragment (LF) and abnormal cell divisions (ACDs) has influenced the correlation between live birth rate and number of blastomeres detected on day 2 by conventional scoring. Methods This study included 578 embryos cultured in time lapse...

  5. Plasmid with Colistin Resistance Gene mcr-1 in Extended-Spectrum-β-Lactamase-Producing Escherichia coli Strains Isolated from Pig Slurry in Estonia.

    Science.gov (United States)

    Brauer, Age; Telling, Kaidi; Laht, Mailis; Kalmus, Piret; Lutsar, Irja; Remm, Maido; Kisand, Veljo; Tenson, Tanel

    2016-11-01

    A plasmid carrying the colistin resistance gene mcr-1 was isolated from a pig slurry sample in Estonia. The gene was present on a 33,311-bp plasmid of the IncX4 group. mcr-1 is the only antibiotic resistance gene on the plasmid, with the other genes mainly coding for proteins involved in conjugative DNA transfer (taxA, taxB, taxC, trbM, and the pilX operon). The plasmid pESTMCR was present in three phylogenetically very different Escherichia coli strains, suggesting that it has high potential for horizontal transfer. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  6. The Salmonella genomic island 1 is specifically mobilized in trans by the IncA/C multidrug resistance plasmid family.

    Science.gov (United States)

    Douard, Gregory; Praud, Karine; Cloeckaert, Axel; Doublet, Benoît

    2010-12-20

    The Salmonella genomic island 1 (SGI1) is a Salmonella enterica-derived integrative mobilizable element (IME) containing various complex multiple resistance integrons identified in several S. enterica serovars and in Proteus mirabilis. Previous studies have shown that SGI1 transfers horizontally by in trans mobilization in the presence of the IncA/C conjugative helper plasmid pR55. Here, we report the ability of different prevalent multidrug resistance (MDR) plasmids including extended-spectrum β-lactamase (ESBL) gene-carrying plasmids to mobilize the multidrug resistance genomic island SGI1. Through conjugation experiments, none of the 24 conjugative plasmids tested of the IncFI, FII, HI2, I1, L/M, N, P incompatibility groups were able to mobilize SGI1 at a detectable level (transfer frequency IncA/C incompatibility group. Several conjugative IncA/C MDR plasmids as well as the sequenced IncA/C reference plasmid pRA1 of 143,963 bp were shown to mobilize in trans SGI1 from a S. enterica donor to the Escherichia coli recipient strain. Depending on the IncA/C plasmid used, the conjugative transfer of SGI1 occurred at frequencies ranging from 10(-3) to 10(-6) transconjugants per donor. Of particular concern, some large IncA/C MDR plasmids carrying the extended-spectrum cephalosporinase bla(CMY-2) gene were shown to mobilize in trans SGI1. The ability of the IncA/C MDR plasmid family to mobilize SGI1 could contribute to its spread by horizontal transfer among enteric pathogens. Moreover, the increasing prevalence of IncA/C plasmids in MDR S. enterica isolates worldwide has potential implications for the epidemic success of the antibiotic resistance genomic island SGI1 and its close derivatives.

  7. Broad-Host-Range IncP-1 plasmids and their resistance potential

    Directory of Open Access Journals (Sweden)

    Magdalena ePopowska

    2013-03-01

    Full Text Available The plasmids of the incompatibility group IncP-1, also called IncP, as extrachromosomal genetic elements can transfer and replicate virtually in all Gram-negative bacteria. They are composed of backbone genes that encode a variety of essential functions and accessory genes that have implications for human health and environmental bioremediation. Broad-host-range IncP plasmids are known to spread genes between distinct phylogenetic groups of bacteria. These genes often code for resistances to a broad spectrum of antibiotics, heavy metals and quaternary ammonium compounds used as disinfectants. The backbone of these plasmids carries modules that enable them to effectively replicate, move to a new host via conjugative transfer and to be stably maintained in bacterial cells. The adaptive, resistance and virulence genes are mainly located on mobile genetic elements integrated between the functional plasmid backbone modules. Environmental studies have demonstrated the wide distribution of IncP-like replicons in manure, soils and wastewater treatment plants. They also are present in strains of pathogenic or opportunistic bacteria, which can be a cause for concern, because they may encode multiresistance. Their broad distribution suggests that IncP plasmids play a crucial role in bacterial adaptation by utilizing horizontal gene transfer. This review summarizes the variety of genetic information and physiological functions carried by IncP plasmids, which can contribute to the spread of antibiotic and heavy metal resistance while also mediating the process of bioremediation of pollutants. Due to the location of the resistance genes on plasmids with a broad host range and the presence of transposons carrying these genes it seems that the spread of these genes would be possible and quite hazardous in infection control. Future studies are required to determine the level of risk of the spread of resistance genes located on these plasmids.

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

    Science.gov (United States)

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

    2017-01-01

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

  9. Genomic and Functional Characterization of qnr-Encoding Plasmids from Municipal Wastewater Biosolid Klebsiella pneumoniae Isolates.

    Science.gov (United States)

    Kaplan, Ella; Sela, Noa; Doron-Faigenboim, Adi; Navon-Venezia, Shiri; Jurkevitch, Edouard; Cytryn, Eddie

    2015-01-01

    Municipal wastewater treatment facilities are considered to be "hotspots" for antibiotic resistance, since they conjoin high densities of environmental and fecal bacteria with selective pressure in the form of sub-therapeutic concentrations of antibiotics. Discharged effluents and biosolids from these facilities can disseminate antibiotic resistant genes to terrestrial and aquatic environments, potentially contributing to the increasing global trend in antibiotic resistance. This phenomenon is especially pertinent when resistance genes are associated with mobile genetic elements such as conjugative plasmids, which can be transferred between bacterial phyla. Fluoroquinolones are among the most abundant antibiotic compounds detected in wastewater treatment facilities, especially in biosolids, where due to their hydrophobic properties they accumulate to concentrations that may exceed 40 mg/L. Although fluoroquinolone resistance is traditionally associated with mutations in the gyrA/topoisomerase IV genes, there is increasing evidence of plasmid-mediated quinolone resistance, which is primarily encoded on qnr genes. In this study, we sequenced seven qnr-harboring plasmids from a diverse collection of Klebsiella strains, isolated from dewatered biosolids from a large wastewater treatment facility in Israel. One of the plasmids, termed pKPSH-11XL was a large (185.4 kbp), multi-drug resistance, IncF-type plasmid that harbored qnrB and 10 additional antibiotic resistance genes that conferred resistance to five different antibiotic families. It was highly similar to the pKPN3-like plasmid family that has been detected in multidrug resistant clinical Klebsiella isolates. In contrast, the six additional plasmids were much smaller (7-9 Kbp) and harbored a qnrS -type gene. These plasmids were highly similar to each other and closely resembled pGNB2, a plasmid isolated from a German wastewater treatment facility. Comparative genome analyses of pKPSH-11XL and other pKPN3-like

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

    Directory of Open Access Journals (Sweden)

    Summers David K

    2010-02-01

    importance of plasmids for lateral gene transfer (including chromosomal sequences to distinct genera.

  11. Direct measurement of heat transfer rates and coefficients in freezing processes by the use of heat flux sensors

    Energy Technology Data Exchange (ETDEWEB)

    Amarante, A.; Lanoiselle, J.L.; Ramirez, A.

    2003-10-01

    Heat exchange is often complex to assess in freezing equipment. Either the extensive calculation procedures based on product time-temperature data, or the lack of accurate thermophysical properties, or even the non-uniform processing conditions in industrial equipment, results in increased difficulty in calculating accurate heat exchange parameters. The present study aims to solve this kind of problem by introducing the use of heat flux sensors (or fluxmeters) for an online measurement of heat exchange parameters during freezing processes. Since food products often have irregular, moist and greasy surfaces, bad attachment of the sensors can lead to low accuracy in heat flux measurement. First, a technique was improved in this particular and a numerical procedure based on matching the experimental and simulated temperature histories was used to calibrate the sensors attached to Tylose gels submitted to freezing and thawing cycles. Following this, the sensors were applied directly to a vegetable product undergoing freezing in a static freezer to measure the instantaneous product heat release rate and the local heat transfer coefficient. A fluxmeter-plastic transducer was also developed and used, coupled to an anemometer to map axially and transversally the local effective heat transfer coefficient and air speed profiles in a Super-Contact freezing tunnel. Results were compared with numerical simulations and showed good agreement. Irregular air speed distribution and low efficiency heat transfer zones were accurately detected, providing information for equipment optimization. (author)

  12. Multi-rate mass transfer modeling of two-phase flow in highly heterogeneous fractured and porous media

    Science.gov (United States)

    Tecklenburg, Jan; Neuweiler, Insa; Carrera, Jesus; Dentz, Marco

    2016-05-01

    We study modeling of two-phase flow in highly heterogeneous fractured and porous media. The flow behaviour is strongly influenced by mass transfer between a highly permeable (mobile) fracture domain and less permeable (immobile) matrix blocks. We quantify the effective two-phase flow behavior using a multirate rate mass transfer (MRMT) approach. We discuss the range of applicability of the MRMT approach in terms of the pertinent viscous and capillary diffusion time scales. We scrutinize the linearization of capillary diffusion in the immobile regions, which allows for the formulation of MRMT in the form of a non-local single equation model. The global memory function, which encodes mass transfer between the mobile and the immobile regions, is at the center of this method. We propose two methods to estimate the global memory function for a fracture network with given fracture and matrix geometry. Both employ a scaling approach based on the known local memory function for a given immobile region. With the first method, the local memory function is calculated numerically, while the second one employs a parametric memory function in form of truncated power-law. The developed concepts are applied and tested for fracture networks of different complexity. We find that both physically based parameter estimation methods for the global memory function provide predictive MRMT approaches for the description of multiphase flow in highly heterogeneous porous media.

