WorldWideScience

Sample records for affecting plasmid production

  1. Factors affecting plasmid production in Escherichia coli from a resource allocation standpoint

    Directory of Open Access Journals (Sweden)

    Cunningham Drew S

    2009-05-01

    Full Text Available Abstract Background Plasmids are being reconsidered as viable vector alternatives to viruses for gene therapies and vaccines because they are safer, non-toxic, and simpler to produce. Accordingly, there has been renewed interest in the production of plasmid DNA itself as the therapeutic end-product of a bioprocess. Improvement to the best current yields and productivities of such emerging processes would help ensure economic feasibility on the industrial scale. Our goal, therefore, was to develop a stoichiometric model of Escherichia coli metabolism in order to (1 determine its maximum theoretical plasmid-producing capacity, and to (2 identify factors that significantly impact plasmid production. Results Such a model was developed for the production of a high copy plasmid under conditions of batch aerobic growth on glucose minimal medium. The objective of the model was to maximize plasmid production. By employing certain constraints and examining the resulting flux distributions, several factors were determined that significantly impact plasmid yield. Acetate production and constitutive expression of the plasmid's antibiotic resistance marker exert negative effects, while low pyruvate kinase (Pyk flux and the generation of NADPH by transhydrogenase activity offer positive effects. The highest theoretical yield (592 mg/g resulted under conditions of no marker or acetate production, nil Pyk flux, and the maximum allowable transhydrogenase activity. For comparison, when these four fluxes were constrained to wild-type values, yields on the order of tens of mg/g resulted, which are on par with the best experimental yields reported to date. Conclusion These results suggest that specific plasmid yields can theoretically reach 12 times their current experimental maximum (51 mg/g. Moreover, they imply that abolishing Pyk activity and/or transhydrogenase up-regulation would be useful strategies to implement when designing host strains for plasmid

  2. 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.

  3. Plasmid genes required for microcin B17 production.

    Science.gov (United States)

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

    1985-09-01

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

  4. Production and pharmaceutical formulation of plasmid DNA vaccines

    NARCIS (Netherlands)

    van der Heijden, I.

    2013-01-01

    Research leading to the thesis ‘Production and pharmaceutical formulation of plasmid DNA vaccines‘ can be divided into two parts. The first part describes the development of a Good Manufacturing Practice (GMP) compliant plasmid DNA production process of pDNA vaccines for the treatment of Human papil

  5. Resveratrol production in bioreactor: Assessment of cell physiological states and plasmid segregational stability

    Directory of Open Access Journals (Sweden)

    Margarida S. Afonso

    2015-03-01

    Full Text Available Resveratrol is a plant secondary metabolite commonly found in peanuts and grapevines with significant health benefits. Recombinant organisms can produce large amounts of resveratrol and, in this work, Escherichia coli BW27784 was used to produce resveratrol in bioreactors while monitoring cell physiology and plasmid stability through flow cytometry and real-time qPCR, respectively. Initially, the influence of culture conditions and precursor addition was evaluated in screening assays and the data gathered was used to perform the bioreactor assays, allowing the production of 160 μg/mL of resveratrol. Cellular physiology and plasmid instability affected the final resveratrol production, with lower viability and plasmid copy numbers associated with lower yields. In sum, this study describes new tools to monitor the bioprocess, evaluating the effect of culture conditions, and its correlation with cell physiology and plasmid segregational stability, in order to define a viable and scalable bioprocess to fulfill the need for larger quantities of resveratrol.

  6. Transcriptional regulation is affected by subnuclear targeting of reporter plasmids to PML nuclear bodies.

    Science.gov (United States)

    Block, Gregory J; Eskiw, Christopher H; Dellaire, Graham; Bazett-Jones, David P

    2006-12-01

    Whereas the PML protein has been reported to have both transcriptional coactivator and corepressor potential, the contribution of the PML nuclear body (PML NB) itself to transcriptional regulation is not well understood. Here we demonstrate that plasmid DNA artificially tethered to PML or the PML NB-targeting domain of Sp100 is preferentially localized to PML NBs. Using the tethering technique, we targeted a simian virus 40 promoter-driven luciferase reporter plasmid to PML NBs, resulting in the repression of the transgene transcriptional activity. Conversely, the tethering of a cytomegalovirus promoter-containing reporter plasmid resulted in activation. Targeting a minimal eukaryotic promoter did not affect its activity. The expression of targeted promoters could be modulated by altering the cellular concentration of PML NB components, including Sp100 and isoforms of the PML protein. Finally, we demonstrate that ICP0, the promiscuous herpes simplex virus transactivator, increases the level of transcriptional activation of plasmid DNA tethered to the PML NB. We conclude that when PML NB components are artificially tethered to reporter plasmids, the PML NB contributes to the regulation of the tethered DNA in a promoter-dependent manner. Our findings demonstrate that transient transcription assays are sensitive to the subnuclear localization of the transgene plasmid.

  7. Polar Fixation of Plasmids during Recombinant Protein Production in Bacillus megaterium Results in Population Heterogeneity.

    Science.gov (United States)

    Münch, Karin M; Müller, Johannes; Wienecke, Sarah; Bergmann, Simone; Heyber, Steffi; Biedendieck, Rebekka; Münch, Richard; Jahn, Dieter

    2015-09-01

    During the past 2 decades, Bacillus megaterium has been systematically developed for the gram-per-liter scale production of recombinant proteins. The plasmid-based expression systems employed use a xylose-controlled promoter. Protein production analyses at the single-cell level using green fluorescent protein as a model product revealed cell culture heterogeneity characterized by a significant proportion of less productive bacteria. Due to the enormous size of B. megaterium, such bistable behavior seen in subpopulations was readily analyzed by time lapse microscopy and flow cytometry. Cell culture heterogeneity was not caused simply by plasmid loss: instead, an asymmetric distribution of plasmids during cell division was detected during the exponential-growth phase. Multicopy plasmids are generally randomly distributed between daughter cells. However, in vivo and in vitro experiments demonstrated that under conditions of strong protein production, plasmids are retained at one of the cell poles. Furthermore, it was found that cells with accumulated plasmids and high protein production ceased cell division. As a consequence, the overall protein production of the culture was achieved mainly by the subpopulation with a sufficient plasmid copy number. Based on our experimental data, we propose a model whereby the distribution of multicopy plasmids is controlled by polar fixation under protein production conditions. Thereby, cell lines with fluctuating plasmid abundance arise, which results in population heterogeneity. Our results provide initial insights into the mechanism of cellular heterogeneity during plasmid-based recombinant protein production in a Bacillus species.

  8. Effects of medium composition on the production of plasmid DNA vector potentially for human gene therapy

    Institute of Scientific and Technical Information of China (English)

    XU Zhi-nan; SHEN Wen-he; CHEN Hao; CEN Pei-lin

    2005-01-01

    Plasmid vector is increasingly applied to gene therapy or gene vaccine. The production of plasmid pCMV-AP3 for cancer gene therapy was conducted in a modified MBL medium using a recombinant E. coli BL21 system. The effects of different MMBL components on plasmid yield, cell mass and specific plasmid DNA productivity were evaluated on shake-flask scale. The results showed that glucose was the optimal carbon source. High plasmid yield (58.3 mg/L) was obtained when 5.0 g/L glucose was added to MMBL. Glycerol could be chosen as a complementary carbon source because of the highest specific plasmid productivity (37.9 mg DNA/g DCW). After tests of different levels of nitrogen source and inorganic phosphate, a modified MMBL medium was formulated for optimal plasmid production. Further study showed that the initial acetate addition (less than 4.0 g/L) in MMBL improved plasmid production significantly, although it inhibited cell growth. The results will be useful for large-scale plasmid production using recombinant E. coli system.

  9. Construction of the plasmid-free strain for human growth hormone production.

    Science.gov (United States)

    Schulga, A A; Mechev, P V; Kirpichnikov, M P; Skryabin, K G; Deyev, S M

    2016-01-01

    The E. coli strain, overproducing human growth hormone (hGH) was made by integration of the hGH gene under the control of T7 promoter into the chromosomal LacZ gene of BL21(DE3) via lambda Red recombineering. The strain gave higher productivity (50 mg·L(-1)·OD550(-1)) and better growth characteristics than the corresponding strain in which the same hGH expression cassette was placed in a plasmid. The protein produced by the plasmid-free strain was purified and characterized to be hGH. The results demonstrates that a plasmid-free recombinant strain having a single-copy gene expression cassette in the chromosome could provide better gene activity regulation, higher productivity, superior growth characteristics, as well as more stringent control of the gene sequence invariance than a plasmid-based strain.

  10. 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.

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

    Science.gov (United States)

    Berzin, Vel; Kiriukhin, Michael; Tyurin, Michael

    2013-03-01

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

  12. ColE1-plasmid production in Escherichia coli: Mathematical Simulation and Experimental Validation

    Directory of Open Access Journals (Sweden)

    Inga eFreudenau

    2015-09-01

    Full Text Available Plasmids have become very important as pharmaceutical gene vectors in the fields of gene therapy and genetic vaccination in the last years. In this study, we present a dynamic model to simulate the ColE1-like plasmid replication control, once for a DH5α-strain carrying a low copy plasmid (DH5α-pSUP 201-3 and once for a DH5α-strain carrying a high copy plasmid (DH5α-pCMV-lacZ by using ordinary differential equations (ODE and the MATLAB software. The model includes the plasmid replication control by two regulatory RNA molecules (RNAI and RNAII as well as the replication control by uncharged tRNA molecules. To validate the model, experimental data like RNAI- and RNAII concentration, plasmid copy number (PCN, and growth rate for three different time points in the exponential phase were determined. Depending on the sampled time point, the measured RNAI and RNAII concentrations for DH5α-pSUP 201-3 reside between 6 ±0.7 to 34 ±7 RNAI molecules per cell and 0.44 ±0.1 to 3 ±0.9 RNAII molecules per cell. The determined plasmid copy numbers (PCN averaged between 46 ±26 to 48 ±30 plasmids per cell. The experimentally determined data for DH5α-pCMV-lacZ reside between 345 ±203 to 1086 ±298 RNAI molecules per cell and 22 ±2 to 75 ±10 RNAII molecules per cell with an averaged PCN of 1514 ±1301 to 5806 ±4828 depending on the measured time point. As the model was shown to be consistent with the experimentally determined data, measured at three different time points within the growth of the same strain, we performed predictive simulations concerning the effect of uncharged tRNA molecules on the ColE1-like plasmid replication control. The hypothesis is that these tRNA molecules would have an enhancing effect on the plasmid production. The in silico analysis predicts that uncharged tRNA molecules would indeed increase the pDNA production.

  13. ColE1-Plasmid Production in Escherichia coli: Mathematical Simulation and Experimental Validation

    Science.gov (United States)

    Freudenau, Inga; Lutter, Petra; Baier, Ruth; Schleef, Martin; Bednarz, Hanna; Lara, Alvaro R.; Niehaus, Karsten

    2015-01-01

    Plasmids have become very important as pharmaceutical gene vectors in the fields of gene therapy and genetic vaccination in the past years. In this study, we present a dynamic model to simulate the ColE1-like plasmid replication control, once for a DH5α-strain carrying a low copy plasmid (DH5α-pSUP 201-3) and once for a DH5α-strain carrying a high copy plasmid (DH5α-pCMV-lacZ) by using ordinary differential equations and the MATLAB software. The model includes the plasmid replication control by two regulatory RNA molecules (RNAI and RNAII) as well as the replication control by uncharged tRNA molecules. To validate the model, experimental data like RNAI- and RNAII concentration, plasmid copy number (PCN), and growth rate for three different time points in the exponential phase were determined. Depending on the sampled time point, the measured RNAI- and RNAII concentrations for DH5α-pSUP 201-3 reside between 6 ± 0.7 and 34 ± 7 RNAI molecules per cell and 0.44 ± 0.1 and 3 ± 0.9 RNAII molecules per cell. The determined PCNs averaged between 46 ± 26 and 48 ± 30 plasmids per cell. The experimentally determined data for DH5α-pCMV-lacZ reside between 345 ± 203 and 1086 ± 298 RNAI molecules per cell and 22 ± 2 and 75 ± 10 RNAII molecules per cell with an averaged PCN of 1514 ± 1301 and 5806 ± 4828 depending on the measured time point. As the model was shown to be consistent with the experimentally determined data, measured at three different time points within the growth of the same strain, we performed predictive simulations concerning the effect of uncharged tRNA molecules on the ColE1-like plasmid replication control. The hypothesis is that these tRNA molecules would have an enhancing effect on the plasmid production. The in silico analysis predicts that uncharged tRNA molecules would indeed increase the plasmid DNA production. PMID:26389114

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

    Science.gov (United States)

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

    2017-01-01

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

  15. 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.

  16. In situ NIR spectroscopy monitoring of plasmid production processes effect of producing strain, medium composition and the cultivation strategy

    OpenAIRE

    Lopes, Marta B.; Gonçalves, Geisa A. L.; Felício-Silva, Daniel; Prather, Kristala L. J.; Monteiro, Gabriel; Prazeres, Duarte M. F.; Calado, Cecília Ribeiro da Cruz

    2015-01-01

    BACKGROUNDWhile the pharmaceutical industry keeps an eye on plasmid DNA production for new generation gene therapies, real-time monitoring techniques for plasmid bioproduction are as yet unavailable. This work shows the possibility of in situ monitoring of plasmid production in Escherichia coli cultures using a near infrared (NIR) fiber optic probe. RESULTSPartial least squares (PLS) regression models based on the NIR spectra were developed for predicting bioprocess critical variables su...

  17. Advances in host and vector development for the production of plasmid DNA vaccines.

    Science.gov (United States)

    Mairhofer, Juergen; Lara, Alvaro R

    2014-01-01

    Recent developments in DNA vaccine research provide a new momentum for this rather young and potentially disruptive technology. Gene-based vaccines are capable of eliciting protective immunity in humans to persistent intracellular pathogens, such as HIV, malaria, and tuberculosis, for which the conventional vaccine technologies have failed so far. The recent identification and characterization of genes coding for tumor antigens has stimulated the development of DNA-based antigen-specific cancer vaccines. Although most academic researchers consider the production of reasonable amounts of plasmid DNA (pDNA) for immunological studies relatively easy to solve, problems often arise during this first phase of production. In this chapter we review the current state of the art of pDNA production at small (shake flasks) and mid-scales (lab-scale bioreactor fermentations) and address new trends in vector design and strain engineering. We will guide the reader through the different stages of process design starting from choosing the most appropriate plasmid backbone, choosing the right Escherichia coli (E. coli) strain for production, and cultivation media and scale-up issues. In addition, we will address some points concerning the safety and potency of the produced plasmids, with special focus on producing antibiotic resistance-free plasmids. The main goal of this chapter is to make immunologists aware of the fact that production of the pDNA vaccine has to be performed with as much as attention and care as the rest of their research.

  18. Production of plasmid-encoding NDM-1 in clinical Raoultella ornithinolytica and Leclercia adecarboxylata from China.

    Science.gov (United States)

    Sun, Fengjun; Yin, Zhe; Feng, Jiao; Qiu, Yefeng; Zhang, Defu; Luo, Wenbo; Yang, Huiying; Yang, Wenhui; Wang, Jie; Chen, Weijun; Xia, Peiyuan; Zhou, Dongsheng

    2015-01-01

    Raoultella ornithinolytica YNKP001 and Leclercia adecarboxylata P10164, which harbor conjugative plasmids pYNKP001-NDM and pP10164-NDM, respectively, were isolated from two different Chinese patients, and their complete nucleotide sequences were determined. Production of NDM-1 enzyme by these plasmids accounts for the carbapenem resistance of these two strains. This is the first report of bla NDM in L. adecarboxylata and third report of this gene in R. ornithinolytica. pYNKP001-NDM is very similar to the IncN2 NDM-1-encoding plasmids pTR3, pNDM-ECS01, and p271A, whereas pP10164-NDM is similar to the IncFIIY bla NDM-1-carrying plasmid pKOX_NDM1. The bla NDM-1 genes of pYNKP001-NDM and pP10164-NDM are embedded in Tn125-like elements, which represent two distinct truncated versions of the NDM-1-encoding Tn125 prototype observed in pNDM-BJ01. Flanking of these two Tn125-like elements by miniature inverted repeat element (MITE) or its remnant indicates that MITE facilitates transposition and mobilization of bla NDM-1 gene contexts.

  19. Production of plasmid-encoding NDM-1 in clinical Raoultella ornithinolytica and Leclercia adecarboxylata from China

    Directory of Open Access Journals (Sweden)

    Dongsheng eZhou

    2015-05-01

    Full Text Available Raoultella ornithinolytica YNKP001 and Leclercia adecarboxylata P10164, harboring conjugative plasmids pYNKP001-NDM and pP10164-NDM with determination of complete nucleotide sequences, respectively, were isolated from two different Chinese patients. Production of NDM-1 enzyme by these plasmids accounts for carbapenem resistance of these two strains. This is the first report of blaNDM in L. adecarboxylata and the third report of this gene in R. ornithinolytica. pYNKP001-NDM is very similar to the IncN2 NDM-1-encoding plasmids pTR3, pNDM-ECS01 and p271A, while pP10164-NDM is similar to the IncFIIY blaNDM-1-carrying plasmid pKOX_NDM1. The blaNDM-1 genes of pYNKP001-NDM and pP10164-NDM are embedded in Tn125-like elements, which represent two distinct truncated versions of the prototype NDM-1-encoding Tn125 as observed in pNDM-BJ01. Flanking of these two Tn125-like elements by miniature inverted repeat element (MITE or its remnant denotes MITE felicitates transposition and mobilization of blaNDM-1 gene contexts.

  20. 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...

  1. 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...

  2. Production and purification of plasmid DNA vaccines: is there scope for further innovation?

    Science.gov (United States)

    Xenopoulos, Alex; Pattnaik, Priyabrata

    2014-12-01

    The demand for plasmid DNA (pDNA) has vastly increased over the past decade in response to significant advances that have been made in its application for gene therapy and vaccine development. Plasmid DNA-based vaccines are experiencing a resurgence due to success with prime-boost immunization strategies. The challenge has always been poor productivity and delivery of pDNA. Plasmid DNA-based vaccines have traditionally required milligram scale of GMP-grade product for vaccination due to the relatively low efficacy and duration of gene expression. However, efforts to increase pDNA vaccine effectiveness are evolving in genetic manipulations of bacterial host, improvements in product recovery and innovative delivery methods. This review summarizes recent advances in large-scale pDNA vaccine manufacturing, ranging from upstream processing, downstream processing and formulation, as such information is usually not available to the scientific community. The article will highlight technology gaps and offer insight on further scope of innovation.

  3. 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

  4. 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...

  5. Plasmid and Host Strain Characteristics of Escherichia coli Resistant to Extended-Spectrum Cephalosporins in the Norwegian Broiler Production.

    Science.gov (United States)

    Mo, Solveig Sølverød; Slettemeås, Jannice Schau; Berg, Einar Sverre; Norström, Madelaine; Sunde, Marianne

    2016-01-01

    Escherichia coli resistant to extended-spectrum cephalosporins have been detected in the Norwegian broiler production, despite the fact that antimicrobial agents are rarely used. The genetic mechanism responsible for cephalosporin resistance is mainly attributed to the presence of the blaCMY-2 gene encoding a plasmid-mediated AmpC-beta-lactamase (pAmpC). The aim of this study was to characterize and compare blaCMY-2 containing Escherichia coli isolated from the intestinal flora of broilers and retail chicken meat (fillets) to identify possible successful clones and/or resistance plasmids widespread in the Norwegian broiler production. Methods used included PCR based phylotyping, conjugation experiments, plasmid replicon typing, pulsed-field gel electrophoresis, multiple locus variable-number tandem-repeats analysis and whole genome sequencing. The nucleotide sequence of an IncK plasmid carrying blaCMY-2 was determined. Intestinal isolates displayed a higher degree of genetic diversity than meat isolates. A cluster of genetically related isolates belonging to ST38, phylogroup D, carrying blaCMY-2 containing IncK plasmids was identified. Furthermore, genes encoding plasmid stability systems (relBE/stbDE and pndAC) were identified on the IncK plasmid. Single nucleotide polymorphism (SNP) analysis of a subset of isolates confirmed a close genetic relationship within the two most prevalent STs. The IncK plasmids within these two STs also shared a high degree of similarity. Cephalosporin-resistant E. coli with the same genetic characteristics have been identified in the broiler production in other European countries, and the IncK plasmid characterized in this study showed close homology to a plasmid isolated from retail chicken meat in the Netherlands. The results indicate that both clonal expansion and horizontal transfer of blaCMY-2 containing plasmids contribute to dissemination of cephalosporin resistant E. coli in the broiler production. The presence of plasmid

  6. 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

  7. 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

  8. Implementation of an Automated High-Throughput Plasmid DNA Production Pipeline.

    Science.gov (United States)

    Billeci, Karen; Suh, Christopher; Di Ioia, Tina; Singh, Lovejit; Abraham, Ryan; Baldwin, Anne; Monteclaro, Stephen

    2016-12-01

    Biologics sample management facilities are often responsible for a diversity of large-molecule reagent types, such as DNA, RNAi, and protein libraries. Historically, the management of large molecules was dispersed into multiple laboratories. As methodologies to support pathway discovery, antibody discovery, and protein production have become high throughput, the implementation of automation and centralized inventory management tools has become important. To this end, to improve sample tracking, throughput, and accuracy, we have implemented a module-based automation system integrated into inventory management software using multiple platforms (Hamilton, Hudson, Dynamic Devices, and Brooks). Here we describe the implementation of these systems with a focus on high-throughput plasmid DNA production management.

  9. A Transient Three-plasmid Expression System for the Production of Hepatocytes Targeting Retroviral Vectors

    Institute of Scientific and Technical Information of China (English)

    Peng QI; Jinxiang HAN; Yanqin LU; Chuanxi WANG; Bo ZHU

    2007-01-01

    Targeting of retroviral vectors to specific cells was attempted through modifying the surface protein of the murine leukemia viruses (MLVs), but in many cases the protein function was affected, and it is difficult to achieve the targeted delivery. In this study, we have tried to engineer ecotropic Moloney murine leukemia viruses (MoMLV)-based retroviral vectors to transduce hepatocytes. A chimeric envelope (Env)expression plasmid was constructed containing the hepatitis B virus PreS2 peptide fused to aa +1 at the Nterminus of Env. Following simultaneous transfection of pgag-pol, pLEGFP and chimeric env plasmids into 293T cells, helper-free retrovirus stocks with the titer of approximately 104 infectious units/ml were achieved at 48 h post-transfection. These pseudotype vectors showed the normal host range of retrovirus, infecting host NIH 3T3 cells, although the efficiency was reduced compared with that of virions carrying wild-type ecotropic MoMLV envelope. In addition, the resultant pseudotype viruses could transduce human hepatoma cells mediated by polymerized human serum albumin with relatively high titers in comparison with those transductions without polymerized human serum albumin. This approach can be used to target hepatocytes selectively.

  10. Affective Productions of Mathematical Experience

    Science.gov (United States)

    Walshaw, Margaret; Brown, Tony

    2012-01-01

    In underscoring the affective elements of mathematics experience, we work with contemporary readings of the work of Spinoza on the politics of affect, to understand what is included in the cognitive repertoire of the Subject. We draw on those resources to tell a pedagogical tale about the relation between cognition and affect in settings of…

  11. Engineering of Escherichia coli strains for plasmid biopharmaceutical production: scale-up challenges.

    Science.gov (United States)

    Gonçalves, Geisa A L; Prather, Kristala L J; Monteiro, Gabriel A; Prazeres, Duarte M F

    2014-05-19

    Plasmid-based vaccines and therapeutics have been making their way into the clinic in the last years. The existence of cost-effective manufacturing processes capable of delivering high amounts of high-quality plasmid DNA (pDNA) is essential to generate enough material for trials and support future commercialization. However, the development of pDNA manufacturing processes is often hampered by difficulties in predicting process scale performance of Escherichia coli cultivation on the basis of results obtained at lab scale. This paper reports on the differences observed in pDNA production when using shake flask and bench-scale bioreactor cultivation of E. coli strains MG1655ΔendAΔrecA and DH5α in complex media with 20 g/L of glucose. MG1655ΔendAΔrecA produced 5-fold more pDNA (9.8 mg/g DCW) in bioreactor than in shake flask (1.9 mg/g DCW) and DH5α produced 4-fold more pDNA (8 mg/g DCW) in bioreactor than in shake flask (2 mg/g DCW). Accumulation of acetate was also significant in shake flasks but not in bioreactors, a fact that was attributed to a lack of control of pH.

  12. How Employee Turnover Affects Productivity

    DEFF Research Database (Denmark)

    Eriksen, Bo

    Research on employee turnover suggests that turnover results in negative organization-level outcomes. This paper provides a firm-level analysis of the impact of the in- and outflows of human resources on productivity and how the presence of organizational slack resources moderates the effects...

  13. Production of 2-ketoisocaproate with Corynebacterium glutamicum strains devoid of plasmids and heterologous genes.

    Science.gov (United States)

    Vogt, Michael; Haas, Sabine; Polen, Tino; van Ooyen, Jan; Bott, Michael

    2015-03-01

    2-Ketoisocaproate (KIC), the last intermediate in l-leucine biosynthesis, has various medical and industrial applications. After deletion of the ilvE gene for transaminase B in l-leucine production strains of Corynebacterium glutamicum, KIC became the major product, however, the strains were auxotrophic for l-isoleucine. To avoid auxotrophy, reduction of IlvE activity by exchanging the ATG start codon of ilvE by GTG was tested instead of an ilvE deletion. The resulting strains were indeed able to grow in glucose minimal medium without amino acid supplementation, but at the cost of lowered growth rates and KIC production parameters. The best production performance was obtained with strain MV-KICF1, which carried besides the ilvE start codon exchange three copies of a gene for a feedback-resistant 2-isopropylmalate synthase, one copy of a gene for a feedback-resistant acetohydroxyacid synthase and deletions of ltbR and iolR encoding transcriptional regulators. In the presence of 1 mM l-isoleucine, MV-KICF1 accumulated 47 mM KIC (6.1 g l(-1)) with a yield of 0.20 mol/mol glucose and a volumetric productivity of 1.41 mmol KIC l(-1)  h(-1). Since MV-KICF1 is plasmid free and lacks heterologous genes, it is an interesting strain for industrial application and as platform for the production of KIC-derived compounds, such as 3-methyl-1-butanol.

  14. How Employee Turnover Affects Productivity

    DEFF Research Database (Denmark)

    Eriksen, Bo

    Research on employee turnover suggests that turnover results in negative organization-level outcomes. This paper provides a firm-level analysis of the impact of the in- and outflows of human resources on productivity and how the presence of organizational slack resources moderates the effects...... of employee turnover. Drawing on a unique longitudinal dataset of 2,926 Danish manufacturing firms that combine individual-level data with firm-level data, the paper shows that job turnover has a substantial negative effect on total productivity but that the firm’s size, its capital intensity, and its age...... moderate this effect so that the negative consequences of employee turnover are less severe for larger, older and capital intensive firms. These moderating variables indicate the presence of slack resources in the firm, and thus that the accumulation of slack reduces the efficiency losses from employee...

  15. Bicistronic expression plasmid for the rapid production of recombinant fused proteins in Escherichia coli.

    Science.gov (United States)

    Yero, Daniel; Pajón, Rolando; Niebla, Olivia; Sardiñas, Gretel; Vivar, Isbel; Perera, Yasser; García, Darien; Delgado, Maité; Cobas, Karem

    2006-04-01

    In the post-genomic era, every aspect of the production of proteins must be accelerated. In this way, several vectors are currently exploited for rapid production of recombinant proteins in Escherichia coli. N-terminal fusions to the first 47 amino acids of the LpdA (dihydrolipoamide dehydrogenase A) protein of Neisseria meningitidis have been shown to increase the expression of recombinant proteins. Consequently, we have constructed a modified N-terminal LpdA fusion vector, introducing the blue/white colony selection by exploiting a bicistronic gene organization. In the new vector, the sequence encoding the first 47 amino acids of meningococcal LpdA and the alpha-peptide sequence of beta-galactosidase were connected via a ribosome-binding site, and two MCSs (multiple cloning sites) were located surrounding the latter, allowing efficient cloning by colour selection of recombinants. The vector was also improved with the addition of a C-terminal polyhistidine tag, and an EKS (enterokinase recognition sequence) immediately after the LpdA fusion sequence. The new plasmid was employed in the expression and purification of six different bacterial polypeptides. One of these recombinant proteins, P6 protein from Haemophilus influenzae, was used as a model and its N-terminal fusion sequence was totally removed from the recombinant version after incubation with the enterokinase protease, while the polyhistidine tail successfully allowed the purification of the unfused protein from the protease reaction. Two completely new neisserial vaccine candidates, NMB0088 and NMB1126 proteins, were cloned, expressed and purified using this system. To our knowledge, this constitutes the first report of the cloning and expression of these proteins in E. coli.

  16. Factors Affecting the Productivity of Government Workers

    Directory of Open Access Journals (Sweden)

    Jerry P. Haenisch

    2012-01-01

    Full Text Available While there have been a variety of studies concerning government worker motivation and productivity, few, if any, studies have focused specifically on state government workers’ perceptions about what factors affect their productivity. With more than 5 million workers employed by state governments in the United States, any improvement in state workplace productivity could have significant financial and service impact for society. In this study, state government workers identified those factors perceived as most affecting their workplace productivity. Data were collected through a survey offered to state government workers in the state of Wyoming. Factor analysis was used to derive key productivity factors from survey responses. The results indicate that state government workers appreciate having freedom and autonomy, like their jobs and the sense of achievement, and welcome teamwork, but feel limited by poor supervision and management, poor communications, and insufficient budgets and staffing. To improve productivity, the workers would eliminate bureaucracy, supervise better, and improve communication.

  17. Does Labour Diversity affect Firm Productivity?

    DEFF Research Database (Denmark)

    Parrotta, Pierpaolo; Pozzoli, Dario; Pytlikova, Mariola

    that labor diversity in education significantly enhances a firm's value added. Conversely, diversity in ethnicity and demographics induces negative effects on firm productivity. Hence, the negative effects, coming from communication and integration costs connected to a more culturally and demographically......Using a matched employer-employee dataset, we analyze how workforce diversity in cultural background, education and demographic characteristics affects productivity of firms in Denmark. Implementing a structural estimation of the firms' production function (Ackerberg et al., 2006) we find...

  18. Enhancing functional production of a chaperone-dependent lipase in Escherichia coli using the dual expression cassette plasmid

    Directory of Open Access Journals (Sweden)

    Quyen Thi Dinh

    2012-03-01

    Full Text Available Abstracts Background The lipase subfamilies I.1 and I.2 show more than 33% homology in the amino acid sequences and most members share another common property that their genes are clustered with the secondary genes whose protein products are required for folding the lipase into an active conformation and secretion into the culture medium. In previous studies, the lipase (LipA and its chaperone (LipB from Ralstonia sp. M1 were overexpressed in E. coli and the lipase was successfully refolded in vitro. The purpose of this study was to enhance the production of the active lipase LipA from Ralstonia sp. M1 in the heterologous host E. coli without in vitro refolding process, using two-plasmid co-expression systems and dual expression cassette plasmid systems. Results To produce more active lipase from Ralstonia sp. M1 in E. coli without in vitro refolding process but with the help of overexpression of the chaperone (LipB1 and LipB3 corresponding to 56-aa truncated and 26-aa truncated chaperone LipB, six different expression systems including 2 two-plasmid co-expression systems (E. coli BL21/pELipABa + pELipB1k and BL21/pELipABa + pELipB3k and 4 dual expression cassette plasmid systems (BL21/pELipAB-LipB1a, BL21/pELipAB-LipB3a, BL21/pELipA-LipB1a, and BL21/pELipA-LipB3a were constructed. The two-plasmid co-expression systems (E. coli BL21/pELipABa + pELipB1k and BL21/pELipABa + pELipB3k produced the active lipase at a level of 4 times as high as the single expression cassette plasmid system E. coli BL21/pELipABa did. For the first time, the dual expression cassette plasmid systems BL21/pELipAB-LipB1a and BL21/pELipAB-LipB3a yielded 29- and 19-fold production of the active lipase in comparison with the single expression cassette plasmid system E. coli BL21/pELipABa, respectively. Although the lipase amount was equally expressed in all these expression systems (40% of total cellular protein and only a small fraction of the overexpressed lipase was

  19. The marketing implications of affective product design.

    Science.gov (United States)

    Seva, Rosemary R; Duh, Henry Been-Lirn; Helander, Martin G

    2007-11-01

    Emotions are compelling human experiences and product designers can take advantage of this by conceptualizing emotion-engendering products that sell well in the market. This study hypothesized that product attributes influence users' emotions and that the relationship is moderated by the adherence of these product attributes to purchase criteria. It was further hypothesized that the emotional experience of the user influences purchase intention. A laboratory study was conducted to validate the hypotheses using mobile phones as test products. Sixty-two participants were asked to assess eight phones from a display of 10 phones and indicate their emotional experiences after assessment. Results suggest that some product attributes can cause intense emotional experience. The attributes relate to the phone's dimensions and the relationship between these dimensions. The study validated the notion of integrating affect in designing products that convey users' personalities.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    1986-07-01

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

  1. Improving putrescine production by Corynebacterium glutamicum by fine-tuning ornithine transcarbamoylase activity using a plasmid addiction system.

    Science.gov (United States)

    Schneider, Jens; Eberhardt, Dorit; Wendisch, Volker F

    2012-07-01

    Corynebacterium glutamicum shows a great potential for the production of the polyamide monomer putrescine (1,4-diaminobutane). Previously, we constructed the putrescine-producing strain PUT1 by deletion of argF, the gene for ornithine transcarbamoylase (OTC), and argR, encoding the L-arginine repressor, combined with heterologous expression of the Escherichia coli gene for L-ornithine decarboxylase SpeC. As a consequence of argF deletion, this strain requires supplementation of L-arginine and shows growth-decoupled putrescine production. To avoid costly supplementation with L-arginine and the strong feedback inhibition of the key enzyme N-acetylglutamate kinase (ArgB) by L-arginine, a plasmid addiction system for low-level argF expression was developed. By fine-tuning argF expression through modifications of the promoter, the translational start codon and/or the ribosome binding site, high productivity and titer could be obtained. OTC activity varied almost thousandfold between 960 and 1 mU mg⁻¹ resulting in putrescine yields on glucose from less than 0.001 up to 0.26 g g⁻¹, the highest yield in bacteria reported to date. The most promising strain, designated PUT21, was characterized comprehensively. PUT21 strain grew with a rate of 0.19 h⁻¹ in mineral salt medium without the need for L-arginine supplementation and produced putrescine with a yield of 0.16 g g⁻¹ glucose at a volumetric productivity of 0.57 g L⁻¹ h⁻¹ and a specific productivity of 0.042 g g⁻¹ h⁻¹. The carbon balance suggested that no major unidentified by-product was produced. Compared to the first-generation strain PUT1, the putrescine yield observed with PUT21 was increased by 60%. In fed-batch cultivation with C. glutamicum PUT21, a putrescine titer of 19 g L⁻¹ at a volumetric productivity of 0.55 g L⁻¹ h⁻¹ and a yield of 0.16 g g⁻¹ glucose could be achieved. Moreover, while plasmid segregation of the initial strain required antibiotic selection

  2. 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.

  3. Pathogenicity of Vibrio anguillarum serogroup O1 strains compared to plasmids, outer membrane protein profiles and siderophore production

    DEFF Research Database (Denmark)

    Pedersen, K.; Gram, Lone; Austin, D.A.;

    1997-01-01

    The virulence of 18 strains of Vibrio anguillarum serogroup 01 was compared to plasmid content, expression of siderophores and outer membrane proteins. All strains, irrespective of plasmid content, produced siderophores and inducible outer membrane proteins under iron-limited conditions. Only...... strains that carried the 67 kbp virulence plasmid or derivatives of it produced the outer membrane protein, OM2. All virulent strains harboured the 67 kbp plasmid or derivatives of it, indicating its importance for virulence. However, some strains carrying the virulence plasmid or a derivative of it...

  4. Factors Affecting Sugarcane Production in Pakistan

    Directory of Open Access Journals (Sweden)

    Adnan Nazir

    2013-05-01

    Full Text Available This study was undertaken to identify the factors affecting sugarcane production in Pakistan. Data were collected from 387 sugarcane growers from Sindh, Punjab and NWFP province. Data were collected during the period 2007-08. The study reveals that the costs of inputs of sugarcane i.e. urea, DAP, FYM, land preparation, seed and its application, weeding and cost of irrigation were the important factors which influenced on the returns of sugarcane growers. The effectiveness was examined by using the Cobb-Douglas production function; MVP and allocative efficiency were calculated. The coefficient of multiple determinations R2 was 0.9249, which indicated that 92% variation in the cost of inputs was explained by all explanatory variables and the adjusted R2 was 92%. The F-value was 666.94 and was highly significant at 5% level of significance, indicating that the regression model was well fitted. The high prices of inputs, low price of output, delay in payments and lack of scientific knowledge were the major problems in sugarcane production. In order to enhance the productivity of sugarcane in the country, government should solve the identified problems to increase the income of sugarcane growers.

  5. Neonatal intramuscular injection of plasmid encoding glucagon-like peptide-1 affects anxiety behaviour and expression of the hippocampal glucocorticoid receptor in adolescent rats

    Indian Academy of Sciences (India)

    Huitao Fan; Lina Wang; Feng Guo; Shi Wei; Ruqian Zhao

    2010-03-01

    Early-life endocrine intervention may programme hippocampal glucocorticoid receptor (GR) expression and cause psychiatric disorders in later life. Glucagon-like peptide-1 (GLP-1) has been implicated in the regulation of neuroendocrine and behavioural responses, but it is yet to be determined whether and how neonatal GLP-1 overexpression may modify hippocampal GR expression and thus programme adolescent behaviour in rats. Two-dayold pups were injected intramuscularly with vacant plasmid (VP) or plasmid DNA encoding secretory GLP-1 (GP). Anxiety-related behaviour was assessed in the elevated plus maze (EPM) test at 8 weeks of age. Plasma corticosterone levels were measured with enzyme immunoassay (EIA). Protein and mRNA levels were determined by western blot and real-time polymerase chain reaction (PCR), respectively. The DNA methylation status of the GR exon 17 promoter was determined by bisulphate sequencing PCR (BSP). GP rats exhibited anxiolytic behaviour compared with their VP counterparts. Hippocampal GLP-1 receptor (GLP-1R) and GR mRNA expression were significantly elevated in GP rats without a significant difference in plasma corticosterone. Significant reduction in DNA methyltransferase 1 (DNMT1) expression was observed in GP rats disconnected with alterations in DNA methylation of the GR exon 17 promoter. Nevertheless, mRNA expression of nerve growth factor-inducible protein A (NGFI-A) was significantly elevated in GP rats. These results suggest that neonatal intramuscular injection of plasmid DNA encoding GLP-1 affects anxiety behaviour in adolescent rats, probably through NGFI-A-activated upregulation of hippocampal GR expression.

  6. Transmissible Plasmid Containing Salmonella enterica Heidelberg Isolates Modulate Cytokine Production During Early Stage of Interaction with Intestinal Epithelial Cells.

    Science.gov (United States)

    Gokulan, Kuppan; Khare, Sangeeta; Williams, Katherine; Foley, Steven L

    2016-08-01

    The variation in cytokine production during bacterial invasion of human intestinal epithelial cells (IECs) is a contributing factor for progression of the infection. A few Salmonella enterica Heidelberg strains isolated from poultry products harbor transmissible plasmids (TPs), including those that encode a type-IV secretion system. Earlier, we showed that these TPs are responsible for increased virulence during infection. This study examines the potential role of these TPs in cytokine production in IECs. This study showed that S. Heidelberg strains containing TPs (we refer as virulent strains) caused decreased interleukin (IL)-10 production in IECs after 1 h infection. The virulent strains induced a high level of tumor necrosis factor-α production under identical conditions. The virulent strains of S. Heidelberg also altered the production of IL-2, IL-17, and granulocyte macrophage colony-stimulating factor compared to an avirulent strain. As a part of infection, bacteria cross the epithelial barrier and encounter intestinal macrophages. Hence, we examined the cytotoxic mechanism of strains of S. Heidelberg in macrophages. Scanning electron microscopy showed cell necrosis occurs during the early stage of infection. In conclusion, virulent S. Heidelberg strains were able to modify the host cytokine profile during the early stages of infection and also caused necrosis in macrophages.

  7. A potential new gene (tccA) on IncP plasmid RK2 and transposon Tn1721: relationship of its product to the TrwC relaxase/helicase of IncW plasmid R388.

    Science.gov (United States)

    Jovanovic, O S; Figurski, D H

    1997-01-01

    Computational analysis of the fully sequenced 60-kb genome of broad-host-range IncP alpha plasmid RK2 revealed a previously unreported potential protein-coding sequence, an 80-codon open reading frame (tccA), located in the region between the vegetative origin of replication (oriV) and the tetR gene of the tetracycline resistance determinant. The coding region is also present in the transposon Tn1721 tet region, which is nearly identical to the tet region of RK2. Remarkably, the predicted polypeptide product of the coding region displays 56% identity and 72% similarity with the C-terminal domain of the TrwC relaxase/helicase protein of IncW plasmid R388.

  8. 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...

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

    Directory of Open Access Journals (Sweden)

    Mourad, Kacem

    2007-06-01

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

  10. Thermostable DNA ligase-mediated PCR production of circular plasmid (PPCP) and its application in directed evolution via in situ error-prone PCR.

    Science.gov (United States)

    Le, Yilin; Chen, Huayou; Zagursky, Robert; Wu, J H David; Shao, Weilan

    2013-08-01

    Polymerase chain reaction (PCR) is a powerful method to produce linear DNA fragments. Here we describe the Tma thermostable DNA ligase-mediated PCR production of circular plasmid (PPCP) and its application in directed evolution via in situ error-prone PCR. In this thermostable DNA ligase-mediated whole-plasmid amplification method, the resultant DNA nick between the 5' end of the PCR primer and the extended newly synthesized DNA 3' end of each PCR cycle is ligated by Tma DNA ligase, resulting in circular plasmid DNA product that can be directly transformed. The template plasmid DNA is eliminated by 'selection marker swapping' upon transformation. When performed under an error-prone condition with Taq DNA polymerase, PPCP allows one-step construction of mutagenesis libraries based on in situ error-prone PCR so that random mutations are introduced into the target gene without altering the expression vector plasmid. A significant difference between PPCP and previously published methods is that PPCP allows exponential amplification of circular DNA. We used this method to create random mutagenesis libraries of a xylanase gene and two cellulase genes. Screening of these libraries resulted in mutant proteins with desired properties, demonstrating the usefulness of in situ error-prone PPCP for creating random mutagenesis libraries for directed evolution.