  13. Fluorescence resonance energy transfer (FRET) in chemistry and biology: Non-Förster distance dependence of the FRET rate

    Indian Academy of Sciences (India)

    Sangeeta Saini; Harjinder Singh; Biman Bagchi

    2006-01-01

    Fluorescence resonance energy transfer (FRET) is a popular tool to study equilibrium and dynamical properties of polymers and biopolymers in condensed phases and is now widely used in conjunction with single molecule spectroscopy. In the data analysis, one usually employs the Förster expression which predicts (1/6) distance dependence of the energy transfer rate. However, critical analysis shows that this expression can be of rather limited validity in many cases. We demonstrate this by explicitly considering a donor-acceptor system, polyfluorene (PF6)-tetraphenylporphyrin (TPP), where the size of both donor and acceptor is comparable to the distance separating them. In such cases, one may expect much weaker distance (as 1/2 or even weaker) dependence. We have also considered the case of energy transfer from a dye to a nanoparticle. Here we find 1/4 distance dependence at large separations, completely different from Förster. We also discuss recent application of FRET to study polymer conformational dynamics.

  14. Adaptation of the "Dynamic Method" for measuring the specific respiration rate in oxygen transfer systems through diffusion membrane.

    Science.gov (United States)

    Pamboukian, Marilena Martins; Pereira, Carlos Augusto; Augusto, Elisabeth de Fatima Pires; Tonso, Aldo

    2011-12-01

    Monitoring the specific respiration rate (Q(O2)) is a valuable tool to evaluate cell growth and physiology. However, for low Q(O2) values the accuracy may depend on the measurement methodology, as it is the case in animal cell culture. The widely used "Dynamic Method" imposes serious difficulties concerning oxygen transfer cancellation, especially through membrane oxygenation. This paper presents an improved procedure to this method, through an automated control of the gas inlet composition that can minimize the residual oxygen transfer driving force during the Q(O2) measurement phase. The improved technique was applied to animal cell cultivation, particularly three recombinant S2 (Drosophila melanogaster) insect cell lines grown in a membrane aeration bioreactor. The average measurements of the proposed method reached 98% of stationary liquid phase balance method, taken as a reference, compared to 21% when the traditional method was used. Furthermore, this methodology does not require knowledge of the volumetric transfer coefficient k(L)a, which may vary during growth.

  15. Multi-rate mass transfer modeling of two-phase flow in highly heterogeneous fractured and porous media

    CERN Document Server

    Tecklenburg, Jan; Carrera, Jesus; Dentz, Marco

    2016-01-01

    We study modeling of two-phase flow in highly heterogeneous fractured and porous media. The flow behaviour is strongly influenced by mass transfer between a highly permeable (mobile) fracture domain and less permeable (immobile) matrix blocks. We quantify the effective two-phase flow behaviour using a multirate rate mass transfer (MRMT) approach. We discuss the range of applicability of the MRMT approach in terms of the pertinent viscous and capillary diffusion time scales. We scrutinize the linearization of capillary diffusion in the immobile regions, which allows for the formulation of MRMT in the form of a non-local single equation model. The global memory function, which encodes mass transfer between the mobile and the immobile regions, is at the center of this method. We propose two methods to estimate the global memory function for a fracture network with given fracture and matrix geometry. Both employ a scaling approach based on the known local memory function for a given immobile region. With the firs...

  16. Protocol for Evaluating the Permissiveness of Bacterial Communities Toward Conjugal Plasmids by Quantification and Isolation of Transconjugants

    DEFF Research Database (Denmark)

    Klümper, Uli; Dechesne, Arnaud; Smets, Barth F.

    2014-01-01

    The transfer of conjugal plasmids is the main bacterial process of horizontal gene transfer to potentially distantly related bacteria. These extrachromosomal, circular DNA molecules host genes that code for their own replication and transfer to other organisms. Because additional accessory genes ...

  17. Bioaugmentation of DDT-contaminated soil by dissemination of the catabolic plasmid pDOD.

    Science.gov (United States)

    Gao, Chunming; Jin, Xiangxiang; Ren, Jingbei; Fang, Hua; Yu, Yunlong

    2015-01-01

    A plasmid transfer-mediated bioaugmentation method for the enhancement of dichlorodiphenyltrichloroethane (DDT) degradation in soil was developed using the catabolic plasmid pDOD from Sphingobacterium sp. D-6. The pDOD plasmid could be transferred to soil bacteria, such as members of Cellulomonas, to form DDT degraders and thus accelerate DDT degradation. The transfer efficiency of pDOD was affected by the donor, temperature, moisture, and soil type. Approximately 50.7% of the DDT in the contaminated field was removed 210 days after the application of Escherichia coli TG I (pDOD-gfp). The results suggested that seeding pDOD into soil is an effective bioaugmentation method for enhancing the degradation of DDT.

  18. Influence of methanol/sorbitol co-feeding rate on pAOX1 induction in a Pichia pastoris Mut+ strain in bioreactor with limited oxygen transfer rate.

    Science.gov (United States)

    Carly, F; Niu, H; Delvigne, F; Fickers, P

    2016-04-01

    High Pichia pastoris biomass density could be obtained using high co-feeding rate of methanol and sorbitol in a fed-batch or continuous culture, while further higher feeding rate finally leads to oxygen limitation in bioreactor. In the literature, there is lack of report about AOX1 promoter regulation with regard to dissolved oxygen level (DO). Therefore, in this work, chemostat cultures were performed to investigate the cell growth, metabolism and regulation of the AOX1 promoter (pAOX1) regarding co-feeding rate of optimized methanol/sorbitol mixture (methanol fraction 0.60 C-mol/C-mol) using a P. pastoris Mut+/pAOX1-lacZ strain. The oxygen transfer rates (OTR) in bioreactor were kept in the range of typical values of large bioreactor, i.e., 4-8 g/(L h) if DO equals 30 % saturation or 5-10 g/(L h) if DO nears zero. For DO >0, an increase of the carbon fed led to an increase of pAOX1 induction. By contrast, when dissolved oxygen was completely depleted, methanol accumulated, causing a 30 % decrease of pAOX1 induction. However, this decrease is more likely to be lined to methanol accumulation than to low level of dissolved oxygen (Methanol/sorbitol co-feeding allowed cells to adapt to oxygen transient limitations that often occur at industrial scale with reduced effect on pAOX1 induction. The optimal feeding rate tested here was 6.6 mmol C (DCW h)(-1) at an OTR of 8.28 g O2(L h)(-1) with over fivefold pAOX1 induction (probably directly associated with target protein productivity) compared with previous work.

  19. Heat and mass transfer analogies for evaporation models at high evaporation rate

    OpenAIRE

    Trontin, P.; Villedieu, P.

    2014-01-01

    International audience; In the framework of anti and deicing applications, heated liquid films can appear above the ice thickness, or directly above the wall. Then, evaporation plays a major role in the Messinger balance and evaporated mass has to be predicted accurately. Unfortunately, it appears that existing models under-estimate evaporation at high temperature. In this study, different evaporation models at high evaporation rates are studied. The different hypothesis on which these models...

  20. Rates and Equilibrium of CuA to heme a electron transfer in Paracoccus denitrificans cytochrome c oxidase

    DEFF Research Database (Denmark)

    Farver, Ole; Grell, Ernst; Ludwig, Bernd

    2006-01-01

    A reoxidation were identical within experimental error and independent of the enzyme concentration and its degree of reduction, demonstrating that a fast intramolecular electron equilibration is taking place between CuA and heme a. The rate constants for CuA --> heme a ET and the reverse heme a --> CuA process......Intramolecular electron transfer between CuA and heme a in solubilized bacterial (Paracoccus denitrificans) cytochrome c oxidase was investigated by pulse radiolysis. CuA, the initial electron acceptor, was reduced by 1-methylnicotinamide radicals in a diffusion-controlled reaction, as monitored...... by absorption changes at 825 nm, followed by partial restoration of the absorption and paralleled by an increase in the heme a absorption at 605 nm. The latter observations indicate partial reoxidation of the CuA center and the concomitant reduction of heme a. The rate constants for heme a reduction and Cu...