  11. Use of FabV-Triclosan Plasmid Selection System for Efficient Expression and Production of Recombinant Proteins in Escherichia coli.

    Directory of Open Access Journals (Sweden)

    Syed A Ali

    Full Text Available Maintenance of recombinant plasmid vectors in host bacteria relies on the presence of selection antibiotics in the growth media to suppress plasmid -free segregants. However, presence of antibiotic resistance genes and antibiotics themselves is not acceptable in several applications of biotechnology. Previously, we have shown that FabV-Triclosan selection system can be used to select high and medium copy number plasmid vectors in E. coli. Here, we have extended our previous work and demonstrated that expression vectors containing FabV can be used efficiently to express heterologous recombinant proteins in similar or better amounts in E. coli host when compared with expression vectors containing β-lactamase. Use of small amount of non-antibiotic Triclosan as selection agent in growth medium, enhanced plasmid stability, applicability in various culture media, and compatibility with other selection systems for multiple plasmid maintenance are noteworthy features of FabV-Triclosan selection system.

  12. Thermostable and site-specific DNA binding of the gene product ORF56 from the Sulfolobus islandicus plasmid pRN1, a putative archael plasmid copy control protein

    Science.gov (United States)

    Lipps, Georg; Stegert, Mario; Krauss, Gerhard

    2001-01-01

    There is still a lack of information on the specific characteristics of DNA-binding proteins from hyperthermophiles. Here we report on the product of the gene orf56 from plasmid pRN1 of the acidophilic and thermophilic archaeon Sulfolobus islandicus. orf56 has not been characterised yet but low sequence similarily to several eubacterial plasmid-encoded genes suggests that this 6.5 kDa protein is a sequence-specific DNA-binding protein. The DNA-binding properties of ORF56, expressed in Escherichia coli, have been investigated by EMSA experiments and by fluorescence anisotropy measurements. Recombinant ORF56 binds to double-stranded DNA, specifically to an inverted repeat located within the promoter of orf56. Binding to this site could down-regulate transcription of the orf56 gene and also of the overlapping orf904 gene, encoding the putative initiator protein of plasmid replication. By gel filtration and chemical crosslinking we have shown that ORF56 is a dimeric protein. Stoichiometric fluorescence anisotropy titrations further indicate that ORF56 binds as a tetramer to the inverted repeat of its target binding site. CD spectroscopy points to a significant increase in ordered secondary structure of ORF56 upon binding DNA. ORF56 binds without apparent cooperativity to its target DNA with a dissociation constant in the nanomolar range. Quantitative analysis of binding isotherms performed at various salt concentrations and at different temperatures indicates that approximately seven ions are released upon complex formation and that complex formation is accompanied by a change in heat capacity of –6.2 kJ/mol. Furthermore, recombinant ORF56 proved to be highly thermostable and is able to bind DNA up to 85°C. PMID:11160922

  13. Effect of excessive cadmium chloride on the plasmids of E. coli HB 101 in vivo

    Institute of Scientific and Technical Information of China (English)

    1999-01-01

    After Escherichia coli HB101 with plasmid pWH58, pWH98, or pTBa5 were cultered respectively in amp LB broth which contained 50 mg/L CdCl2 constantly for 24 h, these plasmids were isolated from E. coli, and the effect of excessive CdCl2 on the E. coli HB101 and plasmid DNA was studied by surveying the growth of E. coli HB101 and plasmid, argarose gel electrophoresis and analysis of restriction fragment length polymorphism (RFLP) of plasmids, and plasmid transformation. The results showed that 50 mg/L CdCl2 treatment lagged the growth of E. coli HB101 for at least 4h, but after grown for 24h there were not significant differences in the growths of E. coli HB101s and the productions of plasmids between the treatment and control. These results implified that E. coli HB101 have induced adaptability to cadmium stress and excessive CdCl2 did not inhibit the replication and amp+ gene's expression of plasmid DNA in vivo of E. coli significantly. 50 mg/L CdCl2 treatment for 24 hours might cause the sequence's change of plasmid DNA, but could not lead to the random breakage of plasmid DNA strands. Moreover, after 50 mg/L of CdCl2 treatment in vivo the transformation activities of plasmid did not altered, implied excessive CdCl2 could not affect the superhelical structure of plasmid and also not break the loop of plasmid DNA evidently.

  14. 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.

  15. Isolation and characterization of Rhizobium meliloti mutants affected in exopolysaccharide production.

    Science.gov (United States)

    Rodríguez-Navarro, D N; Palomares, A J; Casadesús, J

    1991-06-01

    Rhizobium meliloti mutants affected in the production of exopolysaccharide (EPS) were isolated after N-methyl-N'-nitro-N-nitrosoguanidine mutagenesis. The mutants were classified into three phenotypic classes: (I) Exo-, rough mutants lacking exopolysaccharide; (II) Exos (for "small") which form tiny, compact colonies and synthesize reduced amounts of EPS; and (III) Exoc (for "constitutive"), hypermucoid mutants which overproduce EPS. Hypermucoid strains showed increased resistance to desiccation. All the mutants were able to nodulate, although a significant decrease in infectivity degree and/or competitiveness was found in rough and compact strains. Two mutants proved to be deficient in nitrogen fixation. Complementation analysis with cloned R. meliloti exo genes could not be applied to the study of these Fix- mutants because introduction of plasmids derived from cosmid vector pLAFR1 caused loss of nodulating ability. However, complementation of calcofluor staining and EPS production was observed. Complementation with certain exo genes also caused a marked increase in motility.

  16. Parameters Affecting Solvent Production by Clostridium pasteurianum

    OpenAIRE

    Dabrock, Birgit; Bahl, Hubert; Gottschalk, Gerhard

    1992-01-01

    The effect of pH, growth rate, phosphate and iron limitation, carbon monoxide, and carbon source on product formation by Clostridium pasteurianum was determined. Under phosphate limitation, glucose was fermented almost exclusively to acetate and butyrate independently of the pH and growth rate. Iron limitation caused lactate production (38 mol/100 mol) from glucose in batch and continuous culture. At 15% (vol/vol) carbon monoxide in the atmosphere, glucose was fermented to ethanol (24 mol/100...

  17. Does Labor Diversity Affect Firm Productivity?

    DEFF Research Database (Denmark)

    Parrotta, Pierpaolo; Pozzoli, Dario; Pytlikova, Mariola

    Using an employer-employee dataset, we analyze how diversity in cultural background, skills and demographic characteristics a?ects total factor productivity (TFP) of ?rms in Denmark. Implementing structural estimation of ?rms' production function, we ?nd evidence that labor diversity in skills/ed......-average trade openness, giving support to the hypothesis that an ethnically diverse workforce provides information and access to global markets....... and integration costs connected to a more demographically and culturally diverse workforce, counteract the positive e?ects of diversity on ?rm TFP, coming from creativity and knowledge spillovers. However, we ?nd that ethnic diversity is valuable for ?rms operating in industries characterized by above...

  18. Farmer's attitudes affect piglet production parameters

    OpenAIRE

    Kauppinen, T; Valros, A.; Vesala, K.M.

    2010-01-01

    Our results show that farmers' attitudes count: treating the animals humanely, investing in a favourable environment, and having a positive attitude towards new information and scientific research is associated with an above-average productivity on piglet farms. These attitudes, when implemented and concretized in practice, also benefit the animals through a higher standard of welfare.

  19. Productive Parasites: Thinking of Noise as Affect

    Directory of Open Access Journals (Sweden)

    Marie Thompson

    2012-11-01

    Full Text Available Drawing from Michel Serres’ notion of the parasite, this article provides a nuanced explanation for noise that distinguishes itself from prevailing negative narratives, which often seek to define noise as unwanted, undesirable or damaging sound. Such narratives have left noise vulnerable to moralising polemics, which construct silence and noise as a dichotomy between the past and present, natural and cultural, relaxing and disturbing, and, fundamentally, good and bad. This article facilitates a reconsideration of noise’s ethical connotations by proposing the notion of noise as affect.

  20. High-throughput plasmid construction using homologous recombination in yeast: its mechanisms and application to protein production for X-ray crystallography.

    Science.gov (United States)

    Mizutani, Kimihiko

    2015-01-01

    Homologous recombination is a system for repairing the broken genomes of living organisms by connecting two DNA strands at their homologous sequences. Today, homologous recombination in yeast is used for plasmid construction as a substitute for traditional methods using restriction enzymes and ligases. This method has various advantages over the traditional method, including flexibility in the position of DNA insertion and ease of manipulation. Recently, the author of this review reported the construction of plasmids by homologous recombination in the methanol-utilizing yeast Pichia pastoris, which is known to be an excellent expression host for secretory proteins and membrane proteins. The method enabled high-throughput construction of expression systems of proteins using P. pastoris; the constructed expression systems were used to investigate the expression conditions of membrane proteins and to perform X-ray crystallography of secretory proteins. This review discusses the mechanisms and applications of homologous recombination, including the production of proteins for X-ray crystallography.

  1. Spread of Extended Spectrum Cephalosporinase-Producing Escherichia coli Clones and Plasmids from Parent Animals to Broilers and to Broiler Meat in a Production Without Use of Cephalosporins

    DEFF Research Database (Denmark)

    Agersø, Yvonne; Jensen, Jacob Dyring; Hasman, Henrik

    2014-01-01

    Objectives: This study investigated the occurrence of extended spectrum cephalosporinase (ESC)–producing Escherichia coli in a broiler production with no cephalosporin use and a low use of antimicrobials in general. Furthermore, it investigated whether the current consumption of aminopenicillins....... Isolates with blaCMY-2 were subtyped by pulsed-field gel electrophoresis (PFGE), phylotyping, and antimicrobial susceptibility testing. Selected isolates were used as donors in filter-mating experiments, multilocus sequence typing (MLST), and plasmid replicons were typed. Aminopenicillin use at the farm...... selects for ESC-producing E. coli and whether certain clones or plasmids spread from imported parent flocks to the meat. Materials and Methods: ESC-producing E. coli was isolated using MacConkey broth with 1 mg/L of ceftriaxone. ESC genes were identified using polymerase chain reaction and sequencing...

  2. Effects of a recombinant gene expression on ColE1-like plasmid segregation in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Ivanov Ivan

    2011-03-01

    Full Text Available Abstract Background Segregation of expression plasmids leads to loss of recombinant DNA from transformed bacterial cells due to the irregular distribution of plasmids between the daughter cells during cell division. Under non-selective conditions this segregational instability results in a heterogeneous population of cells, where the non-productive plasmid-free cells overgrow the plasmid-bearing cells thus decreasing the yield of recombinant protein. Amongst the factors affecting segregational plasmid instability are: the plasmid design, plasmid copy-number, host cell genotype, fermentation conditions etc. This study aims to investigate the influence of transcription and translation on the segregation of recombinant plasmids designed for constitutive gene expression in Escherichia coli LE392 at glucose-limited continuous cultivation. To this end a series of pBR322-based plasmids carrying a synthetic human interferon-gamma (hIFNγ gene placed under the control of different regulatory elements (promoter and ribosome-binding sites were used as a model. Results Bacterial growth and product formation kinetics of transformed E. coli LE392 cells cultivated continuously were described by a structured kinetic model proposed by Lee et al. (1985. The obtained results demonstrated that both transcription and translation efficiency strongly affected plasmid segregation. The segregation of plasmid having a deleted promoter did not exceed 5% after 190 h of cultivation. The observed high plasmid stability was not related with an increase in the plasmid copy-number. A reverse correlation between the yield of recombinant protein (as modulated by using different ribosome binding sites and segregational plasmid stability (determined by the above model was also observed. Conclusions Switching-off transcription of the hIFNγ gene has a stabilising effect on ColE1-like plasmids against segregation, which is not associated with an increase in the plasmid copy

  3. Measuring and adapting behavior during product interaction to influence affect

    NARCIS (Netherlands)

    Alonso, M.B.; Hummels, C.C.M.; Keyson, D.V.; Hekkert, P.P.M.

    2011-01-01

    Sometimes, the way in which we interact with products implicitly communicates how we feel. Based on previous studies on how emotions can be detected and communicated via product interaction, we discuss how an interactive product could influence affect by responding and changing behaviors expressing

  4. Emergence of co-production of plasmid-mediated AmpC beta-lactamase and ESBL in cefoxitin-resistant uropathogenic Escherichia coli.

    Science.gov (United States)

    Ghosh, B; Mukherjee, M

    2016-09-01

    Plasmid-mediated AmpC (pAmpC) and ESBL co-production was detected in Escherichia coli a major etiologic agent of urinary tract infection. Isolates resistant to cefoxitin by CLSI methodology were tested for pAmpC beta-lactamase using phenylboronic acid and ESBLs by combined disk diffusion method. pAmpC/ESBL genes were characterized by PCR and sequencing. Transconjugation experiments were done to study the transfer of pAmpC and ESBL production from clinical isolates as donor to E. coli J53 AziR as recipient. Incompatibility groups of transmissible plasmids were classified by PCR-based replicon typing (PBRT). Among 148 urine culture positive isolates, E. coli was reported in 39.86 % (59/148), with 93.22 % (55/59) of cefoxitin resistance. pAmpC production was detected in 25, with varied distribution of blaCMY-2 and blaDHA-1type genes alone (n = 13 and 7 respectively) or in combination (n = 5). ESBL co-production was observed in 88 % (22/25) of pAmpC producing isolates with predominance of blaTEM (n = 20). Twenty-three transconjugants showed transmission of pAmpC-and ESBL-resistant genes with co-carriage of blaCMY-2 and blaTEM (n = 15) in plasmids of IncF type (n = 9) being predominant, followed by IncI1 (n = 4) and IncH1 (n = 2) in combination. All clinical isolates were clonally diverse. Resistance against different beta-lactams in uropathogenic E. coli has been an emerging concern in resource- poor countries such as India. Knowledge on the occurrence of AmpC beta-lactamases and ESBL amongst this pathogen and its transmission dynamics may aid in hospital infection control.

  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. 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.

  7. Inhibition on the production of collagen type Ⅰ, Ⅲ of activated hepatic stellate cells by antisense TIMP-1 recombinant plasmid

    Institute of Scientific and Technical Information of China (English)

    Wen-Bin Liu; Chang-Qing Yang; Wei Jiang; Yi-Qing Wang; Jing-Sheng Guo; Bo-Ming He; Ji-Yao Wang

    2003-01-01

    AIM: To investigate the inhibition effects on the productionof collagen type I, Ⅲ secreted by activated rat hepatic stellatecells (rHSCs) by antisense tissue inhibitors of metalloproteinase1 (TIMP-1) recombinant plasmid through elevating interstitialcollagenase activity.METHODS: rHSCs were extracted from normal rat liverby pronase and collagenase digestion and purified bycentrifugal elutriation, and were cultured on plastic dishesuntil they were activated to a myofibroblastic phenotypeafter 7-10 days. RT-Nest-PCR and gene recombinanttechniques were used to construct the rat antisense TIMP-1 recombinant plasmids which can express in eucaryoticcells. The recombinant plasmid and the pcDNA3 emptyplasmid were transfected in rHSCs by Effectene (QIAGEN)separately. Cells were selected after growing in DMEMcontaining 400 μg/ml G418 for 2-3 weeks. Expression ofexogenous gene was assessed by Northern blot, andexpression oflIMP-1 in rHSCs was determined by Northernblot and Western blot. We tested the interstitial collagenaseactivity with FITC-labled type I collagen as substrate.Ultimately, we quantified the type Ⅰ, Ⅲ collagen byWestern blot.RESULTS: The exogenous antisense TIMP-1 recombinantplasmid could be expressed in rHSCs well, which couldblock the expression of TIMP-1 greatly, the ratio of TIMP-1/GAPDH was 0.67, 2.41, and 2.97 separately at mRNAlevel (P<0.05); the ratio of TIMP-1/β-actin was 0.31, 0.98and 1.32 separately at protein level (P<0.05); It mightelevate active and latent interstitial collagenase activity,the collagenase activity was 0.3049, 0.1411 and 0.1196respectively. (P<0.05), which led to promotion thedegradation of type Ⅰ, Ⅲ collagen, the ratio of collagen I/β-actin was 0.63, 1.78 and 1.92 separately (P<0.05); andthe ratio of collagen Ⅲ/β-actin was 0.59, 1.81 and 1.98separately (P<0.05).CONCLUSION: These data shows that the antisense TIMP-1 recombinant plasmid has the inhibitory effects on theproduction of type Ⅰ, Ⅲ collagens

  8. Effect of low temperature on stability of theta-type plasmids in Carnobacterium maltaromaticum.

    Science.gov (United States)

    Bohaychuk, Valerie M; van Belkum, Marco J; Stiles, Michael E; McMullen, Lynn M

    2008-03-01

    The heterologous production of useful peptides such as bacteriocins by lactic acid bacteria (LAB) has been studied for use in the biopreservation of foods. Recombinant plasmids can suffer drawbacks such as segregational instability affecting the production of these peptides in certain environments such as absence of selective pressure or low temperature. The link between growth temperature characteristics of parental strains and stability of theta-type plasmids at a low temperature was investigated. The growth of four parental strains at 4 degrees C and stability of five derivative theta-type plasmids transformed into Carnobacterium maltaromaticum UAL26 at 25 and 4 degrees C were determined. Two plasmids (pCD11 and pCaT) derived from psychrotrophic LAB and plasmid, pHW800, from Enterococcus faecium 226 with unknown growth temperature characteristics, had excellent stability when strains were grown at 4 degrees C. Plasmids (pTRKH2 and pUCB820) derived from LAB that did not grow at refrigeration temperatures were not stable at 4 degrees C. When a DNA fragment from pCD11 containing 22-bp repeats, a putative replication initiation site, and the gene for the RepA protein was inserted into pTRKH2, the resulting derivative plasmid was 100% stable at 4 degrees C.

  9. 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

  10. Relations between affective music and speech: Evidence from dynamics of affective piano performance and speech production

    Directory of Open Access Journals (Sweden)

    Xiaoluan eLiu

    2015-07-01

    Full Text Available This study compares affective piano performance with speech production from the perspective of dynamics: unlike previous research, this study uses finger force and articulatory effort as indexes reflecting the dynamics of affective piano performance and speech production respectively. Moreover, for the first time physical constraints such as piano fingerings and speech articulatory distance are included due to their potential contribution to different patterns of dynamics. A piano performance experiment and speech production experiment were conducted in four emotions: anger, fear, happiness and sadness. The results show that in both piano performance and speech production, anger and happiness generally have high dynamics while sadness has the lowest dynamics, with fear in the middle. Fingerings interact with fear in the piano experiment and articulatory distance interacts with anger in the speech experiment, i.e., large physical constraints produce significantly higher dynamics than small physical constraints in piano performance under the condition of fear and in speech production under the condition of anger. Using production experiments, this study firstly supports previous perception studies on relations between affective music and speech. Moreover, this is the first study to show quantitative evidence for the importance of considering motor aspects such as dynamics in comparing music performance and speech production in which motor mechanisms play a crucial role.

  11. Relations between affective music and speech: evidence from dynamics of affective piano performance and speech production.

    Science.gov (United States)

    Liu, Xiaoluan; Xu, Yi

    2015-01-01

    This study compares affective piano performance with speech production from the perspective of dynamics: unlike previous research, this study uses finger force and articulatory effort as indexes reflecting the dynamics of affective piano performance and speech production respectively. Moreover, for the first time physical constraints such as piano fingerings and speech articulatory constraints are included due to their potential contribution to different patterns of dynamics. A piano performance experiment and speech production experiment were conducted in four emotions: anger, fear, happiness and sadness. The results show that in both piano performance and speech production, anger and happiness generally have high dynamics while sadness has the lowest dynamics. Fingerings interact with fear in the piano experiment and articulatory constraints interact with anger in the speech experiment, i.e., large physical constraints produce significantly higher dynamics than small physical constraints in piano performance under the condition of fear and in speech production under the condition of anger. Using production experiments, this study firstly supports previous perception studies on relations between affective music and speech. Moreover, this is the first study to show quantitative evidence for the importance of considering motor aspects such as dynamics in comparing music performance and speech production in which motor mechanisms play a crucial role.

  12. 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 ...

  13. Product Meaning, Affective Use Evaluation, and Transfer: A Preliminary Study

    Directory of Open Access Journals (Sweden)

    Sacha Helfenstein

    2005-01-01

    Full Text Available The first part of this investigation explored the multidimensional nature of product meaning, referring to the variety of connotations and functions a consumer associates with a particular product category. The subsequent experiment examined the moderation effects of product meaning and other attributes of the user on (a the affective evaluation of an obstructed use interaction, and (b the transfer of emotion between devices presented as being either of the same or a different brand. Although the failure experience essentially caused frustration, this reaction varied substantially among consumers depending on product meaning, age, and gender. The results also showed that the emotion dimensions of pleasure, arousal, and dominance were affected in distinct ways, and that, in addition to the consumer variables, transfer was mainly dependent on the brand relation. Nevertheless, user frustration did not cause general brand aversion, indicating that poor designs do not unconditionally threaten the customer relationship.

  14. Product design enhancement using apparent usability and affective quality.

    Science.gov (United States)

    Seva, Rosemary R; Gosiaco, Katherine Grace T; Santos, Ma Crea Eurice D; Pangilinan, Denise Mae L

    2011-03-01

    In this study, apparent usability and affective quality were integrated in a design framework called the Usability Perception and Emotion Enhancement Model (UPEEM). The UPEEM was validated using structural equation modeling (SEM). The methodology consists of four phases namely product selection, attribute identification, design alternative generation, and design alternative evaluation. The first stage involved the selection of a product that highly involves the consumer. In the attribute identification stage, design elements of the product were identified. The possible values of these elements were also determined for use in the experimentation process. Design of experiments was used to identify how the attributes will be varied in the design alternative stage and which of the attributes significantly contribute to affective quality, apparent usability, and desirability in the design evaluation stage. Results suggest that product attributes related to form are relevant in eliciting intense affect and perception of usability in mobile phones especially those directly related to functionality and aesthetics. This study considered only four product attributes among so many due to the constraints of the research design employed. Attributes related to aesthetic perception of a product enhance apparent usability such as those related to dimensional ratios.

  15. Stress responses and replication of plasmids in bacterial cells

    Directory of Open Access Journals (Sweden)

    Wegrzyn Alicja

    2002-05-01

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

  16. The affecting of perceived product quality and perceived risk on perceived product value

    OpenAIRE

    Pisnik Korda, Aleksandra

    2015-01-01

    Perceived value is an extremely important concept in marketing and many authors have dealt with it in the recent years. There are numerous factors affecting perceived product value and we focused on three: perceived product price, perceived product quality and perceived risk. In this paper we present two of them: perceived product quality and perceived risk. Perceived product value is the difference between value in use and price. Value in use is a mix of benefits, which a customer is likely ...

  17. Loss of plasmids containing cloned inserts coding for novobiocin resistance or novobiocin sensitivity in Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

    Setlow, J.K.; Spikes, D.; Ledbetter, M.

    1984-06-01

    Plasmids pNov1 and pNov1s, coding for resistance and sensitivity to novobiocin, respectively, were readily lost from wild-type Haemophilus influenzae but retained in a strain lacking an inducible defective prophage. The plasmid loss could be partly or wholly eliminated by a low-copy-number mutation in the plasmid or by the presence of certain antibiotic resistance markers in the host chromosome. Release of both phage HP1c1, measured by plaque assay, and defective phage, measured by electron microscopy, was increased when the plasmids were present. The frequency of recombination between pNov1 and the chromosome, causing the plasmid to be converted to pNov1s, could under some circumstances be decreased from the normal 60 to 70% to below 10% by the presence of a kanamycin resistance marker in the chromosome. This suggested that a gene product coded for by the plasmid, the expression of which was affected by the kanamycin resistance marker, was responsible for the high recombination frequency. Evidence was obtained from in vitro experiments that the gene product was a gyrase.

  18. Factors affecting the dairy industry's products export in Iran

    Directory of Open Access Journals (Sweden)

    Aazam Yazdaninasab

    2016-09-01

    Full Text Available Export dairy products as an important part of non-oil exports can play an important role in improving the economic situation of Iran. Therefore, in this study factors affecting the exports of dairy products in the period 2002-2014 will be discussed. The results indicate a trend of increasing exports of dairy products during the 13 year study. This reflects the fact that the dairy industry of great potential capacity to provide part of the non-oil revenues in the agricultural subsector. In this study, the dependent variable was the amount of exports of dairy products. Independent variables included: investment in large dairy factories, the price of goods and services consumed by this sector, GDP, exchange rate, export prices for dairy products. The results showed that the effect of all independent variables on the dependent variable is positive and rising: such as investment and exchange rate. So that with an increase of 10 percent each of the indicators the country's exports of dairy products will be increase. The original proposal of the present study is: the use of policies and financial instruments and non-financial, such as paying attention to the comparative advantages of export, focusing on the right target export markets in order to support the country's dairy industry to increase production and exports.

  19. Food and predators affect egg production in song sparrows.

    Science.gov (United States)

    Zanette, Liana; Clinchy, Michael; Smith, James N M

    2006-10-01

    Although the possibility that food and predators may interact in limiting avian populations has long been recognized, there have been few attempts to test this experimentally in the field. We conducted a manipulative food addition experiment on the demography of Song Sparrows (Melospiza melodia) across sites that varied in predator abundance, near Victoria, British Columbia, Canada, over three consecutive breeding seasons. We previously showed that food and predators had interactive effects on annual reproductive success (young fledged per female). Here, we report the effects on egg production. Our results show that food limits the total number of eggs laid over the breeding season ("total egg production") and that interactive food and predator effects, including food effects on nest predation, determine how those eggs are "parceled out" into different nests. Food addition alone significantly affected total egg production, and there was no significant interannual variability in this result. At the same time, both food and predators affected the two determinants of total egg production: "clutch number" (total number of clutches laid) and average clutch size. Both clutch number and size were affected by a food x predator x year interaction. Clutch number was lower at low-predator locations because there was less nest predation and thus less renesting. Food addition also significantly reduced nest predation, but there was significant interannual variation in this effect. This interannual variation was responsible for the food x predator x year interactions because the larger the effect of food on nest predation in a given year, the smaller was the effect of food on clutch number; and the smaller the effect of food on clutch number, the larger was the effect of food on clutch size. Potential predator and year effects on total egg production were thus cancelled out by an inverse relationship between clutch number and clutch size. We suggest that combined food and

  20. Research Progress in Large-scale Production of Plasmid DNA for Gene Therapy%基因治疗用质粒DNA生产面临的问题及研究进展

    Institute of Scientific and Technical Information of China (English)

    胡春生; 张庆林; 张通

    2011-01-01

    Plasmid DNA encoding treatment gene for pharmaceutical applications was approved into the clinical trials.A pre-requisite to the success of plasmid-based therapies is the development of effective production of plasmid DNA.However, at present, there are several problems and bottlenecks in large-scale production of pharmaceutical plasmid DNA.Such as vector construction, cell lysis, bacteria chromosome DNA removing,bacteria endotoxin removing and quality-control in production process.The downstream for the large-scale production of plasmid DNA, limitations and the strategies used to obtain a final product that meets specifications was reviewed.%基因治疗已成为21世纪一些重大疾病的有效治疗策略,目前携带治疗基因的重组质粒已作为基因药物进入临床研究.对用于基因治疗的生物制品的生产与质量控制都有相当严格的要求.虽然已建立大规模符合药学规格的质粒DNA生产工艺,能满足临床需求,但在这些生产工艺中还存在一些难以克服的瓶颈,如:载体构建、细胞裂解、细菌染色体DNA去除、细菌内毒素去除、生产过程中质量控制等.就近年来大规模生产临床用质粒DNA遇到的相关问题及解决方案作一综述.

  1. Do ICTs Affect Workforce Productivity in Egyptian Industrial Organizations?

    Directory of Open Access Journals (Sweden)

    M. Elsaadani

    2014-06-01

    Full Text Available The present study aims to investigate the influence of Information Communication Technologies-ICTs’ dimensions (Information Technology (IT, Management Information System (MIS, Office automation (OA, Intranet and Internet on workforce productivity for a group of industrial organizations in Alexandria - Egypt. The population of the study included managers and staff members working in different areas related to ICTs in selected industrial organizations at various managerial levels. A descriptive-statistical combined research study was conducted. Simple random sampling was used for the selection of the participating industrial organization. A questionnaire was used as the data collection method. Expert comments were used to check the validity of study instrument, and the reliability of questions was calculated as 79% using Cronbach’s Alpha coefficient. Single variable t-test, Friedman and variance analysis tests were used for the analysis. Study findings revealed that the specified dimensions of ICTs positively affect workforce productivity of industrial organizations in Alexandria - Egypt.

  2. Reproductive biology traits affecting productivity of sour cherry

    Directory of Open Access Journals (Sweden)

    Milica Fotiric Aksic

    2013-01-01

    Full Text Available The objective of this work was to evaluate variability in reproductive biology traits and the correlation between them in genotypes of 'Oblačinska' sour cherry (Prunus cerasus. High genetic diversity was found in the 41 evaluated genotypes, and significant differences were observed among them for all studied traits: flowering time, pollen germination, number of fruiting branches, production of flower and fruit, number of flowers per bud, fruit set, and limb yield efficiency. The number of fruiting branches significantly influenced the number of flower and fruit, fruit set, and yield efficiency. In addition to number of fruiting branches, yield efficiency was positively correlated with fruit set and production of flower and fruit. Results from principal component analysis suggested a reduction of the reproductive biology factors affecting yield to four main characters: number and structure of fruiting branches, flowering time, and pollen germination. Knowledge of the reproductive biology of the 'Oblačinska' genotypes can be used to select the appropriate ones to be grown or used as parents in breeding programs. In this sense, genotypes II/2, III/9, III/13, and III/14 have very good flower production and satisfactory pollen germination.

  3. Genome-based genetic tool development for Bacillus methanolicus: theta- and rolling circle-replicating plasmids for inducible gene expression and application to methanol-based cadaverine production

    Directory of Open Access Journals (Sweden)

    Marta Irla

    2016-09-01

    Full Text Available Bacillus methanolicus is a thermophilic methylotroph able to overproduce amino acids from methanol, a substrate not used for human or animal nutrition. Based on our previous RNA-seq analysis a mannitol inducible promoter and a putative mannitol activator gene mtlR were identified. The mannitol inducible promoter was applied for controlled gene expression using fluorescent reporter proteins and a flow cytometry analysis, and improved by changing the -35 promoter region and by co-expression of the mtlR regulator gene. For independent complementary gene expression control, the heterologous xylose-inducible system from B. megaterium was employed and a two-plasmid gene expression system was developed. Four different replicons for expression vectors were compared with respect to their copy number and stability. As an application example, methanol-based production of cadaverine was shown to be improved from 11.3 g/L to 17.5 g/L when a heterologous lysine decarboxylase gene cadA was expressed from a theta-replicating rather than a rolling-circle replicating vector. The current work on inducible promoter systems and compatible theta- or rolling circle-replicating vectors is an important extension of the poorly developed B. methanolicus genetic toolbox, valuable for genetic engineering and further exploration of this bacterium.

  4. Genome-Based Genetic Tool Development for Bacillus methanolicus: Theta- and Rolling Circle-Replicating Plasmids for Inducible Gene Expression and Application to Methanol-Based Cadaverine Production

    Science.gov (United States)

    Irla, Marta; Heggeset, Tonje M. B.; Nærdal, Ingemar; Paul, Lidia; Haugen, Tone; Le, Simone B.; Brautaset, Trygve; Wendisch, Volker F.

    2016-01-01

    Bacillus methanolicus is a thermophilic methylotroph able to overproduce amino acids from methanol, a substrate not used for human or animal nutrition. Based on our previous RNA-seq analysis a mannitol inducible promoter and a putative mannitol activator gene mtlR were identified. The mannitol inducible promoter was applied for controlled gene expression using fluorescent reporter proteins and a flow cytometry analysis, and improved by changing the -35 promoter region and by co-expression of the mtlR regulator gene. For independent complementary gene expression control, the heterologous xylose-inducible system from B. megaterium was employed and a two-plasmid gene expression system was developed. Four different replicons for expression vectors were compared with respect to their copy number and stability. As an application example, methanol-based production of cadaverine was shown to be improved from 11.3 to 17.5 g/L when a heterologous lysine decarboxylase gene cadA was expressed from a theta-replicating rather than a rolling-circle replicating vector. The current work on inducible promoter systems and compatible theta- or rolling circle-replicating vectors is an important extension of the poorly developed B. methanolicus genetic toolbox, valuable for genetic engineering and further exploration of this bacterium.

  5. Knowledge Elecitation for Factors Affecting Taskforce Productivity using a Questionnaire

    CERN Document Server

    Sohail, Muhammad

    2009-01-01

    In this paper we present the process of Knowledge Elicitation through a structured questionnaire technique. This is an effort to depict a problem domain as Investigation of factors affecting taskforce productivity. The problem has to be solved using the expert system technology. This problem is the very first step how to acquire knowledge from the domain experts. Knowledge Elicitation is one of the difficult tasks in knowledge base formation which is a key component of expert system. The questionnaire was distributed among 105 different domain experts of Public and Private Organizations (i.e. Education Institutions, Industries and Research etc) in Pakistan. A total 61 responses from these experts were received. All the experts were well qualified, highly experienced and has been remained the members for selection committees a number of times for different posts. Facts acquired were analyzed from which knowledge was extracted and elicited. A standard shape was given to the questionnaire for further research as...

  6. Factors Affecting Adoption of Recommended Cauliflower Production Technology in Nepal

    Directory of Open Access Journals (Sweden)

    Pankaj Raj Dhital

    2016-06-01

    Full Text Available Binary logit regression model of econometrics was used to identify the factors affecting adoption of recommended agricultural technology by the commercial farmers of Nepal. A survey was carried out in 2012 in 120 households from Kavre district, Nepal. The objective was to appraise factors affecting adoption of recommended technology of cauliflower, finding out the level of adoption, identifying the constraints of cauliflower cultivation, assessing the perceived level of satisfaction and studying the relationship of certain selected variables on the adoption. Education, Occupation, contact with Personal Localite sources of information, Group membership and Experience were the most influencing factors for adoption of recommended technology. Though, other factors were not strong enough to contribute significantly but indirectly influence the adoption decision of farmers as combined effects. Occupation was found as negatively associated. Landholding size was strongly and positively influencing among the non significant factors. The level of satisfaction and the constraints of cauliflower production were also identified. Transfer of technology will be effective if and only if the client adopts the technology. For an effective transfer of technology, it is better to go for activities for the welfare of the socioeconomic factors of the farmers.

  7. 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.

  8. Factors affecting production rates of cosmogenic nuclides in extraterrestrial matter

    Energy Technology Data Exchange (ETDEWEB)

    Reedy, R.C., E-mail: reedy@psi.edu

    2015-10-15

    Good production rates are needed for cosmic-ray-produced nuclides to interpret their measurements. Rates depend on many factors, especially the pre-atmospheric object’s size, the location of the sample in that object (such as near surface or deep inside), and the object’s bulk composition. The bulk composition affects rates, especially in objects with very low and very high iron contents. Extraterrestrial materials with high iron contents usually have higher rates for making nuclides made by reactions with energetic particles and lower rates for the capture of thermal neutrons. In small objects and near the surface of objects, the cascade of secondary neutrons is being developed as primary particles are being removed. Deep in large objects, that secondary cascade is fully developed and the fluxes of primary particles are low. Recent work shows that even the shape of an object in space has a small but measureable effect. Work has been done and continues to be done on better understanding those and other factors. More good sets of measurements in meteorites with known exposure geometries in space are needed. With the use of modern Monte Carlo codes for the production and transport of particles, the nature of these effects have been and is being studied. Work needs to be done to improve the results of these calculations, especially the cross sections for making spallogenic nuclides.

  9. Initial pH of medium affects organic acids production but do not affect phosphate solubilization

    Directory of Open Access Journals (Sweden)

    Leandro M. Marra

    2015-06-01

    Full Text Available The pH of the culture medium directly influences the growth of microorganisms and the chemical processes that they perform. The aim of this study was to assess the influence of the initial pH of the culture medium on the production of 11 low-molecular-weight organic acids and on the solubilization of calcium phosphate by bacteria in growth medium (NBRIP. The following strains isolated from cowpea nodules were studied: UFLA03-08 (Rhizobium tropici, UFLA03-09 (Acinetobacter sp., UFLA03-10 (Paenibacillus kribbensis, UFLA03-106 (Paenibacillus kribbensis and UFLA03-116 (Paenibacillus sp.. The strains UFLA03-08, UFLA03-09, UFLA03-10 and UFLA03-106 solubilized Ca3(PO42 in liquid medium regardless of the initial pH, although without a significant difference between the treatments. The production of organic acids by these strains was assessed for all of the initial pH values investigated, and differences between the treatments were observed. Strains UFLA03-09 and UFLA03-10 produced the same acids at different initial pH values in the culture medium. There was no correlation between phosphorus solubilized from Ca3(PO42 in NBRIP liquid medium and the concentration of total organic acids at the different initial pH values. Therefore, the initial pH of the culture medium influences the production of organic acids by the strains UFLA03-08, UFLA03-09, UFLA03-10 and UFLA03-106 but it does not affect calcium phosphate solubilization.

  10. Deep sequencing revealed genome-wide single-nucleotide polymorphism and plasmid content of Erwinia amylovora strains isolated in Middle Atlas, Morocco.

    Science.gov (United States)

    Hannou, Najat; Mondy, Samuel; Planamente, Sara; Moumni, Mohieddine; Llop, Pablo; López, María; Manceau, Charles; Barny, Marie-Anne; Faure, Denis

    2013-10-01

    Erwinia amylovora causes economic losses that affect pear and apple production in Morocco. Here, we report comparative genomics of four Moroccan E. amylovora strains with the European strain CFBP1430 and North-American strain ATCC49946. Analysis of single nucleotide polymorphisms (SNPs) revealed genetic homogeneity of Moroccan's strains and their proximity to the European strain CFBP1430. Moreover, the collected sequences allowed the assembly of a 65 kpb plasmid, which is highly similar to the plasmid pEI70 harbored by several European E. amylovora isolates. This plasmid was found in 33% of the 40 E. amylovora strains collected from several host plants in 2009 and 2010 in Morocco.

  11. Transformation of Haemophilus influenzae by plasmid RSF0885

    Energy Technology Data Exchange (ETDEWEB)

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

    1981-12-01

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

  12. Use of sperm plasmid DNA lipofection combined with REMI (restriction enzyme-mediated insertion) for production of transgenic chickens expressing eGFP (enhanced green fluorescent protein) or human follicle-stimulating hormone.

    Science.gov (United States)

    Harel-Markowitz, Eliane; Gurevich, Michael; Shore, Laurence S; Katz, Adi; Stram, Yehuda; Shemesh, Mordechai

    2009-05-01

    Linearized p-eGFP (plasmid-enhanced green fluorescent protein) or p-hFSH (plasmid human FSH) sequences with the corresponding restriction enzyme were lipofected into sperm genomic DNA. Sperm transfected with p-eGFP were used for artificial insemination in hens, and in 17 out of 19 of the resultant chicks, the exogenous DNA was detected in their lymphocytes as determined by PCR and expressed in tissues as determined by (a) PCR, (b) specific emission of green fluorescence by the eGFP, and (c) Southern blot analysis. A complete homology was found between the Aequorea Victoria eGFP DNA and a 313-bp PCR product of extracted DNA from chick blood cells. Following insemination with sperm lipofected with p-hFSH, transgenic offspring were obtained for two generations as determined by detection of the transgene for human FSH (PCR) and expression of the gene (RT-PCR and quantitative real-time PCR) and the presence of the protein in blood (radioimmunoassay). Data demonstrate that lipofection of plasmid DNA with restriction enzyme is a highly efficient method for the production of transfected sperm to produce transgenic offspring by direct artificial insemination.

  13. Knowledge of Repetitions Range Affects Force Production in Trained Females

    Directory of Open Access Journals (Sweden)

    Israel Halperin, Saied J. Aboodarda, Fabien A. Basset, David G. Behm

    2014-12-01

    Full Text Available Most studies have examined pacing strategies with cyclical activities (running and cycling. It has been demonstrated that males employ different pacing strategies during repeated maximal voluntary contractions (MVCs dependent upon a known endpoint. Since different fatiguing mechanisms have been identified between the genders, it is not known if females use comparable pacing strategies. The purpose of this study was to examine if informing female subjects regarding the number of MVCs to perform would affect force and electromyography (EMG. Twenty well-trained females completed 3 fatiguing protocols in a randomized order. In the control condition participants were informed they would perform twelve MVCs and then actually completed twelve. In the unknown condition they were not told how many MVCs to perform but were stopped after twelve. In the deception condition they were initially informed to perform 6 MVCs, but after the 6th MVC they were asked to perform a few more MVCs and were stopped after twelve. During the first 6 MVCs, forces in the deception condition were greater compared to the unknown (p = 0.021, ES = 0.65, 5% and control (p = 0.022, ES = 0.42, 3% conditions. No differences were found between conditions in the last 6 MVCs. A main effect for repetitions showed force deficits during the first 6 MVCs (p = 0.000, ES = 1.81, 13% and last 6 MVCs (p = 0.05, ES = 0.34, 3%. No differences were found between conditions in biceps and triceps EMG. However, EMG decreased during the first 6 MVCs for biceps (p = 0.001, ES = 1.0, 14% and triceps (p = 0.001, ES = 0.76, 14% across conditions. No differences were found in the last 6 MVCs. The anticipation of performing fewer MVCs led to increased force, whereas no endpoint led to decreased force production.

  14. Emotions, affects and the production of social life.

    Science.gov (United States)

    Fox, Nick J

    2015-06-01

    While many aspects of social life possess an emotional component, sociology needs to explore explicitly the part emotions play in producing the social world and human history. This paper turns away from individualistic and anthropocentric emphases upon the experience of feelings and emotions, attending instead to an exploration of flows of 'affect' (meaning simply a capacity to affect or be affected) between bodies, things, social institutions and abstractions. It establishes a materialist sociology of affects that acknowledges emotions as a part, but only a part, of a more generalized affective flow that produces bodies and the social world. From this perspective, emotions are not a peculiarly remarkable outcome of the confluence of biology and culture, but part of a continuum of affectivity that links human bodies to their physical and social environment. This enhances sociological understanding of the part emotions play in shaping actions and capacities in many settings of sociological concern.

  15. 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.