  1. NBTI-Aware Transient Fault Rate Analysis Method for Logic Circuit Based on Probability Voltage Transfer Characteristics

    Directory of Open Access Journals (Sweden)

    Zhiming Yang

    2016-01-01

    Full Text Available The reliability of Very Large Scale Integration (VLSI circuits has become increasingly susceptible to transient faults induced by environmental noise with the scaling of technology. Some commonly used fault tolerance strategies require statistical methods to accurately estimate the fault rate in different parts of the logic circuit, and Monte Carlo (MC simulation is often applied to complete this task. However, the MC method suffers from impractical computation costs due to the size of the circuits. Furthermore, circuit aging effects, such as negative bias temperature instability (NBTI, will change the characteristics of the circuit during its lifetime, leading to a change in the circuit’s noise margin. This change will increase the complexity of transient fault rate estimation tasks. In this paper, an NBTI-aware statistical analysis method based on probability voltage transfer characteristics is proposed for combinational logic circuit. This method can acquire accurate fault rates using a discrete probability density function approximation process, thus resolving the computation cost problem of the MC method. The proposed method can also consider aging effects and analyze statistical changes in the fault rates. Experimental results demonstrate that, compared to the MC simulation, our method can achieve computation times that are two orders of magnitude shorter while maintaining an error rate less than 9%.

  2. A macroscale mixture theory analysis of deposition and sublimation rates during heat and mass transfer in dry snow

    Directory of Open Access Journals (Sweden)

    A. C. Hansen

    2015-09-01

    Full Text Available The microstructure of a dry alpine snowpack is a dynamic environment where microstructural evolution is driven by seasonal density profiles and weather conditions. Notably, temperature gradients on the order of 10–20 K m−1, or larger, are known to produce a faceted snow microstructure exhibiting little strength. However, while strong temperature gradients are widely accepted as the primary driver for kinetic growth, they do not fully account for the range of experimental observations. An additional factor influencing snow metamorphism is believed to be the rate of mass transfer at the macroscale. We develop a mixture theory capable of predicting macroscale deposition and/or sublimation in a snow cover under temperature gradient conditions. Temperature gradients and mass exchange are tracked over periods ranging from 1 to 10 days. Interesting heat and mass transfer behavior is observed near the ground, near the surface, as well as immediately above and below dense ice crusts. Information about deposition (condensation and sublimation rates may help explain snow metamorphism phenomena that cannot be accounted for by temperature gradients alone. The macroscale heat and mass transfer analysis requires accurate representations of the effective thermal conductivity and the effective mass diffusion coefficient for snow. We develop analytical models for these parameters based on first principles at the microscale. The expressions derived contain no empirical adjustments, and further, provide self consistent values for effective thermal conductivity and the effective diffusion coefficient for the limiting cases of air and solid ice. The predicted values for these macroscale material parameters are also in excellent agreement with numerical results based on microscale finite element analyses of representative volume elements generated from X-ray tomography.

  3. A macroscale mixture theory analysis of deposition and sublimation rates during heat and mass transfer in snow

    Directory of Open Access Journals (Sweden)

    A. C. Hansen

    2015-03-01

    Full Text Available The microstructure of a dry alpine snowpack is a dynamic environment where microstructural evolution is driven by seasonal density profiles and weather conditions. Notably, temperature gradients on the order of 10–20 K m−1, or larger, are known to produce a faceted snow microstructure exhibiting little strength. However, while strong temperature gradients are widely accepted as the primary driver for kinetic growth, they do not fully account for the range of experimental observations. An additional factor influencing snow metamorphism is believed to be the rate of mass transfer at the macroscale. We develop a mixture theory capable of predicting macroscale deposition and/or sublimation in a snow cover under temperature gradient conditions. Temperature gradients and mass exchange are tracked over periods ranging from 1 to 10 days. Interesting heat and mass transfer behavior is observed near the ground, near the surface, as well as immediately above and below dense ice crusts. Information about deposition (condensation and sublimation rates may help explain snow metamorphism phenomena that cannot be accounted for by temperature gradients alone. The macroscale heat and mass transfer analysis requires accurate representations of the thermal conductivity and the effective mass diffusion coefficient for snow. We develop analytical models for these parameters based on first principles at the microscale. The expressions derived contain no empirical adjustments, and further, provide self consistent values for thermal conductivity and the effective diffusion coefficient for the limiting cases of air and solid ice. The predicted values for these macroscale material parameters are also in excellent agreement with numerical results based on microscale finite element analyses of representative volume elements generated from X-ray tomography.

  4. Processing of Nonconjugative Resistance Plasmids by Conjugation Nicking Enzyme of Staphylococci

    Energy Technology Data Exchange (ETDEWEB)

    Pollet, Rebecca M.; Ingle, James D.; Hymes, Jeff P.; Eakes, Thomas C.; Eto, Karina Yui; Kwong, Stephen M.; Ramsay, Joshua P.; Firth, Neville; Redinbo, Matthew R.; Christie, P. J.

    2016-01-04

    Antimicrobial resistance inStaphylococcus aureuspresents an increasing threat to human health. This resistance is often encoded on mobile plasmids, such as pSK41; however, the mechanism of transfer of these plasmids is not well understood. In this study, we first examine key protein-DNA interactions formed by the relaxase enzyme, NES, which initiates and terminates the transfer of the multidrug resistance plasmid pSK41. Two loops on the NES protein, hairpin loops 1 and 2, form extensive contacts with the DNA hairpin formed at theoriTregion of pSK41, and here we establish that these contacts are essential for proper DNA cleavage and religation by the full 665-residue NES proteinin vitro. Second, pSK156 and pCA347 are nonconjugativeStaphylococcus aureusplasmids that contain sequences similar to theoriTregion of pSK41 but differ in the sequence predicted to form a DNA hairpin. We show that pSK41-encoded NES is able to bind, cleave, and religate theoriTsequences of these nonconjugative plasmidsin vitro. Although pSK41 could mobilize a coresident plasmid harboring its cognateoriT, it was unable to mobilize plasmids containing the pSK156 and pCA347 variantoriTmimics, suggesting that an accessory protein like that previously shown to confer specificity in the pWBG749 system may also be involved in transmission of plasmids containing a pSK41-likeoriT. These data indicate that the conjugative relaxase intransmechanism recently described for the pWBG749 family of plasmids also applies to the pSK41 family of plasmids, further heightening the potential significance of this mechanism in the horizontal transfer of staphylococcal plasmids.

    IMPORTANCEUnderstanding the

  5. Preferences of subfertile women regarding elective single embryo transfer : additional in vitro fertilization cycles are acceptable, lower pregnancy rates are not

    NARCIS (Netherlands)

    Twisk, Moniek; van der Veen, Fulco; Repping, Sjoerd; Heineman, Maas-Jan; Korevaar, Johanna C.; Bossuyt, Patrick M. M.

    2007-01-01

    With identical pregnancy rates after elective single embryo transfer (ET) and double ET strategies consisting of three cycles of IVF or intracytoplasmic sperm injection (ICSI) plus transfers of thawed/frozen embryos if available, 46% of the women undergoing IVF/ICSI favor elective single ET. If elec

  6. Characterization of Antimicrobial Resistance Dissemination across Plasmid Communities Classified by Network Analysis

    Directory of Open Access Journals (Sweden)

    Akifumi Yamashita

    2014-04-01

    Full Text Available The global clustering of gene families through network analysis has been demonstrated in whole genome, plasmid, and microbiome analyses. In this study, we carried out a plasmidome network analysis of all available complete bacterial plasmids to determine plasmid associations. A blastp clustering search at 100% aa identity cut-off and sharing at least one gene between plasmids, followed by a multilevel community network analysis revealed that a surprisingly large number of the plasmids were connected by one largest connected component (LCC, with dozens of community sub-groupings. The LCC consisted mainly of Bacilli and Gammaproteobacteria plasmids. Intriguingly, horizontal gene transfer (HGT was noted between different phyla (i.e., Staphylococcus and Pasteurellaceae, suggesting that Pasteurellaceae can acquire antimicrobial resistance (AMR genes from closely contacting Staphylococcus spp., which produce the external supplement of V-factor (NAD. Such community network analysis facilitate displaying possible recent HGTs like a class 1 integron, str and tet resistance markers between communities. Furthermore, the distribution of the Inc replicon type and AMR genes, such as the extended-spectrum ß-lactamase (ESBL CTX-M or the carbapenemases KPC NDM-1, implies that such genes generally circulate within limited communities belonging to typical bacterial genera. Thus, plasmidome network analysis provides a remarkable discriminatory power for plasmid-related HGT and evolution.