  16. Discourse Goals Affect the Process and Product of Nominal Metaphor Production.

    Science.gov (United States)

    Utsumi, Akira; Sakamoto, Maki

    2015-10-01

    Although a large number of studies have addressed metaphor comprehension, only a few attempts have so far been made at exploring the process of metaphor production. Therefore, in this paper, we address the problem of how people generate nominal metaphors or identify an apt vehicle for a given topic of nominal metaphors. Specifically, we examine how the process and product of metaphor production differ between two discourse goals of metaphor, namely an explanatory purpose (e.g., to clarify) and a literary purpose (e.g., to aesthetically pleasing). Experiment 1 analyzed the metaphors (or vehicles) generated in the metaphor production task, and demonstrated that people identified more prototypical exemplars of the property to be attributed to the topic as a vehicle for explanatory metaphors than for literary metaphors. In addition, it was found that metaphors generated for the explanatory purpose were more apt and conventional, and had high topic-vehicle similarity than those generated for the literary purpose, while metaphors generated for the literary purpose were more familiar and imageable than those for the explanatory purpose. Experiment 2 used a priming paradigm to assess the online availability of prototypical and less prototypical members of the topic property during metaphor production. The result was that both prototypical and less prototypical members were activated in producing literary metaphors, while neither members were activated in the production of explanatory metaphors. These findings indicate that the process of metaphor production is affected by discourse goals of metaphor; less prototypical members of the category are searched for a vehicle during the production of literary metaphors, and thus literary metaphors are generated with less prototypical vehicles than explanatory metaphors.

  17. 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.

  18. 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

  19. Reassessing culture media and critical metabolites that affect adenovirus production.

    Science.gov (United States)

    Shen, Chun Fang; Voyer, Robert; Tom, Roseanne; Kamen, Amine

    2010-01-01

    Adenovirus production is currently operated at low cell density because infection at high cell densities still results in reduced cell-specific productivity. To better understand nutrient limitation and inhibitory metabolites causing the reduction of specific yields at high cell densities, adenovirus production in HEK 293 cultures using NSFM 13 and CD 293 media were evaluated. For cultures using NSFM 13 medium, the cell-specific productivity decreased from 3,400 to 150 vp/cell (or 96% reduction) when the cell density at infection was increased from 1 to 3 x 10(6) cells/mL. In comparison, only 50% of reduction in the cell-specific productivity was observed under the same conditions for cultures using CD 293 medium. The effect of medium osmolality was found critical on viral production. Media were adjusted to an optimal osmolality of 290 mOsm/kg to facilitate comparison. Amino acids were not critical limiting factors. Potential limiting nutrients including vitamins, energy metabolites, bases and nucleotides, or inhibitory metabolites (lactate and ammonia) were supplemented to infected cultures to further investigate their effect on the adenovirus production. Accumulation of lactate and ammonia in a culture infected at 3 x 10(6) cells/mL contributed to about 20% reduction of the adenovirus production yield, whereas nutrient limitation appeared primarily responsible for the decline in the viral production when NSFM 13 medium was used. Overall, the results indicate that multiple factors contribute to limiting the specific production yield at cell densities beyond 1 x 10(6) cells/mL and underline the need to further investigate and develop media for better adenoviral vector productions.

  20. [Nourseothricin (streptothricin) inactivated by plasmid pIE 636-encoded acetyltransferase: detection of N-acetyl-beta-lysine in the inactivated product].

    Science.gov (United States)

    Seltmann, G

    1985-12-01

    Nourseothricin (streptothricin) can be inactivated by an acetyl transferase synthesized by E. coli strains containing plasmid pIE 636. Nourseothricin inactivated in the presence of 14C-acetyl-coenzyme A was purified and submitted to partial acidic hydrolysis. By electrophoresis of the hydrolysate a 14C-containing substance moving only slowly towards the cathode could be isolated. This substance after complete hydrolysis yields only unlabelled beta-lysine.

  1. Statistical frequency in perception affects children's lexical production.

    Science.gov (United States)

    Richtsmeier, Peter T; Gerken, LouAnn; Goffman, Lisa; Hogan, Tiffany

    2009-06-01

    Children's early word production is influenced by the statistical frequency of speech sounds and combinations. Three experiments asked whether this production effect can be explained by a perceptual learning mechanism that is sensitive to word-token frequency and/or variability. Four-year-olds were exposed to nonwords that were either frequent (presented 10 times) or infrequent (presented once). When the frequent nonwords were spoken by the same talker, children showed no significant effect of perceptual frequency on production. When the frequent nonwords were spoken by different talkers, children produced them with fewer errors and shorter latencies. The results implicate token variability in perceptual learning.

  2. Cloning of affecting pyruvate decarboxylase gene in the production bioethanol of agricultural waste in the E.coli bacteria

    Directory of Open Access Journals (Sweden)

    Masome Zeinali

    2016-09-01

    Full Text Available Introduction: Ethanol made by a biomass is one of the useful strategies in terms of economic and environmental and as a clean and safe energy to replace fossil fuels considered and examined. Materials and methods: In this study, key enzyme in the production of ethanol (Pyruvate decarboxylase from Zymomonas mobilis bacteria was isolated and cloned at E. coli bacteria by freeze and thaw method. For gene cloning, we used specific primers of pdc and PCR reaction and then pdc gene isolated and pET 28a plasmid double digested with (Sal I and Xho I enzymes. Digestion Products were ligated by T4 DNA ligase in 16 °C for 16 hours. Results: Results of bacteria culture showed that a few colonies containing pET 28a plasmid could grow. Result of colony pcr of pdc gene with specific primers revealed 1700 bp bands in 1% agarose gel electrophoresis. The results of PCR with T7 promotor forward primer and pdc revers primer have proved the accurate direction of integration of pdc gene into plasmid and revealed 1885 bp band. Double digestion of recombinant plasmid with SalI and XhoI enzymes revealed same bands. Finally, RT showed the expected band of 1700 bp that implies the desired gene expression in the samples. Discussion and conclusion: Due to the increased production of ethanol via pyruvate decarboxylase gene cloning in expression plasmids with a strong promoter upstream of the cloning site can conclude that, pyruvate decarboxylase cloning as a key gene would be useful and according to beneficial properties of E. coli bacteria, transfering the gene to bacteria appears to be reasonable.

  3. Analysis of Management Behavior Assessments and Affect on Productivity

    Science.gov (United States)

    2005-06-10

    associated with the affects of management behavior on employee beliefs and attitudes; include Herzberg’s Two Factor Theory , Vroom’s Expectancy and Equity...achieved, when recognized, when given additional responsibility, and when advanced in job position. Vroom’s Expectancy and Equity Theories of Motivation...degree or higher. The results of the study proved the validity of the Vroom and Yetton theory , but the employee assessments of management decision-making

  4. Temperature can interact with landscape factors to affect songbird productivity.

    Science.gov (United States)

    Cox, W Andrew; Thompson, Frank R; Reidy, Jennifer L; Faaborg, John

    2013-04-01

    Increased temperatures and more extreme weather patterns associated with global climate change can interact with other factors that regulate animal populations, but many climate change studies do not incorporate other threats to wildlife in their analyses. We used 20 years of nest-monitoring data from study sites across a gradient of habitat fragmentation in Missouri, USA, to investigate the relative influence of weather variables (temperature and precipitation) and landscape factors (forest cover and edge density) on the number of young produced per nest attempt (i.e., productivity) for three species of songbirds. We detected a strong forest cover × temperature interaction for the Acadian Flycatcher (Empidonax virescens) on productivity. Greater forest cover resulted in greater productivity because of reduced brood parasitism and increased nest survival, whereas greater temperatures reduced productivity in highly forested landscapes because of increased nest predation but had no effect in less forested landscapes. The Indigo Bunting (Passerina cyanea) exhibited a similar pattern, albeit with a marginal forest cover × temperature interaction. By contrast, productivity of the Northern Cardinal (Cardinalis cardinalis) was not influenced by landscape effects or temperature. Our results highlight a potential difficulty of managing wildlife in response to global change such as habitat fragmentation and climate warming, as the habitat associated with the greatest productivity for flycatchers was also that most negatively influenced by high temperatures. The influence of high temperatures on nest predation (and therefore, nest predators) underscores the need to acknowledge the potential complexity of species' responses to climate change by incorporating a more thorough consideration of community ecology in the development of models of climate impacts on wildlife.

  5. Sex of littermate twin affects lifetime ewe productivity

    Science.gov (United States)

    Ewe productivity is synonymous with annual litter-weight weaned (LWW) per ewe exposed to rams for breeding, and LWW is largely a function of number of lambs born (NLB) and weaned (NLW). Selecting for LWW should increase litter size and numbers of ewe-ram co-twins. Thus, we used historical records to...

  6. A Content Analysis of Factors Affecting New Product Development Process

    Directory of Open Access Journals (Sweden)

    Eda Atilgan-Inan

    2010-07-01

    Full Text Available The objective of this study is to review the international marketing literature on new product development process and compare the changes in the important factors in the process with the changes in the management approaches. For this purpose, the articles in three international marketing journals were selected and “new product development” and “new product performance” were searched for in the abstracts. After grouping the variables in the process, they were compared with the perspectives of management in the related periods. The results indicated that organizational factors have always been important for new product development process, which is in line with the nature of the innovation process. But the emphasis on internal factors has increased in the 21st century which is congruent with the change in management perspective foregrounding resource based view. The study differs from the similar literature review studies on the point that it deals with the topic from international marketing perspective. Therefore, R&D and other marketing studies are not included in the review and the study proposes the important factors from international firms’ point of view.

  7. Affected functional networks associated with sentence production in classic galactosemia.

    Science.gov (United States)

    Timmers, Inge; van den Hurk, Job; Hofman, Paul Am; Zimmermann, Luc Ji; Uludağ, Kâmil; Jansma, Bernadette M; Rubio-Gozalbo, M Estela

    2015-08-07

    Patients with the inherited metabolic disorder classic galactosemia have language production impairments in several planning stages. Here, we assessed potential deviations in recruitment and connectivity across brain areas responsible for language production that may explain these deficits. We used functional magnetic resonance imaging (fMRI) to study neural activity and connectivity while participants carried out a language production task. This study included 13 adolescent patients and 13 age- and gender-matched healthy controls. Participants passively watched or actively described an animated visual scene using two conditions, varying in syntactic complexity (single words versus a sentence). Results showed that patients recruited additional and more extensive brain regions during sentence production. Both groups showed modulations with syntactic complexity in left inferior frontal gyrus (IFG), a region associated with syntactic planning, and in right insula. In addition, patients showed a modulation with syntax in left superior temporal gyrus (STG), whereas the controls did not. Further, patients showed increased activity in right STG and right supplementary motor area (SMA). The functional connectivity data showed similar patterns, with more extensive connectivity with frontal and motor regions, and restricted and weaker connectivity with superior temporal regions. Patients also showed higher baseline cerebral blood flow (CBF) in right IFG and trends towards higher CBF in bilateral STG, SMA and the insula. Taken together, the data demonstrate that language abnormalities in classic galactosemia are associated with specific changes within the language network. These changes point towards impairments related to both syntactic planning and speech motor planning in these patients.

  8. The FRIABLE1 gene product affects cell adhesion in Arabidopsis.

    Directory of Open Access Journals (Sweden)

    Lutz Neumetzler

    Full Text Available Cell adhesion in plants is mediated predominantly by pectins, a group of complex cell wall associated polysaccharides. An Arabidopsis mutant, friable1 (frb1, was identified through a screen of T-DNA insertion lines that exhibited defective cell adhesion. Interestingly, the frb1 plants displayed both cell and organ dissociations and also ectopic defects in organ separation. The FRB1 gene encodes a Golgi-localized, plant specific protein with only weak sequence similarities to known proteins (DUF246. Unlike other cell adhesion deficient mutants, frb1 mutants do not have reduced levels of adhesion related cell wall polymers, such as pectins. Instead, FRB1 affects the abundance of galactose- and arabinose-containing oligosaccharides in the Golgi. Furthermore, frb1 mutants displayed alteration in pectin methylesterification, cell wall associated extensins and xyloglucan microstructure. We propose that abnormal FRB1 action has pleiotropic consequences on wall architecture, affecting both the extensin and pectin matrices, with consequent changes to the biomechanical properties of the wall and middle lamella, thereby influencing cell-cell adhesion.

  9. Methylaplysinopsin and other marine natural products affecting neurotransmission.

    Science.gov (United States)

    Taylor, K M; Baird-Lambert, J A; Davis, P A; Spence, I

    1981-01-01

    Methylaplysinopsin is a novel marine natural product that, after oral administration, prevented the effects of tetrabenazine in mice and rats. Methylaplysinopsin was a short-acting inhibitor of monoamine oxidase activity with greatest potency when serotonin was the substrate studied. The brain concentration of serotonin in the mouse was increased by methylaplysinopsin over the same time course as monoamine oxidase inhibition ex vivo. Methylaplysinopsin was also a weak inhibitor of the neuronal uptake of [3H]serotonin and a potentiator of the K+-induced release of [3H]serotonin from prelabeled synaptosomes. The predicted potentiation of serotonergic neurotransmission was supported by initial neurophysiological studies in an identified serotonergic pathway in the central nervous system of Aplysia. Two other studies on the pharmacology of marine natural products are reviewed. The majority of polyhalogenated monoterpenes isolated from red algae had central nervous system depressant properties. The exception is plocamadiene A, which caused, in mice, a reversible spastic paresis lasting up to 72 hours after oral administration. The severe muscle spasm was antagonized by diazepam. The final study discussed is the effect of a variety of marine natural products on the synthesis, neuronal uptake, and metabolism of GABA. Their selectivity is discussed with regard to the effects on metabolic respiration, and the correlation of neurochemical and neurophysiological effects on these marine substances.

  10. Factors Affecting the Production of Vietnamese Tones: A Study of American Learners

    Science.gov (United States)

    Nguyen, Hanh thi; Macken, Marlys A.

    2008-01-01

    This study investigates factors that affect the accuracy of tone production by American students of Vietnamese as a second language (L2). Nine hypotheses are examined, each of which isolates a factor expected to affect production accuracy: (a) task type, (b) the position of a tone in a clause, (c) discourse distance between a model provided by a…

  11. The ssbL gene harbored by the ColV plasmid of an Escherichia coli neonatal meningitis strain is an auxiliary virulence factor boosting the production of siderophores through the shikimate pathway.

    Science.gov (United States)

    Lemaître, Chloé; Bidet, Philippe; Benoist, Jean-François; Schlemmer, Dimitri; Sobral, Elsa; d'Humières, Camille; Bonacorsi, Stéphane

    2014-04-01

    The ability to capture iron is a challenge for most bacteria. The neonatal meningitis Escherichia coli strain S88 possesses several iron uptake systems, notably including siderophores. Transcriptional analysis of the ColV plasmid pS88 has shown strong induction of a previously undescribed gene with low identity to three E. coli chromosomal genes encoding phospho-2-dehydro-3-deoxyheptonate aldolases involved in aromatic amino acid and catecholate/phenolate siderophore biosynthesis through the shikimate pathway. Here, we investigated the role of this gene, ssbLp (ssbL carried on the plasmid), in siderophore biosynthesis and, consequently, in S88 virulence. We constructed an S88 mutant designated S88 ΔssbLp, which exhibited reduced growth under low-iron conditions compared to the wild-type strain. Liquid chromatography-mass spectroscopy analysis of culture supernatants showed that the mutant secreted significantly smaller amounts of enterobactin, salmochelin SX, and yersiniabactin than the wild-type strain. The mutant was also less virulent in a neonatal rat sepsis model, with significantly lower bacteremia and mortality. Supplementation with chorismate, the final product of the shikimate pathway, restored the wild-type phenotype in vitro. In a collection of human extraintestinal E. coli isolates, we found that ssbL was present only in strains harboring the iro locus, encoding salmochelins, and was located either on the chromosome or on plasmids. Acquisition of the iro locus has been accompanied by acquisition of the auxiliary gene ssbL, which boosts the metabolic pathway essential for catecholate/phenolate siderophore biosynthesis and could represent potential therapeutic targets.

  12. How hollow melanosomes affect iridescent colour production in birds.

    Science.gov (United States)

    Eliason, Chad M; Bitton, Pierre-Paul; Shawkey, Matthew D

    2013-09-22

    Developmental constraints and trade-offs can limit diversity, but organisms have repeatedly evolved morphological innovations that overcome these limits by expanding the range and functionality of traits. Iridescent colours in birds are commonly produced by melanin-containing organelles (melanosomes) organized into nanostructured arrays within feather barbules. Variation in array type (e.g. multilayers and photonic crystals, PCs) is known to have remarkable effects on plumage colour, but the optical consequences of variation in melanosome shape remain poorly understood. Here, we used a combination of spectrophotometric, experimental and theoretical methods to test how melanosome hollowness--a morphological innovation largely restricted to birds--affects feather colour. Optical analyses of hexagonal close-packed arrays of hollow melanosomes in two species, wild turkeys (Meleagris gallopavo) and violet-backed starlings (Cinnyricinclus leucogaster), indicated that they function as two-dimensional PCs. Incorporation of a larger dataset and optical modelling showed that, compared with solid melanosomes, hollow melanosomes allow birds to produce distinct colours with the same energetically favourable, close-packed configurations. These data suggest that a morphological novelty has, at least in part, allowed birds to achieve their vast morphological and colour diversity.

  13. OVARIAN HORMONE PRODUCTION AFFECTED BY AMYGDALIN ADDITION IN VITRO

    Directory of Open Access Journals (Sweden)

    Marek Halenár

    2015-02-01

    Full Text Available Amygdalin, a natural substance, is a cyanogenic glycoside occurring in the seeds of apricots and bitter almonds. It is a controversial anti-tumor compound that has been used as an alternative cancer drug for many years. Amygdalin is composed of two molecules of glucose, one of benzaldehyde, which induces an analgesic action, and one of hydrocyanic acid, which is an anti-neoplastic compound. This in vitro study was performed to evaluate the possible impact of amygdalin (1, 10, 100, 1000, 10 000 μg/mL on the secretory activity of granulosa cells (GCs from porcine cyclic ovaries. The release of progesterone and estradiol-17β by GCs were evaluated by ELISA. In our study, the noticeable changes in estradiol-17β release by ovarian GCs were determined after the amygdalin addition. Amygdalin, at the highest dose (10 000 μg/mL, significantly (P≤0.05 stimulated the release of estradiol-17β by GCs, in comparison to the untreated control cells. On the contrary, no significant (P≥0.05 changes in the progesterone release by GCs caused by amygdalin addition were observed. In conclusion, obtained results showed that the amygdalin application (various doses to ovarian GCs caused a dose-dependent stimulation of the estradiol-17β release, but not progesterone, and its possible modulatory impact on the steroid production in porcine ovaries.

  14. 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

  15. DOMINANT FACTORS AFFECTING SEAWEED (Gracilaria verrucosa PRODUCTION IN ACID SULFATE SOILS-AFFECTED PONDS OF LUWU REGENCY, INDONESIA

    Directory of Open Access Journals (Sweden)

    Akhmad Mustafa

    2010-12-01

    Full Text Available Most of brackish water ponds used for seaweed (Gracilaria verrucosa culture in Luwu Regency, South Sulawesi, Indonesia are constructed on acid sulfate soil. Despite this inevitable condition, opportunities remain open to increase the seaweed production. The research was conducted to study the dominant factors that affect the seaweed production in ASS-affected ponds of Luwu Regency. As a dependent variable in this research is seaweed production. Independent variables were grouped into: (a farmer status factor, consisting of 9 variables; (b pond condition factor, consisting of 8 variables; (c pond management factor, consisting of 29 variables; (d soil quality factor, consisting of 17 variables and (e water quality factor, consisting of 11 variables. Multiple regression with dummy variable was used to analyze the data in prediction dependent variable. Results show that the average of seaweed production in ASS-affected pond of Luwu Regency is 11,000 kg dry/ha/year. Seaweed production can be increased through: (a decreasing dosage of urea and KCl and increasing dosage and frequency of fertilizer containing phosphate; (b increasing water depth in the pond and decreasing percentage of water exchange,(c conducting remediation to increase the soil pH and decreasing the concentration of Fe in the water, (d increasing stocking density of milkfish to decrease the epiphyte population and (e increasing the frequency of the farmer to attend trainings.

  16. Does the Unemployement Benefit Institution Affect the Productivity of Workers? Evidence from a Field Experiment

    NARCIS (Netherlands)

    Blanco, M.; Dalton, P.S.; Vargas, J.F.

    2013-01-01

    Abstract: We investigate whether and how the type of unemployment bene t institution affects productivity. We designed a field experiment to compare workers' productivity under a welfare system, where the unemployed receive an unconditional monetary transfer, with their productivity under a workfare

  17. 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

  18. 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.

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

    Science.gov (United States)

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

    2012-09-07

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

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

    OpenAIRE

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

    2012-01-01

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

  1. Investigation of diversity of plasmids carrying the blaTEM-52 gene

    DEFF Research Database (Denmark)

    Bielak, Eliza Maria; Bergenholtz, Rikke D.; Jørgensen, Mikael Skaanning;

    2011-01-01

    OBJECTIVES: To investigate the diversity of plasmids that carry blaTEM-52 genes among Escherichia coli and Salmonella enterica originating from animals, meat products and humans. METHODS: A collection of 22 blaTEM-52-encoding plasmids was characterized by restriction fragment length polymorphism...... of self-transfer to a plasmid-free E. coli recipient. CONCLUSIONS: The blaTEM-52 gene found in humans could have been transmitted on transferable plasmids originating from animal sources. Some of the blaTEM-52 plasmids carry replicons that differ from the classical ones. Two novel replicons were detected...

  2. Types of embodiment in design: the embodied foundations of meaning and affect in product design

    NARCIS (Netherlands)

    Rompay, van Thomas J.L.; Ludden, Geke D.S.

    2015-01-01

    Since the introduction of the embodied cognition framework in cognitive and social psychology, numerous studies have demonstrated links between everyday bodily experiences and product evaluations involving meaning portrayal and affect. To date, however, no comprehensive framework for understanding e

  3. Expression analysis of the spi gene in the pock-forming plasmid pSA1.1 from Streptomyces azureus and localization of its product during differentiation.

    Science.gov (United States)

    Doi, Katusmi; Ohyama, Yukiko; Yokoyama, Eiji; Nishiyama, Takashi; Fujino, Yasuhiro; Nagayoshi, Yuko; Ohshima, Toshihisa; Ogata, Seiya

    2012-08-01

    The sporulation inhibitory gene spi in the pock-forming conjugative plasmid pSA1.1 of Streptomyces azureus was introduced into cells via a high or low copy number vector to examine the effect of gene dosage on the growth of Streptomyces lividans TK24 as a host. In transformants carrying a high spi copy number, nutrient mycelial growth was inhibited, as was morphological differentiation from substrate mycelium to aerial mycelium on solid media. The degree of inhibition depended on the spi gene dosage, but the presence of pSA1.1 imp genes, which encode negative repressor proteins for spi, relieved the inhibition. Confocal images of Spi tagged with enhanced green fluorescent protein in cells on solid media revealed that spi expression was initiated at the time of elongation of substrate mycelium, that its expression increased dramatically at septation in aerial hyphae, and that the expression was maximal during prespore formation. Expression of spi covered the whole of the hyphae, and the level of expression at the tip of the hyphae during prespore formation was about sixfold greater than during substrate mycelial growth and threefold greater than during aerial mycelial growth. Thus, localized expression of spi at particular times may inhibit sporulation until triggering imp expression to repress its inhibitory effects.

  4. 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....

  5. Productivity affects the density-body mass relationship of soil fauna communities

    NARCIS (Netherlands)

    Comor, V.N.R.; Thakur, M.P.; Berg, M.P.; Bie, de S.; Prins, H.H.T.; Langevelde, van F.

    2014-01-01

    The productivity of ecosystems and their disturbance regime affect the structure of animal communities. However, it is not clear which trophic levels benefit the most from higher productivity or are the most impacted by disturbance. The density-body mass (DBM) relationship has been shown to reflect

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

    Science.gov (United States)

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

    2013-02-01

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

  7. Study on affecting factors of collaborative product development based on collaboration hierarchy model

    Institute of Scientific and Technical Information of China (English)

    ZHANG Xiaodong; LI Yingzi; ZHANG Zhiqiang

    2007-01-01

    Aiming at the levels of collaborative degree in web-based product development,a collaboration hierarchy model of this product development is developed in this paper.Based on the model,the affecting factors on collaboration levels are analyzed systematically from many aspects,such as technology,organization and business.A gap analysis method is studied in detail,and is applied in a real project.The application shows that it can solve the diverse problems of collaborative product development effectively,and help enterprises find out the critical factors that affect the collaboration.

  8. 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...

  9. Does the Unemployement Benefit Institution Affect the Productivity of Workers? Evidence from a Field Experiment

    OpenAIRE

    2013-01-01

    Abstract: We investigate whether and how the type of unemployment bene t institution affects productivity. We designed a field experiment to compare workers' productivity under a welfare system, where the unemployed receive an unconditional monetary transfer, with their productivity under a workfare system, where the transfer is received conditional on the unemployed spending some time on ancillary activities. First, we fi nd that having an unemployment bene fit institution, regardless of whe...

  10. 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.

  11. Productive Love Promotion Via Affective Technology: An Approach Based On Social Psychology And Philosophy

    Directory of Open Access Journals (Sweden)

    Ramon Solves Pujol

    2010-01-01

    Full Text Available This paper proposes the use of social psychological and philosophical foundations for designing affective technology that promotes the experience of love. The adopted theoretical basis is the concept of productive love, which is heavily based on Enrich Fromm but also includes theories and scientific findings of numerous psychoanalysts, social psychologists, and philosophers. We conducted a review of the theory about the nature of love and found that social psychological and philosophical approaches differ regarding peoples' understandings. The findings were used to elaborate eight principles of productive love. Based on these principles, we derived criteria for designing affective technology when the objective is to promote productive love. We reviewed the existent studies on affective technologies and implemented the criteria into a system design, the Pictures' Call. A prototype of the system was pretested to illustrate how productive love technology could be based on established criteria.

  12. Factors affecting industrial wood, material production yield in Turkey’s natural beech forests

    Directory of Open Access Journals (Sweden)

    Atilla Atik

    2014-07-01

    Full Text Available The objectives of the present study are to determine the most important factors affecting industrial wood material production yield in natural oriental beech forests in Turkey using a multifaceted approach and to help entrepreneurs consider these factors to develop more sensitive and realistic production plans. In Günye Forest Management in Bartın province of the West Black Sea Region of Turkey, 41 production units were chosen as the study area. The 1277 ha study area was included in the 2007 and 2010 production management plan. The general state of the stand, natural stand structure, and production methods and tools are the factors thought most strongly affect industrial wood material production yield; 26 variables representing these factors were evaluated in the study. Through multidimensional statistical analyses, including main components, factor and regression  analysis, we found that the most important factors affecting production yield were fertility, aspect of land, skidding method, stand structure, skidding distance, growing stock, transportation and harmful abiotic factors. Production units were divided into three groups based on yield rates and the 26 variables, using discriminate analysis. From the results of the study, a sample model can be developed to help forest managers predict and plan annual industrial wood production more sensitively and realistically.

  13. 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.

  14. 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...

  15. A comparison of the growth responses following intramuscular GHRH plasmid administration versus daily growth hormone injections in young pigs

    Science.gov (United States)

    The efficacy of daily porcine growth hormone (GH) injections versus plasmid-driven porcine GH-releasing hormone (pGHRH) production to promote growth was assessed. Ten-day-old piglets were injected intramuscularly with 0.1, 1, or 3 mg pGHRH, or a control plasmid followed by electroporation. Plasmid c...

  16. Conservation of plasmids among Escherichia coli K1 isolates of diverse origins.

    Science.gov (United States)

    Mercer, A A; Morelli, G; Heuzenroeder, M; Kamke, M; Achtman, M

    1984-12-01

    Escherichia coli K1 isolates of various O types were previously assigned to different clonal groups. Members of the two clones defined by membrane pattern 9 (MP9) and serotypes O18:K1 and O1:K1 had been found to be very similar to each other. The plasmid contents of these bacteria confirmed this conclusion. Both groups carried a self-transmissible plasmid of the FI incompatibility group that coded for colicin production and a major outer membrane protein called the plasmid-coded protein (PCP). The size of this plasmid varied from 76 to 96 megadaltons, but restriction endonuclease digestion and DNA heteroduplex analysis revealed that these plasmids were highly related. O18:K1 bacteria of MP6 had previously been determined to represent a subclone, related to but different from O18:K1 MP9 bacteria. These MP6 bacteria carried a different, smaller IncFI plasmid which did not code for colicin production or the PCP protein. This smaller plasmid was primarily related to the larger plasmid within the regions of DNA encoding incompatibility, replication, and conjugation. O1:K1 bacteria of MP5 contained other unrelated plasmids in agreement with the previous conclusion that they are unrelated to O1:K1 bacteria of MP9. The bacteria examined had been isolated from two continents over a time span of 38 years, and the results attest to conservative inheritance of plasmids within bacteria of common descent.

  17. Conservation of plasmids among Escherichia coli K1 isolates of diverse origins.

    Science.gov (United States)

    Mercer, A A; Morelli, G; Heuzenroeder, M; Kamke, M; Achtman, M

    1984-01-01

    Escherichia coli K1 isolates of various O types were previously assigned to different clonal groups. Members of the two clones defined by membrane pattern 9 (MP9) and serotypes O18:K1 and O1:K1 had been found to be very similar to each other. The plasmid contents of these bacteria confirmed this conclusion. Both groups carried a self-transmissible plasmid of the FI incompatibility group that coded for colicin production and a major outer membrane protein called the plasmid-coded protein (PCP). The size of this plasmid varied from 76 to 96 megadaltons, but restriction endonuclease digestion and DNA heteroduplex analysis revealed that these plasmids were highly related. O18:K1 bacteria of MP6 had previously been determined to represent a subclone, related to but different from O18:K1 MP9 bacteria. These MP6 bacteria carried a different, smaller IncFI plasmid which did not code for colicin production or the PCP protein. This smaller plasmid was primarily related to the larger plasmid within the regions of DNA encoding incompatibility, replication, and conjugation. O1:K1 bacteria of MP5 contained other unrelated plasmids in agreement with the previous conclusion that they are unrelated to O1:K1 bacteria of MP9. The bacteria examined had been isolated from two continents over a time span of 38 years, and the results attest to conservative inheritance of plasmids within bacteria of common descent. Images PMID:6094355

  18. Swarm-Optimization-Based Affective Product Design Illustrated by a Mobile Phone Case-Study

    Directory of Open Access Journals (Sweden)

    Koffka Khan

    2012-05-01

    Full Text Available This paper presents a new approach of user-oriented design for transforming users’ perception into product elements design. An experimental study on mobile phones is conducted to examine how product form and product design parameters affect consumer’s perception of a product. The concept of Kansei Engineering is used to extract the experimental samples as a data base for neural networks (NNs with particle swarm optimization (PSO analysis. The result of numerical analysis suggests that mobile phone makers need to focus on particular design parameters to attract specific target user groups, in addition to product forms. This paper demonstrates the advantage of using KE-PSO for determining the optimal combination of product design parameters. Based on the analysis, we can use KE-PSO to suggest customers’ preferences for mobile phone design attributes that would be considered optimal by various user groups of all surveyed. They can be used for improvement and development of new future products.

  19. Genes influencing milk production traits predominantly affect one of four biological pathways

    Directory of Open Access Journals (Sweden)

    Goddard Michael

    2008-01-01

    Full Text Available Abstract In this study we introduce a method that accounts for false positive and false negative results in attempting to estimate the true proportion of quantitative trait loci that affect two different traits. This method was applied to data from a genome scan that was used to detect QTL for three independent milk production traits, Australian Selection Index (ASI, protein percentage (P% and fat percentage corrected for protein percentage (F% – P%. These four different scenarios are attributed to four biological pathways: QTL that (1 increase or decrease total mammary gland production (affecting ASI only; (2 increase or decrease lactose synthesis resulting in the volume of milk being changed but without a change in protein or fat yield (affecting P% only; (3 increase or decrease protein synthesis while milk volume remains relatively constant (affecting ASI and P% in the same direction; (4 increase or decrease fat synthesis while the volume of milk remains relatively constant (affecting F% – P% only. The results indicate that of the positions that detected a gene, most affected one trait and not the others, though a small proportion (2.8% affected ASI and P% in the same direction.

  20. Factors Affecting Pheromone Production by the Pepper Weevil, Anthonomus eugenii Cano (Coleoptera: Curculionidae) and Collection Efficiency

    OpenAIRE

    Eller, Fred; Palmquist,Debra

    2014-01-01

    Several factors affecting pheromone production by male pepper weevils, Anthonomus eugenii Cano (Coleoptera: Curculionidae) as well as collection efficiency were investigated. Factors studied included: porous polymer adsorbents (Tenax versus Super Q), male age, time of day, male density, and male diet. Super Q was found to be a superior adsorbent for the male-produced alcohols and geranic acid as well as the plant-produced E-β-ocimene. Pheromone production increased with male age up to about a...

  1. Factors Affecting Pheromone Production by the Pepper Weevil, Anthonomus eugenii Cano (Coleoptera: Curculionidae) and Collection Efficiency

    OpenAIRE

    Eller, Fred J.; Debra E. Palmquist

    2014-01-01

    Several factors affecting pheromone production by male pepper weevils, Anthonomus eugenii Cano (Coleoptera: Curculionidae) as well as collection efficiency were investigated. Factors studied included: porous polymer adsorbents (Tenax versus Super Q), male age, time of day, male density, and male diet. Super Q was found to be a superior adsorbent for the male-produced alcohols and geranic acid as well as the plant-produced E-β-ocimene. Pheromone production increased with male age up to about ...

  2. Metaphor priming in sentence production: concrete pictures affect abstract language production.

    Science.gov (United States)

    Sato, Manami; Schafer, Amy J; Bergen, Benjamin K

    2015-03-01

    People speak metaphorically about abstract concepts-for instance, a person can be "full of love" or "have a lot of love to give." Over the past decade, research has begun to focus on how metaphors are processed during language comprehension. Much of this work suggests that understanding a metaphorical expression involves activating brain and body systems involved in perception and motor control. However, no research to date has asked whether the same is true while speakers produce language. We address this gap using a sentence production task. Its results demonstrate that visually activating a concrete source domain can trigger the use of metaphorical language drawn from that same concrete domain, even in sentences that are thematically unrelated to the primes, a metaphorical priming effect. This effect suggests that conceptual metaphors play a part in language production. It also shows that activation in the perceptual system that is not part of an intended message can nevertheless influence sentence formulation.

  3. ALOX5 gene variants affect eicosanoid production and response to fish oil supplementation

    Science.gov (United States)

    The objective of this study was to determine whether 5-lipoxygenase (ALOX5) gene variants associated with cardiovascular disease affect eicosanoid production by monocytes. The study was a randomized, double-masked, parallel intervention trial with fish oil (5.0 g of fish oil daily, containing 2.0 g ...

  4. [Localization of denitrification genes in plasmid DNA of bacteria Azospirillum brasilense].

    Science.gov (United States)

    Petrova, L P; Varshalomidze, O É; Shelud'ko, A V; Katsy, E I

    2010-07-01

    In 85-Mda plasmid (p85) of plant-associated bacteria Azospirillum brasilense Sp245 model strain, the genes encoding copper-containing nitrite reductase (nirK); heterodimeric NO-reductase (norCB); NorQ and NorD proteins affecting synthesis and (or) activation of NirK and (or) NO-reductase (norQD); catalytic subunit I ofcytochrom c oxidase (CccoN); presumable NO sensor carrying two hemeerythrine domains (orf181); and an enzyme required for synthesis of presumable NO antagonist, homocystein (metC) were identified. In the same region of p85, orf293 encoding transcriptional regulator of LysR type, orf208 whose protein product carries a formylmethanofuran dehydrogenase subunit E domain, and an orf164-encoding conservative secretory protein with unknown function were also found. Localization of a set of denitrification genes in the plasmid DNA A. brasilense Sp245 adjacent to IS elements ISAzba1 and ISAzba2 indicates potential mobility of these genes and high probability of their horizontal transfer among populations of rhizospheric bacteria. A site homologous to p85 nirK-orf208-orf181 genes was detected in the 115 kb plasmid of A. brasilense Sp7 type strain.

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

    Science.gov (United States)

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

    2015-02-01

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

  6. Antifungal activity of different natural dyes against traditional products affected fungal pathogens

    Institute of Scientific and Technical Information of China (English)

    R Mari selvam; AJA Ranjit Singh; K Kalirajan

    2012-01-01

    Objective: In the present study to evaluate the anti fungal activity of natural dyes against traditional products affected fungal pathogens. Methods: Many traditional craft products affected fungal pathogens were isolated using potato dextrose agar medium. The isolated fungus were identified by morphological and microscopically characterization using Alexopolus manual. 50μl of Turmeric, Terminalli, Guava and Henna natural dyes were poured into the wells of the culture plates. If antifungal activity was present on the plates, it was indicated by an inhibition zone surrounding the well containing the natural dye. Result: At a dose level of 50μl of terminalli dye was able to inhibit the growth of all the fungi tested. The absorbance rate of natural dyes analyzed by UV Spectrophotometer. The absorbance rate is high in terminalli (2.266) and turmeric (2.255). Conclusions: Natural dyes were bound with traditional products to give good colour and good antimicrobial activity against isolated fungal pathogens.

  7. Plasmid containing a DNA ligase gene from Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

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

    1984-05-01

    A ligase gene from Haemophilus influenzae was cloned into the shuttle vector pDM2. Although the plasmid did not affect X-ray sensitivity, it caused an increase in UV sensitivity of the wild-type but not excision-defective H. influenzae and a decrease in UV sensitivity of the rec-1 mutant. 14 references, 2 figures.

  8. Construction of Recombinant Plasmid Containing S. Mutans F-ATPase β Subunit Gene

    Institute of Scientific and Technical Information of China (English)

    YU Dan-ni; JIANG Li

    2005-01-01

    objective: construct a homologous recombinant plasmid which was expected to be transformed into S. mutans Methods: a region at the 5' terminus of the S. mutans F-ATPase β subunit gene was amplified by PCR, the PCR product was inserted into vector pVA891, yielding recombinant plasmid. Results: the DNA sequence of the recombinant plasmid was identified correct in whole by restriction endonuclease and DNA sequence techniques. Conclusion: the recombinant plasmid of S. mutans DNA was cloned in effect ,it may assist in construction of homologues recombinant mutant.

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

    Directory of Open Access Journals (Sweden)

    Guss Adam M

    2012-05-01

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

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2012-01-01

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

  11. Efficient production of lycopene in Saccharomyces cerevisiae by expression of synthetic crt genes from a plasmid harboring the ADH2 promoter.

    Science.gov (United States)

    Bahieldin, Ahmed; Gadalla, Nour O; Al-Garni, Saleh M; Almehdar, Hussein; Noor, Samah; Hassan, Sabah M; Shokry, Ahmed M; Sabir, Jamal S M; Murata, Norio

    2014-03-01

    Lycopene is an effective antioxidant proposed as a possible treatment for some cancers and other degenerative human conditions. This study aims at generation of a yeast strain (Saccharomyces cerevisiae) of efficient productivity of lycopene by overexpressing synthetic genes derived from crtE, crtB and crtI genes of Erwinia uredovora. These synthetic genes were constructed in accordance with the preferred codon usage in S. cerevisiae but with no changes in amino acid sequences of the gene products. S. cerevisiae cells were transformed with these synthetic crt genes, whose expression was regulated by the ADH2 promoter, which is de-repressed upon glucose depletion. The RT-PCR and Western blotting analyses indicated that the synthetic crt genes were efficiently transcribed and translated in crt-transformed S. cerevisiae cells. The highest level of lycopene in one of the transformed lines was 3.3mglycopene/g dry cell weight, which is higher than the previously reported levels of lycopene in other microorganisms transformed with the three genes. These results suggest the excellence of using the synthetic crt genes and the ADH2 promoter in generation of recombinant S. cerevisiae that produces a high level of lycopene. The level of ergosterol was reversely correlated to that of lycopene in crt-transformed S. cerevisiae cells, suggesting that two pathways for lycopene and ergosterol syntheses compete for the use of farnesyl diphosphate.

  12. Factors affecting Small-Scale Coffee Production in Githunguri District, Kenya

    Directory of Open Access Journals (Sweden)

    Margaret Njeri Gathura

    2013-09-01

    Full Text Available This study was carried out to determine the factors affecting small scale coffee production in Kenya. The establishments under study were small-scale coffee farms in Githunguri District.It was to determine whether marketing factors, finances, government policies and physical and human resources affect coffee production in Githunguri District. Primary sources included use of questionnaires, observation and interviews. Secondary sources included desk research, library research on journals, text books and factory publications. The target population was over 700,000 small-scale coffee producers in Kenya out of which the accessible population of 10,000 producers drawn from Githunguri District in Kiambu County was selected which a sample size of 120 respondents was sampled. Stratified sampling technique was employed to compare views among coffee producers from the various coffee societies in the area. Data analysis was both qualitative and quantitative using descriptive statistics. Data presentation was in form of tables to help interpret findings and generate conclusions that aided solutions to identified problems. The research established that marketing factors, finances, government policies and physical and human resources greatly affected coffee production. The study recommended that the government should encourage coffee production by formulating favorable marketing factors and other policies and provide finances to small scale coffee producers. Producers on the other hand should strive to provide conducive working environment to their workers so as to sustain them in their farms. This will help to improve coffee yields and quality.

  13. Genetic Factors in Rhizobium Affecting the Symbiotic Carbon Costs of N2 Fixation and Host Plant Biomass Production

    DEFF Research Database (Denmark)

    Skøt, L.; Hirsch, P. R.; Witty, J. F.

    1986-01-01

    The effect of genetic factors in Rhizobium on host plant biomass production and on the carbon costs of N2 fixation in pea root nodules was studied. Nine strains of Rhizobium leguminosarum were constructed, each containing one of three symbiotic plasmids in combination with one of three different...... with the background of B151. The relationship between nitrogenase activity, carbon costs of N2 fixation and host plant biomass production is discussed....... the lowest carbon costs of N2 fixation (7.10–8.10 μmol C/μmol N2), but shoot dry weight of those plants was also smaller than that of plants nodulated by strains with the background of B151 or JI8400. Nodules formed by these two strain types had carbon costs of N2 fixation varying between 11.26 and 13...

  14. 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.

  15. Diversity and stability of plasmids from glycopeptide resistant Enterococcus faecium isolated from pigs in Denmark

    DEFF Research Database (Denmark)

    Hasman, H.; Villadsen, A. G.; Aarestrup, Frank Møller

    2005-01-01

    was seen at the end of the 7-year period, coinciding with the ban in 1998 of the macrolide tylosin as growth promoter for pig production. The stability of the plasmid in its original host was compared with stability of the same plasmid in BM4105RF, when both strains were maintained in liquid cultures...

  16. Nucleotide sequence analysis of the lactococcal EPS plasmid pNZ4000

    NARCIS (Netherlands)

    Kranenburg, van R.; Kleerebezem, M.; Vos, de W.M.

    2000-01-01

    The complete 42180-bp nucleotide sequence of the mobilization plasmid pNZ4000, coding for exopolysaccharide (EPS) production in Lactococcus lactis, was determined. This plasmid contains a region involved in EPS biosynthesis, four functional replicons, a region containing mobilization genes, and thre

  17. Pig slurry characteristics, nutrient balance and biogas production as affected by separation and acidification

    DEFF Research Database (Denmark)

    Sommer, S G; Hjorth, Maibritt; Leahy, J J

    2015-01-01

    Animal slurry is separated in order to avoid excessive nitrogen, phosphorus and potassium (NPK) fertilization of crops in the field. To enhance fertilizer efficiency further, slurry and its separation products may be acidified, for instance in animal houses. The current study quantified the effects...... of these treatments, both individually and in combination, on fertilizer efficiency, energy production and heavy metal accumulation as a result of manure management. Acidification increased the availability of N to plants in the manure applied, and provided a better match between plant-available NPK in the manure...... and separation fraction applied to fields and crop need. Total biogas production was not affected by separation, whereas acidification reduced biogas production because the process was inhibited by a low pH and a high sulphur concentration. The amount of copper applied per hectare in the liquid manure...

  18. Quality and safety aspects of meat products as affected by various physical manipulations of packaging materials.

    Science.gov (United States)

    Lee, Keun Taik

    2010-09-01

    This article explores the effects of physically manipulated packaging materials on the quality and safety of meat products. Recently, innovative measures for improving quality and extending the shelf-life of packaged meat products have been developed, utilizing technologies including barrier film, active packaging, nanotechnology, microperforation, irradiation, plasma and far-infrared ray (FIR) treatments. Despite these developments, each technology has peculiar drawbacks which will need to be addressed by meat scientists in the future. To develop successful meat packaging systems, key product characteristics affecting stability, environmental conditions during storage until consumption, and consumers' packaging expectations must all be taken into consideration. Furthermore, the safety issues related to packaging materials must also be taken into account when processing, packaging and storing meat products.

  19. Persistence of Antibiotic Resistance Plasmids in Biofilms

    Science.gov (United States)

    2014-10-01

    wounds, facilitates the persistence of MDR plasmids in Acinetobacter baumannii , a problematic wound pathogen. Moreover, we have shown that plasmids...which plasmid persistence can improve in Acinetobacter baumannii and other wound pathogens when grown in biofilm environments. This project has the... Acinetobacter * baumannii ,!Klebsiella*pneumoniae,!Enterobacter*sp.,! and! Escherichia* coli! (Eardley! et! al.,! 2011;! Gaynes! &! Edwards,! 2005;! Murray

  20. 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.

  1. Plasmid profile in oral Fusobacterium nucleatum from humans and Cebus apella monkeys

    Directory of Open Access Journals (Sweden)

    Paula Marcia O.

    2003-01-01

    Full Text Available Fusobacterium nucleatum is a strict anaerobe and is indigenous of the human oral cavity. This organism is commonly recovered from different monomicrobial and mixed infections in humans and animals. In this study, the plasmid profile, the plasmid stability and the penicillin-resistance association in oral F. nucleatum isolated from periodontal patients, healthy subjects and Cebus apella monkeys were evaluated. Forty-five F. nucleatum strains from patients, 38 from healthy subjects and seven from C. apella were identified and analyzed. Plasmid extraction was performed in all the isolated strains. These elements were found in 26.7% strains from patients and one strain from C. apella. Strains from healthy subjects did not show any plasmid. Most of strains showed two plasmid bands ranging from 4 to 16 Kb, but digestions with endonucleases showed that they belonged to a single plasmid. The plasmid profile was similar and stable in human and monkey strains. Also, plasmids were classified into three groups according to size. Two strains were positive to beta-lactamase production and no plasmid DNA-hybridization with a beta-lactamase gene probe was observed, suggesting a chromosomal resistance.

  2. 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.

  3. Fermentation conditions that affect clavulanic acid production in Streptomyces clavuligerus: a systematic review

    Directory of Open Access Journals (Sweden)

    Hooi-Leng eSer

    2016-04-01

    Full Text Available The β-lactamase inhibitor, clavulanic acid is frequently used in combination with β-lactam antibiotics to treat a wide spectrum of infectious diseases. Clavulanic acid prevents drug resistance by pathogens against these β-lactam antibiotics by preventing the degradation of the β-lactam ring, thus ensuring eradication of these harmful microorganisms from the host. This systematic review provides an overview on the fermentation conditions that affect the production of clavulanic acid in the firstly described producer, Streptomyces clavuligerus. A thorough search was conducted using predefined terms in several electronic databases (PubMed, Medline, ScienceDirect, EBSCO, from database inception to June 30th 2015. Studies must involve wild-type Streptomyces clavuligerus, and full texts needed to be available. A total of 29 eligible articles were identified. Based on the literature, several factors were identified that could affect the production of clavulanic acid in S. clavuligerus. The addition of glycerol or other vegetable oils (e.g. olive oil, corn oil could potentially affect clavulanic acid production. Furthermore, some amino acids such as arginine and ornithine, could serve as potential precursors to increase clavulanic acid yield. The comparison of different fermentation systems revealed that fed-batch fermentation yields higher amounts of clavulanic acid as compared to batch fermentation, probably due to the maintenance of substrates and constant monitoring of certain entities (such as pH, oxygen availability, etc.. Overall, these findings provide vital knowledge and insight that could assist media optimization and fermentation design for clavulanic acid production in S. clavuligerus.

  4. Fermentation Conditions that Affect Clavulanic Acid Production in Streptomyces clavuligerus: A Systematic Review.

    Science.gov (United States)

    Ser, Hooi-Leng; Law, Jodi Woan-Fei; Chaiyakunapruk, Nathorn; Jacob, Sabrina Anne; Palanisamy, Uma Devi; Chan, Kok-Gan; Goh, Bey-Hing; Lee, Learn-Han

    2016-01-01

    The β-lactamase inhibitor, clavulanic acid is frequently used in combination with β-lactam antibiotics to treat a wide spectrum of infectious diseases. Clavulanic acid prevents drug resistance by pathogens against these β-lactam antibiotics by preventing the degradation of the β-lactam ring, thus ensuring eradication of these harmful microorganisms from the host. This systematic review provides an overview on the fermentation conditions that affect the production of clavulanic acid in the firstly described producer, Streptomyces clavuligerus. A thorough search was conducted using predefined terms in several electronic databases (PubMed, Medline, ScienceDirect, EBSCO), from database inception to June 30th 2015. Studies must involve wild-type Streptomyces clavuligerus, and full texts needed to be available. A total of 29 eligible articles were identified. Based on the literature, several factors were identified that could affect the production of clavulanic acid in S. clavuligerus. The addition of glycerol or other vegetable oils (e.g., olive oil, corn oil) could potentially affect clavulanic acid production. Furthermore, some amino acids such as arginine and ornithine, could serve as potential precursors to increase clavulanic acid yield. The comparison of different fermentation systems revealed that fed-batch fermentation yields higher amounts of clavulanic acid as compared to batch fermentation, probably due to the maintenance of substrates and constant monitoring of certain entities (such as pH, oxygen availability, etc.). Overall, these findings provide vital knowledge and insight that could assist media optimization and fermentation design for clavulanic acid production in S. clavuligerus.

  5. Do non-native plant species affect the shape of productivity-diversity relationships?

    Science.gov (United States)

    Drake, J.M.; Cleland, E.E.; Horner-Devine, M. C.; Fleishman, E.; Bowles, C.; Smith, M.D.; Carney, K.; Emery, S.; Gramling, J.; Vandermast, D.B.; Grace, J.B.

    2008-01-01

    The relationship between ecosystem processes and species richness is an active area of research and speculation. Both theoretical and experimental studies have been conducted in numerous ecosystems. One finding of these studies is that the shape of the relationship between productivity and species richness varies considerably among ecosystems and at different spatial scales, though little is known about the relative importance of physical and biological mechanisms causing this variation. Moreover, despite widespread concern about changes in species' global distributions, it remains unclear if and how such large-scale changes may affect this relationship. We present a new conceptual model of how invasive species might modulate relationships between primary production and species richness. We tested this model using long-term data on relationships between aboveground net primary production and species richness in six North American terrestrial ecosystems. We show that primary production and abundance of non-native species are both significant predictors of species richness, though we fail to detect effects of invasion extent on the shapes of the relationship between species richness and primary production.

  6. Phenolic compounds affect production of pyocyanin, swarming motility and biofilm formation of Pseudomonas aeruginosa

    Institute of Scientific and Technical Information of China (English)

    Aylin Ugurlu; Aysegul Karahasan Yagci; Seyhan Ulusoy; Burak Aksu; Gulgun Bosgelmez-Tinaz

    2016-01-01

    Objective: To investigate the effects of plant-derived phenolic compounds (i.e. caffeic acid, cinnamic acid, ferulic acid and vanillic acid) on the production of quorum sensing regulated virulence factors such as pyocyanin, biofilm formation and swarming motility of Pseudomonas aeruginosa (P. aeruginosa) isolates. Methods: Fourteen clinical P. aeruginosa isolates obtained from urine samples and P. aeruginosa PA01 strain were included in the study. The antibacterial effects of phenolic compounds were screened by well diffusion assay. Pyocyanin and biofilm ac-tivity were measured from culture supernatants and the absorbance values were measured using a spectrophotometer. Swarming plates supplemented with phenolic acids were point inoculated with P. aeruginosa strains and the ability to swarm was determined by measuring the distance of swarming from the central inoculation site. Results: Tested phenolic compounds reduced the production of pyocyanin and biofilm formation without affecting growth compared to untreated cultures. Moreover, these compounds blocked about 50% of biofilm production and swarming motility in P. aeruginosa isolates. Conclusions: We may suggest that if swarming and consecutive biofilm formation could be inhibited by the natural products as shown in our study, the bacteria could not attach to the surfaces and produce chronic infections. Antimicrobials and natural products could be combined and the dosage of antimicrobials could be reduced to overcome antimicrobial resistance and drug side effects.

  7. Phenolic compounds affect production of pyocyanin, swarming motility and biofilm formation of Pseudomonas aeruginosa

    Institute of Scientific and Technical Information of China (English)

    Aylin Ugurlu; Aysegul Karahasan Yagci; Seyhan Ulusoy; Burak Aksu; Gulgun Bosgelmez-Tinaz

    2016-01-01

    Objective: To investigate the effects of plant-derived phenolic compounds(i.e. caffeic acid, cinnamic acid, ferulic acid and vanillic acid) on the production of quorum sensing regulated virulence factors such as pyocyanin, biofilm formation and swarming motility of Pseudomonas aeruginosa(P. aeruginosa) isolates.Methods: Fourteen clinical P. aeruginosa isolates obtained from urine samples and P. aeruginosa PA01 strain were included in the study. The antibacterial effects of phenolic compounds were screened by well diffusion assay. Pyocyanin and biofilm activity were measured from culture supernatants and the absorbance values were measured using a spectrophotometer. Swarming plates supplemented with phenolic acids were point inoculated with P. aeruginosa strains and the ability to swarm was determined by measuring the distance of swarming from the central inoculation site.Results: Tested phenolic compounds reduced the production of pyocyanin and biofilm formation without affecting growth compared to untreated cultures. Moreover, these compounds blocked about 50% of biofilm production and swarming motility in P. aeruginosa isolates.Conclusions: We may suggest that if swarming and consecutive biofilm formation could be inhibited by the natural products as shown in our study, the bacteria could not attach to the surfaces and produce chronic infections. Antimicrobials and natural products could be combined and the dosage of antimicrobials could be reduced to overcome antimicrobial resistance and drug side effects.

  8. Gyrase-dependent stabilization of pSC101 plasmid inheritance by transcriptionally active promoters.

    Science.gov (United States)

    Beaucage, S L; Miller, C A; Cohen, S N

    1991-09-01

    The pSC101 plasmid encodes a cis-acting genetic locus termed par that ensures the stable inheritance of plasmids in a population of dividing cells. In the absence of selection, par-defective plasmids are lost rapidly from the bacterial population. We report here that the stability of par-deleted pSC101 derivatives is restored by introducing certain adventitious bacterial promoters onto the plasmid. Stabilization requires active transcription from the inserted promoter and is affected by the site and orientation of the insertion, the length of the nascent transcript and DNA gyrase activity. While a promotor-associated overall increase in negative superhelicity of plasmid DNA was observed, stabilized inheritance appeared to be dependent on localized rather than generalized supercoiling. Our demonstration that promoter-induced DNA supercoiling can mimic the effects of the pSC101 par locus provides evidence that the previously reported superhelicity-generating effects of par are intrinsic to its function.

  9. TLR-mediated NF-kB-dependent cytokine production is differently affected by HIV therapeutics

    DEFF Research Database (Denmark)

    Melchjorsen, Jesper; Paludan, Søren Riis; Mogensen, Trine

      Pathogen-recognizing Toll-like receptors 2 (TLR2) and TLR4 are known to recognize a number of pathogens, including E.Coli, S. Pneumonia and N. Meningococcus. We have studied whether a number of HIV therapeutics affect immediate proinflammatory cytokine responses in cell cultures. Preliminary...... results suggest an opposing effect of the drugs Tenofovir and Retrovir (AZT) on TLR-mediated production of NF-kappaB-dependent proinflammatory cytokines. We present data on the mechanisms behind the drug-mediated remodeling of innate immune activation and how the drugs effect early host...

  10. Does Mixed Reimbursement Schemes Affect Hospital Activity and Productivity? An Analysis of the Case of Denmark

    DEFF Research Database (Denmark)

    Hansen, Xenia Brun; Bech, Mickael; Jakobsen, Mads Leth;

    2013-01-01

    The majority of public hospitals in Scandinavia are reimbursed through a mixture of two prospective reimbursement schemes, block grants (a fixed amount independent of the number of patients treated) and activity-based financing (ABF). This article contributes theoretically to the existing...... whether different incentives affects the performance of hospitals regarding activity and productivity differently. Information on Danish reimbursement schemes has been collected from documents provided by the regional governments and through interviews with regional administrations. The data cover...... and region we show that there have not been any significant changes in the number of hospital discharges for any of the regions from 2007 to 2010 within any of the treatment groups....

  11. An Affective Prototyping Approach for the User-Satisfying Assistive Products

    DEFF Research Database (Denmark)

    Lai, Yoke Chin

    2013-01-01

    This paper outlines the key concepts of a study whose main aim is to devise an integrated design method for efficiently designing usable assistive products. The study grounds on Affective Design principle formulated for user’s needs, emotions and experience handling in an attempt to outline...... an integrated approach so that the key concept of participatory design could be practiced. Incorporating the maturing Augmented Reality technology is expected to create a design setting that could motivate active end-user involvement from the start throughout the product design process. With the aids...... of ontological modeling approach, the design setting is envisioned suitable for disabled and elderly end users participation and to support capturing the abstract but valuable user’s real needs that are influenced by emotions and experiences. The ontology model will outline the basis of a design knowledge...

  12. Factors affecting release of ethanol vapour in active modified atmosphere packaging systems for horticultural products

    Directory of Open Access Journals (Sweden)

    Weerawate Utto

    2014-04-01

    Full Text Available The active modified atmosphere packaging (active MAP system , which provides interactive postharvest control , using ethanol vapour controlled release, is one of the current interests in the development of active packaging for horticultural products. A number of published research work have discussed the relationship between the effectiveness of ethanol vapour and its concentration in the package headspace, including its effect on postharvest decay and physiological controls. This is of importance because a controlled release system should release and maintain ethanol vapour at effective concentrations during the desired storage period. A balance among the mass transfer processes of ethanol vapour in the package results in ethanol vapour accumulation in the package headspace. Key factors affecting these processes include ethanol loading, packaging material, packaged product and storage environment (temperature and relative h umidity. This article reviews their influences and discusses future work required to better understand their influences on ethanol vapour release and accumulations in active MAP.

  13. How do strategic decisions and operative practices affect operating room productivity?

    Science.gov (United States)

    Peltokorpi, Antti

    2011-12-01

    Surgical operating rooms are cost-intensive parts of health service production. Managing operating units efficiently is essential when hospitals and healthcare systems aim to maximize health outcomes with limited resources. Previous research about operating room management has focused on studying the effect of management practices and decisions on efficiency by utilizing mainly modeling approach or before-after analysis in single hospital case. The purpose of this research is to analyze the synergic effect of strategic decisions and operative management practices on operating room productivity and to use a multiple case study method enabling statistical hypothesis testing with empirical data. 11 hypotheses that propose connections between the use of strategic and operative practices and productivity were tested in a multi-hospital study that included 26 units. The results indicate that operative practices, such as personnel management, case scheduling and performance measurement, affect productivity more remarkably than do strategic decisions that relate to, e.g., units' size, scope or academic status. Units with different strategic positions should apply different operative practices: Focused hospital units benefit most from sophisticated case scheduling and parallel processing whereas central and ambulatory units should apply flexible working hours, incentives and multi-skilled personnel. Operating units should be more active in applying management practices which are adequate for their strategic orientation.

  14. Factors Affecting Pheromone Production by the Pepper Weevil, Anthonomus eugenii Cano (Coleoptera: Curculionidae) and Collection Efficiency.

    Science.gov (United States)

    Eller, Fred J; Palmquist, Debra E

    2014-11-18

    Several factors affecting pheromone production by male pepper weevils, Anthonomus eugenii Cano (Coleoptera: Curculionidae) as well as collection efficiency were investigated. Factors studied included: porous polymer adsorbents (Tenax versus Super Q), male age, time of day, male density, and male diet. Super Q was found to be a superior adsorbent for the male-produced alcohols and geranic acid as well as the plant-produced E-β-ocimene. Pheromone production increased with male age up to about age 15 days old and then tapered off. Male pepper weevils produced the highest amount of pheromone between noon and 2 pm (i.e., 4 to 6 h after "lights on") and were producing ca. 800 ng/h during this period. Thereafter, pheromone production decreased and was extremely low during the scotophase (i.e., ca. 12 ng/h). Male pepper weevil density had a significant effect on both release rate and pheromone composition. Pheromone production on a per male basis was highest for individual males and the percentage of geranic acid in the blend was lowest for individual males. Male pepper weevils produced only extremely low amounts of pheromone when feeding on artificial diet; however, they produced very high amounts when on fresh peppers. Together, this information will be useful in designing better attractant lures for pepper weevils.

  15. Factors Affecting Pheromone Production by the Pepper Weevil, Anthonomus eugenii Cano (Coleoptera: Curculionidae and Collection Efficiency

    Directory of Open Access Journals (Sweden)

    Fred J. Eller

    2014-11-01

    Full Text Available Several factors affecting pheromone production by male pepper weevils, Anthonomus eugenii Cano (Coleoptera: Curculionidae as well as collection efficiency were investigated. Factors studied included: porous polymer adsorbents (Tenax versus Super Q, male age, time of day, male density, and male diet. Super Q was found to be a superior adsorbent for the male-produced alcohols and geranic acid as well as the plant-produced E-β-ocimene. Pheromone production increased with male age up to about age 15 days old and then tapered off. Male pepper weevils produced the highest amount of pheromone between noon and 2 pm (i.e., 4 to 6 h after “lights on” and were producing ca. 800 ng/h during this period. Thereafter, pheromone production decreased and was extremely low during the scotophase (i.e., ca. 12 ng/h. Male pepper weevil density had a significant effect on both release rate and pheromone composition. Pheromone production on a per male basis was highest for individual males and the percentage of geranic acid in the blend was lowest for individual males. Male pepper weevils produced only extremely low amounts of pheromone when feeding on artificial diet; however, they produced very high amounts when on fresh peppers. Together, this information will be useful in designing better attractant lures for pepper weevils.

  16. Evaluation of Factors Affecting Uricase Production by the Screened Wild/Natural Microbes

    Directory of Open Access Journals (Sweden)

    Harinath Dwivedi

    2012-01-01

    Full Text Available Gout is a metabolic disorder due to the deposition of uric acid crystals within articular or periarticular tissues. Uricase (urate oxidase catalyzes the oxidation of less water soluble uric acid (7 mg/dl to a compound allantoin which is more water soluble(11g/L at 40°C resulting into the ease of excretion of uric acid. The objective of the work was to develop a new method for screening of microbes for uricase production and estimation of uricase thereof. This was achieved by utilizing the fact that uric acid dissolves on being acted upon by uricase. The proposed method is a novel, inexpensive, simple and sensitive technique for screening and estimation of uricase. Biomass and uricase production at different stages of microbial growth curve for the uricase producing microbe was studied. Effects of different medium components affecting uricase production by microbes were studied using Placket Burman statistical design. Addition of uric acid in the nutrient medium was found to be effective in increasing the uricase production by microbes growing in the medium.

  17. Innovation Barriers and Enablers that Affect Productivity in Uganda Building Industry

    Directory of Open Access Journals (Sweden)

    Henry Mwanaki Alinaitwe, , and

    2007-06-01

    Full Text Available The construction industry has of recent been blamed for lack of innovation. Lack of innovation in the industry is believed to be responsible for the decreasing or stagnant levels of productivity in comparison with other industries. This paper reviews the major barriers and enablers to innovation in general. Propositions were made about the factors that affect innovation in the construction industry which were then formulated into a questionnaire. A survey was made on building contractors in Uganda, a developing country, targeting those with financial strength, large in size, and with high capacity to carry out big projects. The identified factors were then ranked and correlated. The level of training in science, engineering and technical education, and the level of research and development at the industry level are looked at as the greatest innovation enablers in building that will drive forward labour productivity. The size of the domestic market and the level of security are the worst innovation barriers that lead to low productivity in the building industry in Uganda. Contractors, policy makers and the government should address the identified factors in order to improve productivity.

  18. Factors affecting the milk production of dairy cattle in northern rural areas of Bangladesh

    Directory of Open Access Journals (Sweden)

    M.R. Begum

    2014-12-01

    Full Text Available A cross sectional study was conducted to observe the factors affecting the productive performance of dairy cattle from northern rural areas of Bangladesh during July and September 2013. Data of 105 cows, 85 (80.95% from local and 20 (19.05% cows from cross breed, were randomly selected for the study. A binary logistic regression, expressed by odds ratio with 95% confidence interval, was done to determine the association of daily milk production categorized into ≤ 2 and > 2 liters (L, based on median, with the significant explanatory variables of body weight, age at first calving, lactation period, vitamin use, type of floor and milking person. The result demonstrated that the probability of milk production of >2 L was 6.16, 4.5, 20.65 and 5.7 times higher from the with animal body weight of >140 kg, age at first calving of >36 m, lactation period of >8 m and vitamin use than that of body weight of 140 kg, age at first calving of ≤36 m, lactation period of ≤ 8 m, and not vitamin used respectively. The chance of milk production of > 2 L was 0.25 and 0.22 times lower for mud floor, and owner milking than that of brick floor and gowala (professional milking person respectively.

  19. Factors affecting exocellular polysaccharide production by Lactobacillus delbrueckii subsp. bulgaricus grown in a chemically defined medium.

    Science.gov (United States)

    Petry, S; Furlan, S; Crepeau, M J; Cerning, J; Desmazeaud, M

    2000-08-01

    We developed a chemically defined medium (CDM) containing lactose or glucose as the carbon source that supports growth and exopolysaccharide (EPS) production of two strains of Lactobacillus delbrueckii subsp. bulgaricus. The factors found to affect EPS production in this medium were oxygen, pH, temperature, and medium constituents, such as orotic acid and the carbon source. EPS production was greatest during the stationary phase. Composition analysis of EPS isolated at different growth phases and produced under different fermentation conditions (varying carbon source or pH) revealed that the component sugars were the same. The EPS from strain L. delbrueckii subsp. bulgaricus CNRZ 1187 contained galactose and glucose, and that of strain L. delbrueckii subsp. bulgaricus CNRZ 416 contained galactose, glucose, and rhamnose. However, the relative proportions of the individual monosaccharides differed, suggesting that repeating unit structures can vary according to specific medium alterations. Under pH-controlled fermentation conditions, L. delbrueckii subsp. bulgaricus strains produced as much EPS in the CDM as in milk. Furthermore, the relative proportions of individual monosaccharides of EPS produced in pH-controlled CDM or in milk were very similar. The CDM we developed may be a useful model and an alternative to milk in studies of EPS production.

  20. Colostrum and milk production of sows is affected by dietary conjugated linoleic acid

    DEFF Research Database (Denmark)

    Krogh, Uffe; Flummer, Christine; Jensen, Søren Krogh;

    2012-01-01

    (cis-9, trans-11 and trans-10, cis-12) from day 108 of gestation until weaning (4 wk after parturition) to evaluate whether dietary CLA affects the yield and composition of colostrum, time for initiation of milk production, and sow milk yield. Sows fed CLA tended to produce more colostral fat (6.3 vs...... was more than double in the CLA group (17.6 vs. 7.8%, respectively; P = 0.04). Copious milk production was initiated 33 h (CLA) and 34 h (CON) after parturition and was not affected by dietary treatments (P = 0.41). Sow milk yield was improved by the CLA treatment from days 7 to 14 of lactation (P = 0.......03). Weight at birth (1.40 kg for both groups; P = 0.98) and at weaning [8.2 kg (CLA) and 8.0 kg (CON); P = 0.52] was not statistically different. In conclusion, colostrum yield was inhibited but milk yield was stimulated by dietary inclusion of cis-9, trans-11 and trans-10, cis-12 CLA and indicates that sow...

  1. Mutations affecting extracellular protease production in the filamentous fungus Aspergillus nidulans.

    Science.gov (United States)

    Katz, M E; Flynn, P K; vanKuyk, P A; Cheetham, B F

    1996-04-10

    The extracellular proteases of Aspergillus nidulans are known to be regulated by carbon, nitrogen and sulphur metabolite repression. In this study, a mutant with reduced levels of extracellular protease was isolated by screening for loss of halo production on milk plates. Genetic analysis of the mutant showed that it contains a single, recessive mutation, in a gene which we have designated xprE, located on chromosome VI. The xprE1 mutation affected the production of extracellular proteases in response to carbon, nitrogen and, to a lesser extent, sulphur limitation. Three reversion mutations, xprF1, xprF2 and xprG1, which suppress xprE1, were characterised. Both xprF and xprG map to chromosome VII but the two genes are unlinked. The xprF1, xprF2 and xprG1 mutants showed high levels of milk-clearing activity on medium containing milk as a carbon source but reduced growth on a number of nitrogen sources. Evidence is presented that the xprE1 and xprG1 mutations alter expression of more than one protease and affect levels of alkaline protease gene mRNA.

  2. Estrogenic octylphenol affects seminal fluid production and its biochemical composition of eelpout (Zoarces viviparus).

    Science.gov (United States)

    Rasmussen, Tina H; Korsgaard, Bodil

    2004-10-01

    Estrogenic chemicals such as alkylphenols (APs) have been shown to disrupt the reproductive system of male fish. In the present study, the effects of the estrogenic chemical octylphenol (100 microg g(-1)) and 17 beta-estradiol on the semen production and the biochemical composition of the seminal fluid of the viviparous eelpout (Zoarces viviparus) were investigated at the time of spawning. After 10 days of octylphenol or estrogen treatment, vitellogenin (Vtg) synthesis was induced as indicated by increased plasma vitellogenin concentration. In accordance with the increased vitellogenin concentration, hepatosomatic index (HSI), total protein concentration, and total calcium concentration were also increased, and free amino acids concentration was decreased in blood plasma. Octylphenol treatment caused a decrease in the gonadosomatic index (GSI) and the milt volume and an increase in the spermatocrit. The histological examination revealed that octylphenol affected the normal lobular structure, including the Sertoli cells. In the majority of the octylphenol-treated fish, trapped sperm cells were observed in parts of the seminiferous lobules and the sperm ducts. The biochemical composition of the seminal fluid was also affected by the octylphenol or estrogen. The seminal plasma concentrations of magnesium, calcium, and total protein were elevated, and the concentration of free amino acids was reduced in the treated fish. This study indicates that octylphenol inhibits the seminal fluid production and changes its biochemical composition in eelpouts.

  3. Dietary PUFA Intervention Affects Fatty Acid- and Micronutrient Profiles of Beef and Related Beef Products

    Directory of Open Access Journals (Sweden)

    Dirk Dannenberger

    2013-07-01

    Full Text Available The study investigated the dietary impact of 18:3n-3 vs. 18:2n-6 on fatty acid- and micronutrient concentration of beef muscle and the extent of diet- and processing-induced changes of lipid- and micronutrient concentrations of beef products made thereof (German Corned beef (GCB, tea sausage spread (TSS, scalded sausage (SS. Beef and beef products were obtained from German Holstein bulls which either received a control diet consisting of maize silage and concentrate with soybean meal (41%, or an experimental diet of grass silage and concentrate plus rapeseed cake (12% and linseed oil (3%. The study revealed that upon an 18:3n-3 vs. 18:2n-6 intervention the amounts of 18:3n-3, EPA and Σn-3 LC-PUFA were significantly increased by 2.6, 2.3 and 1.7 fold, respectively. Experimental diet significantly increased β-carotene contents, and the γ-tocopherol contents were decreased. During beef processing, n-3 PUFA from beef were found to be product-specifically transferred into the corresponding beef products. 18:3n-3 and Σn-3 LC-PUFA contents were found to be 1.4 and 1.5 times higher in GCB from grass silage- than maize silage-fed bulls. The trace element contents in GCB (iron, copper, zinc, selenium were not affected by the diet; however γ-tocopherol contents were decreased by experimental diet. In conclusion, dietary n-3 PUFA were completely transferred into beef products unaffected by beef processing conditions.

  4. New product, familiar taste: Effects of slogans on cognitive and affective responses to an unknown food product among food neophobics and neophilics

    NARCIS (Netherlands)

    Fenko, A.; Leufkens, Jean-Marie; Hoof, van J.J.

    2015-01-01

    Food neophobia is considered a potential barrier for the introduction of new food products. This study investigated how advertising slogans could influence cognitive and affective responses to a new product in food neophobics and food neophilics. An unknown dairy product was used to examine the effe

  5. 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.

  6. 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.

  7. Nitrogen and phosphorus fertilization negatively affects strigolactone production and exudation in sorghum.

    Science.gov (United States)

    Yoneyama, Kaori; Xie, Xiaonan; Kisugi, Takaya; Nomura, Takahito; Yoneyama, Koichi

    2013-11-01

    Strigolactones (SLs) are essential host recognition signals for both root parasitic plants and arbuscular mycorrhizal fungi, and SLs or their metabolites function as a novel class of plant hormones regulating shoot and root architecture. Our previous study indicated that nitrogen (N) deficiency as well as phosphorus (P) deficiency in sorghum enhanced root content and exudation of 5-deoxystrigol, one of the major SLs produced by sorghum. In the present study, we examined how N and P fertilization affects SL production and exudation in sorghum plants subjected to short- (5 days) or long-term (10 days) N or P deficiency and demonstrated their common and distinct features. The root contents and exudation of SLs in the N- or P-deficient sorghum plants grown for 6, 12 or 24 h with or without N or P fertilization were quantified by LC-MS/MS. In general, without fertilization, root contents and exudation of SLs stayed at similar levels at 6 and 12 h and then significantly increased at 24 h. The production of SLs responded more quickly to P fertilization than the secretion of SLs, while regulation of SL secretion began earlier after N fertilization. It is suggested that sorghum plants regulate SL production and exudation when they are subjected to nutrient deficiencies depending on the type of nutrient and degree of deficiency.

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

    Energy Technology Data Exchange (ETDEWEB)

    Balganesh, M.; Setlow, J.K.

    1984-05-14

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

  9. Thiazide diuretics affect osteocalcin production in human osteoblasts at the transcription level without affecting vitamin D3 receptors.

    Science.gov (United States)

    Lajeunesse, D; Delalandre, A; Guggino, S E

    2000-05-01

    Besides their natriuretic and calciuretic effect, thiazide diuretics have been shown to decrease bone loss rate and improve bone mineral density. Clinical evidence suggests a specific role of thiazides on osteoblasts, because it reduces serum osteocalcin (OC), an osteoblast-specific protein, yet the mechanisms implicated are unknown. We therefore investigated the role of hydrochlorothiazide (HCTZ) on OC production by the human osteoblast-like cell line MG-63. HCTZ dose-dependently (1-100 microM) inhibited 1,25-dihydroxyvitamin D3 [1,25(OH)2D3]-induced OC release by these cells (maximal effect, -40-50% and p ethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) only partly prevented the inhibitory effect of the diuretic on OC secretion (maximal effect, -22.5+/-6.9%), suggesting that thiazide-dependent Ca2+ influx is not sufficient to elicit the inhibition of OC secretion. Because OC production is strictly dependent on the presence of 1,25(OH)2D3 in human osteoblasts, we next evaluated the possible role of HCTZ on vitamin D3 receptors (VDR) at the mRNA and protein levels. Both Northern and Western blot analyses showed no effect of HCTZ (1-100 microM) on VDR levels. The presence of EGTA in the culture media reduced slightly the VDR mRNA levels under basal condition but this was not modified in the presence of increasing levels of HCTZ. The OC gene promoter also is under the control of transcription factors such as Yin Yang 1 (YY1) and cFOS. Western blot analysis revealed no changes in YY1 levels in response to HCTZ either in the presence or in the absence of 0.5 mM EGTA in the culture media. In contrast, HCTZ induced a dose-dependent increase in cFOS levels (p production by HCTZ could explain its preventive role in bone loss rate.

  10. 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.

  11. The sensory quality of meat, game, poultry, seafood and meat products as affected by intense light pulses: a systematic review

    OpenAIRE

    2015-01-01

    The effect of intense light pulses (ILP) on sensory quality of 16 different varieties of meat, meat products, game, poultry and seafood are reviewed. Changes induced by ILP are animal species, type of meat product and fluences applied dependent. ILP significantly deteriorates sensory quality of cooked meat products. It causes less change in the sensory properties of dry cured than cooked meat products while fermented sausage is least affected. The higher fluence applied significantly changes ...

  12. The receptor for advanced glycation end products (RAGE) affects T cell differentiation in OVA induced asthma.

    Science.gov (United States)

    Akirav, Eitan M; Henegariu, Octavian; Preston-Hurlburt, Paula; Schmidt, Ann Marie; Clynes, Raphael; Herold, Kevan C

    2014-01-01

    The receptor for glycation end products (RAGE) has been previously implicated in shaping the adaptive immune response. RAGE is expressed in T cells after activation and constitutively in T cells from patients with diabetes. The effects of RAGE on adaptive immune responses are not clear: Previous reports show that RAGE blockade affects Th1 responses. To clarify the role of RAGE in adaptive immune responses and the mechanisms of its effects, we examined whether RAGE plays a role in T cell activation in a Th2 response involving ovalbumin (OVA)-induced asthma in mice. WT and RAGE deficient wild-type and OT-II mice, expressing a T cell receptor specific for OVA, were immunized intranasally with OVA. Lung cellular infiltration and T cell responses were analyzed by immunostaining, FACS, and multiplex bead analyses for cytokines. RAGE deficient mice showed reduced cellular infiltration in the bronchial alveolar lavage fluid and impaired T cell activation in the mediastinal lymph nodes when compared with WT mice. In addition, RAGE deficiency resulted in reduced OT-II T cell infiltration of the lung and impaired IFNγ and IL-5 production when compared with WT mice and reduced infiltration when transferred into WT hosts. When cultured under conditions favoring the differentiation of T cells subsets, RAGE deficient T cells showed reduced production of IFNγ but increased production of IL-17. Our data show a stimulatory role for RAGE in T activation in OVA-induced asthma. This role is largely mediated by the effects of RAGE on T cell proliferation and differentiation. These findings suggest that RAGE may play a regulatory role in T cell responses following immune activation.

  13. Environmental factors influencing trace house gas production in permafrost-affected soils

    Science.gov (United States)

    Walz, Josefine; Knoblauch, Christian; Böhme, Luisa; Pfeiffer, Eva-Maria

    2016-04-01

    The permafrost-carbon feedback has been identified as a major feedback mechanism to climate change. Soil organic matter (SOM) decomposition in the active layer and thawing permafrost is an important source of atmospheric carbon dioxide (CO2) and methane (CH4). Decomposability and potential CO2 and CH4 production are connected to the quality of SOM. SOM quality varies with vegetation composition, soil type, and soil depth. The regulating factors affecting SOM decomposition in permafrost landscapes are not well understood. Here, we incubated permafrost-affected soils from a polygonal tundra landscape in the Lena Delta, Northeast Siberia, to examine the influence of soil depth, oxygen availability, incubation temperature, and fresh organic matter addition on trace gas production. CO2 production was always highest in topsoil (0 - 10 cm). Subsoil (10 - 50 cm) and permafrost (50 - 90 cm) carbon did not differ significantly in their decomposability. Under anaerobic conditions, less SOM was decomposed than under aerobic conditions. However, in the absence of oxygen, CH4 can also be formed, which has a substantially higher warming potential than CO2. But, within the four-month incubation period (approximate period of thaw), methanogenesis played only a minor role with CH4 contributing 1-30% to the total anaerobic carbon release. Temperature and fresh organic matter addition had a positive effect on SOM decomposition. Across a temperature gradient (1, 4, 8°C) aerobic decomposition in topsoil was less sensitive to temperature than in subsoil or permafrost. The addition of labile plant organic matter (13C-labelled Carex aquatilis, a dominant species in the region) significantly increased overall CO2 production across different depths and temperatures. Partitioning the total amount of CO2 in samples amended with Carex material into SOM-derived CO2 and Carex-derived CO2, however, revealed that most of the additional CO2 could be assigned to the organic carbon from the amendment

  14. Fine Mapping QTLs Affecting Milk Production Traits on BTA6 in Chinese Holstein with SNP Markers

    Institute of Scientific and Technical Information of China (English)

    LIU Rui; SUN Dong-xiao; WANG Ya-chun; YU Ying; ZHANG Yi; CHEN Hui-yong; ZHANG Qin; ZHANG Sheng-li; ZHANG Yuan

    2013-01-01

    Our previous studies demonstrated that the region around markers BMS470 and BMS1242 on BTA6 showed a linkage to 305-d milk yield and composition traits in the Chinese Holstein population. We herein focused on such narrow region to fine map milk production QTLs with 15 SNPs across 25 Mb with each SNP in 1 Mb within most regions in a Chinese Holstein population with daughter design. 1 449 Holstein cows and 11 sires were genotyped for such SNPs by using TaqMan probe and RFLP assays. Multipoint linkage analysis across family revealed a QTL affecting milk yield between PPARGC1A C4075T and SLC34A2 T1713C. Meanwhile, within family analysis found three milk yield QTLs (two in CR T60984131G-CEP135 C501T and one in PDLIM5 A106C-OPN T3907, a fat yield QTL in UGDH T1670C-CR T60984131G region, and two protein yield QTLs in TBC1D1 G501C-UGDH T1670C and PPARGC1A C4075T-SLC34A2 T1713C, respectively. Associations between aforementioned significant SNP markers and milk production traits were further implemented. We found significant associations of PPARGC1A C4075T, SLC34A2 T1713C with milk yield (P<0.05, P<0.01, P<0.01), UGDH T1670C, and CR T60984131G with fat yield (P<0.01, P<0.01), and PPARGC1A C4075T, SLC34A2 T1713C, UGDH T1670C and OPN T3907 with protein yield (P<0.01, P<0.01, P<0.01, P<0.01). Our findings implied that QTLs affecting milk production traits on BTA6 were pleictropism or multigenic effect and PPARGC1A and OPN may be the causal mutations behind milk production QTLs on BTA6 in the Chinese Holstein population.

  15. Decreased summer drought affects plant productivity and soil carbon dynamics in a Mediterranean woodland

    Directory of Open Access Journals (Sweden)

    M. F. Cotrufo

    2011-09-01

    Full Text Available Precipitation patterns are expected to change in the Mediterranean region within the next decades, with projected decreases in total rainfall and increases in extreme events. We manipulated precipitation patterns in a Mediterranean woodland, dominated by Arbutus unedo L., to study the effects of changing precipitation regimes on above-ground net primary production (ANPP and soil C dynamics, specifically plant-derived C input to soil and soil respiration (SR. Experimental plots were exposed to either a 20 % reduction of throughfall or to water addition targeted at maintaining soil water content above a minimum of 10 % v/v. Treatments were compared to control plots which received ambient precipitation. Enhanced soil moisture during summer months highly stimulated annual stem primary production, litter fall, SR and net annual plant-derived C input to soil which on average increased by 130 %, 26 %, 58 % and 220 %, respectively, as compared to the control. In contrast, the 20 % reduction in throughfall (equivalent to 10 % reduction in precipitation did not significantly change soil moisture at the site, and therefore did not significantly affect ANPP or SR. We conclude that minor changes (around 10 % reduction in precipitation amount are not likely to significantly affect ANPP or soil C dynamics in Mediterranean woodlands. However, if summer rain increases, C cycling will significantly accelerate but soil C stocks are not likely to be changed in the short-term. More studies involving modelling of long-term C dynamics are needed to predict if the estimated increases in soil C input under wet conditions is going to be sustained and if labile C is being substituted to stable C, with a negative effect on long-term soil C stocks.

  16. Quality of shrimp analogue product as affected by addition of modified potato starch.

    Science.gov (United States)

    Remya, S; Basu, S; Venkateshwarlu, G; Mohan, C O

    2015-07-01

    The present study was aimed to investigate the effects of addition of modified potato starch on the biochemical and textural properties of shrimp analogue/imitation shrimp, a popular value-added product prepared from surimi. Three batches of shrimp analogues were prepared with 0 % (NPS), 50 % (CPS) and 100 % (MPS) of modified starch incorporation and various quality attributes were monitored at regular intervals during frozen storage (-20 °C). Loss of myofibrillar protein was least for the shrimp analogue sample added with 100 % modified potato starch. The expressible moisture content of MPS (2.48 %) was less affected by long term storage compared to CPS (3.38 %) and NPS (3.99 %). During extended low temperature storage, the textural quality of sea food analogue was highly influenced by the type of starch added to it. The percentage of modified potato starch added to shrimp analogue significantly (p ≤ 0.05) affected its hardness and fracturability. MPS samples did not show significant changes in hardness during storage as compared to other two samples. Springiness of shrimp analogue increased 2.57, 1.5 and 1.77 times with the storage period for samples with NPS, CPS and MPS, respectively. Addition of modified potato starch improved the sensory quality and textural properties of shrimp analogue and reduced the quality degradation during frozen storage as compared to NPS which contained only native potato starch.