  7. 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

  8. Mechanism of acquisition of chromosomal markers by plasmids in Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

    Setlow, J.K.; Cabrera-Juarez, E.; Griffin, K.

    1984-11-01

    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 pNova. 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 than from labeled rec-2 recipients, in which no recombination took place. EcoRI digestion of pNov1 divided the DNA into a 1.7-kilobase-pair fragment containing the novobiocin resistance marker and a 13-kilobase-pair fragment containing all of the original vector and considerable portions homologous to the chromosome. Transformation by the large fragment alone resulted in a plasmid the size of the original pNov1. The hypothesis to explain the data is that genetic transfer from chromosome to plasmid took place by a copy choice mechanism.

  9. 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.

  10. Replicon typing of plasmids carrying blaCTX-M-15 among Enterobacteriaceae isolated at the environment, livestock and human interface.

    Science.gov (United States)

    Zurfluh, Katrin; Glier, Melinda; Hächler, Herbert; Stephan, Roger

    2015-07-15

    One of the currently most important antibiotic resistance mechanisms in Enterobacteriaceae is based on the production of ESBL enzymes that inactivate β-lactam antibiotics including cephalosporins and monobactams by hydrolyzing their β-lactam ring. In humans, the most prevalent ESBL enzyme type is encoded by blaCTX-M-15. CTX-M-15 producing enterobacterial strains were also frequently isolated from environmental samples including surface water and freshwater fish. Plasmids, which can be grouped in different plasmid incompatibility types, play a key role in the horizontal spread of these multidrug resistance genes. The purpose of this study was to investigate the diversity of plasmids that carry blaCTX-M-15 genes among Enterobacteriaceae isolated at the environment, livestock and human interface. In total, 81 blaCTX-M-15-harboring isolates collected between 2009 and 2014 were tested for its ability to transfer blaCTX-M-15 by conjugation. These plasmids were further typed. Transfer of a single blaCTX-M-15-harboring plasmid was observed in 32 (39.5%) of the isolates. The most frequent replicon types detected among these plasmids are IncF-type plasmids (n=12) (mostly multi replicon plasmids with a combination of following replicons: IncFII, IncFIA and IncFIB), followed by IncI1 (n=8), IncK (n=3) and IncR (n=1). A noticeable number of plasmids (n=8) could not be assigned to any of the tested replicon types. Knowledge about the plasmid types circulating in bacterial populations is indispensable for understanding epidemiological dynamics and for establishing intervention strategies to stop further dissemination of particular plasmids.

  11. Novel assay to measure the plasmid mobilizing potential of mixed microbial communities

    DEFF Research Database (Denmark)

    Klümper, Uli; Droumpali, Ariadni; Dechesne, Arnaud;

    2014-01-01

    compare the transfer frequency of a mobilizable plasmid to that of a mobilizing and conjugal plasmid measured for a model strain and for the assayed community. With Pseudomonas putida carrying the gfp-tagged mobilizable RSF1010 plasmid as donor strain, we conducted solid surface mating experiments...... community for RP4 (at 1.16x10-4 transconjugants per recipient (T/R)) was similar to that previously measured for soil microbial communities. RSF1010 was mobilized by the model community at a frequency of 1.16x10-5 T/R, only one order of magnitude lower than its permissiveness to RP4. This mobilization...... frequency is unexpectedly high considering that (i) mobilization requires the presence of mobilizing conjugal plasmids within the permissive fraction of the recipients; (ii) in pure culture experiments with P. putida retromobilization of RSF1010 through RP4 only took place in approximately half...

  12. Genomic and functional characterization of qnr-encoding plasmids from municipal wastewater biosolid Klebsiella pneumoniae isolates

    Directory of Open Access Journals (Sweden)

    Ella eKaplan

    2015-12-01

    Full Text Available Municipal wastewater treatment facilities are considered to be hotspots for antibiotic resistance since they conjoin high densities of environmental and fecal bacteria with selective pressure in the form of sub-therapeutic concentrations of antibiotics. Discharged effluents and biosolids from these facilities can disseminate antibiotic resistant genes to terrestrial and aquatic environments, potentially contributing to the increasing global trend in antibiotic resistance. This phenomenon is especially pertinent when resistance genes are associated with mobile genetic elements such as conjugative plasmids, which can be transferred between bacterial phyla. Fluoroquinolones are among the most abundant antibiotic compounds detected in wastewater treatment facilities, especially in biosolids, where due to their hydrophobic properties they accumulate to concentrations that may exceed 40 mg/L. Although fluoroquinolone resistance is traditionally associated with mutations in the gyrA/topoisomerase IV genes, there is increasing evidence of plasmid-mediated quinolone resistance, which is primarily encoded on qnr genes. In this study, we sequenced seven qnr-harboring plasmids from a diverse collection of Klebsiella strains, isolated from dewatered biosolids from a large wastewater treatment facility in Israel. One of the plasmids, termed pKPSH-11XL was a large (185.4 kbp, multi-drug resistance, IncF-type plasmid that harbored qnrB and 10 additional antibiotic resistance genes that conferred resistance to 5 different antibiotic families. It was highly similar to the pKPN3-like plasmid family that has been detected in multidrug resistant clinical Klebsiella isolates. In contrast, the six additional plasmids were much smaller (7-9Kbp and harbored a qnrS -type gene. These plasmids were highly similar to each other and closely resembled pGNB2, a plasmid isolated from a German wastewater treatment facility. Comparative genome analyses of pKPSH-11XL and other p

  13. 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

  14. Effect of the Brazilian conditional cash transfer and primary health care programs on the new case detection rate of leprosy.

    Science.gov (United States)

    Nery, Joilda Silva; Pereira, Susan Martins; Rasella, Davide; Penna, Maria Lúcia Fernandes; Aquino, Rosana; Rodrigues, Laura Cunha; Barreto, Mauricio Lima; Penna, Gerson Oliveira

    2014-11-01

    Social determinants can affect the transmission of leprosy and its progression to disease. Not much is known about the effectiveness of welfare and primary health care policies on the reduction of leprosy occurrence. The aim of this study is to evaluate the impact of the Brazilian cash transfer (Bolsa Família Program-BFP) and primary health care (Family Health Program-FHP) programs on new case detection rate of leprosy. We conducted the study with a mixed ecological design, a combination of an ecological multiple-group and time-trend design in the period 2004-2011 with the Brazilian municipalities as unit of analysis. The main independent variables were the BFP and FHP coverage at the municipal level and the outcome was new case detection rate of leprosy. Leprosy new cases, BFP and FHP coverage, population and other relevant socio-demographic covariates were obtained from national databases. We used fixed-effects negative binomial models for panel data adjusted for relevant socio-demographic covariates. A total of 1,358 municipalities were included in the analysis. In the studied period, while the municipal coverage of BFP and FHP increased, the new case detection rate of leprosy decreased. Leprosy new case detection rate was significantly reduced in municipalities with consolidated BFP coverage (Risk Ratio 0.79; 95% CI  =0.74-0.83) and significantly increased in municipalities with FHP coverage in the medium (72-95%) (Risk Ratio 1.05; 95% CI  =1.02-1.09) and higher coverage tertiles (>95%) (Risk Ratio 1.12; 95% CI  =1.08-1.17). At the same time the Family Health Program had been effective in increasing the new case detection rate of leprosy in Brazil, the Bolsa Família Program was associated with a reduction of the new case detection rate of leprosy that we propose reflects a reduction in leprosy incidence.

  15. Effect of the Brazilian conditional cash transfer and primary health care programs on the new case detection rate of leprosy.

    Directory of Open Access Journals (Sweden)

    Joilda Silva Nery

    2014-11-01

    Full Text Available Social determinants can affect the transmission of leprosy and its progression to disease. Not much is known about the effectiveness of welfare and primary health care policies on the reduction of leprosy occurrence. The aim of this study is to evaluate the impact of the Brazilian cash transfer (Bolsa Família Program-BFP and primary health care (Family Health Program-FHP programs on new case detection rate of leprosy.We conducted the study with a mixed ecological design, a combination of an ecological multiple-group and time-trend design in the period 2004-2011 with the Brazilian municipalities as unit of analysis. The main independent variables were the BFP and FHP coverage at the municipal level and the outcome was new case detection rate of leprosy. Leprosy new cases, BFP and FHP coverage, population and other relevant socio-demographic covariates were obtained from national databases. We used fixed-effects negative binomial models for panel data adjusted for relevant socio-demographic covariates. A total of 1,358 municipalities were included in the analysis. In the studied period, while the municipal coverage of BFP and FHP increased, the new case detection rate of leprosy decreased. Leprosy new case detection rate was significantly reduced in municipalities with consolidated BFP coverage (Risk Ratio 0.79; 95% CI  =0.74-0.83 and significantly increased in municipalities with FHP coverage in the medium (72-95% (Risk Ratio 1.05; 95% CI  =1.02-1.09 and higher coverage tertiles (>95% (Risk Ratio 1.12; 95% CI  =1.08-1.17.At the same time the Family Health Program had been effective in increasing the new case detection rate of leprosy in Brazil, the Bolsa Família Program was associated with a reduction of the new case detection rate of leprosy that we propose reflects a reduction in leprosy incidence.