  17. Low-dose plasmid DNA treatment increases plasma vasopressin and regulates blood pressure in experimental endotoxemia

    Directory of Open Access Journals (Sweden)

    Malardo Thiago

    2012-11-01

    Full Text Available Abstract Background Although plasmid DNA encoding an antigen from pathogens or tumor cells has been widely studied as vaccine, the use of plasmid vector (without insert as therapeutic agent requires further investigation. Results Here, we showed that plasmid DNA (pcDNA3 at low doses inhibits the production of IL-6 and TNF-α by lipopolysaccharide (LPS-stimulated macrophage cell line J774. These findings led us to evaluate whether plasmid DNA could act as an anti-inflammatory agent in a Wistar rat endotoxemia model. Rats injected simultaneously with 1.5 mg/kg of LPS and 10 or 20 μg of plasmid DNA had a remarkable attenuation of mean arterial blood pressure (MAP drop at 2 hours after treatment when compared with rats injected with LPS only. The beneficial effect of the plasmid DNA on MAP was associated with decreased expression of IL-6 in liver and increased concentration of plasma vasopressin (AVP, a known vasoconstrictor that has been investigated in hemorrhagic shock management. No difference was observed in relation to nitric oxide (NO production. Conclusion Our results demonstrate for the first time that plasmid DNA vector at low doses presents anti-inflammatory property and constitutes a novel approach with therapeutic potential in inflammatory diseases.

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

    Science.gov (United States)

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

    2013-01-14

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

  19. Antibiotic-resistance and plasmids in Staphylococcus-hyicus isolated from pigs with exudative eperdermitis and from healthy pigs

    DEFF Research Database (Denmark)

    Wegener, Henrik Caspar; Schwarz, S.

    1993-01-01

    A total of 100 S. hyicus strains isolated from healthy piglets and piglets with exudative epidermitis originating from 100 different herds was examined for drug-resistance and prevalence of plasmids. Resistance to macrolide/lincosamide antibiotics could be related to plasmids in 55 (93%) of the 59...... resistant strains: A plasmid of 2.4 kb mediating resistance to macrolides and lincosamides was observed in 25 strains, and a plasmid of 11.5 kb mediating resistance to both macrolides/lincosamides and tetracycline was observed in 30 strains. A plasmid with a molecular weight of 4.5 kb was shown by curing......% of the strains isolated from healthy piglets. Despite its higher prevalence in strains from piglets with EE, the 11.5 kb plasmid could not be shown to encode production of capsule or exfoliative substances: factors which might play a role in the development of exudative epidermitis in piglets....

  20. Selection of barley grain affects ruminal fermentation, starch digestibility, and productivity of lactating dairy cows.

    Science.gov (United States)

    Silveira, C; Oba, M; Yang, W Z; Beauchemin, K A

    2007-06-01

    The objective of this study was to evaluate the effects of 2 lots of barley grain cultivars differing in expected ruminal starch degradation on dry matter (DM) intake, ruminal fermentation, ruminal and total tract digestibility, and milk production of dairy cows when provided at 2 concentrations in the diet. Four primiparous ruminally cannulated (123 +/- 69 d in milk; mean +/- SD) and 4 multiparous ruminally and duodenally cannulated (46 +/- 14 d in milk) cows were used in a 4 x 4 Latin Square design with a 2 x 2 factorial arrangement of treatments with 16-d periods. Primiparous and multiparous cows were assigned to different squares. Treatments were 2 dietary starch concentrations (30 vs. 23% of dietary DM) and 2 lots of barley grain cultivars (Xena vs. Dillon) differing in expected ruminal starch degradation. Xena had higher starch concentration (58.7 vs. 50.0%) and greater in vitro 6-h starch digestibility (78.0 vs. 73.5%) compared with Dillon. All experimental diets were formulated to supply 18.3% crude protein and 20.0% forage neutral detergent fiber. Dry matter intake and milk yield were not affected by treatment. Milk fat concentration (3.55 vs. 3.29%) was greater for cows fed Dillon compared with Xena, but was not affected by dietary starch concentration. Ruminal starch digestion was greater for cows fed high-starch diets compared with those fed low-starch diets (4.55 vs. 2.49 kg/d), and tended to be greater for cows fed Xena compared with those fed Dillon (3.85 vs. 3.19 kg/d). Ruminal acetate concentration was lower, and propionate concentration was greater, for cows fed Xena or high-starch diets compared with cows fed Dillon or low-starch diets, respectively. Furthermore, cows fed Xena or high-starch diets had longer duration that ruminal pH was below 5.8 (6.6 vs. 4.0 and 6.4 vs. 4.2 h/d) and greater total tract starch digestibility (94.3 vs. 93.0 and 94.3 vs. 93.0%) compared with cows fed Dillon or low-starch diets, respectively. These results

  1. Factors affecting wild rabbit production in extensive breeding enclosures: how can we optimise efforts?

    Directory of Open Access Journals (Sweden)

    José Guerrero-Casado

    2013-09-01

    Full Text Available The declining rabbit population in the Iberian Peninsula has led hunters and authorities to rear rabbits in captivity systems for their subsequent release. One alternative method to intensive rabbitry systems is the use of extensive breeding enclosures, since they produce animals of greater quality for hunting and conservation purposes. However, some of the factors that affect rabbit production in breeding enclosures are still unknown. The present study used partial least squares regression (PLSR to analyse the effects of plot size, scrub cover, slope, initial rabbit abundance, the resources needed to dig warrens, predation and proximity to other enclosures on rabbit abundance. The results of our study show a positive effect of the number of other fenced plots within a radius of 3 km, a positive relationship with the availability of optimal resources for building warrens and a positive influence of intermediate values of scrub cover. According to our results, to maximise rabbit production in the enclosures it would be advisable to concentrate the restocking effort by ensuring that the restocking plots are close to each other, thus avoiding isolated enclosures. Furthermore, the selection of plots with an appropriate scrub cover and high availability of elements that favour the construction of warrens, such as large stones, sloping land or tall shrubs, may optimise results.

  2. Klebsiella pneumoniae yfiRNB operon affects biofilm formation, polysaccharide production and drug susceptibility.

    Science.gov (United States)

    Huertas, Mónica G; Zárate, Lina; Acosta, Iván C; Posada, Leonardo; Cruz, Diana P; Lozano, Marcela; Zambrano, María M

    2014-12-01

    Klebsiella pneumoniae is an opportunistic pathogen important in hospital-acquired infections, which are complicated by the rise of drug-resistant strains and the capacity of cells to adhere to surfaces and form biofilms. In this work, we carried out an analysis of the genes in the K. pneumoniae yfiRNB operon, previously implicated in biofilm formation. The results indicated that in addition to the previously reported effect on type 3 fimbriae expression, this operon also affected biofilm formation due to changes in cellulose as part of the extracellular matrix. Deletion of yfiR resulted in enhanced biofilm formation and an altered colony phenotype indicative of cellulose overproduction when grown on solid indicator media. Extraction of polysaccharides and treatment with cellulase were consistent with the presence of cellulose in biofilms. The enhanced cellulose production did not, however, correlate with virulence as assessed using a Caenorhabditis elegans assay. In addition, cells bearing mutations in genes of the yfiRNB operon varied with respect to the WT control in terms of susceptibility to the antibiotics amikacin, ciprofloxacin, imipenem and meropenem. These results indicated that the yfiRNB operon is implicated in the production of exopolysaccharides that alter cell surface characteristics and the capacity to form biofilms--a phenotype that does not necessarily correlate with properties related with survival, such as resistance to antibiotics.

  3. Developmental and communicative factors affecting VOT production in English and Arabic bilingual and monolingual speakers

    Science.gov (United States)

    Khattab, Ghada

    2001-05-01

    VOT patterns were investigated in the production of three Lebanese-English bilinguals' aged 5, 7, and 10, six aged-matched monolingual controls from the bilinguals' immediate communities, and the parents of bilinguals and monolinguals. The aim was to examine the extent to which children exposed to two languages acquire separate VOT patterns for each language and to determine the factors that affect such acquisition. Results showed that VOT patterns for each bilingual child differed significantly across the two languages. But while the contrast in English resembled a monolingual-like model, that for Arabic exhibited persisting developmental features; explanations were offered in terms of the relationship between input and complexity of voicing lead production. Evidence was used from developmental changes that were noted for two of the bilingual subjects over a period of 18 months. English code-switches produced by the bilinguals during Arabic sessions exhibited different VOT patterns from those produced during English sessions, which underlined the importance of taking the language context into consideration. Finally, results from monolinguals and bilinguals showed that the short lag categories for the two languages were different despite a degree of overlap. Such findings require finer divisions of the three universal VOT categories to account for language-specific patterns.

  4. Optimization of factors affecting the production of biodiesel from crude palm kernel oil and ethanol

    Directory of Open Access Journals (Sweden)

    David K. Kuwornoo, Julius C. Ahiekpor

    2010-07-01

    Full Text Available Biodiesel, an alternative diesel fuel made from renewable sources such as vegetable oils and animal fats, has been identified by government to play a key role in the socio-economic development of Ghana. The utilization of biodiesel is expected to be about 10% of the total liquid fuel mix of the country by the year 2020. Despite this great potential and the numerous sources from which biodiesel could be developed in Ghana, studies on the sources of biodiesel and their properties as a substitute for fossil diesel have tended to be limited to Jatropha oil. This paper, however, reports the parameters that influences the production of biodiesel from palm kernel oil, one of the vegetable oils obtained from oil palm which is the highest vegetable oil source in Ghana. The parameters studied are; mass ratio of ethanol to oil, reaction temperature, catalyst concentration, and reaction time using completely randomized 24 factorial design. Results indicated that ethanol to oil mass ratio, catalyst concentration and reaction time were the most important factors affecting the ethyl ester yield. There was also an interaction effect between catalyst and time and ethanol- oil ratio and time on the yield. Accordingly, the optimal conditions for the production of ethyl esters from crude palm kernel oil were determined as; 1:5 mass ratio of ethanol to oil, 1% catalyst concentration by weight of oil, 90 minutes reaction time at a temperature of 30 oC.

  5. Trinitrotoluene and mandarin peels selectively affect lignin-modifying enzyme production in white-rot basidiomycetes.

    Science.gov (United States)

    Kachlishvili, Eva; Asatiani, Mikheil; Kobakhidze, Aza; Elisashvili, Vladimir

    2016-01-01

    Five white-rot basidiomycetes (WRB) species have been evaluated for their potential to tolerate and to degrade 0.2 mM 2, 4, 6-trinitrotoluene (TNT) as well as to produce laccase and manganese peroxidase (MnP) in presence of this xenobiotic. The tested fungal strains produced laccase in both glycerol and mandarin peels-containing media, whereas in the glycerol-containing medium only Cerrena unicolor strains and Trametes versicolor BCC 775 secreted MnP. Replacement of glycerol by milled mandarin peels 3- to 45-fold increased laccase activity, promoted C. unicolor strains and T. versicolor MnP secretion and induced this enzyme production by Fomes fomentarius BCC 38 and Funalia trogii BCC 146. Differential response of the WRB strains to the TNT addition was observed. In particular, laccase activity of C. unicolor increased 2- to 3-fold in both media whereas no stimulation of the laccase production was revealed in cultivation of F. fomentarius. TNT practically did not affect the MnP activity. Two strains of C. unicolor followed by T. versicolor producing laccase and MnP almost completely removed 0.2 mM TNT from the synthetic medium. Increase of TNT concentration from 0 to 0.4 mM in the mandarin peels-based medium and from 0 to 0.3 mM in the glycerol-containing medium stimulated C. unicolor BCC 300 laccase production from 92.4 to 240.7 U/ml and from 17.1 to 48.6 U/ml, respectively. This strain has been resistant to the TNT high concentration and has ability to remove 85 % of initial 0.3 mM TNT content during 6 days of the submerged cultivation.

  6. Soil hydraulic properties affected by topsoil thickness in cultivated switchgrass and corn-soybean rotation production systems

    Science.gov (United States)

    Loss of productive topsoil by soil erosion over time can reduce the productive capacity of soil and can significantly affect soil hydraulic properties. This study evaluated the effects of reduced topsoil thickness and perennial switchgrass (Panicum virgatum L.) versus corn (Zea mays L.)/soybean [Gly...

  7. Stress responses in pathogenic Yersinia enterocolitica with reference to the stability of the virulence plasmid in food

    Science.gov (United States)

    Yersinia enterocolitica has been associated with food-borne illness, most often due the ingestion of pork products. The pathogenic effects induced by a Y. enterocolitica infection are caused by the interplay of chromosomal genes and a virulence plasmid, pYV. Generally, the plasmid is lost during g...

  8. 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.

  9. Diversity and stability of Plasmids from glycopeptide-resistant Enterococcus faecium (GRE) isolated from pigs in Denmark

    DEFF Research Database (Denmark)

    Hasman, Henrik; Villadsen, A.G.; Aarestrup, Frank Møller

    2005-01-01

    was seen at the end of the 7-year period, coinciding with the ban in 1998 of the macrolide tylosin as growth promoter for pig production. The stability of the plasmid in its original host was compared with stability of the same plasmid in BM4105RF, when both strains were maintained in liquid cultures...

  10. Characterisation of TNF block haplotypes affecting the production of TNF and LTA.

    Science.gov (United States)

    Tan, J H; Temple, S E L; Kee, C; Waterer, G W; Tan, C R T; Gut, I; Price, P

    2011-02-01

    Polymorphisms in the central major histocompatibility complex (MHC) (particularly TNF and adjacent genes) associate with several immunopathological diseases and with susceptibility to pneumonia. The MHC is characterised by strong linkage disequilibrium (LD), so identification of loci affecting disease must be based on haplotypes. We have defined 31 tumour necrosis factor (TNF) block haplotypes (denoted FV1-31) in Caucasians, Asians and Australian Aboriginals. This study correlates the carriage of TNF block haplotypes with TNF and lymphotoxin alpha (LTA) protein production by peripheral blood mononuclear cells from 205 healthy Caucasian subjects, following in vitro stimulation with Streptococcus pneumoniae (S. pneumoniae; gram-positive bacteria), Escherichia coli (E. coli; gram-negative bacteria) or TNF over 4, 8 and 24 h. Fifteen haplotypes were present at >1%, accounting for 94.5% of the cohort. The haplotypes were grouped into five families based on common alleles. Following stimulation, cells from carriers of the FV10 haplotype (family 2) produced less LTA compared with non-FV10 carriers. Carriers of the FV18 haplotype (family 4) produced more LTA than other donors. Induction of TNF by S. pneumoniae following 24 h stimulation was also greater in donors with FV18. The FV18 haplotype associated with the 44.1 MHC ancestral haplotype (HLA-A2, -C5, -B44, -DRB1*0401 and -DQB1*0301) that has few disease associations. FV16 occurred in the 8.1 MHC haplotype (HLA-A2, B8, DR3) that is associated with multiple immunopathological diseases. FV16 did not affect TNF or LTA levels. The findings suggest that many genetic variations critical in vivo are not effectively modelled by short-term cultures.

  11. Physical processes affecting availability of dissolved silicate for diatom production in the Arabian Sea

    Science.gov (United States)

    Young, David K.; Kindle, John C.

    1994-01-01

    A passive tracer to represent dissolved silicate concentrations, with biologically realistic uptake kinetics, is successfully incorporated into a three-dimensional, eddy-resolving, ocean circulation model of the Indian Ocean. Hypotheses are tested to evaluate physical processes which potentially affect the availability of silicate for diatom production in the Arabian Sea. An alternative mechanism is offered to the idea that open ocean upwelling is primarily responsible for the high, vertical nutrient flux and consequent large-scale phytoplankton bloom in the northwestern Arabian Sea during the southwest monsoon. Model results show that dissolved silicate in surface waters available for uptake by diatoms is primarily influenced by the intensity of nearshore upwelling from soutwest monsoonal wind forcing and by the offshore advective transport of surface waters. The upwelling, which in the model occurs within 200 +/- 50 km of the coast, appears to be a result of a combination of coastal upwelling, Elkman pumping, and divergence of the coastal flow as it turns offshore. Localized intensifications of silicate concentrations appear to be hydrodynamically driven and geographically correlated to coastal topographic features. The absence of diatoms in sediments of the eastern Arabian Basin is consistent with modeled distributional patterns of dissolved silicate resulting from limited westward advection of upwelled coastal waters from the western continental margin of India and rapid uptake of available silicate by diatoms. Concentrations of modeled silicate become sufficiently low to become unavailable for diatom production in the eastern Arabian Sea, a region between 61 deg E and 70 deg E at 8 deg N on the south, with the east and west boundaries converging on the north at approximately 67 deg E, 20 deg N.

  12. Denitrifying bacterial communities affect current production and nitrous oxide accumulation in a microbial fuel cell.

    Directory of Open Access Journals (Sweden)

    Ariadna Vilar-Sanz

    Full Text Available The biocathodic reduction of nitrate in Microbial Fuel Cells (MFCs is an alternative to remove nitrogen in low carbon to nitrogen wastewater and relies entirely on microbial activity. In this paper the community composition of denitrifiers in the cathode of a MFC is analysed in relation to added electron acceptors (nitrate and nitrite and organic matter in the cathode. Nitrate reducers and nitrite reducers were highly affected by the operational conditions and displayed high diversity. The number of retrieved species-level Operational Taxonomic Units (OTUs for narG, napA, nirS and nirK genes was 11, 10, 31 and 22, respectively. In contrast, nitrous oxide reducers remained virtually unchanged at all conditions. About 90% of the retrieved nosZ sequences grouped in a single OTU with a high similarity with Oligotropha carboxidovorans nosZ gene. nirS-containing denitrifiers were dominant at all conditions and accounted for a significant amount of the total bacterial density. Current production decreased from 15.0 A · m(-3 NCC (Net Cathodic Compartment, when nitrate was used as an electron acceptor, to 14.1 A · m(-3 NCC in the case of nitrite. Contrarily, nitrous oxide (N2O accumulation in the MFC was higher when nitrite was used as the main electron acceptor and accounted for 70% of gaseous nitrogen. Relative abundance of nitrite to nitrous oxide reducers, calculated as (qnirS+qnirK/qnosZ, correlated positively with N2O emissions. Collectively, data indicate that bacteria catalysing the initial denitrification steps in a MFC are highly influenced by main electron acceptors and have a major influence on current production and N2O accumulation.

  13. Accurate determination of plasmid copy number of flow-sorted cells using droplet digital PCR.

    Science.gov (United States)

    Jahn, Michael; Vorpahl, Carsten; Türkowsky, Dominique; Lindmeyer, Martin; Bühler, Bruno; Harms, Hauke; Müller, Susann

    2014-06-17

    Many biotechnological processes rely on the expression of a plasmid-based target gene. A constant and sufficient number of plasmids per cell is desired for efficient protein production. To date, only a few methods for the determination of plasmid copy number (PCN) are available, and most of them average the PCN of total populations disregarding heterogeneous distributions. Here, we utilize the highly precise quantification of DNA molecules by droplet digital PCR (ddPCR) and combine it with cell sorting using flow cytometry. A duplex PCR assay was set up requiring only 1000 sorted cells for precise determination of PCN. The robustness of this method was proven by thorough optimization of cell sorting, cell disruption, and PCR conditions. When non plasmid-harboring cells of Pseudomonas putida KT2440 were spiked with different dilutions of the expression plasmid pA-EGFP_B, a PCN from 1 to 64 could be accurately detected. As a proof of principle, induced cultures of P. putida KT2440 producing an EGFP-fused model protein by means of the plasmid pA-EGFP_B were investigated by flow cytometry and showed two distinct subpopulations, fluorescent and nonfluorescent cells. These two subpopulations were sorted for PCN determination with ddPCR. A remarkably diverging plasmid distribution was found within the population, with nonfluorescent cells showing a much lower PCN (≤1) than fluorescent cells (PCN of up to 5) under standard conditions.

  14. Assessment of cultivation factors that affect biomass and geraniol production in transgenic tobacco cell suspension cultures.

    Directory of Open Access Journals (Sweden)

    Nikolay Vasilev

    Full Text Available A large-scale statistical experimental design was used to determine essential cultivation parameters that affect biomass accumulation and geraniol production in transgenic tobacco (Nicotiana tabacum cv. Samsun NN cell suspension cultures. The carbohydrate source played a major role in determining the geraniol yield and factors such as filling volume, inoculum size and light were less important. Sucrose, filling volume and inoculum size had a positive effect on geraniol yield by boosting growth of plant cell cultures whereas illumination of the cultures stimulated the geraniol biosynthesis. We also found that the carbohydrates sucrose and mannitol showed polarizing effects on biomass and geraniol accumulation. Factors such as shaking frequency, the presence of conditioned medium and solubilizers had minor influence on both plant cell growth and geraniol content. When cells were cultivated under the screened conditions for all the investigated factors, the cultures produced ∼ 5.2 mg/l geraniol after 12 days of cultivation in shaking flasks which is comparable to the yield obtained in microbial expression systems. Our data suggest that industrial experimental designs based on orthogonal arrays are suitable for the selection of initial cultivation parameters prior to the essential medium optimization steps. Such designs are particularly beneficial in the early optimization steps when many factors must be screened, increasing the statistical power of the experiments without increasing the demand on time and resources.

  15. Cowpea production as affected by dry spells in no-tillage and conventional crop systems

    Directory of Open Access Journals (Sweden)

    Rômulo Magno Oliveira de Freitas

    2013-12-01

    Full Text Available The objective of this study was to evaluate the effect of different periods of water shortage in no-tillage and conventional crop systems on cowpea yield components and grain yield in the Mossoró-RN region. For this, an experiment was conducted using two tillage systems (conventional and no-tillage subjected to periods of irrigation suspension (2; 6; 10; 14; 18 end 22 days, started at flowering (34 days after sowing. Plants were harvested 70 days after sowing, and the studied variables were: Pods length (CV, number of grains per pod (NGV, number of pods per plant (NPP, the hundred grains weight (PCG and grain yield (kg ha-1. The no-tillage system is more productive than the conventional under both irrigation and water stress treatments. The water stress length affected grain yield and all yield components studied in a negative way, except for the hundred grains weight. Among the systems studied, the no-tillage provides higher values for the yield components, except the hundred grains weight.

  16. Biotic and abiotic factors affect green ash volatile production and emerald ash borer adult feeding preference.

    Science.gov (United States)

    Chen, Yigen; Poland, Therese M

    2009-12-01

    The emerald ash borer, Agrilus planipennis Fairmaire (Coleoptera: Buprestidae), is an exotic woodborer first detected in 2002 in Michigan and Ontario and is threatening the ash resource in North America. We examined the effects of light exposure and girdling on green ash (Fraxinus pennsylvanica Marsh) volatile production, and effects of light exposure, girdling, and leaf age on emerald ash borer adult feeding preferences and phototaxis. Green ash seedlings grown under higher light exposure had lower amounts of three individual volatile compounds, (Z)-3-hexenol, (E)-beta-ocimene, and (Z,E)-alpha-farnesene, as well as the total amount of six detected volatile compounds. Girdling did not affect the levels of these volatiles. Emerald ash borer females preferred mature leaves, leaves from girdled trees, and leaves grown in the sun over young leaves, leaves from nongirdled trees, and leaves grown in the shade, respectively. These emerald ash borer preferences were most likely because of physical, nutritional, or biochemical changes in leaves in response to the different treatments. Emerald ash borer females and males showed positive phototaxis in laboratory arenas, a response consistent with emerald ash borer preference for host trees growing in sunlight.

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

    Science.gov (United States)

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

    1998-06-05

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

  18. Antibiotic-Resistant Extended Spectrum ß-Lactamase- and Plasmid-Mediated AmpC-Producing Enterobacteriaceae Isolated from Retail Food Products and the Pearl River in Guangzhou, China

    Science.gov (United States)

    Ye, Qinghua; Wu, Qingping; Zhang, Shuhong; Zhang, Jumei; Yang, Guangzhu; Wang, Huixian; Huang, Jiahui; Chen, Mongtong; Xue, Liang; Wang, Juan

    2017-01-01

    We conducted a survey in 2015 to evaluate the presence of extended spectrum β-lactamase (ESBL)- and plasmid-mediated AmpC-producing Enterobacteriaceae in retail food and water of the Pearl River in Guangzhou, China, as well as their antibiotic resistance profiles. Samples (88 fresh food samples and 43 water samples) from eight different districts were analyzed by direct plating and after enrichment. Multidrug-resistant strains were found in 41.7 and 43.4% of food and water samples, respectively. ESBLs were found in 3.4 and 11.6% of food and water samples, respectively, and AmpC producers were found in 13.6 and 16.3% of food and water samples, respectively. Molecular characterization revealed the domination of blaCTX−Mgenes; plasmidic AmpC was of the type DHA-1 both in food and water samples. Thirteen of Fifty one β-lactamase-producing positive isolates were detected to be transconjugants, which readily received the β-lactamase genes conferring resistance to β-lactam antibiotics as well as some non-β-lactam antibiotics. These findings provide evidence that retail food and the river water may be considered as reservoirs for the dissemination of β-lactam antibiotics, and these resistance genes could readily be transmitted to humans through the food chain and water. PMID:28217112

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

    Science.gov (United States)

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

    1988-06-01

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

  20. Influenza Plasmid DNA Vaccines: Progress and Prospects.

    Science.gov (United States)

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

    2015-01-01

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

  1. Population dynamics of dechlorinators and factors affecting the level and products of PCB dechlorination in sediments

    Energy Technology Data Exchange (ETDEWEB)

    Kim, J.S.; Sokol, R.C.; Liu, X.; Bethoney, C.M.; Rhee, G.Y. [State Univ. of New York and New York State Department of Health, Albany, NY (United States)

    1996-12-31

    Microbial dechlorination of polychlorinated biphenyls (PCBs) often stops although a significant number of removable chlorines remain. To determine the reason for the cessation, we investigated the limitation of organic carbon, PCB bioavailability, and inhibition by metabolic products. Enrichment with carbon sources did not induce additional chlorination, indicating the plateau was not due to depletion of organic carbon. The bioavailability was not limiting, since a subcritical micelle concentration of the surfactant, which enhanced desorption without inhibiting dechlorinating microorganisms, failed to lower the plateau. Neither was it due to accumulation of metabolites, since no additional dechlorination was detected when plateau sediments were incubated with fresh medium. Similarly, dechlorination was not inhibited in freshly spiked sediment slurries. Dechlorination ended up at the same level with nearly identical congener profiles, regardless of treatment. These results indicate that cessation of dechlorination was due to the accumulation of daughter congeners, which cannot be used as electron acceptors by microbes. To determine whether the decreasing availability affected the microorganisms, we determined the population dynamics of dechlorinators using the most probable number technique. The growth dynamics of the dechlorinators mirrored the time course of dechlorination. It started when the population increased by two orders of magnitude. Once dechlorination stopped the dechlorinating population also began to decrease. When dechlorinators were inoculated into PCB-free sediments, the population decreased over time. The decrease of the population as dechlorination ceased confirms that the diminishing availability of congeners was the reason for the incomplete dechlorination. Recent findings have shown that a second phase of dechlorination of certain congeners can occur after a long lag. 45 refs., 8 figs.

  2. Bone Formation is Affected by Matrix Advanced Glycation End Products (AGEs) In Vivo.

    Science.gov (United States)

    Yang, Xiao; Mostafa, Ahmed Jenan; Appleford, Mark; Sun, Lian-Wen; Wang, Xiaodu

    2016-10-01

    Advanced glycation end products (AGEs) accumulate in bone extracellular matrix as people age. Although previous evidence shows that the accumulation of AGEs in bone matrix may impose significant effects on bone cells, the effect of matrix AGEs on bone formation in vivo is still poorly understood. To address this issue, this study used a unique rat model with autograft implant to investigate the in vivo response of bone formation to matrix AGEs. Fluorochrome biomarkers were sequentially injected into rats to label the dynamic bone formation in the presence of elevated levels of matrix AGEs. After sacrificing animals, dynamic histomorphometry was performed to determine mineral apposition rate (MAR), mineralized surface per bone surface (MS/BS), and bone formation rate (BFR). Finally, nanoindentation tests were performed to assess mechanical properties of newly formed bone tissues. The results showed that MAR, MS/BS, and BFR were significantly reduced in the vicinity of implant cores with high concentration of matrix AGEs, suggesting that bone formation activities by osteoblasts were suppressed in the presence of elevated matrix AGEs. In addition, MAR and BFR were found to be dependent on the surrounding environment of implant cores (i.e., cortical or trabecular tissues). Moreover, MS/BS and BFR were also dependent on how far the implant cores were away from the growth plate. These observations suggest that the effect of matrix AGEs on bone formation is dependent on the biological milieu around the implants. Finally, nanoindentation test results indicated that the indentation modulus and hardness of newly formed bone tissues were not affected by the presence of elevated matrix AGEs. In summary, high concentration of matrix AGEs may slow down the bone formation process in vivo, while imposing little effects on bone mineralization.

  3. TOL plasmid carriage enhances biofilm formation and increases extracellular DNA content in Pseudomonas putida KT2440

    DEFF Research Database (Denmark)

    D'Alvise, Paul; Sjoholm, O.R.; Yankelevich, T.;

    2010-01-01

    : TOL carriage leads to more extracellular DNA (eDNA) in pellicles and biofilms. Pellicles were dissolved by DNase I treatment. Enhanced cell lysis due to plasmid carriage was ruled out as the mechanism for eDNA release. We report, for the first time, that carriage of a conjugative plasmid leads......Adherent growth of Pseudomonas putida KT2440 with and without the TOL plasmid (pWWO) at the solid-liquid and air-liquid interface was examined. We compared biofilm formation on glass in flow cells, and assayed pellicle (air-liquid interface biofilm) formation in stagnant liquid cultures by confocal...... laser scanning microscopy. The TOL-carrying strains formed pellicles and thick biofilms, whereas the same strains without the plasmid displayed little adherent growth. Microscopy using fluorescent nucleic acid-specific stains revealed differences in the production of extracellular polymeric substances...

  4. Plasmid and chromosome segregation in prokaryotes

    DEFF Research Database (Denmark)

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

    2000-01-01

    Recent major advances in the understanding of prokaryotic DNA segregation have been achieved by using fluorescence microscopy to visualize the localization of cellular components. Plasmids and bacterial chromosomes are partitioned in a highly dynamic fashion, suggesting the presence of a mitotic......-like apparatus in prokaryotes. The identification of chromosomal homologues of the well-characterized plasmid partitioning genes indicates that there could be a general mechanism of bacterial DNA partitioning. Udgivelsesdato: July 1...

  5. Protein diversity confers specificity in plasmid segregation.

    Science.gov (United States)

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

    2005-04-01

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

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

    Directory of Open Access Journals (Sweden)

    Xiankai Liu

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

  7. [Effects of plasmid pKM101 on the expression of Escherichia coli and Salmonella typhimurium genes under ultraviolet irradiation].

    Science.gov (United States)

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

    2003-01-01

    The study focused on plasmid pKM101, which is a necessary component of the short-term test of Eim's system (Salmonella-microsome test), to detect the potential carcinogens through their mutagen activity. We found a previously unknown feature of the plasmid to enhance the expression of certain plasmid and chromosome genes. The purpose of the present study was to examine and specify the role of operon mucAB responsible for the mutation properties of the plasmid in activating the expression of bacterial genes. An ultraviolet-induction examination of bacterial genes, with the mutants of plasmid pKM101 affecting operon mucAB being used, showed that the function of genes mucAB did activate, but, on the contrary, suppressed the induction of genes elt (i.e. of genes controlling the formation of LT-toxin of Escherichia coli) and of sfiA (SOS-regulated gen E. col controlling the cell division.

  8. Plasmids in the driving seat: The regulatory RNA Rcd gives plasmid ColE1 control over division and growth of its E. coli host.

    Science.gov (United States)

    Gaimster, Hannah; Summers, David

    2015-03-01

    Regulation by non-coding RNAs was found to be widespread among plasmids and other mobile elements of bacteria well before its ubiquity in the eukaryotic world was suspected. As an increasing number of examples was characterised, a common mechanism began to emerge. Non-coding RNAs, such as CopA and Sok from plasmid R1, or RNAI from ColE1, exerted regulation by refolding the secondary structures of their target RNAs or modifying their translation. One regulatory RNA that seemed to swim against the tide was Rcd, encoded within the multimer resolution site of ColE1. Required for high fidelity maintenance of the plasmid in recombination-proficient hosts, Rcd was found to have a protein target, elevating indole production by stimulating tryptophanase. Rcd production is up-regulated in dimer-containing cells and the consequent increase in indole is part of the response to the rapid accumulation of dimers by over-replication (known as the dimer catastrophe). It is proposed that indole simultaneously inhibits cell division and plasmid replication, stopping the catastrophe and allowing time for the resolution of dimers to monomers. The idea of a plasmid-mediated cell division checkpoint, proposed but then discarded in the 1980s, appears to be enjoying a revival.

  9. Biological control of soilborne diseases in organic potato production as affected by varying environmental conditions

    Science.gov (United States)

    Soilborne diseases are persistent problems in potato production and alternative management practices are needed, particularly in organic production, where control options are limited. Selected biocontrol organisms, including two naturally-occurring hypovirulent strains of Rhizoctonia solani (Rhs1a1 ...

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

    Science.gov (United States)

    Kado, Clarence I

    2014-10-01

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

  11. Situated embodied cognition: monitoring orientation cues affects product evaluation and choice

    NARCIS (Netherlands)

    J. Eelen; S. Dewitte; L. Warlop

    2013-01-01

    Consumers generally prefer products that are easy to interact with. In three studies, we show that this preference arises from the fit between product orientation and monitored situational constraints. Flexible right-handers, who monitor situational constraints, recall product orientations better an

  12. Serious overestimation in quantitative PCR by circular (supercoiled plasmid standard: microalgal pcna as the model gene.

    Directory of Open Access Journals (Sweden)

    Yubo Hou

    Full Text Available Quantitative real-time PCR (qPCR has become a gold standard for the quantification of nucleic acids and microorganism abundances, in which plasmid DNA carrying the target genes are most commonly used as the standard. A recent study showed that supercoiled circular confirmation of DNA appeared to suppress PCR amplification. However, to what extent to which different structural types of DNA (circular versus linear used as the standard may affect the quantification accuracy has not been evaluated. In this study, we quantitatively compared qPCR accuracies based on circular plasmid (mostly in supercoiled form and linear DNA standards (linearized plasmid DNA or PCR amplicons, using proliferating cell nuclear gene (pcna, the ubiquitous eukaryotic gene, in five marine microalgae as a model gene. We observed that PCR using circular plasmids as template gave 2.65-4.38 more of the threshold cycle number than did equimolar linear standards. While the documented genome sequence of the diatom Thalassiosira pseudonana shows a single copy of pcna, qPCR using the circular plasmid as standard yielded an estimate of 7.77 copies of pcna per genome whereas that using the linear standard gave 1.02 copies per genome. We conclude that circular plasmid DNA is unsuitable as a standard, and linear DNA should be used instead, in absolute qPCR. The serious overestimation by the circular plasmid standard is likely due to the undetected lower efficiency of its amplification in the early stage of PCR when the supercoiled plasmid is the dominant template.

  13. Terroir et vignoble: how the farming management can affect the production of a quality wine

    Science.gov (United States)

    Gallo, Alba; Bini, Claudio

    2016-04-01

    Italian wine is one of the most exported wine in the world. The particular climate, the soil characteristics and other several factors have contributed to this success. Italy is located in the temperate belt, with a suitable climate for grapevine cultivation. For this reason, all regions in Italy produce wine, first of all the Veneto region, with 8.569.000 hl of wine in 2011. Wine quality derives from the perfect interaction among climate, morphology, soil and plant, i.e. the terroir. So, knowledge of the land characteristics, together with cultivation techniques and management, is essential to understand this interaction and the typicality of the wine. For example, large utilization of fertilizers and pesticides may determine accumulation of toxic substances in soil and possible translocation to the food chain. For this reason, metal contamination of soils and plants becomes a main issue in agricultural production. Therefore, our attention was focused on the determination of soil quality of the Prosecco DOCG (controlled and guaranteed denomination of origin) area, particularly in Conegliano. Conegliano is a town located in Veneto, in the province of Treviso, known for its wine. This wine variety is regulated by the Conegliano-Valdobbiadene production Consortium, to protect both consumers and producers. The goals of this research are: evaluation of trace metal content (Al ,Cd, Cr, Cu, Fe, Mg, Mn, Ni, P, Pb, V and Zn) in soils and possible uptake by grape leaves; estimation of biological soil quality (QBS-ar index); analysis of oxidative stress in dandelion (Taraxacum officinale) and grape leaves, by the Lipid peroxidation test (LPO test). Results concerning trace metal concentration show: i) a high content of Al, Mg and P in soils, and ii) high concentration of Al, Cu, Fe and Zn in grape leaves. High contents of Al in topsoil are consistent with the high concentration of organic matter. Instead, high Al contents in subsoil are related to clay. Mg and P are usually

  14. 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.

  15. Plasmid selection in Escherichia coli using an endogenous essential gene marker

    Directory of Open Access Journals (Sweden)

    Good Liam

    2008-08-01

    Full Text Available Abstract Background Antibiotic resistance genes are widely used for selection of recombinant bacteria, but their use risks contributing to the spread of antibiotic resistance. In particular, the practice is inappropriate for some intrinsically resistant bacteria and in vaccine production, and costly for industrial scale production. Non-antibiotic systems are available, but require mutant host strains, defined media or expensive reagents. An unexplored concept is over-expression of a host essential gene to enable selection in the presence of a chemical inhibitor of the gene product. To test this idea in E. coli, we used the growth essential target gene fabI as the plasmid-borne marker and the biocide triclosan as the selective agent. Results The new cloning vector, pFab, enabled selection by triclosan at 1 μM. Interestingly, pFab out-performed the parent pUC19-ampicillin system in cell growth, plasmid stability and plasmid yield. Also, pFab was toxic to host cells in a way that was reversed by triclosan. Therefore, pFab and triclosan are toxic when used alone but in combination they enhance growth and plasmid production through a gene-inhibitor interaction. Conclusion The fabI-triclosan model system provides an alternative plasmid selection method based on essential gene over-expression, without the use of antibiotic-resistance genes and conventional antibiotics.

  16. A global analysis of fine root production as affected by soil nitrogen and phosphorus.

    Science.gov (United States)

    Yuan, Z Y; Chen, Han Y H

    2012-09-22

    Fine root production is the largest component of belowground production and plays substantial roles in the biogeochemical cycles of terrestrial ecosystems. The increasing availability of nitrogen (N) and phosphorus (P) due to human activities is expected to increase aboveground net primary production (ANNP), but the response of fine root production to N and P remains unclear. If roots respond to nutrients as ANNP, fine root production is anticipated to increase with increasing soil N and P. Here, by synthesizing data along the nutrient gradient from 410 natural habitats and from 469 N and/or P addition experiments, we showed that fine root production increased in terrestrial ecosystems with an average increase along the natural N gradient of up to 0.5 per cent with increasing soil N. Fine root production also increased with soil P in natural conditions, particularly at P soil types. The global average increases in fine root production are lower than those of ANNP, indicating that above- and belowground counterparts are coupled, but production allocation shifts more to aboveground with higher soil nutrients. Our results suggest that the increasing fertilizer use and combined N deposition at present and in the future will stimulate fine root production, together with ANPP, probably providing a significant influence on atmospheric CO(2) emissions.

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

    OpenAIRE

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

    2011-01-01

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

  18. Molecular characterization of Syrian date palm cultivars using plasmid-like DNA markers.

    Science.gov (United States)

    Haider, N; Nabulsi, I

    2012-02-01

    Date palm (Phoenix dactylifera L.) is one of the most important domesticated fruit trees in the Near East and North African countries. This tree has been, for several decades, in serious threat of being completely destroyed by the "Bayoud" disease caused by Fusarium oxysporum f. sp. albedinis. In this study, 18 Syrian date palm cultivars and four male trees were analyzed according to the identity of mitochondrial plasmid-like DNAs. A PCR strategy that employs plasmid-like DNAs-specific primer pair was used. These primers amplify a product of either 373-bp or 265-bp that corresponds to the S-(Bayoud-susceptible) or the R-plasmid (Bayoud-resistant), respectively. Generated data revealed that only six cultivars ('Medjool', 'Ashrasi', 'Gish Rabi', 'Khineze', and yellow- and red-'Kabkab') have the S-plasmid, suggesting their susceptibility to the fusariosis, while the remaining 12 cultivars and the four male trees contain the R-plasmid, suggesting their resistance to the fusariosis. The PCR process applied here has been proved efficient for the rapid screening for the presence of the S and R DNAs in Syrian date palm. PCR markers developed in this study could be useful for the screening of date palm lines growing in the field. The availability of such diagnostic tool for plasmid characterization in date palm would also be of great importance in establishing propagation and breeding programs of date palm in Syria.

  19. Recombinational instability of F' plasmids in Escherichia coli K-12: localization of fre-sites.

    Science.gov (United States)

    Bresler, S E; Krivonogov, S V; Lanzov, V A

    1981-01-01

    The F' plasmids ORF-1 (purE+ tsxs proC+ lac+) and F'14 (argE+ metB+ ilv+) contain active regions of recombination, fre I and fre II correspondingly. The plasmid ORF-1 is stable in recF- cells (i.e., with the RecBC pathway of recombination) and decays in rec+ cells (RecBCF pathway) giving two types of product: F+ and plasmid pCK-1 (tsxs proC+ lac+) containing part of the initial DNA. They are extremely instable in the presence of the RecF pathway, (recBC- sbcB-), yielding F+ and plasmid pCK-2 (proC+ lac+). The instability of plasmids depends on a region of homology between the chromosome and the episome. The instability of ORF-1 shows the participation of IS3 elements (alpha 1 beta 3 and alpha 3 beta 1) in the recA, recF-dependent recombinational decay and allows localization of two active sites on the chromosome: fre I1 between purE and tsx markers and fre I2 between tsx and proC. The plasmid F'14, in accordance with published data, is able to yield F+ cells by recA-independent recombination. But eventually this plasmid may undergo a recA, recF-dependent decay. Genetic analysis of these events allows localization of an active point of recombination, freII1, between argE and metB. Another active point is localized inside the F factor. The recA-dependent decay of plasmid F-14 is also excluded on the RecBC pathway (recF- strains).