  16. Transport and transfer rates in the waters of the Continental Shelf. Annual report

    Energy Technology Data Exchange (ETDEWEB)

    Biscaye, P.E.

    1978-07-01

    The present contract year has been one of transition from an emphasis on field work and sample gathering to the predominance of sample and data analysis and the formulation of testable hypotheses concerning specific processes in the New York Bight. We have begun to understand the seasonal transition in the role of phytoplankton vs. grazing zooplankton in forming the particles on which some reactive pollutants are removed. Using natural radioactive tracers we have estimated the removal rates of reactive metals from the surface waters and these range over an order of magnitude from most rapid nearshore to least rapid over the upper continental slope. Once removed nearshore, however, these tracers, and the pollutants for which they proxy, do not remain permanently in the sediments but appear to be remobilized (probably by oxidation) during the winter and are reintroduced into the water column. Work on transport and mixing processes of pollutants which are or behave like those in solution has continued along several fronts. Hydrographic data on the structure of the water column continues to give a description of the system that is crucial to understanding geochemical and biological processes which affect pollutants. Hydrographic characterization of water masses from the data sets of cruises has resulted in hypotheses concerning the renewal of shelf water by direct exchange between shelf and upper slope water.

  17. Rate constants for 1,5- and 1,6-hydrogen atom transfer reactions of mono-, di-, and tri-aryl-substituted donors, models for hydrogen atom transfers in polyunsaturated fatty acid radicals.

    Science.gov (United States)

    DeZutter, Christopher B; Horner, John H; Newcomb, Martin

    2008-03-06

    Rate constants for 1,5- and 1,6-hydrogen atom transfer reactions in models of polyunsaturated fatty acid radicals were measured via laser flash photolysis methods. Photolyses of PTOC (pyridine-2-thioneoxycarbonyl) ester derivatives of carboxylic acids gave primary alkyl radicals that reacted by 1,5-hydrogen transfer from mono-, di-, and tri-aryl-substituted positions or 1,6-hydrogen transfer from di- and tri-aryl-substituted positions to give UV-detectable products. Rate constants for reactions in acetonitrile at room temperature ranged from 1 x 10(4) to 4 x 10(6) s(-1). The activation energies for a matched pair of 1,5- and 1,6-hydrogen atom transfers giving tri-aryl-substituted radicals were approximately equal, as were the primary kinetic isotope effects, but the 1,5-hydrogen atom transfer reaction was 1 order of magnitude faster at room temperature than the 1,6-hydrogen atom transfer reaction due to a less favorable entropy of activation for the 1,6-transfer reaction. Solvent effects on the rate constants for the 1,5-hydrogen atom transfer reaction of the 2-[2-(diphenylmethyl)phenyl]ethyl radical at ambient temperature were as large as a factor of 2 with the reaction increasing in rate in lower polarity solvents. Hybrid density functional theory computations for the 1,5- and 1,6-hydrogen atom transfers of the tri-aryl-substituted donors were in qualitative agreement with the experimental results.

  18. The Gas Transfer through Polar Sea ice experiment: Insights into the rates and pathways that determine geochemical fluxes

    Science.gov (United States)

    Lovely, A.; Loose, B.; Schlosser, P.; McGillis, W.; Zappa, C.; Perovich, D.; Brown, S.; Morell, T.; Hsueh, D.; Friedrich, R.

    2015-12-01

    Sea ice is a defining feature of the polar marine environment. It is a critical domain for marine biota and it regulates ocean-atmosphere exchange, including the exchange of greenhouse gases such as CO2 and CH4. In this study, we determined the rates and pathways that govern gas transport through a mixed sea ice cover. N2O, SF6, 3He, 4He, and Ne were used as gas tracers of the exchange processes that take place at the ice-water and air-water interfaces in a laboratory sea ice experiment. Observation of the changes in gas concentrations during freezing revealed that He is indeed more soluble in ice than in water; Ne is less soluble in ice, and the larger gases (N2O and SF6) are mostly excluded during the freezing process. Model estimates of gas diffusion through ice were calibrated using measurements of bulk gas content in ice cores, yielding gas transfer velocity through ice (kice) of ˜5 × 10-4 m d-1. In comparison, the effective air-sea gas transfer velocities (keff) ranged up to 0.33 m d-1 providing further evidence that very little mixed-layer ventilation takes place via gas diffusion through columnar sea ice. However, this ventilation is distinct from air-ice gas fluxes driven by sea ice biogeochemistry. The magnitude of keff showed a clear increasing trend with wind speed and current velocity beneath the ice, as well as the combination of the two. This result indicates that gas transfer cannot be uniquely predicted by wind speed alone in the presence of sea ice.

  19. The functional transfer of genes from the mitochondria to the nucleus: the effects of selection, mutation, population size and rate of self-fertilization.

    Science.gov (United States)

    Brandvain, Yaniv; Wade, Michael J

    2009-08-01

    The transfer of mitochondrial genes to the nucleus is a recurrent and consistent feature of eukaryotic genome evolution. Although many theories have been proposed to explain such transfers, little relevant data exist. The observation that clonal and self-fertilizing plants transfer more mitochondrial genes to their nuclei than do outcrossing plants contradicts predictions of major theories based on nuclear recombination and leaves a gap in our conceptual understanding how the observed pattern of gene transfer could arise. Here, with a series of deterministic and stochastic simulations, we show how epistatic selection and relative mutation rates of mitochondrial and nuclear genes influence mitochondrial-to-nuclear gene transfer. Specifically, we show that when there is a benefit to having a mitochondrial gene present in the nucleus, but absent in the mitochondria, self-fertilization dramatically increases both the rate and the probability of gene transfer. However, absent such a benefit, when mitochondrial mutation rates exceed those of the nucleus, self-fertilization decreases the rate and probability of transfer. This latter effect, however, is much weaker than the former. Our results are relevant to understanding the probabilities of fixation when loci in different genomes interact.

  20. Modelling of Radionuclides Transfer and Ambient Dose Rates in Fukushima Forest Ecosystems

    Energy Technology Data Exchange (ETDEWEB)

    Calmon, P.; Gonze, M.A.; Mourlon, C.; Simon-Cornu, M. [Institute of Radiation Protection and Nuclear Safety, CE Cadarache-Bat 153, BP3 - 13115 St-Paul-lez- Durance cedex (France)

    2014-07-01

    the precise different deposition sequences and the meteorological conditions. Moreover, the characteristics of Japanese cedar forests seem to be very specific and the foliar absorption parameter has to be adjusted, in link with the very particular foliage of this tree species. The model is able to reproduce quite well the caesium deposition in soil, but also the throughfall and the stem flow that occurred during the six months after the accident. Several ambient dose rate calculations have been also tested against measurements in various stations of evergreen (Japanese cedar, spruces and pines) and deciduous forests in eastern Fukushima Prefecture, where deposition essentially occurred as humid and 85% of the total deposit was on the soil in the 3 months following the accident. The ambient dose rates are well correlated with the caesium concentrations in the soil, and even more in the tree canopy. (authors)

  1. Hydrocarbon mineralization in sediments and plasmid incidence in sediment bacteria from the campeche bank.

    Science.gov (United States)

    Leahy, J G; Somerville, C C; Cunningham, K A; Adamantiades, G A; Byrd, J J; Colwell, R R

    1990-06-01

    Rates of degradation of radiolabeled hydrocarbons and incidence of bacterial plasmid DNA were investigated in sediment samples collected from the Campeche Bank, Gulf of Mexico, site of an offshore oil field containing several petroleum platforms. Overall rates of mineralization of [C]hexadecane and [C]phenanthrene measured for sediments were negligible; <1% of the substrate was converted to CO(2) in all cases. Low mineralization rates are ascribed to nutrient limitations and to lack of adaptation by microbial communities to hydrocarbon contaminants. Plasmid frequency data for sediment bacteria similarly showed no correlation with proximity to the oil field, but, instead, showed correlation with water column depth at each sampling site. Significant differences between sites were observed for proportion of isolates carrying single or multiple plasmids and mean number of plasmids per isolate, each of which increased as a function of depth.

  2. Theoretical prediction of the effect of heat transfer parameters on cooling rates of liquid-filled plastic straws used for cryopreservation of spermatozoa.

    Science.gov (United States)

    Sansinen, M; Santos, M V; Zaritzky, N; Baez, R; Chirife, J

    2010-01-01

    Heat transfer plays a key role in cryopreservation of liquid semen in plastic straws. The effect of several parameters on the cooling rate of a liquid-filled polypropylene straw when plunged into liquid nitrogen was investigated using a theoretical model. The geometry of the straw containing the liquid was assimilated as two concentric finite cylinders of different materials: the fluid and the straw; the unsteady-state heat conduction equation for concentric cylinders was numerically solved. Parameters studied include external (convection) heat transfer coefficient (h), the thermal properties of straw manufacturing material and wall thickness. It was concluded that the single most important parameter affecting the cooling rate of a liquid column contained in a straw is the external heat transfer coefficient in LN2. Consequently, in order to attain maximum cooling rates, conditions have to be designed to obtain the highest possible heat transfer coefficient when the plastic straw is plunged in liquid nitrogen.

  3. Plasmids of psychrophilic and psychrotolerant bacteria and their role in adaptation to cold environments.

    Science.gov (United States)

    Dziewit, Lukasz; Bartosik, Dariusz

    2014-01-01

    Extremely cold environments are a challenge for all organisms. They are mostly inhabited by psychrophilic and psychrotolerant bacteria, which employ various strategies to cope with the cold. Such harsh environments are often highly vulnerable to the influence of external factors and may undergo frequent dynamic changes. The rapid adjustment of bacteria to changing environmental conditions is crucial for their survival. Such "short-term" evolution is often enabled by plasmids-extrachromosomal replicons that represent major players in horizontal gene transfer. The genomic sequences of thousands of microorganisms, including those of many cold-active bacteria have been obtained over the last decade, but the collected data have yet to be thoroughly analyzed. This report describes the results of a meta-analysis of the NCBI sequence databases to identify and characterize plasmids of psychrophilic and psychrotolerant bacteria. We have performed in-depth analyses of 66 plasmids, almost half of which are cryptic replicons not exceeding 10 kb in size. Our analyses of the larger plasmids revealed the presence of numerous genes, which may increase the phenotypic flexibility of their host strains. These genes encode enzymes possibly involved in (i) protection against cold and ultraviolet radiation, (ii) scavenging of reactive oxygen species, (iii) metabolism of amino acids, carbohydrates, nucleotides and lipids, (iv) energy production and conversion, (v) utilization of toxic organic compounds (e.g., naphthalene), and (vi) resistance to heavy metals, metalloids and antibiotics. Some of the plasmids also contain type II restriction-modification systems, which are involved in both plasmid stabilization and protection against foreign DNA. Moreover, approx. 50% of the analyzed plasmids carry genetic modules responsible for conjugal transfer or mobilization for transfer, which may facilitate the spread of these replicons among various bacteria, including across species boundaries.

  4. Plasmid as a measure of microbial degradation capacity for 2,4-dichlorophenoxyacetic acid.

    Science.gov (United States)

    Chong, Nyuk-Min; Chang, Hung-Wei

    2009-02-01

    The purpose of this research was to pursuit the quantification of microbial degradation capacity for 2,4-dichlorophenoxyacetic acid (2,4-D) by detecting and quantifying a prominent 2,4-D degradation encoding plasmid. Batch reactor acclimation, de-acclimation, and re-acclimation tests were conducted during which periods the courses of 2,4-D dissipation and plasmid evolution were quantitatively measured. Pure cultures of bacterial strains were detected to give rise to a plasmid approximately the size of 90 kb after acclimation. The 90 kb plasmid content of Arthrobacter sp. increased when degradation occurred after acclimation, with a rate that corresponded closely to the degradation rate. During de-acclimation, plasmid content declined exponentially at a half-life of approximately 3.5 days. Re-acclimation saw a renewed induction of plasmid, but substrate consumption limited the rise of plasmid to a level much lower than after the first acclimation. This research recommends a method for measuring the microbial degradation capability for a xenobiotic.

  5. Characterization of a cfr-Carrying Plasmid from Porcine Escherichia coli That Closely Resembles Plasmid pEA3 from the Plant Pathogen Erwinia amylovora.

    Science.gov (United States)

    Zhang, Rongmin; Sun, Bin; Wang, Yang; Lei, Lei; Schwarz, Stefan; Wu, Congming

    2015-11-02

    The multiresistance gene cfr was found in two porcine Escherichia coli isolates, one harboring it on the conjugative 33,885-bp plasmid pFSEC-01, the other harboring it in the chromosomal DNA. Sequence analysis of pFSEC-01 revealed that a 6,769-bp fragment containing the cfr gene bracketed by two IS26 elements was inserted into a plasmid closely related to pEA3 from the plant pathogen Erwinia amylovora, suggesting that pFSEC-01 may be transferred between different bacterial genera of both animal and plant origin.

  6. Complete genetic analysis of plasmids carrying mcr-1 and other resistance genes in an Escherichia coli isolate of animal origin.

    Science.gov (United States)

    Li, Ruichao; Xie, Miaomiao; Lv, Jingzhang; Wai-Chi Chan, Edward; Chen, Sheng

    2017-03-01

    To investigate the genetic features of three plasmids recovered from an MCR-1 and ESBL-producing Escherichia coli strain, HYEC7, and characterize the transmission mechanism of mcr-1 . The genetic profiles of three plasmids were determined by PCR, S1-PFGE, Southern hybridization and WGS analysis. The ability of the mcr-1 -bearing plasmid to undergo conjugation was also assessed. The mcr-1 -bearing transposon Tn 6330 was characterized by PCR and DNA sequencing. Complete sequences of three plasmids were obtained. A non-conjugative phage P7-like plasmid, pHYEC7- mcr1 , was found to harbour the mcr-1 -bearing transposon Tn 6330 , which could be excised from the plasmid by generating a circular intermediate harbouring mcr-1 and the IS Apl1 element. The insertion of the circular intermediate into another plasmid, pHYEC7-IncHI2, could form pHNSHP45-2, the original IncHI2-type mcr-1 -carrying plasmid that was reported. The third plasmid, pHYEC7-110, harboured two replicons, IncX1 and IncFIB, and comprised multiple antimicrobial resistance mobile elements, some of which were shared by pHYEC7-IncHI2. The Tn 6330 element located in the phage-like plasmid pHYEC7- mcr1 could be excised from the plasmid and formed a circular intermediate that could be integrated into plasmids containing the IS Apl1 element. This phenomenon indicated that Tn 6330 is a key element responsible for widespread dissemination of mcr-1 among various types of plasmids and bacterial chromosomes. The dissemination rate of such an element may be further enhanced upon translocation into phage-like vectors, which may also be transmitted via transduction events.

  7. Reduced Ectopic Pregnancy Rate on Day 5 Embryo Transfer Compared with Day 3: A Meta-Analysis

    Science.gov (United States)

    Tang, Rong; Ding, Lingling; Yan, Lei; Chen, Zi-Jiang

    2017-01-01

    Objective To compare the risk of ectopic pregnancy (EP) after embryo transfer on day 3(D3-ET) and day 5(D5-ET). Design Meta-analysis Patients Women with pregnancy resulting from in vitro undergoing in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) Result(s) Twenty-two studies were identified through research conducted using the PubMed, Embase, and Cochrane databases and ClinicalTrials.gov. All studies were conducted prior to October 2016. Adding the reproductive data from our center, a total of 143 643 pregnancies were reviewed(D3-ET: n = 62027,D5-ET:n = 81616). A lower EP rate was found in women undergoing D5-ET than in those undergoing D3-ET [relative risk (RR), 0.67;95% confidence interval (CI), 0.54–0.85;143643 pregnancies in 23 studies; I2 = 67%]. These results were validated in subgroups of fresh embryo-transfer (Fre-ET) cycles [RR, 0.78; 95%CI, 0.69–0.88; 91 871 pregnancies in 21 studies; I2 = 29%] and frozen-thawed embryo-transfer (Fro-ET) cycles [RR, 0.43; 95%CI, 0.36–0.51; 51 772 pregnancies in 10 studies; I2 = 33%]. After separating out the randomized controlled trials (RCTs), a significant difference was found in the retrospective studies in both subgroups [both Fre-ET (RR,0.78;95% CI 0.69–0.88);91182 pregnancies in 14 studies; I2 = 45%] and Fro-ET(RR,0.43;95% CI 0.36–0.51; 51751pregnancies in 9 studies;I2 = 33%)], while the RCTs showed no statistical significance for Fre-ET cycles[RR,0.86;95% CI 0.32–2.26); 689 pregnancies in 7 studies; I2 = 0%]. Conclusion(s) The present study indicates that D5-ET reduces the risk for EP in cycles that use IVF or ICSI, compared with D3-ET. It suggests that D5-ET may be a better choice for decreasing the EP rate in assisted reproductive technology. Further high-quality randomized controlled trials are anticipated. PMID:28121989

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

    Science.gov (United States)

    Crespo, M D; Altermann, E; Olson, J; Miller, W G; Chandrashekhar, K; Kathariou, S

    2016-07-01

    In Campylobacter spp., resistance to the antimicrobials 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; 44,095nt) harboring tet(O) was identified in C. jejuni strain 11601MD, which was isolated from the jejunum of a turkey produced conventionally in North Carolina. Analysis of the p11601MD sequence revealed the presence of a high-GC content cassette with four genes that included tet(O) and a putative aminoglycoside transferase gene (aphA-3) highly similar to kanamycin resistance determinants. Several genes putatively involved in conjugative transfer were also identified on the plasmid. These findings will contribute to a better understanding of the distribution of potentially self-mobilizing plasmids harboring antibiotic resistance determinants in Campylobacter spp. from turkeys and other sources.