  20. Analyzing factors Affecting Consumers’ Attitude & Intention to Purchase Counterfeit products of luxury Brands In clothing Industry

    Directory of Open Access Journals (Sweden)

    Abdolhamid Ebrahimi

    2012-01-01

    Full Text Available One of the world trends growing alarmingly is Producing, distributing & Consuming counterfeit goods of Credible & luxury brands & one of the industries facing this crisis increasingly is clothing industry. Based on this, the present study aims to analyze & identify factors influencing Consumers’ attitude toward counterfeit products & intention to purchase such products in clothing industry that has been done in Sari, Mazandaran. And in it, these Factors effect on attitude toward counterfeit products & then the impact of attitude toward counterfeit products on intention to buy such products have been measured. The study method is a Survey type & in order to collect study data, a questionnaire designed with a combination of different available resources in literature has been used. To achieve the study goals, 384 individuals who purchased counterfeit products intentionally were chosen from Sari. To analyze data & test hypotheses, Study model & questionnaire, Cronbach's alpha coefficient tests, Structural Equations Modeling (SEM, Confirmatory Factorial Analysis (CFA & one-way Variance analysis (ANONA have been applied & two Software Packages SPSS & LISREL were used. Through reviewing the literature available & analyzing experts’ views, generally, 11 variables have been identified: Personal gratification, Value consciousness, Price–quality inference, Social Effect, Brand prestige, Brand Loyalty, Ethical Issues, Risk averseness, Subjective norm, perceived risk, Brand consciousness, as the influencing factors on attitude toward counterfeit products. Study Findings have indicated that the factors as Personal gratification, Value consciousness, Price–quality inference, Social Effect, Ethical Issues, Subjective norm, perceived risk, Brand consciousness, have meaningful effect on attitude to counterfeit products & the impact of Brand prestige, Brand Loyalty, Risk averseness, was not meaningfull on view about counterfeit products. Besides, the

  1. Analyzing factors Affecting Consumers’ Attitude & Intention to Purchase Counterfeit products of luxury Brands In clothing Industry

    Directory of Open Access Journals (Sweden)

    abdolhamid ebrahimi

    2012-12-01

    Full Text Available One of the world trends growing alarmingly is Producing, distributing & Consuming counterfeit goods of Credible & luxury brands & one of the industries facing this crisis increasingly is clothing industry. Based on this, the present study aims to analyze & identify factors influencing Consumers’ attitude toward counterfeit products & intention to purchase such products in clothing industry that has been done in Sari, Mazandaran. And in it, these Factors effect on attitude toward counterfeit products & then the impact of attitude toward counterfeit products on intention to buy such products have been measured. The study method is a Survey type & in order to collect study data, a questionnaire designed with a combination of different available resources in literature has been used. To achieve the study goals, 384 individuals who purchased counterfeit products intentionally were chosen from Sari. To analyze data & test hypotheses, Study model & questionnaire, Cronbach's alpha coefficient tests, Structural Equations Modeling (SEM, Confirmatory Factorial Analysis (CFA & one-way Variance analysis (ANONA have been applied & two Software Packages SPSS & LISREL were used. Through reviewing the literature available & analyzing experts’ views, generally, 11 variables have been identified: Personal gratification, Value consciousness, Price–quality inference, Social Effect, Brand prestige, Brand Loyalty, Ethical Issues, Risk averseness, Subjective norm, perceived risk, Brand consciousness, as the influencing factors on attitude toward counterfeit products. Study Findings have indicated that the factors as Personal gratification, Value consciousness, Price–quality inference, Social Effect, Ethical Issues, Subjective norm, perceived risk, Brand consciousness, have meaningful effect on attitude to counterfeit products & the impact of Brand prestige, Brand Loyalty, Risk averseness, was not meaningfull on view about counterfeit products. Besides, the

  2. They Raise Them Differently Up North - Different Production Practices in Australian Growing Areas May Affect Mycorrhizae

    Science.gov (United States)

    This report briefly summarizes the results from a survey to determine cultural blueberry practices and production differences used in New South Wales (NSW), Victoria, Tasmania, and Queensland. The purpose of the survey was to collect information on a number of production practices that had been s...

  3. 29 CFR 784.138 - Perishable state of the aquatic product as affecting exemption.

    Science.gov (United States)

    2010-07-01

    ... such nonperishable products as fish oil and fish meal, or canned seafood, are not within the exemption... or manufacturing operations on products previously rendered nonperishable, such as refining fish oil or handling fish meal in connection with the manufacture of feeds. Further specific examples...

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2012-01-15

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

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

    Science.gov (United States)

    Yu, Mingrui; Du, Yinming; Jiang, Wenyan; Chang, Wei-Lun; Yang, Shang-Tian; Tang, I-Ching

    2012-01-01

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

  6. The prediction of the methane production in landfill affected by the temperature

    Directory of Open Access Journals (Sweden)

    Zhou Lianying

    2015-01-01

    Full Text Available The functional relationship between the generation rate coefficient and temperature was developed for quantitative prediction of the temperature effect in this paper. The methane production of the landfill was predicted under the condition of the seasonal variation. The results showed that considering the temperature effect, the methane production is higher than the methane production without temperature effect of the 0.14×106 m3~0.28×106m3.With the depth increasing, the effect of the atmospheric temperature fluctuation on the temperature change of the landfill was weakened. The temperature has a significant effect on the methane production in the landfill. The temperature effect should be considered when simulate the long time effect of the landfill methane production.

  7. Does species richness affect fine root biomass and production in young forest plantations?

    DEFF Research Database (Denmark)

    Domisch, Timo; Finér, Leena; Dawud, Seid Muhie;

    2015-01-01

    species composition from fine root biomass samples with the near-infrared reflectance spectroscopy method. We did not observe higher biomass or production in mixed stands compared to monocultures. Neither did we observe any differences in tree root length or fine root turnover. One reason for this could......Tree species diversity has been reported to increase forest ecosystem above-ground biomass and productivity, but little is known about below-ground biomass and production in diverse mixed forests compared to single-species forests. For testing whether species richness increases below-ground biomass...... and production and thus complementarity between forest tree species in young stands, we determined fine root biomass and production of trees and ground vegetation in two experimental plantations representing gradients in tree species richness. Additionally, we measured tree fine root length and determined...

  8. Distribution and abundance of predators that affect duck production--prairie pothole region

    Science.gov (United States)

    Sargeant, A.B.; Greenwood, R.J.; Sovada, M.A.; Shaffer, T.L.

    1993-01-01

    During 1983-88, the relative abundance of 18 species and species-groups of mammalian and avian predators affecting duck production in the prairie pothole region was determined in 33 widely scattered study areas ranging in size from 23-26 km2. Accounts of each studied species and species-group include habitat and history, population structure and reported densities, and information on distribution and abundance from the present study. Index values of undetected, scarce, uncommon, common, or numerous were used to rate abundance of nearly all species in each study area. Principal survey methods were livetrapping of striped skunks (Mephitis mephitis) and Franklin's ground squirrels (Spermophilus franklinii), systematic searches for carnivore tracks in quarter sections (0.65 km2), daily records of sightings of individual predator species, and systematic searches for occupied nests of tree-nesting avian predators. Abundances of predators in individual areas were studied 1-3 years.The distribution and abundance of predator species throughout the prairie pothole region have undergone continual change since settlement of the region by Europeans in the late 1800's. Predator populations in areas we studied differed markedly from those of pristine times. The changes occurred from habitat alterations, human-inflicted mortality of predators, and interspecific relations among predator species. Indices from surveys of tracks revealed a decline in the abundance of red foxes (Vulpes vulpes) and an albeit less consistent decline in the abundance of raccoons (Procyon lotor) with an increase in the abundance of coyotes (Canis latrans). Records of locations of occupied nests revealed great horned owls (Bubo virginianus) and red-tailed hawks (Buteo jamaicensis) tended to nest 0.5 km apart, and American crows (Corvus brachyrhynchos) tended to avoid nesting 0.5 km of nests of red-tailed hawks. Excluding large gulls, for which no measurements of abundance were obtained, the number of

  9. How can farming intensification affect the environmental impact of milk production?

    Science.gov (United States)

    Bava, L; Sandrucci, A; Zucali, M; Guerci, M; Tamburini, A

    2014-07-01

    The intensification process of the livestock sector has been characterized in recent decades by increasing output of product per hectare, increasing stocking rate, including more concentrated feed in the diet, and improving the genetic merit of the breeds. In dairy farming, the effects of intensification on the environmental impact of milk production are not completely clarified. The aim of the current study was to assess the environmental impacts of dairy production by a life cycle approach and to identify relations between farming intensity and environmental performances expressed on milk and land units. A group of 28 dairy farms located in northern Italy was involved in the study; data collected during personal interviews of farmers were analyzed to estimate emissions (global warming potential, acidification, and eutrophication potentials) and nonrenewable source consumption (energy and land use). The environmental impacts of milk production obtained from the life cycle assessment were similar to those of other recent studies and showed high variability among the farms. From a cluster analysis, 3 groups of farms were identified, characterized by different levels of production intensity. Clusters of farms showed similar environmental performances on product basis, despite important differences in terms of intensification level, management, and structural characteristics. Our study pointed out that, from a product perspective, the most environmentally friendly way to produce milk is not clearly identifiable. However, the principal component analysis showed that some characteristics related to farming intensification, such as milk production per cow, dairy efficiency, and stocking density, were negatively related to the impacts per kilogram of product, suggesting a role of these factors in the mitigation strategy of environmental burden of milk production on a global scale. Considering the environmental burden on a local perspective, the impacts per hectare were

  10. 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.

  11. Production and characterization of chicken origin Shigella dysenteriae ghosts with double bacteriolysis plasmids%双重溶菌质粒制备鸡痢疾志贺菌菌壳的研究

    Institute of Scientific and Technical Information of China (English)

    蒋大伟; 刘军; 张瑞安; 夏力亮; 孙洋; 祝令伟; 冯书章; 许兰菊

    2013-01-01

    In order to improve lysis rate and stability of chicken origin Shigella dysenteriae ghosts,recombinant strain of Shigella with double bacteriolytic plasmids was constructed and its characteristics was studied. Two recombinant bacteriolysis plasmid pBV220: :E and pBBR IMCS-E were co-transformed into the Shigella strain isolated from chicken to produce recombinant strain Sd(pBV220::E /pBBRIMCS-E). The recombinant strain Sd(pBV220::E /pBBRIMCS-E) was grown at 28℃ with shaking until mid log-phase,followed by incubation at 42℃ to induce the expression of gene E and produce Shigella dysenteriae ghosts. The D600 value of bacterial cultures was monitored over 20 h at 30 min intervals. The morphology of chicken origin Shigella dysenteriae ghosts was observed by electron microscope. The recombinant strains Sd(pBV220 : : E/pB-BRIMCS-E) generated chicken origin Shigella dysenteriae ghosts after inducing at 42℃. The ghosts retained basic cell morphology,with the lysis rate of 99. 999 9%. The chicken origin Shigella dysenteriae ghosts were generated successfully with double bacteriolysis plasmids, which laid foundation for future development of bacterial ghost vaccine against Shigella infection in chicken.%通过将2种不同的重组溶菌质粒转化至鸡痢疾志贺菌中,构建含有双重溶菌质粒的志贺菌重组菌株,以提高该菌菌壳制备的裂解率和稳定性,并对其特性进行研究.用重组溶菌质粒pBV220::E和pBBRIMCS-E共同转化到鸡痢疾志贺菌,构建志贺菌重组菌株Sd(pBV220::E/pBBRI MCS-E),重组菌株于28 C培养至D600值为0.5时升至42℃继续培养,间隔30 min检测菌液D600值,进行重组菌株的溶菌裂解动力学比较试验,绘制裂解曲线图并计算出裂解率,制备菌壳并对其形态进行电镜观察.重组菌株Sd(pBV220::E/pBBRIMCS-E)经42℃诱导后可形成菌壳且保留了基本的细胞形态,裂解率达到了99.999 9%.本研究成功制备出鸡痢疾志贺菌菌壳,为研究利用该

  12. How body mass and lifestyle affect juvenile biomass production in placental mammals.

    Science.gov (United States)

    Sibly, Richard M; Grady, John M; Venditti, Chris; Brown, James H

    2014-02-22

    In mammals, the mass-specific rate of biomass production during gestation and lactation, here called maternal productivity, has been shown to vary with body size and lifestyle. Metabolic theory predicts that post-weaning growth of offspring, here termed juvenile productivity, should be higher than maternal productivity, and juveniles of smaller species should be more productive than those of larger species. Furthermore because juveniles generally have similar lifestyles to their mothers, across species juvenile and maternal productivities should be correlated. We evaluated these predictions with data from 270 species of placental mammals in 14 taxonomic/lifestyle groups. All three predictions were supported. Lagomorphs, perissodactyls and artiodactyls were very productive both as juveniles and as mothers as expected from the abundance and reliability of their foods. Primates and bats were unproductive as juveniles and as mothers, as expected as an indirect consequence of their low predation risk and consequent low mortality. Our results point the way to a mechanistic explanation for the suite of correlated life-history traits that has been called the slow-fast continuum.

  13. 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.

  14. Compost amendment of sandy soil affects soil properties and greenhouse tomato productivity

    DEFF Research Database (Denmark)

    Arthur, Emmanuel; Cornelis, W.; Razzaghi, Fatemeh

    2012-01-01

    Sandy soils, with low productivity, could be improved by compost application to sustain crop production. This study aimed to examine the effect of three compost types (vegetable, fruit and yard waste compost, garden waste compost, and spent mushroom compost) on basic properties of a loamy sand...... and greenhouse tomato productivity. Disturbed and intact soil samples were taken from a decade-long compost field experiment on loamy sand with three compost types at application rate of 30 m3 ha-1 yr-1 (7.5 ton ha-1 yr-1). The soils were characterized for chemical and physical properties. Tomato was planted...... in a greenhouse using soil samples from the field and vegetative and yield parameters (plant height, stem diameter, leaf number, and fruit yield), water productivity, and harvest index were evaluated. All compost types significantly increased soil total carbon, total nitrogen, pH, electrical conductivity...

  15. Biogas production as affected by heavy metals in the anaerobic digestion of sludge

    Directory of Open Access Journals (Sweden)

    Hussein I. Abdel-Shafy

    2014-12-01

    The sewage sludge samples were separated from the sewage water of the pilot plant at the National Research Centre, TDC site. The effect of heavy metals on the biogas production of the anaerobic digester was studied. The inhibitory effect on the biogas production and toxic level of metals was determined in this study. The general ranking of heavy metal toxicity appears to be Hg > Cd > Cr (III. The present investigation reveals that heavy metals in addition to the anaerobic digester decreased the biogas production as an indication of efficiency of the process. A significant decrease in gas production and volatile organic matter removal was obtained. It was also noted that an accumulation of organic acid intermediates was obtained as a result of methanogenic bacterial inhibition. This accumulation was limited during the pulse feed of metals. This is due to the rapid poisoning of the active bacterial forms in the digester.

  16. Characteristics affecting management of design information in the production system design process

    OpenAIRE

    Bruch, Jessica; Bellgran, Monica

    2013-01-01

    Although it has been argued that the design of production systems is crucial, there is a general lack of empirical studies analysing and identifying resources and capabilities required for an efficient production system design process. One of these resources is the critical role attributed to design information and one such capability is how the design information is managed. To address this research gap, this paper reports the results from two in-depth case studies in the automotive industry...

  17. Earthworm functional traits and interspecific interactions affect plant nitrogen acquisition and primary production

    NARCIS (Netherlands)

    Andriuzzi, Walter; Schmidt, Olaf; Brussaard, L.; Faber, J.H.; Bolger, T.

    2016-01-01

    We performed a greenhouse experiment to test how the functional diversity of earthworms, the dominant group of soil macro-invertebrates in many terrestrial ecosystems, affects nitrogen cycling and plant growth. Three species were chosen to represent a range of functional traits: Lumbricus terrestris

  18. DID THE 2004 CAP REFORM AFFECT PRODUCTION PRACTICES OF CEREALS? INSIGHTS FROM THE AGRICULTURAL INPUT SUPPLIERS

    Directory of Open Access Journals (Sweden)

    Theodoros MARKOPOULOS

    2015-10-01

    Full Text Available The Mid-term review of the Common Agricultural Policy in 2003/2004 has strengthened the multifunctional role of agriculture by implementing “decoupling”, “modulation” and “cross-compliance” and created a number of significant changes in agricultural production in all EU member states. Specifically, the reform shifted emphasis away from commodity support towards environmental contracts, diversified production practices and rural development. In the case of cereals, a full decoupling was applied in subsidies and integration through rights in the Single Payment Scheme, except rice, which was one of the few crop cases in which part of the subsidy remained coupled, particular in countries with significant production like Greece. Within this context, the present study aims to analyze the impact that the reformed CAP measures had on agricultural production and more specifically variations in production diversification. The novelty of this study is that instead of focusing on the producers, it targeted the agricultural input stores, so as to get better insights of the CAP reform impacts on a larger scale of the regional economy. Accordingly, primary data were collected through personal interviews (structured questionnaire from 209 owners of agricultural input stores in the region of Anatoliki Makedonia and Thraki and were analyzed through multivariate data analysis. The results identify important antecedents for the regional economy and the viability of agricultural input stores, which include factors of the reformed CAP, environmental issues, financial measures and CAP effects on cereal production and marketing.

  19. An experimental study on fermentative H₂ production from food waste as affected by pH.

    Science.gov (United States)

    Cappai, G; De Gioannis, G; Friargiu, M; Massi, E; Muntoni, A; Polettini, A; Pomi, R; Spiga, D

    2014-08-01

    Batch dark fermentation experiments were performed on food waste and mixtures of food waste and wastewater activated sludge to evaluate the influence of pH on biological H2 production and compare the process performance with and without inoculum addition. The effect of a preliminary thermal shock treatment of the inoculum was also investigated as a means to harvest the hydrogenogenic biomass. The best performance in terms of both H2 generation potential and process kinetics was observed at pH=6.5 under all experimental conditions (no inoculum, and untreated or thermally treated inoculum added). H2 production from food waste was found to be feasible even without inoculum addition, although thermal pre-treatment of the inoculum notably increased the maximum production and reduced the lag phase duration. The analysis of the fermentation products indicated that the biological hydrogen production could be mainly ascribed to a mixed acetate/butyrate-type fermentation. However, the presence of additional metabolites in the digestate, including propionate and ethanol, also indicated that other metabolic pathways were active during the process, reducing substrate conversion into hydrogen. The plateau in H2 generation was found to mirror the condition at which soluble carbohydrates were depleted. Beyond this condition, homoacetogenesis probably started to play a role in the degradation process.

  20. PREDICTING OF THE FACTORS AFFECTING CONSUMER BEHAVIOR THE CHOICE OF ORGANIC PRODUCTS BY LOGIT REGRESSION ANALYSIS

    Directory of Open Access Journals (Sweden)

    Selma KARABAŞ

    2012-06-01

    Full Text Available The purpose of this study is to determine consumer behavior towards organic products in Samsun city center and to determine the factors affacting their preferences of organic products. The study was interviewed with 478 consumers living the city center of Samsun. According of the findings, ease of accesibility of organic products, spouse’s educational level, paying extra for the organic produce, aware of food health benefits, considering the harmful effects of conventional produce, having complete knowledge of organic farming and one unit increase household number income to result increase consumption of organic produce. Consumers were not aware of the certification and control process. As a result, easy of accesibility of organic produces in the supermarkets need to be improved.

  1. Does warming affect growth rate and biomass production of shrubs in the High Arctic?

    DEFF Research Database (Denmark)

    Campioli, Matteo; Schmidt, Niels Martin; Albert, Kristian Rost

    2013-01-01

    Few studies have assessed directly the impact of warming on plant growth and biomass production in the High Arctic. Here, we aimed to investigate the impact of 7 years of warming (open greenhouses) on the aboveground relative growth rate (RGR) of Cassiope tetragona and Salix arctica in North......-Eastern Greenland. RGR was assessed for apical (leaves, stem, reproductive organs) and lateral meristems (secondary growth of stem and branches) and accompanied by measures of gross ecosystem production (GEP), branching and tissue carbon (C) concentration. Measurements were based on harvest and biometric methods...... limits the growth of Cassiope but not that of Salix in North-Eastern Greenland. Summer warming thus has the potential to stimulate biomass production in the High Arctic but major species-specific differences are expected....

  2. Dirhinus giffardii (Hymenoptera: Chalcididae, parasitoid affecting Black Soldier Fly production systems in West Africa

    Directory of Open Access Journals (Sweden)

    Emilie Devic

    2015-12-01

    Full Text Available Interest for insect farming is currently growing globally. Conditions in West Africa appear suitable for developing such farming systems that can benefit communities by improving livelihoods, food and feed security or sanitation. In Ghana and Mali, the Black Soldier Fly (Hermetia illucens Linnaeus, 1758 is being produced for waste recycling and animal feed. In a two stages process (egg and larvae production, egg production was hampered by a pupal parasitoid, Dirhinus giffardii Silvestri, 1913 (Hymenoptera: Chalcididae, which reduced future broodstock by almost 72%. This is the first time D. giffardii is reported as a parasitoid of H. illucens pupae and one of the first reports of parasitism in this commercially important fly species. The introduction of precautionary measures is highly recommended for the success of H. illucens production systems in West Africa.

  3. Process parameters affecting the sustainability of fermentative hydrogen production: A short-review

    Directory of Open Access Journals (Sweden)

    Carlos Dinamarca, Rune Bakke

    2011-11-01

    Full Text Available Anaerobic fermentation is regarded as the least energy intensive method for H2 production. Extensive literature on experimental attempts to achieve the highest possible theoretical yield (e.g. 4 mol H2/mol glucose is available. All published steady state, mixed culture studies show yields much lower than the theoretical maximums for the substrates applied. This article summarizes the influence of key process parameters (pH and buffer systems, temperature, H2 partial pressure, feed stock, and reactor configuration on fermentative hydrogen production. The following three requirements for successful Bio-H2 fermentation in mixed cultures are identified: (1 Maintain environmental conditions for the formation of oxidized products; (2 Optimize the relationship between biomass and hydrogen yields; and (3 Maintain unfavorable conditions for hydrogen consuming organisms. Fulfilling these requirements has not yet been achieved in stable continuous cultures, and it may not be achievable do to some fundamental limitation.

  4. Engaged or exhausted—How does it affect dentists’ clinical productivity?

    Directory of Open Access Journals (Sweden)

    Jari J. Hakanen

    2014-06-01

    Full Text Available This cross-sectional study examines whether job burnout (exhaustion and work engagement are associated with the clinical productivity of dentists measured by the amount of paid procedure fees in a single month. We conducted an OLS regression analyses of data on dentists working at municipal health centers in Finland (N = 269; response rate 37%. The results indicated that work engagement was positively associated with the amount of procedure fees and consequently with dentists’ pay level after several work-related and demographic background variables were controlled for. However, exhaustion was not related to productivity after controlling for the impact of other factors.

  5. Detection of genetic variants affecting cattle behaviour and their impact on milk production: a genome-wide association study.

    Science.gov (United States)

    Friedrich, Juliane; Brand, Bodo; Ponsuksili, Siriluck; Graunke, Katharina L; Langbein, Jan; Knaust, Jacqueline; Kühn, Christa; Schwerin, Manfred

    2016-02-01

    Behaviour traits of cattle have been reported to affect important production traits, such as meat quality and milk performance as well as reproduction and health. Genetic predisposition is, together with environmental stimuli, undoubtedly involved in the development of behaviour phenotypes. Underlying molecular mechanisms affecting behaviour in general and behaviour and productions traits in particular still have to be studied in detail. Therefore, we performed a genome-wide association study in an F2 Charolais × German Holstein cross-breed population to identify genetic variants that affect behaviour-related traits assessed in an open-field and novel-object test and analysed their putative impact on milk performance. Of 37,201 tested single nucleotide polymorphism (SNPs), four showed a genome-wide and 37 a chromosome-wide significant association with behaviour traits assessed in both tests. Nine of the SNPs that were associated with behaviour traits likewise showed a nominal significant association with milk performance traits. On chromosomes 14 and 29, six SNPs were identified to be associated with exploratory behaviour and inactivity during the novel-object test as well as with milk yield traits. Least squares means for behaviour and milk performance traits for these SNPs revealed that genotypes associated with higher inactivity and less exploratory behaviour promote higher milk yields. Whether these results are due to molecular mechanisms simultaneously affecting behaviour and milk performance or due to a behaviour predisposition, which causes indirect effects on milk performance by influencing individual reactivity, needs further investigation.

  6. Cultivar, harvest date, and nitrogen fertilization affect production and quality of fall oat

    Science.gov (United States)

    Previous research has shown that oat (Avena sativa L.) has promise as a fall-forage option for dairy producers. In addition, dairy producers often have a recurring need to identify opportunity windows for manure hauling other than before or after production of corn (Zea mays L.). Our objectives were...

  7. Market oriented new product development: How can a means-end chain approach affect the process?

    DEFF Research Database (Denmark)

    Søndergaard, Helle Alsted

    Few disagree on the advantage of market oriented product development. However, can a well-known theory on consumer behaviour be used as a catalyst for achieving it? This paper describes a case study where means-end chain (MEC) approach was introduced to a cross-functional development team at two...

  8. Diversity of condensed tannin structures affects rumen in vitro methane production in sainfoin (Onobrychis viciifolia) accessions

    NARCIS (Netherlands)

    Hatew, B.; Hayot Carbonero, C.; Stringano, E.; Sales, L.F.; Smith, L.M.J.; Mueller-Harvey, I.; Hendriks, W.H.; Pellikaan, W.F.

    2015-01-01

    Sainfoin is a non-bloating temperate forage legume with a moderate-to-high condensed tannin (CT) content. This study investigated whether the diversity of sainfoin accessions in terms of CT structures and contents could be related to rumen in vitro gas and methane (CH4) production and fermentation c

  9. Structural features of condensed tannins affect in vitro ruminal methane production and fermentation characteristics

    NARCIS (Netherlands)

    Huyen, N.T.; Fryganas, C.; Uittenbogaard, G.; Mueller-Harvey, I.; Verstegen, M.W.A.; Hendriks, W.H.; Pellikaan, W.F.

    2016-01-01

    An in vitro study was conducted to investigate the effects of condensed tannin (CT) structural properties, i.e. average polymer size (or mean degree of polymerization), percentage of cis flavan-3-ols and percentage of prodelphinidins in CT extracts on methane (CH4) production and ferme

  10. Statistical analysis of factors affecting crop production in Navrongo, Tono irrigation dam a case study

    Directory of Open Access Journals (Sweden)

    D. JAKPERIK

    2013-02-01

    Full Text Available This study identified the essential factors of production in the Tono irrigation dam of the Upper East Region. The accessibility and patronage of these factors by farmers in this area was studied and how they influence crop production in the Region. A total of two hundred questionnaires designed by the Ministry of Food and Agriculture for farmers in Navrongo were administered. A snowball sampling design was employed to identify farmers on these facilities in the study area. Pearson correlation coefficient, principal component analysis, and subset regression analysis were used to unveil the relevant information in the study. The results revealed a high correlation between the factors of production being studied with each pair having a probability level less than 0.0001. The full general linear model was highly significant (F=662.50, psmaller tahn 0.0001 with only two factors (Farm size and Fertilizer accounting for 98.86% of the total variation in yield. This is a clear indication of multicollinearity and a subset regression analysis was used to identify the best subset that improves yield in the irrigation dam. The best subset comprised of Age, Farm size, seed, and Fertilizer accounting for 97.75% of the total variation in crop production in Navrongo. To enhance yield in Navrongo therefore, high yielding seeds, timely granting of fertilizer credit to farmers who mature enough and responsible with reasonable farm sizes should be encouraged.

  11. Intestinal microbial affects of yeast products on weaned and transport stressed pigs

    Science.gov (United States)

    Study objectives were to determine effects of a commercially available yeast product (XPC, Diamond-V Mills) and stress of transportation on total Enterobacteriaceae, Escherichia coli, coliforms, and Lactobacilli populations in the intestine of weaning pigs. In a RCB design with a 2 x 2 factorial ar...

  12. How forest management affects ecosystem services, including timber production and economic return

    DEFF Research Database (Denmark)

    Duncker, Philipp S.; Raulund-Rasmussen, Karsten; Gundersen, Per;

    2012-01-01

    Forest ecosystems deliver multiple goods and services and, traditionally, forest owners tend to have a high interest in goods in the form of merchantable wood. As a consequence, forest management often aims to increase timber production and economic returns through intervention into natural...

  13. Microvesicle formulations used in topical drugs and cosmetics affect product efficiency, performance and allergenicity

    DEFF Research Database (Denmark)

    Madsen, Jakob Torp; Ejner Andersen, Klaus

    2010-01-01

    Attempts to improve the formulations of topical products are continuing processes (ie, to increase cosmetic performance, enhance effects, and protect ingredients from degradation). The development of micro- and nanovesicular systems has led to the marketing of topical drugs and cosmetics that use...

  14. Soil water infiltration affected by biofuel and grain crop production systems in claypan landscape

    Science.gov (United States)

    The effect of soil management systems on water infiltration is very crucial within claypan landscapes to maximize production as well as minimize environmental risks. The objective of this study was to assess the effect of topsoil thickness on water infiltration in claypan soils for grain and biofuel...

  15. Divergence for residual feed intake of Holstein-Friesian cattle during growth did not affect production and reproduction during lactation.

    Science.gov (United States)

    Macdonald, K A; Thomson, B P; Waghorn, G C

    2016-11-01

    Residual feed intake (RFI) is the difference between actual and predicted dry matter intake (DMI) of individual animals. Recent studies with Holstein-Friesian calves have identified an ~20% difference in RFI during growth (calf RFI) and these groups remained divergent in RFI during lactation. The objective of the experiment described here was to determine if cows selected for divergent RFI as calves differed in milk production, reproduction or in the profiles of BW and body condition score (BCS) change during lactation, when grazing pasture. The cows used in the experiment (n=126) had an RFI of -0.88 and +0.75 kg DM intake/day for growth as calves (efficient and inefficient calf RFI groups, respectively) and were intensively grazed at four stocking rates (SR) of 2.2, 2.6, 3.1 and 3.6 cows/ha on self-contained farmlets, over 3 years. Each SR treatment had equal number of cows identified as low and high calf RFI, with 24, 28, 34 and 40/11 ha farmlet. The cows divergent for calf RFI were randomly allocated to each SR. Although SR affected production, calf RFI group (low or high) did not affect milk production, reproduction, BW, BCS or changes in these parameters throughout lactation. The most efficient animals (low calf RFI) lost similar BW and BCS as the least efficient (high calf RFI) immediately post-calving, and regained similar BW and BCS before their next calving. These results indicate that selection for RFI as calves to increase efficiency of feed utilisation did not negatively affect farm productivity variables (milk production, BCS, BW and reproduction) as adults when managed under an intensive pastoral grazing system.

  16. Studies on Saccharomyces cerevisiae carrying the plasmid pCYG4 related with ammonia assimilation. Batch experiments.

    Science.gov (United States)

    Lima Filho, J L; Ledingham, W M

    1988-10-01

    Batch culture experiments of three different strains of Saccharomyces cerevisiae have been carried out. The first strain was transformed by a plasmid pCYG4, which carries the glutamate dehydrogenase (NADP-GDH, E.C. 1.4.14) gene conferring an 11-fold increase in activity. The second was transformed by the same plasmid, but without NADP-GDH, and the third was the wild type. The specific growth rates of the two recombinant DNA strains were below that of the wild type, which can be related to extra plasmid protein production.

  17. Plasmid mediated quinolone resistance in Enterobacteriaceae

    NARCIS (Netherlands)

    Veldman, K.T.; LS Klinisch Onderzoek Wagenaar

    2014-01-01

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

  18. Enterococcus faecium isolated from Lombo, a Portuguese traditional meat product: characterisation of antibacterial compounds and factors affecting bacteriocin production.

    Science.gov (United States)

    Todorov, S D; Favaro, L; Gibbs, P; Vaz-Velho, M

    2012-12-01

    Strain ST211CH, identified as a strain of Enterococcus faecium, isolated from Lombo produced a bacteriocin that inhibited the growth of Enterococcus spp., Listeria spp., Klebsiella spp., Lactobacillus spp., Pseudomonas spp., Staphylococcus spp. and Streptococcus spp. The mode of action of the bacteriocin named as bacteriocin ST211Ch was bactericidal against Enterococcus faecalis ATCC19443. As determined by Tricine-SDS-PAGE, the approximate molecular mass of the bacteriocin was 8.0 kDa. Loss in antimicrobial activity was recorded after treatment with proteolytic enzymes. Maximum activity of bacteriocin ST211Ch was measured in broth cultures of E. faecium strain ST211Ch after 24 h; thereafter, the activity was reduced. Bacteriocin ST211Ch remained active after exposure to various temperatures and pHs, as well as to Triton X-100, Tween-80, Tween-20, sodium dodecyl sulfate, NaCl, urea and EDTA. Effect of media components on production of bacteriocin ST211Ch was also studied. On the basis of PCR reactions targeting different bacteriocin genes, i.e. enterocins, curvacins and sakacins, no evidences for the presence of these genes in the total DNA of E. faecium strain ST211Ch was obtained. The bacterium most probably produced a bacteriocin different from those mentioned above. Based on the antimicrobial spectrum, stability and mode of action of bacteriocin ST211CH, E. faecium strain ST211Ch might be considered as a potential candidate with beneficial properties for use in biopreservation to control food spoilage bacteria.

  19. Do new cellulolytic enzyme preparations affect the industrial strategies for high solids lignocellulosic ethanol production?

    DEFF Research Database (Denmark)

    Cannella, David; Jørgensen, Henning

    2014-01-01

    Production of ethanol from lignocellulosic materials has a promising market potential, but the process is still only at pilot/demonstration scale due to the technical and economical difficulties of the process. Operating the process at very high solids concentrations (above 20% dry matter—DM) has...... matter conditions. In this work the impact of selected enzyme preparation and processing strategy (SHF, presaccharification and simultaneous saccharification and fermentation—PSSF, and SSF) on final ethanol yield and overall performance was investigated with pretreated wheat straw up to 30% DM....... The experiments revealed that an SSF strategy was indeed better than SHF when applying an older generation enzyme cocktail (Celluclast-Novozym 188). In case of the newer product Cellic CTec 2, SHF resulted in 20% higher final ethanol yield compared to SSF. It was possible to close the mass balance around...

  20. The Major Recessive Calamities Affecting the Wheat Production in Chuxiong Prefecture and Control Countermeasures

    Institute of Scientific and Technical Information of China (English)

    Maochang; YANG

    2014-01-01

    The impact of recessive calamities was analyzed,including seasonal drought,cold injury,dry hot wind and aphid in the wheat production of Chuxiong Prefecture,and the countermeasures that prevented and controlled the recessive calamities in a target-oriented way were proposed,including the improvement of basic farmland,the application of organic manure,the promotion of the breed with high stress resistance,the seedling at suitable date,the improvement of control on fertilizing and watering,the enhancement of management on cultivating and controlling disease in time,and the breeding new variety adaptive to local ecosystem,in order to advance the wheat production in a sustainable way.

  1. How Can Diet Affect the Accumulation of Advanced Glycation End-Products in the Human Body?

    Directory of Open Access Journals (Sweden)

    Axel Guilbaud

    2016-12-01

    Full Text Available The accumulation of advanced glycation end products (AGEs is associated with the complications of diabetes, kidney disease, metabolic disorders and degenerative diseases. It is recognized that the pool of glycation products found in the human body comes not only from an endogenous formation, but also from a dietary exposure to exogenous AGEs. In recent years, the development of pharmacologically-active ingredients aimed at inhibiting endogenous glycation has not been successful. Since the accumulation of AGEs in the human body appears to be progressive throughout life, an early preventive action against glycation could be effective through dietary adjustments or supplementation with purified micronutrients. The present article provides an overview of current dietary strategies tested either in vitro, in vivo or both to reduce the endogenous formation of AGEs and to limit exposure to food AGEs.

  2. Analysis of the Factors Affecting Consumer’s Some Traditional Food Products Preferences by Multidimensional Scaling Method

    Directory of Open Access Journals (Sweden)

    Y. Oraman G. Unakıtan E. Yılmaz B. Başaran

    2011-01-01

    Full Text Available The aim of this study is to evaluate consumer behaviour towards factors affecting purchase decision of some traditional food products and grouping those consumer attitudes. The original data was obtained from results of a survey conducted in 14 different districts in Tekirdag. The survey consisted of face-to-face interviews conducted within a total sample of 166 households randomly. In the study, multidimensional scaling analysis is used for evaluating the effective factors of consumer preferences for traditional products and grouping their preferences. It is found that taste, food safety and freshness have similar effects on consumer preferences for yogurt, molasses and noodles. Price has an important and positive loaded effect on other variables for the three products.

  3. Rethinking Food: How United States Agriculture Production Affects Security Policy and Global Markets

    Science.gov (United States)

    2014-06-13

    of re- exporting , it is assumed some countries (like Argentina did) must have done so for either economic or political gain with the Soviets. Following... Argentina and Brazil began exporting soy meal in 1977. That same year Brazil displaced America as the top soy meal exporter in the world while Argentina ...likely prevent an ideal distribution of food. The United States has been and remains a major exporter of several key commodities giving US production

  4. The interactive effects of affect and shopping goal on information search and product evaluations.

    Science.gov (United States)

    Chen, Fangyuan; Wyer, Robert S; Shen, Hao

    2015-12-01

    Although shoppers often want to evaluate products to make a purchase decision, they can also shop for enjoyment. In each case, the amount of time they spend on shopping and the number of options they consider can depend on the mood they happen to be in. We predicted that mood can signal whether the goal has been attained and when people should stop processing information. When people are primarily motivated to purchase a particular type of product, positive mood signals that they have done enough. Thus, they consider less information if they are happy than if they are unhappy. When people shop for enjoyment, however, positive mood signals that they are still having fun. Thus, they consider more information when they are happy than when they are not. Four experiments among university students (N = 827) examined these possibilities. Experiment 1 provided initial evidence for the interactive effects of mood and goals on search behavior and product evaluation. Other studies examined the implications of this conceptualization for different domains: (a) the relative impact of brand and attribute information on judgments (Experiment 2), (b) gender differences in shopping behavior (Experiment 3), and (c) the number of options that people review in an actual online shopping website (Experiment 4).

  5. Blockage of the pyrimidine biosynthetic pathway affects riboflavin production in Ashbya gossypii.

    Science.gov (United States)

    Silva, Rui; Aguiar, Tatiana Q; Domingues, Lucília

    2015-01-10

    The Ashbya gossypii riboflavin biosynthetic pathway and its connection with the purine pathway have been well studied. However, the outcome of genetic alterations in the pyrimidine pathway on riboflavin production by A. gossypii had not yet been assessed. Here, we report that the blockage of the de novo pyrimidine biosynthetic pathway in the recently generated A. gossypii Agura3 uridine/uracil auxotrophic strain led to improved riboflavin production on standard agar-solidified complex medium. When extra uridine/uracil was supplied, the production of riboflavin by this auxotroph was repressed. High concentrations of uracil hampered this (and the parent) strain growth, whereas excess uridine favored the A. gossypii Agura3 growth. Considering that the riboflavin and the pyrimidine pathways share the same precursors and that riboflavin overproduction may be triggered by nutritional stress, we suggest that overproduction of riboflavin by the A. gossypii Agura3 may occur as an outcome of a nutritional stress response and/or of an increased availability in precursors for riboflavin biosynthesis, due to their reduced consumption by the pyrimidine pathway.

  6. The yeast TUM1 affects production of hydrogen sulfide from cysteine treatment during fermentation.

    Science.gov (United States)

    Huang, Chien-Wei; Walker, Michelle E; Fedrizzi, Bruno; Roncoroni, Miguel; Gardner, Richard C; Jiranek, Vladimir

    2016-12-01

    The undesirable rotten-egg odour of hydrogen sulfide (H2S) produced by yeast shortly after yeast inoculation of grape musts might be an important source of desirable varietal thiols, which contribute to tropical aromas in varieties such as Sauvign-on Blanc. In this study, we observed that Saccharomyces cerevisiae strains produce an early burst of H2S from cysteine. Both Δmet2 and Δmet17 strains produce a larger burst, likely because they are unable to utilise the H2S in the sulfate assimilation pathway. For the first time, we show that TUM1 is partly responsible for the early production of H2S from cysteine. Overex-pressing TUM1 elevated production of H2S, whilst its deletion yields only half of the H2S. We further confirmed that yeast convert cysteine to H2S by analysing growth of mutants lacking components of the transsulfuration pathway. High concent-rations of cysteine overcame this growth block, but required TUM1 Collectively, the data indicate that S. cerevisiae does not convert cysteine to sulfate or sulfite, but rather to sulfide via a novel pathway that requires the action of Tum1p. The findi-ngs of this study may allow the improvement of commercial yeasts through the manipulation of sulfur metabolism that are better suited towards production of fruit-driven styles.

  7. 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.

  8. 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.

  9. Development and application of a general plasmid reference material for GMO screening.

    Science.gov (United States)

    Wu, Yuhua; Li, Jun; Wang, Yulei; Li, Xiaofei; Li, Yunjing; Zhu, Li; Li, Jun; Wu, Gang

    The use of analytical controls is essential when performing GMO detection through screening tests. Additionally, the presence of taxon-specific sequences is analyzed mostly for quality control during GMO detection. In this study, 11 commonly used genetic elements involving three promoters (P-35S, P-FMV35S and P-NOS), four marker genes (Bar, NPTII, HPT and Pmi), and four terminators (T-NOS, T-35S, T-g7 and T-e9), together with the reference gene fragments from six major crops of maize, soybean, rapeseed, rice, cotton and wheat, were co-integrated into the same single plasmid to construct a general reference plasmid pBI121-Screening. The suitability test of pBI121-Screening plasmid as reference material indicated that the non-target sequence on the pBI121-Screening plasmid did not affect the PCR amplification efficiencies of screening methods and taxon-specific methods. The sensitivity of screening and taxon-specific assays ranged from 5 to 10 copies of pBI121-Screening plasmid, meeting the sensitivity requirement of GMO detection. The construction of pBI121-Screening solves the lack of a general positive control for screening tests, thereby reducing the workload and cost of preparing a plurality of the positive control.

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

    OpenAIRE

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

    1995-01-01

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

  11. 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...

  12. Replication of plasmids in gram-negative bacteria.

    OpenAIRE

    1989-01-01

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

  13. 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.

  14. Identification of significant medium components that affect docosahexaenoic acid production by Schizochytrium sp. SW1

    Science.gov (United States)

    Manikan, Vidyah; Hamid, Aidil A.