  9. Replication efficiency of rolling-circle replicon-based plasmids derived from porcine circovirus 2 in eukaryotic cells.

    Science.gov (United States)

    Faurez, Florence; Dory, Daniel; Henry, Aurélie; Bougeard, Stéphanie; Jestin, André

    2010-04-01

    In this study, a method was developed to measure replication rates of rolling-circle replicon-based plasmids in eukaryotic cells. This method is based on the discriminative quantitation of MboI-resistant, non-replicated input plasmids and DpnI-resistant, replicated plasmids. To do so, porcine circovirus type 2 (PCV2) replicon-based plasmids were constructed. These plasmids contained the PCV2 origin of replication, the PCV2 Rep promoter and the PCV2 Rep gene. The results show that the replication rate depends on the length of the PCV2 replicon-based plasmid and not on the respective position of the Rep promoter and the promoter of the gene of interest that encodes the enhanced green fluorescent protein (eGFP). In all cases, it was necessary to add the Rep gene encoded by a plasmid and cotransfected as a replication booster. This method can evaluate the replication potential of replicon-based plasmids quickly and is thereby a promising tool for the development of plasmids for vaccine purposes.

  10. Effect of transfer of one or two in vitro-produced embryos and post-transfer administration of gonadotropin releasing hormone on pregnancy rates of heat-stressed dairy cattle.

    Science.gov (United States)

    Franco, M; Block, J; Jousan, F D; de Castro e Paula, L A; Brad, A M; Franco, J M; Grisel, F; Monson, R L; Rutledge, J J; Hansen, P J

    2006-07-15

    Pregnancy rates following transfer of an in vitro-produced (IVP) embryo are often lower than those obtained following transfer of an embryo produced by superovulation. The purpose of the current pair of experiments was to examine two strategies for increasing pregnancy rates in heat stressed, dairy recipients receiving an IVP embryo. One method was to transfer two embryos into the uterine horn ipsilateral to the CL, whereas the other method involved injection of GnRH at Day 11 after the anticipated day of ovulation. In Experiment 1, 32 virgin crossbred heifers and 26 lactating crossbred cows were prepared for timed embryo transfer by being subjected to a timed ovulation protocol. Those having a palpable CL were randomly selected to receive one (n = 31 recipients) or two (n = 27 recipients) embryos on Day 7 after anticipated ovulation. At Day 64 of gestation, the pregnancy rate tended to be higher (P = 0.07) for cows than for heifers. Heifers that received one embryo tended to have a higher pregnancy rate than those that received two embryos (41% versus 20%, respectively) while there was no difference in pregnancy rate for cows that received one or two embryos (57% versus 50%, respectively). Pregnancy loss between Day 64 and 127 only occurred for cows that received two embryos (pregnancy rate at Day 127=17%). Between Day 127 and term, one animal (a cow with a single embryo) lost its pregnancy. There was no difference in pregnancy rates at Day 127 or calving rates between cows and heifers, but females that received two embryos had lower Day-127 pregnancy rates and calving rates than females that received one embryo (P cows were synchronized for timed embryo transfer as in Experiment 1. Cows received a single embryo in the uterine horn ipsilateral to the ovary containing the CL and received either 100 microg GnRH or vehicle at Day 11 after anticipated ovulation (i.e. 4 days after embryo transfer). There was no difference in pregnancy rate for cows that received the Gn

  11. Ultrasound-enhanced gene transfer: Comparison of contrast agents and ultrasound modalities in vitro and in vivo

    Science.gov (United States)

    Pislaru, Sorin V.; Rajiv, Gulati; Pislaru, Cristina; Kinnick, Randall R.; Singh, Ripudamanjit; Greenleaf, James F.; Simari, Robert D.

    2002-11-01

    Ultrasound (US) enhancement of plasmid-based gene transfer is an emerging technique. Our hypothesis was that two contrast agents (Optison and PESDA), and two US exposure modalities (dedicated continuous wave system and diagnostic scanner) may have different effects. Luciferase plasmid with or without contrast agent was added to vascular smooth muscle cells and endothelial cells, followed by US exposure. Luciferase activity was measured 24 h later. US exposure consistently induced higher transfection rates than all controls. PESDA was superior to Optison in both cell lines. In vitro, continuous wave and diagnostic US were not significantly different. In vivo, Lux and PESDA were injected into skeletal muscles of rats (IM or intra-arterial) followed or not by US exposure. In separate animals, adenovirus encoding for luciferase was injected IM and was not followed by US exposure. Gene transfer efficacy was 8-10 fold higher with US and PESDA than with plasmid alone, but 2 fold lower than with adenovirus. However, as opposed to adenovirus, US-enhanced plasmid gene transfer was highly localized to the injected muscle, with no expression at distal sites. Our results support the hypothesis that contrast agents and exposure modalities are not equivalent with regard to gene transfer efficacy.

  12. Conjugative multi-resistant plasmids in Haihe River and their impacts on the abundance and spatial distribution of antibiotic resistance genes.

    Science.gov (United States)

    Dang, Bingjun; Mao, Daqing; Xu, Yan; Luo, Yi

    2017-03-15

    In this study, five classes of antibiotic resistance genes (ARGs) were quantified in sediment samples of Haihe River, China, with abundance ranging from 1.39 × 10(4) to 1.58 × 10(10) copies/g dry weight. Meanwhile, antibiotic resistant conjugative plasmids were also isolated from these samples through filter mating assays. In total, 202 transconjugants were isolated and tested for their antibiotic resistance phenotypes, among which 26 different types of conjugative plasmids were observed. The majority of these plasmids showed a multi-resistant phenotype and the most prevalent resistance was tetracycline resistance and sulfonamide resistance. Furthermore, we tested the transfer frequencies of these plasmids, determined their genotypes and then compared the plasmid-borne ARGs with their corresponding abundance in Haihe River. Most of the isolated plasmids exhibited high transfer frequencies to the recipient strain Escherichia coli J53. Plasmids isolated from the urban areas of Haihe River have higher transfer frequencies than the rural areas. Results from comprehensive analysis of plasmid genotypes, ARG abundance and plasmid sequencing confirmed that most of the plasmid-borne ARGs were the dominant genes in the Haihe River. Therefore, conjugative plasmids isolated from the Haihe River plays a crucial role in the dissemination, abundance and spatial distribution of ARGs in Haihe River, especially some unfrequent ARGs like blaGES-1. This study will help to increase the knowledge on the conjugative plasmid-mediated ARG propagation in the environment. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. Rate of bitumen film transfer from a quartz surface to an air bubble as observed by optical microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Lelinski, D. [FFE Minerals USA Inc., Salt Lake City, UT (United States); Drelich, J. [Michigan Technological Univ., Houghton, MI (United States). Dept. of Materials Science and Engineering; Miller, J.D. [Utah Univ., Salt Lake City, UT (United States). Dept. of Metallurgical Engineering; Hupka, J. [Gdansk Univ. of Technology, Gdansk (Poland). Dept. of Chemical Technology

    2004-08-01

    Froth flotation is a separation process used to recover oil from oil sands. The two separation steps in froth flotation include the release of oil from mineral matter, and oil flotation from the slurry. During flotation, gas bubbles collide with and attach to oil droplets and to oily soil particles. The flotation of oil-in water emulsions in the absence of solids has been studied extensively. This study focused on liquid/liquid spreading. It was limited to simple systems involving pure single-component liquids. Multi-component oils were only used in a few experiments. In particular, this paper discussed the rate of high viscosity bitumen transfer from a bitumen-coated quartz slide to an air bubble surface that occurred in an aqueous alkaline solution. Bitumen spreading observations were recorded with a video camera attached to a stereoscopic microscope. The objective was to improve hot-water processing technology for bitumen recovery from Utah oil sands. The study showed that during bitumen spreading, a bulk layer followed the formation of thin bitumen films. The spreading velocity was one order of magnitude less than the velocity of the precursor film. The study provided a better understanding of the basic separation techniques encountered in the sequence of oil sand slurry digestion, aeration, gravity separation and flotation. It was concluded that a strong intermolecular attraction exists between the bitumen molecules. This interaction reduces the rate of bitumen spreading at the water surface. 33 refs., 2 tabs., 12 figs.