    2013-11-01

    Central composite design (CCD) was employed to investigate the significance of glucose, yeast extract, MSG and sea salt in affecting the amount of docosahexaenoic acid (DHA) accumulated by a locally isolated strain of Schizochytrium. Design Expert software was used to construct a set of experiments where each medium component mentioned above was varied over three levels. Cultivation was carried out in 250mL flasks containing 50mL of medium, incubated at 30°C with 200 rpm agitation for 96 hours. ANOVA was conducted to identify the influential factors and the level of their significance where factors that scored a probability value of less than 0.05 were considered significant. The level of influence for each independent variable was also interpreted using perturbation whereas pattern of interaction between the factors were interpreted using interaction plots. This experiment revealed that yeast extract and monosodium glutamate have significant influence on DHA accumulation process by Schizochytrium sp. SW1.

  15. Social Factors Affecting Wetlands Utilization for Agriculture in Nigeria: A case study of sawah rice production

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Wetlands have immense poverty-fighting potentials and in Nigeria,more and more people are dependent on wetlands for their livelihoods.To examine the social factors affecting the current status of the wetlands utilization for agriculture in Nigeria,a simple random sampling technique was used to select 200 farmers cultivating wetlands and a structured questionnaire was applied to elicit the information on the social factors.Data collected were described using frequency and percentage and a multiple regression analysis was used to identify significant variables that are determinants of wetland utilization.The results of the analysis showed that significant variables included crop preferences,farming system,culture,taste,land tenure,knowledge of wetland cultivation,perceived suitability,farmers' tribe,location of wetland,and farmers' age.It was concluded with suggestions for the right combination of policies,public awareness,and appropriate farming methods in order to improve wetland utilization in Nigeria.

  16. Do new cellulolytic enzyme preparations affect the industrial strategies for high solids lignocellulosic ethanol production?

    Science.gov (United States)

    Cannella, David; Jørgensen, Henning

    2014-01-01

    Production of ethanol from lignocellulosic materials has a promising market potential, but the process is still only at pilot/demonstration scale due to the technical and economical difficulties of the process. Operating the process at very high solids concentrations (above 20% dry matter-DM) has proven essential for economic feasibility at industrial scale. Historically, simultaneous saccharification and fermentation (SSF) was found to give better ethanol yields compared to separate hydrolysis and fermentation (SHF), but data in literature are typically based on operating the process at low dry matter conditions. In this work the impact of selected enzyme preparation and processing strategy (SHF, presaccharification and simultaneous saccharification and fermentation-PSSF, and SSF) on final ethanol yield and overall performance was investigated with pretreated wheat straw up to 30% DM. The experiments revealed that an SSF strategy was indeed better than SHF when applying an older generation enzyme cocktail (Celluclast-Novozym 188). In case of the newer product Cellic CTec 2, SHF resulted in 20% higher final ethanol yield compared to SSF. It was possible to close the mass balance around cellulose to around 94%, revealing that the most relevant products could be accounted for. One observation was the presence of oxidized sugar (gluconic acid) upon enzymatic hydrolysis with the latest enzyme preparation. Experiments showed gluconic acid formation by recently discovered enzymatic class of lytic polysaccharides monoxygenases (LPMO's) to be depending on the processing strategy. The lowest concentration was achieved in SSF, which could be correlated with less available oxygen due to simultaneous oxygen consumption by the yeast. Quantity of glycerol and cell mass was also depending on the selected processing strategy.

  17. Evaluation of Factors Affecting Polyhydroxyalkanoates Production by Comamonas sp. EB172 Using Central Composite Design

    Directory of Open Access Journals (Sweden)

    Noor Azman Mohd Johar

    2012-09-01

    Full Text Available Aims: Statistical approach, central composite design (CCD was used to investigate the complex interaction among temperature (25-37 °C, initial medium pH (5-9, inoculum size (4-10 % (v/v, concentration of (NH42SO4 (0-1 g/L and concentration of mixed organic acids (5-10 g/L in the production of polyhydroxyalkanoates by Comamonas sp. EB172.Methodology and Results: Mixed organic acids derived from anaerobically treated palm oil mill effluent (POME containing acetic:propionic:butyric (ratio of 3:1:1 were used as carbon source in the batch culture of Comamonas sp. EB172 to produce polyhydoxyalkanoates (PHAs. The analysis of variance (ANOVA showed that all five factors were significantly important in the batch fermentation by shake flask with a P value of less than 0.001. The optimal temperature, initial medium pH, inoculum size, concentration of (NH42SO4 and concentration of mixed organic acids were 30 °C, 7.04, 4.0 % (v/v, 0.01 g/L and 5.05 g/L respectively.Conclusion, significance and impact of study: Optimization of the production medium containing mixed organic acids has improved the PHA production for more than 2 folds. Under optimal condition in the shake flask fermentation, the predicted growth is 2.98 g/L of dry cell weight (DCW with 47.07 wt % of PHA content. The highest yield of PHA was 0.28 g of PHA per g mixed organic acids.

  18. Population structure and fruit production of Pyrus bourgaeana D. are affected by land-use

    Science.gov (United States)

    Arenas-Castro, Salvador; Fernández-Haeger, Juan; Jordano-Barbudo, Diego

    2016-11-01

    The Iberian wild pear (Pyrus bourgaeana D.) is a rare, fleshy-fruited tree restricted to dehesas and evergreen sclerophyllous Mediterranean forests in the southwestern Iberian Peninsula. It produces palatable fruits and leaves attractive to different species groups, playing an important trophic role in the ecological networks of Mediterranean ecosystems. However, the intensification in the traditional land-use linked to these areas could threaten the stability of the wild pear populations in the short/medium-term. In order to determine the population dynamics of this relevant species in relation to the land-use history, we selected two populations (southern Spain) subjected to different land-use management, dehesa (D) and abandoned olive grove (AOG). An analysis of 122 adult trees reported an overall density of 0.6 trees ha-1. The tree age was estimated by tree-rings analysis in all adult trees. Dendrometric parameters, reproductive features, and germination rates were also measured. Regeneration was clearly biased, as evidenced by the truncated age structure. A low correlation (R2 = 34%) between age and DBH (diameter at breast height) (244 cores analysed) showed that diameter seems not to be a reliable predictor of tree age. Trees from AOG populations had significantly-higher values of DBH, height and crown diameter, but were less productive in terms of fruits and seeds. Nested analysis of variance showed significant variation in fruit production, fruit size, dry mass, water content and seed viability. There were also significant differences in masting. No evidence was found to demonstrate that fruit production, seed viability, or germination rate influence the low natural recruitment of this species. These findings indicate that the traditional agrosilvopastoral practices carried out in the study area for decades, and its subsequent intensification, have strongly influenced the ecological structure of the Iberian wild pear populations at the local scale, which

  19. Tetrazolium-based assays for cellular viability: a critical examination of selected parameters affecting formazan production.

    Science.gov (United States)

    Vistica, D T; Skehan, P; Scudiero, D; Monks, A; Pittman, A; Boyd, M R

    1991-05-15

    The hydrogen acceptor 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) is commonly utilized to estimate cellular viability in drug screening protocols. The present investigation was prompted, in part, by observations that reduction of MTT to its colored reaction product, MTT formazan, varied between cell lines and with culture age. A correlation was established between the D-glucose concentration of the culture medium at the time of assay and the production of MTT formazan for cell lines representing seven tumor histologies. A decrease in the concentration of D-glucose from culture medium was accompanied by a decrease in MTT specific activity (MTT formazan/microgram cell protein) for a number of cell lines. Cells which extensively metabolized D-glucose exhibited the greatest reduction in MTT specific activity. Further evidence that the D-glucose concentration of the culture medium played an important role in MTT reduction was provided by experiments which demonstrated that transfer of cells to a glucose-free medium (L-15) was accompanied by an immediate decrease in MTT reduction which was pH independent. These studies suggested that cellular transport and constant metabolism of glucose were required for maximum MTT reduction. Decreases in the cellular concentration of the reduced pyridine nucleotides NADH and NADPH were accompanied by concomitant decreases in MTT formazan production. MTT formazan varied significantly among cell lines in both the kinetics of its formation and the degree of saturability exhibited. Apparent IC50 values for Adriamycin varied, in a cell line-specific manner, with MTT exposure time. These results indicate that MTT specific activity is significantly influenced by a number of parameters and suggest that assay conditions should be established which minimize their effects.

  20. Nutrient availability affects pigment production but not growth in lichens of biological soil crusts

    Science.gov (United States)

    Bowker, M.A.; Koch, G.W.; Belnap, J.; Johnson, N.C.

    2008-01-01

    Recent research suggests that micronutrients such as Mn may limit growth of slow-growing biological soil crusts (BSCs) in some of the drylands of the world. These soil surface communities contribute strongly to arid ecosystem function and are easily degraded, creating a need for new restoration tools. The possibility that Mn fertilization could be used as a restoration tool for BSCs has not been tested previously. We used microcosms in a controlled greenhouse setting to investigate the hypothesis that Mn may limit photosynthesis and consequently growth in Collema tenax, a dominant N-fixing lichen found in BSCs worldwide. We found no evidence to support our hypothesis; furthermore, addition of other nutrients (primarily P, K, and Zn) had a suppressive effect on gross photosynthesis (P = 0.05). We also monitored the growth and physiological status of our microcosms and found that other nutrients increased the production of scytonemin, an important sunscreen pigment, but only when not added with Mn (P = 0.01). A structural equation model indicated that this effect was independent of any photosynthesis-related variable. We propose two alternative hypotheses to account for this pattern: (1) Mn suppresses processes needed to produce scytonemin; and (2) Mn is required to suppress scytonemin production at low light, when it is an unnecessary photosynthate sink. Although Mn fertilization does not appear likely to increase photosynthesis or growth of Collema, it could have a role in survivorship during environmentally stressful periods due to modification of scytonemin production. Thus, Mn enrichment should be studied further for its potential to facilitate BSC rehabilitation. ?? 2008 Elsevier Ltd.

  1. Magnetic nanowires and hyperthermia: How geometry and material affect heat production efficiency

    KAUST Repository

    Contreras, Maria F.

    2015-05-01

    Magnetic hyperthermia, which refers to the production of heat by magnetic nanostructures under an alternating magnetic field (AMF), has been previously investigated with superparamagnetic nanobeads as a cancer therapy method. Magnetic nanowires (NWs) used in hyperthermia can be very promising, as it has been shown that they have a larger magnetic moment per unit of volume compared to the nanobeads. Moreover, Fe NWs proved to have a higher heating efficiency compared to Fe nanobeads, when exposed to an AMF at the same concentration [1].

  2. Evaluation of Factors Affecting Uricase Production by the Screened Wild/Natural Microbes

    OpenAIRE

    Harinath Dwivedi; Kusum Agrawal; Saraf, Shubhini A.

    2012-01-01

    Gout is a metabolic disorder due to the deposition of uric acid crystals within articular or periarticular tissues. Uricase (urate oxidase) catalyzes the oxidation of less water soluble uric acid (7 mg/dl) to a compound allantoin which is more water soluble(11g/L at 40°C) resulting into the ease of excretion of uric acid. The objective of the work was to develop a new method for screening of microbes for uricase production and estimation of uricase thereof. This was achieved by utilizing the ...

  3. TNT transformation products are affected by the growth conditions of Raoultella terrigena.

    Science.gov (United States)

    Claus, Harald; Perret, Nina; Bausinger, Tobias; Fels, Gregor; Preuss, Johannes; König, Helmut

    2007-03-01

    High concentrations of 2,4,6-trinitrotoluene (TNT) and related nitroaromatic compounds are commonly found in soil and groundwater at former explosive plants. The bacterium, Raoultella terrigena strain HB, isolated from a contaminated site, converts TNT into the corresponding amino products. Radio-HPLC analysis with [(14)C]TNT identified aminodinitrotoluene, diaminonitrotoluene and azoxy-dimers as the main metabolites. Transformation rate and the type of metabolites that predominated in the culture medium and within the cells were significantly influenced by the culture conditions. The NAD(P)H-dependent enzymatic reduction of nitro-substituted compounds by cell-free extracts of R. terrigena was evaluated in vitro.

  4. Plasmid Segregation: Spatial Awareness at the Molecular Level

    DEFF Research Database (Denmark)

    Møller-Jensen, Jakob; Gerdes, Kenn

    2007-01-01

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

  5. Microbiological and physicochemical factors affecting Aspergillus section Flavi incidence in Cavendish banana (Musa cavendishii) chips production in Southern Philippines.

    Science.gov (United States)

    Sales, A C; Azanza, P V; Yoshizawa, T

    2005-01-01

    Microbiological and physicochemical factors affecting the incidence of Aspergillus section Flavi in dried Cavendish banana (Musa cavendishii) chips production in Southern Philippines were examined. The average counts of Aspergillus section Flavi (AFC) in fresh and dried Cavendish bananas from 10 production batches of the Philippine Agro-Industrial Development Cooperative in Davao del Norte, Southern Philippines were 1.2 x 10(2) and 1.6 x 10(2) cfu/g, respectively. Isolates from both samples were identified to be Aspergillus flavus based on spore type and conidial structure of isolates. An increasing trend in the AFC of Cavendish bananas was observed during dried banana chips processing. Variability in the AFC between production batches was attributed to differences in aerobic and fungal populations and physicochemical characteristics of the fruits, peel damage of the raw materials, concentration of AFC in the air and food-contact surfaces of the production area, and temperature and relative humidity (RH) conditions of the environment during production and storage. Physicochemical characteristics of Cavendish bananas from the receipt of raw materials up to the first day of drying were within the reported range of values allowing growth and toxin production by aflatoxigenic fungi. Air-borne AFC varied depending on the section of the production area examined. The close proximity of the waste disposal area from the production operation to the preparation, drying and storage areas suggests that cross-contamination, probably air-borne or insect-borne was a likely occurrence. The hands of workers were also identified as AFC sources. Results of this study highlight the need for the development of strategies to control aflatoxigenic fungi and aflatoxin contamination in Philippine dried Cavendish bananas.

  6. Gas exchange rates, plant height, yield components, and productivity of upland rice as affected by plant regulators

    Directory of Open Access Journals (Sweden)

    Rita de Cássia Félix Alvarez

    2012-10-01

    Full Text Available The objective of this work was to evaluate gas exchange rates, plant height, yield components, and productivity of upland rice, as affected by type and application time of plant growth regulators. A randomized block design, in a 4x2 factorial arrangement, with four replicates was used. Treatments consisted of three growth regulators (mepiquat chloride, trinexapac-ethyl, and paclobutrazol, besides a control treatment applied at two different phenological stages: early tillering or panicle primordial differentiation. The experiment was performed under sprinkler-irrigated field conditions. Net CO2 assimilation, stomatal conductance, plant transpiration, and water-use efficiency were measured four times in Primavera upland rice cultivar, between booting and milky grain phenophases. Gas exchange rates were neither influenced by growth regulators nor by application time. There was, however, interaction between these factors on the other variables. Application of trinexapac-ethyl at both tillering and differentiation stages reduced plant height and negatively affected yield components and rice productivity. However, paclobutrazol and mepiquat chloride applied at tillering, reduced plant height without affecting rice yield. Mepiquat chloride acted as a growth stimulator when applied at the differentiation stage, and significantly increased plant height, panicle number, and grain yield of upland rice.

  7. Bioaugmentation of Hydrogenispora ethanolica LX-B affects hydrogen production through altering indigenous bacterial community structure.

    Science.gov (United States)

    Yang, Zhiman; Guo, Rongbo; Shi, Xiaoshuang; He, Shuai; Wang, Lin; Dai, Meng; Qiu, Yanling; Dang, Xiaoxiao

    2016-07-01

    Bioaugmentation can facilitate hydrogen production from complex organic substrates, but it still is unknown how indigenous microbial communities respond to the added bacteria. Here, using a Hydrogenispora ethanolica LX-B (named as LX-B) bioaugmentation experiments, the distribution of metabolites and the responses of indigenous bacterial communities were investigated via batch cultivation (BC) and repeated batch cultivation (RBC). In BC the LX-B/sludge ratio of 0.12 achieved substantial high hydrogen yield, which was over twice that of control. In RBC one-time bioaugmentation and repeated batch bioaugmentation of LX-B resulted in the hydrogen yield that was average 1.2-fold and 0.8-fold higher than that in control, respectively. This improved hydrogen production performance mainly benefited from a shift in composition of the indigenous bacterial community caused by LX-B bioaugmentation. The findings represented an important step in understanding the relationship between bioaugmentation, a shift in bacterial communities, and altered bioreactor performance.

  8. Factors affecting catalysis of copper corrosion products in NDMA formation from DMA in simulated premise plumbing.

    Science.gov (United States)

    Zhang, Hong; Andrews, Susan A

    2013-11-01

    This study investigated the effects of corrosion products of copper, a metal commonly employed in household plumbing systems, on N-nitrosodimethylamine (NDMA) formation from a known NDMA precursor, dimethylamine (DMA). Copper-catalyzed NDMA formation increased with increasing copper concentrations, DMA concentrations, alkalinity and hardness, but decreased with increasing natural organic matter (NOM) concentration. pH influenced the speciation of chloramine and the interactions of copper with DMA. The transformation of monochloramine (NH2Cl) to dichloramine and complexation of copper with DMA were involved in elevating the formation of NDMA by copper at pH 7.0. The inhibiting effect of NOM on copper catalysis was attributed to the rapid consumption of NH2Cl by NOM and/or the competitive complexation of NOM with copper to limit the formation of DMA-copper complexes. Hardness ions, as represented by Ca(2+), also competed with copper for binding sites on NOM, thereby weakening the inhibitory effect of NOM on NDMA formation. Common copper corrosion products also participated in these reactions but in different ways. Aqueous copper released from malachite [Cu2CO3(OH)2] was shown to promote NDMA formation while NDMA formation decreased in the presence of CuO, most likely due to the adsorption of DMA.

  9. Bc、Bt、Ba 质粒研究进展%Plasmids Research Progress of Bc,Bt and Ba

    Institute of Scientific and Technical Information of China (English)

    王品舒; 岳瑾; 乔岩; 董杰; 张金良; 袁志强; 杨建国∗

    2013-01-01

      质粒是蜡样芽胞杆菌群中蜡样芽胞杆菌、苏云金芽胞杆菌和炭疽芽胞杆菌呈现出不同表型特征和病理效果的重要原因。本文对三种细菌质粒 Bc、Bt、Ba 的研究进展进行了综述,重点阐述了质粒与毒素的关系、质粒的接合转移、插入序列、代表性质粒的研究进展,分析了质粒研究在公共卫生安全、生物防治及转基因技术发展与应用中的重要作用,最后展望了质粒研究的方向。%  Plasmids have important role in phenotype characteristics and pathological effects of Bacillus cereus( Bc), Bacillus thuringiensis(Bt) and Bacillus anthracis ( Ba). This paper reviewed the research progress of plasmids of the three kinds of Bacillus, in which we mainly elaborated the relationship between the plasmids and the toxin production, plasmids conjugal transfer, inserted sequence, and typical plasmids. We analyzed the important role of plasmids research in public health security, biological control and transgenic technology. Finally, We prospected the direction of the plasmids research.

  10. [Effect of plasmid pKM101 on the expression of bacterial genes not related to DNa metabolism].

    Science.gov (United States)

    Skavronskaya, A G; Tiganova, I G; Andreeva, I V; Rusina, O Iu

    1999-02-01

    An experimental system ensuring fusion of bacterial genes to the lac operon of the Mu dl(Aplac) phage was used. Fusion operons in which the lac operon was under the control of promoters of the elt gene, responsible for synthesis of the LT toxin, of the tetracyclin-resistance tet gene, and sfiA gene encoding filament production, was studied. Using this experimental system, plasmid pKM101 was shown to be capable of activating the expression of the above Escherichia coli and Salmonella typhimurium genes, which is manifested as the activation of beta-galactosidase synthesis. The activation of the elt gene expression by the pKM101 plasmid was also confirmed in experiments on detecting the LT toxin synthesized by bacteria carrying this plasmid. Effect of the plasmid on the activation of elt operon expression, unlike the effect of this plasmid on mutability, does not depend on the functioning of the lexA and recA genes, i.e., this is not a SOS-regulated process. The mutant plasmid pGW12, a derivative of pKM101, deficient in the mucAB genes responsible for mutagenesis, causes a more pronounced activation of the elt gene than plasmid pKM101.

  11. Organic loading rates affect composition of soil-derived bacterial communities during continuous, fermentative biohydrogen production

    Energy Technology Data Exchange (ETDEWEB)

    Luo, Yonghua; Bruns, Mary Ann [Department of Crop and Soil Sciences, The Pennsylvania State University, University Park, PA 16802 (United States); Zhang, Husen; Salerno, Michael; Logan, Bruce E. [Department of Civil and Environmental Engineering, The Pennsylvania State University, University Park, PA 16802 (United States)

    2008-11-15

    Bacterial community composition during steady-state, fermentative H{sub 2} production was compared across a range of organic loading rates (OLRs) of 0.5-19 g COD l{sup -1} h{sup -1} in a 2-l continuous flow reactor at 30 C. The varied OLRs were achieved with glucose concentrations of 2.5-10 g l{sup -1} and hydraulic retention times of 1-10 h. The synthetic wastewater feed was amended with L-cysteine and maintained at a pH of 5.5. For each run at a given glucose concentration, the reactor was inoculated with an aliquot of well-mixed agricultural topsoil that had been heat-treated to reduce numbers of vegetative cells. At OLRs less than 2 g COD l{sup -1} h{sup -1}, DNA sequences from ribosomal RNA intergenic spacer analysis profiles revealed more diverse and variable populations (Selenomonas, Enterobacter, and Clostridium spp.) than were observed above 2 g COD l{sup -1} h{sup -1} (Clostridium spp. only). An isolate, LYH1, was cultured from a reactor sample (10 g glucose l{sup -1} at a 10-h HRT) on medium containing L-cysteine. In confirming H{sub 2} production by LYH1 in liquid batch culture, lag periods for H{sub 2} production in the presence and absence of L-cysteine were 5 and 50 h, respectively. The 16S rRNA gene sequence of LYH1 indicated that the isolate was a Clostridium sp. affiliated with RNA subcluster Ic, with >99% similarity to Clostridium sp. FRB1. In fluorescent in situ hybridization tests, an oligonucleotide probe complementary to the 16S rRNA of LYH1 hybridized with 90% of cells observed at an OLR of 2 g COD h{sup -1}, compared to 26% of cells at an OLR of 0.5 g COD l{sup -1} h{sup -1}. An OLR of 2 g COD l{sup -1} h{sup -1} appeared to be a critical threshold above which clostridia were better able to outcompete Enterobacteriaceae and other organisms in the mixed soil inoculum. Our results are discussed in light of other biohydrogen studies employing pure cultures and mixed inocula. (author)

  12. UV-B affects photosynthesis, ROS production and motility of the freshwater flagellate, Euglena agilis Carter.

    Science.gov (United States)

    Kottuparambil, Sreejith; Shin, Woongghi; Brown, Murray T; Han, Taejun

    2012-10-15

    The effects of ultraviolet B (UV-B; 295-320 nm) radiation on certain vital physiological (photosynthesis), biochemical (production of reactive oxygen species - ROS) and behavioral (motility and orientation) characteristics were investigated in the unicellular photoautotroph, Euglena agilis Carter. The photosynthetic performance of E. agilis was recorded after exposure of between 15 and 60 min followed by a period of recovery lasting 6-24h under dim light (5-10 μmol photons m(-2) s(-1)). The maximum quantum yield of PS II (F(v)/F(m)) was reduced to 65% and 14% of initial values immediately following 15 and 30 min UV-B exposure, but recovered to 100 and 86% of the initials, respectively. Values of rETR(max) in E. agilis exposed to 15 min UV-B were similar to those of the initials, but a 30 min UV exposure resulted in 75% reduction of rETR(max) with only a 43% recovery as compared with the initial after 24h recovery. After a 60 min UV-B exposure, there were no Chl a fluorescence signals, and hence no F(v)/F(m) or rETR(max). A UV dose-dependent increase in DCFH-DA fluorescence was found in E. agilis cells, reflecting an increase in ROS production. After exposures to UV-B for between 15 and 60 min, the percentages of motile cells in the population decreased to 76, 39 and 15%, respectively. Following 24h in dim light, the percentage of motile cells increased to between 66% and 95% of the initial value. The velocity of non-irradiated cells was 60 μm s(-1), which decreased to 16-35 μm s(-1) immediately following exposure for 15-60 min. After periods of time in dim light (6, 12 and 24h) velocities had recovered to between 44 and 81% of the initial value. In untreated controls, the r-value was 0.23, indicating random movement of E. agilis, but it increased to 0.35 and 0.72 after exposure to UV-B for 30 and 60 min, respectively. There was a tendency towards vertical downward movement of cells proportional to the duration of exposure. The compactness of E. agilis decreased

  13. Characterization of an IncA/C Multidrug Resistance Plasmid in Vibrio alginolyticus.

    Science.gov (United States)

    Ye, Lianwei; Li, Ruichao; Lin, Dachuan; Zhou, Yuanjie; Fu, Aisi; Ding, Qiong; Chan, Edward Wai Chi; Yao, Wen; Chen, Sheng

    2016-05-01

    Cephalosporin-resistant Vibrio alginolyticus was first isolated from food products, with β-lactamases encoded by blaPER-1, blaVEB-1, and blaCMY-2 being the major mechanisms mediating their cephalosporin resistance. The complete sequence of a multidrug resistance plasmid, pVAS3-1, harboring the blaCMY-2 and qnrVC4 genes was decoded in this study. Its backbone exhibited genetic homology to known IncA/C plasmids recoverable from members of the family Enterobacteriaceae, suggesting its possible origin in Enterobacteriaceae.

  14. Plasmid Content of Isolates of Erwinia amylovora from Orchards in Washington and Oregon in the USA

    Science.gov (United States)

    Washington (WA) and Oregon (OR) represent a major pome fruit production region of the United States, and streptomycin-resistant isolates of the fire blight pathogen Erwinia amylovora are common in orchards in this region. We examined the plasmid content of a collection of more than 200 isolates of ...

  15. Seedling quality of common sage (Salvia officinalis L. as affected by seedling production methods

    Directory of Open Access Journals (Sweden)

    Ömer Çalişkan

    2006-09-01

    Full Text Available The study was conducted to determine the effects of four different seedling production methods (i.e., open seedbed, greenhouse seedbed, float system and modified float system on the seedling growth and quality of common sage. Sowing rates were 1 g per m2 in an open seedbed and a greenhouse seedbed system, whereas the sowing rate was one seed per cell in the float and modified float systems. Emergence period, seedling growth period, the fresh and dry weights of aerial parts of seedlings, the fresh and dry weights of roots, seedling height, root height and stem diameter were determined as agronomical observations and measurements. The greenhouse seedling method was found to be superior over the other methods

  16. Factors Affecting the Production of Aromatic Immonium Ions in MALDI 157 nm Photodissociation Studies

    Science.gov (United States)

    DeGraan-Weber, Nick; Ashley, Daniel C.; Keijzer, Karlijn; Baik, Mu-Hyun; Reilly, James P.

    2016-05-01

    Immonium ions are commonly observed in the high energy fragmentation of peptide ions. In a MALDI-TOF/TOF mass spectrometer, singly charged peptides photofragmented with 157 nm VUV light yield a copious abundance of immonium ions, especially those from aromatic residues. However, their intensities may vary from one peptide to another. In this work, the effect of varying amino acid position, peptide length, and peptide composition on immonium ion yield is investigated. Internal immonium ions are found to have the strongest intensity, whereas immonium ions arising from C-terminal residues are the weakest. Peptide length and competition among residues also strongly influence the immonium ion production. Quantum calculations provide insights about immonium ion structures and the fragment ion conformations that promote or inhibit immonium ion formation.

  17. Small doses, big troubles: modeling growth dynamics of organisms affecting microalgal production cultures in closed photobioreactors.

    Science.gov (United States)

    Forehead, Hugh I; O'Kelly, Charles J

    2013-02-01

    The destruction of mass cultures of microalgae by biological contamination of culture medium is a pervasive and expensive problem, in industry and research. A mathematical model has been formulated that attempts to explain contaminant growth dynamics in closed photobioreactors (PBRs). The model simulates an initial growth phase without PBR dilution, followed by a production phase in which culture is intermittently removed. Contaminants can be introduced at any of these stages. The model shows how exponential growth from low initial inocula can lead to "explosive" growth in the population of contaminants, appearing days to weeks after inoculation. Principal influences are contaminant growth rate, PBR dilution rate, and the size of initial contaminant inoculum. Predictions corresponded closely with observed behavior of two contaminants, Uronema sp. and Neoparamoeba sp., found in operating PBRs. A simple, cheap and effective protocol was developed for short-term prediction of contamination in PBRs, using microscopy and archived samples.

  18. Factors affecting production of nonaqueous peracetic acid in tubular packed reactors

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    The synthesis of nonaqueous peracetic acid in acetone by acetaldehyde oxidation was carried out in a tubular packed reactor.The influencing factors of the reacting system including packing material,oxygen carrier,and reactor configuration were investigated.The results show that porous materials are inappropriate for peracetic acid synthesis and only non porous material with appropriate surface area can provide good peracetic acid selectivity and yield.Among the six kinds of packing material investigated,SA-5118 is the best one.As oxidizing gas,pure oxygen is superior to air.The optimum length-to-inner diameter ratio of the reactor is about 40.Under the proper reaction conditions,the highest peracetic acid yield of 84.15% and the highest selectivity of 93.34% can be achieved which indicates that the novel reacting system is effective and economical for nonaqueous peracetic acid production.

  19. The factors affecting the green brand equity of electronic products: Green marketing

    Directory of Open Access Journals (Sweden)

    Narges Delafrooz

    2015-12-01

    Full Text Available This study is intentionally functional, and, from the data collection aspect, is a descriptive survey. The statistical data for the present study includes low-power electronic and electric products in Guilan Province. A total of 384 consumers were chosen using the random cluster sampling method. We have used confirmatory factor analysis and structural equation modeling to analyze the given data. The findings show that there is a significant relationship among brand-perceived quality, green brand image, (GBI and brand value, but not with the green brand-perceived value (GBPV. In addition, the results of the study show that brand credibility has a significant relationship with the GBPV and GBI, but not with the GBPV and GBI, each of which are associated with the green brand in a different way. At the end of this survey, you will read about the intermediate variables, all of which are acceptable, except the intermediate variables of brand credibility and GBI.

  20. Factors Affecting Embryogenic Callus Production and Plant Regeneration in Anther Culture of Bupleurum chinense

    Institute of Scientific and Technical Information of China (English)

    YANG Cheng-min; ZHAO Yu-kai; WEI Jian-he; ZHAO Li-zi; SUI Chun; ZHANG Zheng; CUI Lu-ying

    2011-01-01

    Objective To evaluate the influences of the genotypes,anther developmental stages,and cultural conditions on the efficiency of embryogenic callus induction and plant regeneration in the anthers culture of Bupleurum chinense.Methods The different effects such as four genotypes,plant growth regulators,and temperature condition were compared in the experiments.The histological study was performed with the process of the anther culture.Results The highest inducing rate of embryogenic calli were achieved for the genotypes Zhongcaiyihao(ZCYH),Z4,and Z5 at the early-to middle-uninucleate stages,except for genotype ZPM1 at the tetrad stage.Cold pretreatment increased the production of the embryogenic callus,in which 4-day cold pretreatment improved the production of embryogenic callus from 0% to 2.2% and 5.0% for genotypes ZPM1 and ZCYH,respectively.No embryogenic callus was induced in the medium containing less than 0.75 mg/L 2,4-dichlorophenoxyacetic acid(2,4-D).The highest regeneration rate (34.6%)was obtained in 1/2 MS salts regeneration medium supplemented with 0.1 mg/L 6-benzylmaminopurine (BA).The low concentration of BA was able to promote the embryogenic callus formation and subsequent plantlet regeneration via somatic embryogenesis.Chromosome counting of regenerated plantlets showed mostly diploid plant (2n = 12)with only one haploid plant(n = 6).Because of the low rate of microspore embryo formation,we only tracked the process of embryogenesis from the connective tissue,instead of microspore by histological observations.Conclusion This study establishes an efficient system for embryogenic callus induction and plant regeneration system.This is the first report on the haploid plantlet through the anther culture orB.chinense.

  1. Disruption of quercetin metabolism by fungicide affects energy production in honey bees (Apis mellifera).

    Science.gov (United States)

    Mao, Wenfu; Schuler, Mary A; Berenbaum, May R

    2017-02-13

    Cytochrome P450 monooxygenases (P450) in the honey bee, Apis mellifera, detoxify phytochemicals in honey and pollen. The flavonol quercetin is found ubiquitously and abundantly in pollen and frequently at lower concentrations in honey. Worker jelly consumed during the first 3 d of larval development typically contains flavonols at very low levels, however. RNA-Seq analysis of gene expression in neonates reared for three days on diets with and without quercetin revealed that, in addition to up-regulating multiple detoxifying P450 genes, quercetin is a negative transcriptional regulator of mitochondrion-related nuclear genes and genes encoding subunits of complexes I, III, IV, and V in the oxidative phosphorylation pathway. Thus, a consequence of inefficient metabolism of this phytochemical may be compromised energy production. Several P450s metabolize quercetin in adult workers. Docking in silico of 121 pesticide contaminants of American hives into the active pocket of CYP9Q1, a broadly substrate-specific P450 with high quercetin-metabolizing activity, identified six triazole fungicides, all fungal P450 inhibitors, that dock in the catalytic site. In adults fed combinations of quercetin and the triazole myclobutanil, the expression of five of six mitochondrion-related nuclear genes was down-regulated. Midgut metabolism assays verified that adult bees consuming quercetin with myclobutanil metabolized less quercetin and produced less thoracic ATP, the energy source for flight muscles. Although fungicides lack acute toxicity, they may influence bee health by interfering with quercetin detoxification, thereby compromising mitochondrial regeneration and ATP production. Thus, agricultural use of triazole fungicides may put bees at risk of being unable to extract sufficient energy from their natural food.

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

    Science.gov (United States)

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

    2011-12-23

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

  3. Bacillus cereus and Bacillus thuringiensis spores in Korean rice: prevalence and toxin production as affected by production area and degree of milling.

    Science.gov (United States)

    Kim, Booyoung; Bang, Jihyun; Kim, Hoikyung; Kim, Yoonsook; Kim, Byeong-Sam; Beuchat, Larry R; Ryu, Jee-Hoon

    2014-09-01

    We determined the prevalence of and toxin production by Bacillus cereus and Bacillus thuringiensis in Korean rice as affected by production area and degree of milling. Rough rice was collected from 64 farms in 22 agricultural areas and polished to produce brown and white rice. In total, rice samples were broadly contaminated with B. cereus spores, with no effect of production area. The prevalence and counts of B. cereus spores declined as milling progressed. Frequencies of hemolysin BL (HBL) production by isolates were significantly (P ≤ 0.01) reduced as milling progressed. This pattern corresponded with the presence of genes encoding the diarrheal enterotoxins. The frequency of B. cereus isolates positive for hblC, hblD, or nheB genes decreased as milling progressed. Because most B. cereus isolates from rice samples contained six enterotoxin genes, we concluded that B. cereus in rice produced in Korea is predominantly of the diarrheagenic type. The prevalence of B. thuringiensis in rice was significantly lower than that of B. cereus and not correlated with production area. All B. thuringiensis isolates were of the diarrheagenic type. This study provides information useful for predicting safety risks associated with B. cereus and B. thuringiensis in rough and processed Korean rice.

  4. UV-B affects photosynthesis, ROS production and motility of the freshwater flagellate, Euglena agilis Carter

    Energy Technology Data Exchange (ETDEWEB)

    Kottuparambil, Sreejith [Institute of Green Environmental Research Center, University of Incheon, Incheon, 406 840 (Korea, Republic of); Shin, Woongghi [Department of Biology, Chungnam University, Daejeon, 306 764 (Korea, Republic of); Brown, Murray T. [School of Marine Science and Engineering, Plymouth University, Drake Circus, Plymouth PL4 8AA (United Kingdom); Han, Taejun, E-mail: hanalgae@hanmail.net [Institute of Green Environmental Research Center, University of Incheon, Incheon, 406 840 (Korea, Republic of); Department of Marine Science, University of Incheon, Incheon, 406 840 (Korea, Republic of)

    2012-10-15

    Highlights: Black-Right-Pointing-Pointer We proposed a hypothesis for the UV-B protective/adaptive mechanism in Euglena agilis. After moderate levels of UV-B radiation, ROS plays a signaling role to shut down photosynthetic system for protection against harmful UV radiation. Black-Right-Pointing-Pointer E. agilis exposed to excessive UV appears to become animal-like, investing all its stored energy into movement rather than into sustaining its photosynthetic machinery. Black-Right-Pointing-Pointer This adaptation allows E. agilis to avoid harmful UV and seek a safe place where the organism may regain its photosynthetic capacity for survival. - Abstract: The effects of ultraviolet B (UV-B; 295-320 nm) radiation on certain vital physiological (photosynthesis), biochemical (production of reactive oxygen species - ROS) and behavioral (motility and orientation) characteristics were investigated in the unicellular photoautotroph, Euglena agilis Carter. The photosynthetic performance of E. agilis was recorded after exposure of between 15 and 60 min followed by a period of recovery lasting 6-24 h under dim light (5-10 {mu}mol photons m{sup -2} s{sup -1}). The maximum quantum yield of PS II (F{sub v}/F{sub m}) was reduced to 65% and 14% of initial values immediately following 15 and 30 min UV-B exposure, but recovered to 100 and 86% of the initials, respectively. Values of rETR{sub max} in E. agilis exposed to 15 min UV-B were similar to those of the initials, but a 30 min UV exposure resulted in 75% reduction of rETR{sub max} with only a 43% recovery as compared with the initial after 24 h recovery. After a 60 min UV-B exposure, there were no Chl a fluorescence signals, and hence no F{sub v}/F{sub m} or rETR{sub max}. A UV dose-dependent increase in DCFH-DA fluorescence was found in E. agilis cells, reflecting an increase in ROS production. After exposures to UV-B for between 15 and 60 min, the percentages of motile cells in the population decreased to 76, 39 and 15

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

    Science.gov (United States)

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

    2011-01-01

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

  6. Dietary inclusion of diallyl disulfide, yucca powder, calcium fumarate, an extruded linseed product, or medium-chain fatty acids does not affect methane production in lactating dairy cows.

    Science.gov (United States)

    van Zijderveld, S M; Dijkstra, J; Perdok, H B; Newbold, J R; Gerrits, W J J

    2011-06-01

    Two similar experiments were conducted to assess the effect of diallyl disulfide (DADS), yucca powder (YP), calcium fumarate (CAFU), an extruded linseed product (UNSAT), or a mixture of capric and caprylic acid (MCFA) on methane production, energy balance, and dairy cow performance. In experiment 1, a control diet (CON1) and diets supplemented with 56 mg of DADS/kg of dry matter (DM), 3g of YP/kg of DM, or 25 g of CAFU/kg of DM were evaluated. In experiment 2, an inert saturated fat source in the control diet (CON2) was exchanged isolipidically for an extruded linseed source (100g/kg of DM; UNSAT) or a mixture of C8:0 and C10:0 (MCFA; 20.3g/kg of DM). In experiment 2, a higher inclusion level of DADS (200mg/kg of DM) was also tested. Both experiments were conducted using 40 lactating Holstein-Friesian dairy cows. Cows were adapted to the diet for 12 d and were subsequently kept in respiration chambers for 5 d to evaluate methane production, diet digestibility, energy balance, and animal performance. Feed intake was restricted to avoid confounding effects of possible differences in ad libitum feed intake on methane production. Feed intake was, on average, 17.5 and 16.6 kg of DM/d in experiments 1 and 2, respectively. None of the additives reduced methane production in vivo. Methane production in experiment 1 was 450, 453, 446, and 423 g/d for CON1 and the diets supplemented with DADS, YP, and CAFU, respectively. In experiment 2, methane production was 371, 394, 388, and 386 g/d for CON2 and the diets supplemented with UNSAT, MCFA, and DADS, respectively. No effects of the additives on energy balance or neutral detergent fiber digestibility were observed. The addition of MCFA increased milk fat content (5.38% vs. 4.82% for control) and fat digestibility (78.5% vs. 59.8% for control), but did not affect milk yield or other milk components. The other products did not affect milk yield or composition. Results from these experiments emphasize the need to confirm methane

  7. Maize production as affected by sowing date, plant density and row spacing in the Bolivian Amazon

    Directory of Open Access Journals (Sweden)

    Paolo Casini

    2012-12-01

    Full Text Available The traditional cropping system widespread in the Department of Pando (Bolivian Amazon is the slash and burn methodology. The main crops sowed soon after the slash are maize, rice, cassava and common beans. Two separate field experiments (carried out in 2008-2009 were carried out to determine the agronomic responses of maize to sowing date, plant population and row width. For the first experiment a split-plot design was used. Maize cultivar (Bayo Blando and Perla Pandino was considered as main plots and the date of sowing as subplots. For the second experiment a split-split-plot design was used. Row spacing (0.5, 0.7 and 0.9 m was considered as main plot, maize cultivar (Cubano Amrarillo and Perla Pandino as subplots, and plant density (5.0, 7.5 and 10 plant m-2 as sub-subplots. A significant reduction of grain yield was observed as the date of sowing (DS delayied. Yield reduction of the second DS compared to the first, was 85 and 45% for Perla Pandino and Bayo Blando. The importance of plant density as a function of the correct row spacing is clearly shown. With the row spacing in use in the considered area (0.9 m and with the narrowest (0.5 m, the best yields were obtained with 10 plants m-2 (5.5 t ha-1. The following conclusions can be drawn from the present study: 1. A delay in the sowing date for maize by 15-20 days (compared to sowing ??immediately after the cutting of the virgin forest or the secondary forest strongly reduces grain production. The cultivar Perla Pandino was the most susceptible with a reduction of 85%. Late sowing of maize (mais de socorro, is suitable only if intercropped with other crops in order to protect the soil from erosion. Traditionally, rice and cassava are intercropped with maize, even if common beans or a legume cover crop would be more advisable. 2. The density of maize may be increased up until 10 m-2 in order to achieve the most productive results by using row spacings of 0.5 e 0.9 m, respectively

  8. Factors affecting production of compound A from the interaction of sevoflurane with Baralyme and soda lime.

    Science.gov (United States)

    Fang, Z X; Kandel, L; Laster, M J; Ionescu, P; Eger, E I

    1996-04-01

    Various alkali (e.g., soda lime) convert sevoflurane to CF2=C(CF3)OCH2F, a vinyl ether called "Compound A, " whose toxicity raises concerns regarding the safe administration of sevoflurane via rebreathing circuits. In the present investigation, we measured the sevoflurane degradation and output of Compound A caused by standard (13% water) Baralyme brand absorbent and standard (15% water) soda lime, and Baralyme and soda lime having various water contents (including no water). We used a flow-through system, applying a gas flow rate relative to absorbent volume that roughly equaled the rate/volume found in clinical practice. Both absorbents, at similar water contents, temperatures, and sevoflurane concentrations, produced roughly equal concentrations of Compound A. Dry and nearly dry absorbents produced less Compound A early in exposure to sevoflurane, and more later, than standard absorbents. Increases in temperature and sevoflurane concentration increased output of Compound A. Both absorbents, especially when dry, also destroyed Compound A, the concentration exiting from absorbent resulting from a complex sum of production and destruction. We conclude that the variability of concentrations of Compound A found in clinical practice may be largely explained by the inflow rate used (i.e., by rebreathing), sevoflurane concentration, and absorbent temperature and dryness. The effect of dryness is complex, with fresh dry absorbent destroying Compound A as it is made, and with dry absorbent that has been exposed to sevoflurane for a period of time providing a sometimes unusually high output of Compound A.