  14. The rotation rates of massive stars: the role of binary interaction trough tides, mass transfer and mergers

    CERN Document Server

    de Mink, S E; Izzard, R G; Sana, H; de Koter, A

    2012-01-01

    Rotation is thought to be a major factor in the evolution of massive stars, especially at low metallicity, with consequences for their chemical yields, ionizing flux and final fate. The natal rotation-rate distribution of stars is of high priority given its importance as a constraint on theories of massive star formation and as input for models of stellar populations in the local Universe and at high redshift. Recently, it has become clear that the majority of massive stars interact with a binary companion before they die. We investigate how this affects the distribution of rotation rates. For this purpose, we simulate a massive binary-star population typical for our Galaxy assuming continuous star formation. We find that, because of binary interaction, 20^+5_-10% of all massive main-sequence stars have projected rotational velocities in excess of 200km/s. We evaluate the effect of uncertain input distributions and physical processes and conclude that the main uncertainties are the mass transfer efficiency an...

  15. Endogenous Sensory Discrimination and Selection by a Fast Brain Switch for a High Transfer Rate Brain-Computer Interface.

    Science.gov (United States)

    Xu, Ren; Jiang, Ning; Dosen, Strahinja; Lin, Chuang; Mrachacz-Kersting, Natalie; Dremstrup, Kim; Farina, Dario

    2016-08-01

    In this study, we present a novel multi-class brain-computer interface (BCI) for communication and control. In this system, the information processing is shared by the algorithm (computer) and the user (human). Specifically, an electro-tactile cycle was presented to the user, providing the choice (class) by delivering timely sensory input. The user discriminated these choices by his/her endogenous sensory ability and selected the desired choice with an intuitive motor task. This selection was detected by a fast brain switch based on real-time detection of movement-related cortical potentials from scalp EEG. We demonstrated the feasibility of such a system with a four-class BCI, yielding a true positive rate of  ∼ 80% and  ∼ 70%, and an information transfer rate of  ∼ 7 bits/min and  ∼ 5 bits/min, for the movement and imagination selection command, respectively. Furthermore, when the system was extended to eight classes, the throughput of the system was improved, demonstrating the capability of accommodating a large number of classes. Combining the endogenous sensory discrimination with the fast brain switch, the proposed system could be an effective, multi-class, gaze-independent BCI system for communication and control applications.

  16. Three classes of plasmid (47-63 kb) carry the type B neurotoxin gene cluster of group II Clostridium botulinum.

    Science.gov (United States)

    Carter, Andrew T; Austin, John W; Weedmark, Kelly A; Corbett, Cindi; Peck, Michael W

    2014-08-01

    Pulsed-field gel electrophoresis and DNA sequence analysis of 26 strains of Group II (nonproteolytic) Clostridium botulinum type B4 showed that 23 strains carried their neurotoxin gene cluster on a 47-63 kb plasmid (three strains lacked any hybridization signal for the neurotoxin gene, presumably having lost their plasmid). Unexpectedly, no neurotoxin genes were found on the chromosome. This apparent constraint on neurotoxin gene transfer to the chromosome stands in marked contrast to Group I C. botulinum, in which neurotoxin gene clusters are routinely found in both locations. The three main classes of type B4 plasmid identified in this study shared different regions of homology, but were unrelated to any Group I or Group III plasmid. An important evolutionary aspect firmly links plasmid class to geographical origin, with one class apparently dominant in marine environments, whereas a second class is dominant in European terrestrial environments. A third class of plasmid is a hybrid between the other two other classes, providing evidence for contact between these seemingly geographically separated populations. Mobility via conjugation has been previously demonstrated for the type B4 plasmid of strain Eklund 17B, and similar genes associated with conjugation are present in all type B4 plasmids now described. A plasmid toxin-antitoxin system pemI gene located close to the neurotoxin gene cluster and conserved in each type B4 plasmid class may be important in understanding the mechanism which regulates this unique and unexpected bias toward plasmid-borne neurotoxin genes in Group II C. botulinum type B4.

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

    Science.gov (United States)

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

    2017-05-01

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

  18. A procedure for maintenance of the virulence plasmid (pYV) in Yersinia pestis under culture conditions

    Science.gov (United States)

    The pathogenicity of Yersinia pestis depends on the presence of a virulence plasmid (pYV). The unstable nature of pYV in Y. pestis leads to the eventual outgrowth of pYV less cells due to its higher growth rate. Thus, it was necessary to develop procedures to monitor the presence of the plasmid du...

  19. A procedure for monitoring the presence of the virulence plasmid (pYV) in Yersinia pestis under culture conditions

    Science.gov (United States)

    The pathogenicity of Yersinia pestis depends on the presence of a virulence plasmid (pYV). The unstable nature of pYV in Y. pestis leads to the eventual outgrowth of pYV less cells due its higher growth rate. Thus, it was necessary to develop procedures to monitor the presence of the plasmid durin...

  20. The effects of mass transfer rate limitations and NAPL access heterogeneity on nonaqueous phase liquid tracer tests -- A field example

    Energy Technology Data Exchange (ETDEWEB)

    Burt, R.A.; Wilson, D.J.; Christians, G.L.; Williams, S.P.

    1999-07-01

    Nonaqueous phase liquid (NAPL) tracer tests using combinations of nonpartitioning and partitioning tracers have proven to be an effective means of confirming, delineating, and, under appropriate conditions, quantifying the presence of nonaqueous phase liquids in the subsurface. Some factors that can confound the interpretation of these tests include heterogeneous porosity and permeability distributions, heterogeneous NAPL distributions, diffusion of the tracers into and from low-permeability materials, heterogeneous access to the NAPL by the tracers, and deviations from local equilibrium with respect to mass transfer of the tracers between the aqueous phase and the NAPL phase. NAPL tracer tests conducted with separate injection and extraction wells in an alluvial aquifer at a site near Fort Worth, Texas were affected by these factors with the most notable departures from ideal responses evidently attributable to rate limitations on mass transfer. Positive identification of NAPL presence was achieved by clear separation of the breakthrough curves for partitioning tracers from those of nonpartitioning tracers. A two-dimensional model was used to simulate the observed responses. The model is similar to earlier models of tracer tests of this sort, but includes two significant innovations. First, the model takes into account the kinetics of transport by diffusion of partitioning tracers into and from the NAPL. Second, the model permits the inclusion of porous lenses of low permeability into and from which both partitioning and nonpartitioning tracers can move by diffusion. Visual matching of simulated breakthrough curves to the field data was used to semi-quantitatively estimate the mass of NAPL present. Reasonable but imperfect matches of the simulated breakthrough curves to the field data illustrated the significant effect of a heterogeneous distribution of access to the NAPL by the tracers.

  1. Genomics of high molecular weight plasmids isolated from an on-farm biopurification system.

    Science.gov (United States)

    Martini, María C; Wibberg, Daniel; Lozano, Mauricio; Torres Tejerizo, Gonzalo; Albicoro, Francisco J; Jaenicke, Sebastian; van Elsas, Jan Dirk; Petroni, Alejandro; Garcillán-Barcia, M Pilar; de la Cruz, Fernando; Schlüter, Andreas; Pühler, Alfred; Pistorio, Mariano; Lagares, Antonio; Del Papa, María F

    2016-06-20

    The use of biopurification systems (BPS) constitutes an efficient strategy to eliminate pesticides from polluted wastewaters from farm activities. BPS environments contain a high microbial density and diversity facilitating the exchange of information among bacteria, mediated by mobile genetic elements (MGEs), which play a key role in bacterial adaptation and evolution in such environments. Here we sequenced and characterized high-molecular-weight plasmids from a bacterial collection of an on-farm BPS. The high-throughput-sequencing of the plasmid pool yielded a total of several Mb sequence information. Assembly of the sequence data resulted in six complete replicons. Using in silico analyses we identified plasmid replication genes whose encoding proteins represent 13 different Pfam families, as well as proteins involved in plasmid conjugation, indicating a large diversity of plasmid replicons and suggesting the occurrence of horizontal gene transfer (HGT) events within the habitat analyzed. In addition, genes conferring resistance to 10 classes of antimicrobial compounds and those encoding enzymes potentially involved in pesticide and aromatic hydrocarbon degradation were found. Global analysis of the plasmid pool suggest that the analyzed BPS represents a key environment for further studies addressing the dissemination of MGEs carrying catabolic genes and pathway assembly regarding degradation capabilities.