  9. Radio-transmitters do not affect seasonal productivity of female Golden-winged Warblers

    Science.gov (United States)

    Streby, Henry M.; Peterson, Sean M.; Gesmundo, Callie; Johnson, Michael K.; Fish, Alexander C.; Lehman, Justin A.; Andersen, David E.

    2013-01-01

    Investigating the potential effects of handling and marking techniques on study animals is important for correct interpretation of research results and to effect progress in data-collection methods. Few investigators have compared the reproductive output of radio-tagged and non-radio-tagged songbirds, and no one to date has examined the possible effect of radio-tagging adult songbirds on the survival of their fledglings. In 2011 and 2012, we compared several parameters of reproductive output of two groups of female Golden-winged Warblers (Vermivora chrysoptera) breeding in Minnesota, including 45 females with radio-transmitters and 73 females we did not capture, handle, or mark. We found no difference between groups in clutch sizes, hatching success, brood sizes, length of incubation and nestling stages, fledging success, number of fledglings, or survival of fledglings to independence. Thus, radio-tags had no measurable impact on the productivity of female Golden-winged Warblers. Our results build upon previous studies where investigators have reported no effects of radio-tagging on the breeding parameters of songbirds by also demonstrating no effect of radio-tagging through the post-fledging period and, therefore, the entire breeding season.

  10. Elevated Atmospheric CO2 Affects Ectomycorrhizal Species Abundance and Increases Sporocarp Production under Field Conditions

    Directory of Open Access Journals (Sweden)

    Douglas L. Godbold

    2015-04-01

    Full Text Available Anthropogenic activities during the last century have increased levels of atmospheric CO2. Forest net primary productivity increases in response to elevated CO2, altering the quantity and quality of carbon supplied to the rhizosphere. Ectomycorrhizal fungi form obligate symbiotic associations with the fine roots of trees that mediate improved scavenging for nutrients in exchange for a carbohydrate supply. Understanding how the community structure of ectomycorrhizal fungi is altered by climate change is important to further our understanding of ecosystem function. Betula pendula and Fagus sylvatica were grown in an elevated CO2 atmosphere delivered using free air carbon dioxide enrichment (FACE under field conditions in the U.K., and Picea abies was grown under elevated CO2 in glass domes in the Czech Republic. We used morphotyping and sequencing of the internal transcribed spacer region of the fungal ribosomal operon to study ectomycorrhizal community structure. Under FACE, un-colonised roots tips increased in abundance for Fagus sylvatica, and during 2006, sporocarp biomass of Peziza badia significantly increased. In domes, ectomycorrhizal community composition shifted from short-distance and smooth medium-distance to contact exploration types. Supply and competition for carbon belowground can influence ectomycorrhizal community structure with the potential to alter ecosystem function.

  11. Organic fertilization affecting pests and production of Lippia sidoides Cham. (Verbenacea

    Directory of Open Access Journals (Sweden)

    G.L.D. Leite

    2013-01-01

    Full Text Available The objective of this paper was to study the effect of four levels of organic fertilization (0, 3, 6 and 9 kg m-2 to produce the essential oil of Lippia sidoides Cham. (Verbenacea and its pests. A higher number of red mite Tetranychus sp. (Acari: Tetranychiidae was observed on the abaxial face of the median and basal leaves of L. sidoides plants with eight weeks of age cultivated with 6 kg m-2 of organic fertilization. On the other hand, the smallest number of this mite was noted in control (0 kg m-2. L. sidoides cultivated with 6 kg m-2 of organic fertilization produced a greater quantity of essential oil. The highest number of mealy bug Phenacoccus sp. (Hemiptera: Pseudococcidae was observed on the apical leaves (both leaf surfaces of L. sidoides plants with nine and ten weeks of age cultivated with 3 kg m-2 of organic fertilization. No significant effect of the organic fertilization on Aphis gossypii Glover (Hemiptera: Aphididae was detected. However, a greater number of this aphid and of its predator Scymnus sp. (Coleoptera: Coccinelidae was observed on the abaxial face of the bottom leaves of L. sidoides plants with eight weeks of age. The dose of organic fertilization recommended for the production of essential oil of L. Sidoides is 6 kg m-2, however, one must pay attention to the fact this dosage favors the attack by Tetranychus sp.. And, when needed, pulverization must always be directed to the lower surface of the leaf.

  12. Hydrodynamic Shear Stress Affects Cell Growth and Metabolite Production by Medicinal Mushroom Ganoderma lucidum

    Institute of Scientific and Technical Information of China (English)

    龚海刚; 钟建江

    2005-01-01

    By investigating the shear effect on submerged cultivation of a traditional Chinese medicinal herb Ganoderma lucidum, a relatively high cell concentration of 13.8 g·L-1 by dry mass was obtained in bioreactor at an impeller tip speed (ITS) of 0.51m·s-1. At an ITS of 0.51,1.02 and 1.53m·s-1, a maximal production titer of intracellular polysaccharide was 2.64, 2.20 and 2.28g·L-1 and that of ganoderic acid was 306, 299 and 273g·L-1, respectively. Under these ITSs, the maximal mean projected area of dispersed hypheue was 3.70, 2.54 and 2.13 × 104μm2, and that of pellets was 0.91, 0.67 and 0.55 mm2, respectively. The information obtained is useful for efficient submerged cultivation of mushrooms on a large scale.

  13. Fruit quality and production of Vitis vinifera L. Chardonnay affected by partial defoliation in tropical highlands

    Directory of Open Access Journals (Sweden)

    Jaime Ernesto Peña Olmos

    2015-06-01

    Full Text Available Using a completely randomized design with twotreatments (50% defoliation and a control without defoliationand six replications per treatment, the fruit quality and productionof grape plants subjected to early partial-defoliation in Villa deLeyva – Colombia were determined. Five-year-old Vitis vinifera L.'Chardonnay' plants were used as the plant material, planted in atrellis system with distances of 2.1 x 1 m. The plants were partiallydefoliated biweekly, removing every other recently emerged leaf ineach of the plants subjected to the defoliation treatment. 130 daysafter the start of the pruning, the number of harvested clusterswas determined for each of the evaluated plants along with thenumber of berries per cluster, the fresh weight of the clusters,the average berry weight, the fruit pH and diameter, the TSS, theTTA and the fruit maturity index (MI. The average weights of theclusters and the berries were significantly higher in the plantssubjected to a reduced foliar area. All of the fruit quality parametervalues increased with defoliation with the exception of TTA, whichwas higher in the control plants. Therefore, it was concluded thatpartial-defoliation had a pronounced influence on the fruit qualityand production of the 'Chardonnay' grapes.

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

    Directory of Open Access Journals (Sweden)

    Val F Lanza

    2014-12-01

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

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

    Science.gov (United States)

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

    2014-12-01

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

  16. Alcohol-to-acid ratio and substrate concentration affect product structure in chain elongation reactions initiated by unacclimatized inoculum.

    Science.gov (United States)

    Liu, Yuhao; Lü, Fan; Shao, Liming; He, Pinjing

    2016-10-01

    The objective of the study was to investigate whether the ratio of ethanol to acetate affects yield and product structure in chain elongation initiated by unacclimatized mixed cultures. The effect of varying the substrate concentration, while maintaining the same ratio of alcohol to acid, was also investigated. With a high substrate concentration, an alcohol to acid ratio >2:1 provided sufficient electron donor capacity for the chain elongation reaction. With an ethanol to acetate ratio of 3:1 (300mM total carbon), the highest n-caproate concentration (3033±98mg/L) was achieved during the stable phase of the reaction. A lower substrate concentration (150mM total carbon) gave a lower yield of products and led to reduced carbon transformation efficiency compared with other reaction conditions. The use of unacclimatized inoculum in chain elongation can produce significant amounts of odd-carbon-number carboxylates as a result of protein hydrolysis.

  17. Detection of genetic association and functional polymorphisms of UGDH affecting milk production trait in Chinese Holstein cattle

    Directory of Open Access Journals (Sweden)

    Xu Qing

    2012-11-01

    Full Text Available Abstract Background We previously localized a quantitative trait locus (QTL on bovine chromosome 6 affecting milk production traits to a 1.5-Mb region between BMS483 and MNB-209 via genome scanning followed by fine mapping. Results Totally 15 genes were mapped within such linkage region through bioinformatic analysis of the cattle-human comparative map and bovine genome assembly. Of them, the UDP-glucose dehydrogenase (UGDH was suggested as a potential positional candidate gene for milk production traits based on its corresponding physiological and biochemical functions and genetic effects. By sequencing all the coding exons and the untranslated regions in UGDH with pooled DNA of 8 sires represented the separated families detected in our previous studies, a total of ten SNPs were identified and genotyped in 1417 Holstein cows of 8 separation families. Individual SNP-based association analysis revealed 4 significant associations of SNP Ex1-1, SNP Int3-1, SNP Int5-1, and SNP Ex12-3 with milk yield (P protein percentage (F=4.15; P=0.0418 and fat percentage (F=5.18~7.25; P=0.0072~0.0231. Finally, by using an in vitro expression assay, we demonstrated that the A allele of SNP Ex1-1 and T allele of SNP Ex11-1of UGDH significantly decreases the expression of UGDH by 68.0% at the RNA, and 50.1% at the protein level, suggesting that SNP Ex1-1 and Ex11-1 represent two functional polymorphisms affecting expression of UGDH and may partly contributed to the observed association of the gene with milk production traits in our samples. Conclusions Taken together, our findings strongly indicate that UGDH gene could be involved in genetic variation underlying the QTL for milk production traits.

  18. A geostatistical synthesis study of factors affecting gross primary productivity in various ecosystems of North America

    Directory of Open Access Journals (Sweden)

    V. Yadav

    2010-02-01

    Full Text Available A coupled Bayesian model selection and geostatistical regression modeling approach is adopted for empirical analysis of gross primary productivity (GPP at six AmeriFlux sites, including the Kennedy Space Center Scrub Oak, Vaira Ranch, Tonzi Ranch, Blodgett Forest, Morgan Monroe State Forest, and Harvard Forest sites. The analysis is performed at a continuum of temporal scales ranging from daily to monthly, for a period of seven years. A total of 10 covariates representing environmental stimuli and indices of plant physiology are considered in explaining variations in GPP. Similar to other statistical methods, the proposed approach estimates regression coefficients and uncertainties associated with the covariates in a selected regression model. However, unlike traditional regression methods, the presented approach also estimates the uncertainty associated with the selection of a single "best" model of GPP. In addition, the approach provides an enhanced understanding of how the importance of specific covariates changes with temporal resolutions. An examination of trends in the importance of specific covariates reveals scaling thresholds above or below which covariates become significant in explaining GPP. Results indicate that most sites (especially those with a stronger seasonal cycle exhibit at least one prominent scaling threshold between daily to 20-day temporal scale. This demonstrates that environmental variables that explain GPP at synoptic scales are different from those that capture its seasonality. At shorter time scales, radiation, temperature, and vapor pressure deficit exert most significant influence on GPP at most examined sites. However, at coarser time scales, the importance of these covariates in explaining GPP declines. Overall, unique best models are identified at most sites at the daily scale, whereas multiple competing models are identified at larger time scales. In addition, the selected models are able to explain a larger

  19. A geostatistical synthesis study of factors affecting gross primary productivity in various ecosystems of North America

    Directory of Open Access Journals (Sweden)

    V. Yadav

    2010-09-01

    Full Text Available A coupled Bayesian model selection and geostatistical regression modeling approach is adopted for empirical analysis of gross primary productivity (GPP at six AmeriFlux sites, including the Kennedy Space Center Scrub Oak, Vaira Ranch, Tonzi Ranch, Blodgett Forest, Morgan Monroe State Forest, and Harvard Forest sites. The analysis is performed at a continuum of temporal scales ranging from daily to monthly, for a period of seven years. A total of 10 covariates representing environmental stimuli and indices of plant physiology are considered in explaining variations in GPP. Similarly to other statistical methods, the presented approach estimates regression coefficients and uncertainties associated with the covariates in a selected regression model. Unlike traditional regression methods, however, the approach also estimates the uncertainty associated with the selection of a single "best" model of GPP. In addition, the approach provides an enhanced understanding of how the importance of specific covariates changes with the examined timescale (i.e. temporal resolution. An examination of changes in the importance of specific covariates across timescales reveals thresholds above or below which covariates become important in explaining GPP. Results indicate that most sites (especially those with a stronger seasonal cycle exhibit at least one prominent scaling threshold between the daily and 20-day temporal scales. This demonstrates that environmental variables that explain GPP at synoptic scales are different from those that capture its seasonality. At shorter time scales, radiation, temperature, and vapor pressure deficit exert the most significant influence on GPP at most examined sites. At coarser time scales, however, the importance of these covariates in explaining GPP declines. Overall, unique best models are identified at most sites at the daily scale, whereas multiple competing models are identified at longer time scales.

  20. 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

  1. 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, ...

  2. The signaling protein MucG negatively affects the production and the molecular mass of alginate in Azotobacter vinelandii.

    Science.gov (United States)

    Ahumada-Manuel, Carlos Leonel; Guzmán, Josefina; Peña, Carlos; Quiroz-Rocha, Elva; Espín, Guadalupe; Núñez, Cinthia

    2017-02-01

    Azotobacter vinelandii is a soil bacterium that produces the polysaccharide alginate. In this work, we identified a miniTn5 mutant, named GG9, which showed increased alginate production of higher molecular mass, and increased expression of the alginate biosynthetic genes algD and alg8 when compared to its parental strain. The miniTn5 was inserted within ORF Avin07920 encoding a hypothetical protein. Avin07910, located immediately downstream and predicted to form an operon with Avin07920, encodes an inner membrane multi-domain signaling protein here named mucG. Insertional inactivation of mucG resulted in a phenotype of increased alginate production of higher molecular mass similar to that of mutant GG9. The MucG protein contains a periplasmic and putative HAMP and PAS domains, which are linked to GGDEF and EAL domains. The last two domains are potentially involved in the synthesis and degradation, respectively, of bis-(3'-5')-cyclic dimeric GMP (c-di-GMP), a secondary messenger that has been reported to be essential for alginate production. Therefore, we hypothesized that the negative effect of MucG on the production of this polymer could be explained by the putative phosphodiesterase activity of the EAL domain. Indeed, we found that alanine replacement mutagenesis of the MucG EAL motif or deletion of the entire EAL domain resulted in increased alginate production of higher molecular mass similar to the GG9 and mucG mutants. To our knowledge, this is the first reported protein that simultaneous affects the production of alginate and its molecular mass.

  3. Factors Affecting the Biomass and Lipid Production from Chlorella sp. TISTR 8990 under Mixotrophic Culture

    Directory of Open Access Journals (Sweden)

    Mohammad Ariful HAQUE

    2012-12-01

    Full Text Available Effects of media compositions on biomass and lipid accumulation of the isolate Chlorella sp. TISTR 8990 were investigated under a Plackett-Burman experimental design with mixotrophic cultivation conditions. Under this experimental design there were 15 different runs with ten factors-yeast extract, KH2PO4, MgSO4, FeSO4, MnCl2, CuSO4, Na2MoO4, H3BO3, ZnSO4 and pH. Cultures were grown mixotrophically under 16 h light and 8 h dark regime at 30 ºC for a period of 7 days. During the light regime, the light intensity at the surface of the vessels and agitation speed were set to 67.5 µmol photons m-2s-1 and 150 rpm, respectively. Initial cell concentration was set to an absorbance (A540 of 0.5. For high biomass production (2.2 g/L, run no. 6, the most effective and significant factors were yeast extract, KH2PO4, FeSO4 and ZnSO4 at concentrations 0.3 g/L, 0.3 g/L, 3 mg/L and 0.3 mg/L, respectively. Whereas for high lipid accumulation (19.59 %DCW, run no. 2, these were KH2PO4, pH and yeast extract, at a level of 1.7 g/L, 6.0 and 0.1 g/L, respectively. No significant factors were obtained for higher lipid content. The best treatment for biomass and lipid content was run no. 6, whose medium formula consisted of 0.3 g/L yeast extract, 1.7 g/L KH2PO4, 1.7 g/L MgSO4, 1 mg/L FeSO4, 0.9 mg/L MnCl2, and pH 7.0, together with fixed concentrations of glucose, NaHCO3 and KNO3 at 5 g/L, 0.05 g/L and 0.5 g/L, respectively.

  4. Nucleotide sequences of Herpes Simplex Virus type 1 (HSV-1) affecting virus entry, cell fusion, and production of glycoprotein gB (VP7)

    Energy Technology Data Exchange (ETDEWEB)

    DeLuca, N.; Bzik, D.J.; Bond, V.C.; Person, S.; Snipes, W.

    1982-10-30

    The tsB5 strain of Herpes Simplex Virus type 1 (HSV-1) contains at least two mutations; one mutation specifies the syncytial phenotype and the other confers temperature sensitivity for virus growth. These functions are known to be located between the prototypic map coordinates 0.30 and 0.42. In this study it was demonstrated that tsB5 enters human embryonic lung (HEL) cells more rapidly than KOS, another strain of HSV-1. The EcoRI restriction fragment F from the KOS strain (map coordinates 0.315 to 0.421) was mapped with eight restriction endonucleases, and 16 recombinant plasmids were constructed which contained varying portions of the KOS genome. Recombinant viruses were generated by marker-rescue and marker-transfer cotransfection procedures, using intact DNA from one strain and a recombinant plasmid containing DNA from the other strain. The region of the crossover between the two nonisogenic strains was inferred by the identification of restriction sites in the recombinants that were characteristic of the parental strains. The recombinants were subjected to phenotypic analysis. Syncytium formation, rate of virus entry, and the production of gB were all separable by the crossovers that produced the recombinants. The KOS sequences which rescue the syncytial phenotype of tsB5 were localized to 1.5 kb (map coordinates 0.345 to 0.355), and the temperature-sensitive mutation was localized to 1.2 kb (0.360 to 0.368), giving an average separation between the mutations of 2.5 kb on the 150-kb genome. DNA sequences that specify a functional domain for virus entry were localized to the nucleotide sequences between the two mutations. All three functions could be encoded by the virus gene specifying the gB glycoprotein.

  5. Heterogeneous conditions in dissolved oxygen affect N-glycosylation but not productivity of a monoclonal antibody in hybridoma cultures.

    Science.gov (United States)

    Serrato, J Antonio; Palomares, Laura A; Meneses-Acosta, Angélica; Ramírez, Octavio T

    2004-10-20

    It is known that heterogeneous conditions exist in large-scale animal cell cultures. However, little is known about how heterogeneities affect cells, productivities, and product quality. To study the effect of non-constant dissolved oxygen tension (DOT), hybridomas were subjected to sinusoidal DOT oscillations in a one-compartment scale-down simulator. Oscillations were forced by manipulating the inlet oxygen partial pressure through a feedback control algorithm in a 220-mL bioreactor maintained at a constant agitation. Such temporal DOT oscillations simulate spatial DOT gradients that can occur in large scales. Different oscillation periods, in the range of 800 to 12,800 s (axis of 7% (air saturation) and amplitude of 7%), were tested and compared to constant DOT (10%) control cultures. Oscillating DOT decreased maximum cell concentrations, cell growth rates, and viability indexes. Cultures at oscillating DOT had an increased glycolytic metabolism that was evidenced by a decrease in yield of cells on glucose and an increase in lactate yield. DOT gradients, even several orders of magnitude higher than those expected under practical large-scale conditions, did not significantly affect the maximum concentration of an IgG(1) monoclonal antibody (MAb). The glycosylation profile of the MAb produced at a constant DOT of 10% was similar to that reported in the literature. However, MAb produced under oscillating culture conditions had a higher amount of triantennary and sialylated glycans, which can interfere with effector functions of the antibody. It was shown that transient excursions of hybridomas to limiting DOT, as occurs in deficiently mixed large-scale bioreactors, is important to culture performance as the oscillation period, and thus the time cells spent at low DOT, affected cell growth, metabolism, and the glycosylation pattern of MAb. Such results underline the importance of monitoring protein characteristics for the development of large-scale processes.

  6. Phenotypes and gene expression profiles of Saccharopolyspora erythraea rifampicin-resistant (rif mutants affected in erythromycin production

    Directory of Open Access Journals (Sweden)

    Bicciato Silvio

    2009-03-01

    Full Text Available Abstract Background There is evidence from previous works that bacterial secondary metabolism may be stimulated by genetic manipulation of RNA polymerase (RNAP. In this study we have used rifampicin selection as a strategy to genetically improve the erythromycin producer Saccharopolyspora erythraea. Results Spontaneous rifampicin-resistant (rif mutants were isolated from the parental strain NRRL2338 and two rif mutations mapping within rpoB, S444F and Q426R, were characterized. With respect to the parental strain, S444F mutants exhibited higher respiratory performance and up to four-fold higher final erythromycin yields; in contrast, Q426R mutants were slow-growing, developmental-defective and severely impaired in erythromycin production. DNA microarray analysis demonstrated that these rif mutations deeply changed the transcriptional profile of S. erythraea. The expression of genes coding for key enzymes of carbon (and energy and nitrogen central metabolism was dramatically altered in turn affecting the flux of metabolites through erythromycin feeder pathways. In particular, the valine catabolic pathway that supplies propionyl-CoA for biosynthesis of the erythromycin precursor 6-deoxyerythronolide B was strongly up-regulated in the S444F mutants, while the expression of the biosynthetic gene cluster of erythromycin (ery was not significantly affected. In contrast, the ery cluster was down-regulated ( Conclusion Rifampicin selection is a simple and reliable tool to investigate novel links between primary and secondary metabolism and morphological differentiation in S. erythraea and to improve erythromycin production. At the same time genome-wide analysis of expression profiles using DNA microarrays allowed information to be gained about the mechanisms underlying the stimulatory/inhibitory effects of the rif mutations on erythromycin production.

  7. Characterization of a cryptic plasmid pSFKW33 from Shewanella sp. 33B.

    Science.gov (United States)

    Werbowy, Katarzyna; Cieśliński, Hubert; Kur, Józef

    2009-07-01

    A cryptic plasmid pSFKW33 from psychrotrophic bacterium Shewanella sp. 33B, an isolate from the Gulf of Gdansk (the Baltic Sea), was sequenced and characterized. It is an 8021bp circular molecule with 38% GC content, which shows a distinctive nucleotide sequence without homology to other known plasmids. The nucleotide sequence analysis predicts eight open reading frames. The deduced amino acid sequence of ORF-1 shared significant similarity with the phage integrase protein from Pseudomonas putida GB-1. The ORF2 product showed some similarity with the hypothetical protein Shal_1786 from Shewanella halifaxensis HAW-EB4. The ORF-3 product revealed high amino acid sequence homology with the Escherichia coli replication initiation protein similar to pi protein of R6K plasmid. Five repeat regions (three perfect 22-bp repeats and two imperfect motifs), a putative ribosome binding site, and -10 and -35 promotor sequences were identified upstream of the ORF-3 (rep). The rep module is very similar to several theta replicating iteron family plasmids, suggesting that pSFKW33 replication follows the same course. The ORF-5 product revealed significant identity with the entry exclusion protein 1 of plasmid pIS2 from enteropathogenic strain E. coli 0111:H. The ORF-6 product showed significant identity with the putative transcriptional regulator protein from Shewanella benthica KT99 belonging to HTH-XRE-family protein. The ORF-8 encoded protein that showed some similarity with the hypothetical protein KT99_00146 from Shewanella benthica KT99. The ORF-4 and ORF-7 encoded putative proteins of 189 and 118 amino acid residues, respectively, with no homologies to any known proteins. The absolute copy number of pSFKW33 was estimated to be five copies per chromosome by real-time PCR.

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

    Directory of Open Access Journals (Sweden)

    F BENSALAH

    2003-06-01

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

  9. Chloride stress triggers maturation and negatively affects the postharvest quality of persimmon fruit. Involvement of calyx ethylene production.

    Science.gov (United States)

    Besada, Cristina; Gil, Rebeca; Bonet, Luis; Quiñones, Ana; Intrigliolo, Diego; Salvador, Alejandra

    2016-03-01

    In recent years many hectares planted with persimmon trees in E Spain have been diagnosed with chloride toxicity. An effect of this abiotic stress on fruit quality has been reported in different crops. However, the impact of chloride stress on persimmon fruit quality is unknown. The harvest and postharvest quality of persimmons harvested from trees that manifest different intensities of chloride toxicity foliar symptoms was evaluated herein. Our results revealed that fruits from trees under chloride stress conditions underwent chloride accumulation in the calyx, which was more marked the greater the salt stress intensity trees were exposed to. Increased chloride concentrations in the calyx stimulated ethylene production in this tissue. In the fruits affected by slight and moderate chloride stress, calyx ethylene production accelerated the maturity process, as reflected by increased fruit colour and diminished fruit firmness. In the fruits under severe chloride stress, the high ethylene levels in the calyx triggered autocatalytic ethylene production in other fruit tissues, which led fruit maturity to drastically advance. In these fruits effectiveness of CO2 deastringency treatment was not complete and fruit softening enhanced during the postharvest period. Moreover, chloride stress conditions had a marked effect on reducing fruit weight, even in slightly stressed trees.

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

    Directory of Open Access Journals (Sweden)

    Eric Smit

    1997-01-01

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

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

    OpenAIRE

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

    1992-01-01

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

  12. Affective Urbanism

    DEFF Research Database (Denmark)

    Samson, Kristine

    Urban design and architecture are increasingly used as material and affective strategies for setting the scene, for manipulation and the production of urban life: The orchestration of atmospheres, the framing and staging of urban actions, the programming for contemplation, involvement, play, expe...... affects can be choreographed and designed intentionally or whether it arises from unpredictable circumstances within urbanity itself....

  13. Affective Maps

    DEFF Research Database (Denmark)

    Salovaara-Moring, Inka

    of environmental knowledge production. It uses InfoAmazonia, the databased platform on Amazon rainforests, as an example of affective geo-visualization within information mapping that enhances embodiment in the experience of the information. Amazonia is defined as a digitally created affective (map)space within...

  14. Regional Algal Biofuel Production Potential in the Coterminous United States as Affected by Resource Availability Trade-offs

    Energy Technology Data Exchange (ETDEWEB)

    Venteris, Erik R. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Skaggs, Richard [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Wigmosta, Mark S. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Coleman, Andre M. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

    2014-03-15

    The warm sunny climate and unoccupied arid lands in the American southwest are favorable factors for algae cultivation. However, additional resources affect the overall viability of specific sites and regions. We investigated the tradeoffs between growth rate, water, and CO2 availability and costs for two strains: N. salina and Chlorella sp. We conducted site selection exercises (~88,000 US sites) to produce 21 billion gallons yr-1 (BGY) of renewable diesel (RD). Experimental trials from the National Alliance for Advanced Biofuels and Bio-Products (NAABB) team informed the growth model of our Biomass Assessment Tool (BAT). We simulated RD production by both lipid extraction and hydrothermal liquefaction. Sites were prioritized by the net value of biofuel minus water and flue gas costs. Water cost models for N. salina were based on seawater and high salinity groundwater and for Chlorella, fresh and brackish groundwater. CO2 costs were based on a flue gas delivery model. Selections constrained by production and water were concentrated along the Gulf of Mexico and southeast Atlantic coasts due to high growth rates and low water costs. Adding flue gas constraints increased the spatial distribution, but the majority of sites remained in the southeast. The 21 BGY target required ~3.8 million hectares of mainly forest (41.3%) and pasture (35.7%). Exclusion in favor of barren and scrub lands forced most production to the southwestern US, but with increased water consumption (5.7 times) and decreased economic efficiency (-38%).

  15. Fertilization Affects Biomass Production of Suaeda salsa and Soil Organic Carbon Pool in East Coastal Region of China

    Institute of Scientific and Technical Information of China (English)

    MENG Qing-feng; YANG Jing-song; YAO Rong-jiang; LIU Guang-ming; YU Shi-peng

    2013-01-01

    Land use practice significantly affects soil properties. Soil is a major sink for atmospheric carbon, and soil organic carbon (SOC) is considered as an essential indicator of soil quality. The objective of this study was to assess the effects of N and P applied to Suaeda salsa on biomass production, SOC concentration, labile organic carbon (LOC) concentration, SOC pool and carbon management index (CMI) as well as the effect of the land use practice on soil quality of coastal tidal lands in east coastal region of China. The study provided relevant references for coastal exploitation, tidal land management and related study in other countries and regions. The field experiment was laid out in a randomized complete block design, consisting of four N-fertilization rates (0 (N0), 60 (N1), 120 (N2) and 180 kg ha-1 (N3)), three P-fertilization rates (0 (P0), 70 (P1) and 105 kg ha-1 (P2)) and bare land without vegetation. N and P applied to S. salsa on coastal tidal lands significantly affected biomass production (above-ground biomass and roots), bulk density (ρb), available N and P, SOC, LOC, SOC pool and CMI. Using statistical analysis, significantly interactions in N and P were observed for biomass production and the dominant factor for S. salsa production was N in continuous 2-yr experiments. There were no significant interactions between N and P for SOC concentration, LOC concentration and SOC pool. However, significant interaction was obtained for CMI at the 0-20 cm depth and N played a dominant role in the variation of CMI. There were significant improvements for soil measured attributes and parameters, which suggested that increasing the rates of N and P significantly decreasedρb at the 0-20 cm depth and increased available N and P, SOC, LOC, SOC pool as well as CMI at both the 0-20 and 20-40 cm depth, respectively. By correlation analysis, there were significantly positive correlations between biomass (above-ground biomass and roots) and SOC as well as LOC in

  16. Production of biochar out of organic urban waste to amend salt affected soils in the basin of Mexico

    Science.gov (United States)

    Chavez Garcia, Elizabeth; Siebe, Christina

    2016-04-01

    Biochar is widely recognized as an efficient tool for carbon sequestration and soil fertility. The understanding of its chemical and physical properties, strongly related to the biomass and production conditions, is central to identify the most suitable application of biochar. On the other hand, salt affected soils reduce the value and productivity of extensive areas worldwide. One feasible option to recover them is to add organic amendments, which improve water holding capacity and increase sorption sites for cations as sodium. The former lake Texcoco in the basin of Mexico has been a key area for the control of surface run-off and air quality of Mexico City. However, the high concentrations of soluble salts in their soils do not allow the development of a vegetation cover that protects the soil from wind erosion, being the latter the main cause of poor air quality in the metropolitan area during the dry season. On the other hand, the population of the city produces daily 2000 t of organic urban wastes, which are currently composted. Thus, we tested if either compost or biochar made out of urban organic waste can improve the salt affected soils of former lake Texcoco to grow grass and avoid wind erosion. We examined the physico-chemical properties of biochar produced from urban organic waste under pyrolysis conditions. We also set up a field experiment to evaluate the addition of these amendments into the saline soils of Texcoco. Our preliminary analyses show biochar yield was ca. 40%, it was mainly alkaline (pH: 8-10), with a moderate salt content (electrical conductivity: 0.5-3 mS/cm). We show also results of the initial phase of the field experiment in which we monitor the electrical conductivity, pH, water content, water tension and soil GHG fluxes on small plots amended with either biochar or compost in three different doses.

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

    Science.gov (United States)

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

    2007-09-01

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

  18. 基于色谱法的超螺旋质粒DNA纯化与分析进展%Advances on Purification Process and Analysis of Supercoiled Plasmid DNA Based on Chromatography

    Institute of Scientific and Technical Information of China (English)

    李亮; 柳方方; 宛煜嵩; 金芜军

    2014-01-01

    质粒DNA含有独立复制的遗传结构,是基因工程的常用工具,广泛应用于分子生物学基础研究、农业转基因检测、医疗诊断与基因治疗等领域。质粒DNA的构型一般分为超螺旋、开口环状及线性3种。初步纯化的质粒DNA溶液中常混有3种构型,并且掺杂一定量的蛋白质、RNA、内毒素以及宿主基因组DNA,这些杂质会影响后续的应用,因此质粒DNA需要进一步精细纯化。该文对质粒DNA的新应用领域、基于色谱的精细纯化技术及产物质量分析体系进行了综述,并展望了高纯度质粒DNA精细纯化及产物分析的发展方向。%Plasmid DNA,a common tool of genetic engineering containing the independent genetic structure of the replication,is widely used in basic research of molecular biology,genetically modi-fied detection,medical diagnosis and gene therapy. Plasmid DNA conformations are generally divided into three types:supercoiled,open circle and linear. Crude plasmid DNA often mixes with three configurations,and the amount of protein,RNA,endotoxin with the host genome DNA. These im-purities need to be further refined as they affect subsequent applications. The new applications of plasmid DNA,refined technologies and products based on chromatography system,and quality anal-ysis of purified plasmid DNA is reviewed in this paper. Finally,the trends of technology of high pu-rity plasmid DNA and analysis of refined product are presented.

  19. Persistence of Free Plasmid DNA in Soil Monitored by Various Methods, Including a Transformation Assay

    Science.gov (United States)

    Romanowski, Gerd; Lorenz, Michael G.; Sayler, Gary; Wackernagel, Wilfried

    1992-01-01

    The persistence and stability of free plasmid pUC8-ISP DNA introduced into 10-g samples of various soils and kept at 23°C were monitored over a period of 60 days. The soils were sampled at a plant science farm and included a loamy sand soil (no. 1), a clay soil (no. 2), and a silty clay soil (no. 3). Four different methods allowed monitoring of (i) the production of acid-soluble radioactive material from [3H]thymidine-labeled plasmid DNA, (ii) the decrease of hybridizing nucleotide sequences in slot blot analysis, (iii) the loss of plasmid integrity measured by Southern hybridization, and (iv) the decay of the biological activity as determined by transformation of Ca2+-treated Escherichia coli cells with the DNA extracted from soil. Acid-soluble material was not produced within the first 24 h but then increased to 45% (soil no. 1), 27% (soil no. 2), and 77% (soil no. 3) until the end of incubation. A quite parallel loss of material giving a slot blot hybridization signal was observed. Southern hybridization indicated that after 1 h in the soils, plasmid DNA was mostly in the form of circular and full-length linear molecules but that, depending on the soil type, after 2 to 5 days full-length plasmid molecules were hardly detectable. The transforming activity of plasmid DNA reextracted from the soils followed inactivation curves over 2 to 4 orders of magnitude and dropped below the detection limit after 10 days. The inactivation was slower in soil no. 2 (28.2-h half-life time of the transforming activity of a plasmid molecule) than in soils no. 3 (15.1 h) and no. 1 (9.1 h). The studies provide data on the persistence of free DNA molecules in natural bacterial soil habitats. The data suggest that plasmid DNA may persist long enough to be available for uptake by competent recipient cells in situ. Images PMID:16348772

  20. 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.

  1. 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.

  2. Fermentation temperature affects the antioxidant activity of the enzyme-ripened sufu, an oriental traditional fermented product of soybean.

    Science.gov (United States)

    Huang, Yung-Hsin; Lai, Ying-Jang; Chou, Cheng-Chun

    2011-07-01

    In this study, sufu, a Chinese traditional fermented product of soybean, was prepared by ripening salted tofu cubes in the mash of Aspergillus oryzae-fermented rice-soybean koji possessing various hydrolytic enzymes at 25°C, 37°C and 45°C. Antioxidant activity including 2,2-diphenyl-2-picylhydoxyl (DPPH) radical-scavenging activity, Fe(2+)-chelating ability and reducing power exerted by the methanol extract of sufu was determined and compared with that of the non-fermented tofu extract. It was found that antioxidant activity of the sufu extracts was, generally, higher than the non-fermented tofu extract. Ripening temperature and the duration of ripening period affected the antioxidant activity of the sufu extracts. Taking into account of extraction yields, the sufu product ripened at 45°C for 16 days showed the most profound enhancement in the DPPH radical-scavenging effect and Fe(2+)-iron-chelating ability, which is 3.4 and 11.5 folds, respectively, that noted with the non-fermented tofu.

  3. Oral administration of γ-aminobutyric acid affects heat production in a hot environment in resting humans

    Directory of Open Access Journals (Sweden)

    Miyazawa Taiki

    2012-02-01

    Full Text Available Abstract Background Central administration of γ-amino butyric acid (GABA induces lower body temperature in animals in hot ambient air. However, it is still unknown whether oral GABA administration affects temperature regulation at rest in a hot environment in humans. Therefore, in the present study, we specifically hypothesized that systemic administration of GABA in humans would induce hypothermia in a hot environment and that this response would be observed in association with decreased heat production. Methods Eight male participants drank a 200-ml sports drink with 1 g of GABA (trial G or without GABA (trial C, then rested for 30 minutes in a sitting position in a hot environment (ambient air temperature 33°C, relative humidity 50%. Results We found that changes in esophageal temperature from before drinking the sports drink were lower in trial G than in trial C (-0.046 ± 0.079°C vs 0.001 ± 0.063°C; P 2 vs 47 ± 8 W/m2; P Conclusions In this study, we have demonstrated that a single oral administration of GABA induced a larger decrease in body core temperature compared to a control condition during rest in a hot environment and that this response was concomitant with a decrease in total heat production.

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

    OpenAIRE

    Chattoraj, D; Cordes, K.; Abeles, A

    1984-01-01

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

  5. Bacteriophages Limit the Existence Conditions for Conjugative Plasmids

    Science.gov (United States)

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

    2015-01-01

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

  6. [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.

  7. Genomic saturation mutagenesis and polygenic analysis identify novel yeast genes affecting ethyl acetate production, a non-selectable polygenic trait

    Directory of Open Access Journals (Sweden)

    Tom Den Abt

    2016-03-01

    Full Text Available Isolation of mutants in populations of microorganisms has been a valuable tool in experimental genetics for decades. The main disadvantage, however, is the inability of isolating mutants in non-selectable polygenic traits. Most traits of organisms, however, are non-selectable and polygenic, including industrially important properties of microorganisms. The advent of powerful technologies for polygenic analysis of complex traits has allowed simultaneous identification of multiple causative mutations among many thousands of irrelevant mutations. We now show that this also applies to haploid strains of which the genome has been loaded with induced mutations so as to affect as many non-selectable, polygenic traits as possible. We have introduced about 900 mutations into single haploid yeast strains using multiple rounds of EMS mutagenesis, while maintaining the mating capacity required for genetic mapping. We screened the strains for defects in flavor production, an important non-selectable, polygenic trait in yeast alcoholic beverage production. A haploid strain with multiple induced mutations showing reduced ethyl acetate production in semi-anaerobic fermentation, was selected and the underlying quantitative trait loci (QTLs were mapped using pooled-segregant whole-genome sequence analysis after crossing with an unrelated haploid strain. Reciprocal hemizygosity analysis and allele exchange identified PMA1 and CEM1 as causative mutant alleles and TPS1 as a causative genetic background allele. The case of CEM1 revealed that relevant mutations without observable effect in the haploid strain with multiple induced mutations (in this case due to defective mitochondria can be identified by polygenic analysis as long as the mutations have an effect in part of the segregants (in this case those that regained fully functional mitochondria. Our results show that genomic saturation mutagenesis combined with complex trait polygenic analysis could be used

  8. Yield and nutritional quality of greenhouse lettuce (Lactuca sativa L. as affected by genotype and production methods

    Directory of Open Access Journals (Sweden)

    Govedarica-Lučić Aleksandra

    2014-01-01

    Full Text Available Greenhouse experiments were conducted in winter growing seasons in order to evaluate the effects of genotype and production methods on yield and nutritional quality of lettuce (Lactuca sativa L.. A three-year (2009-2011 study was conducted by randomized block system in a greenhouse without additional heating. The trial included three genotypes of lettuce (Archimedes RZ, Santoro RZ, Kibou RZ. Each row with these genotypes was exposed to the following variants of covering: control-planting on bare soil, mulching before sowing with PE-black foil, agro textile-covering plants after planting with agro textile (17 g, a combination of mulching + agro textile. Throughout of all the three years of the trial, it was continuously evidenced that the genotype “Santoro RZ” had the biggest heads and the highest yield (15.33 kg 10 m-2, which leads to conclusion that the yield of lettuce is a genotype characteristics. Moreover, the nutritional value (ascorbic acid concentration has shown that, depending on the method of production, in average, the combination of mulching + agro textile (26.77 mg 100 g-1 had the highest content while the control variant had significantly lower vitamin C content (21.10 mg 100 g-1. The three-year researches have shown that the production method and genotype significantly affect the nitrate content. An average nitrate content was 2196.33 mg kg-1 on the control variant, and 2526.24 mg kg-1 on agro textile. Leafy lettuce of genotyp „Kibou RZ“ had lower nitrate content (2176.85 mg kg-1 compared to „Archimedes RZ“ (2843.05 mg kg-1 and „Santoro RZ“ (2221.37 mg kg-1. However nitrate concentration in all treatments remained within the European Union’s permissible levels.

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

    Directory of Open Access Journals (Sweden)

    Bahig E.  Deeb

    2009-01-01

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

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

    Science.gov (United States)

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

    2008-07-01

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

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

    Energy Technology Data Exchange (ETDEWEB)

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

    1984-07-01

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

  12. Identification of bla KPC-2 on different plasmids of three Morganella morganii isolates.

    Science.gov (United States)

    Shi, D-S; Wang, W-P; Kuai, S-G; Shao, H-F; Huang, M

    2012-05-01

    Three Morganella morganii strains resistant to carbapenems were recovered from the surgical intensive care unit (SICU) in our hospital. Carbapenemases and extended-spectrum β-lactamases (ESBLs) were respectively detected by the modified Hodge test and the modified Clinical and Laboratory Standards Institute (CLSI) ESBL confirmatory test in all isolates. Amplification of whole-cell and plasmid DNAs extracted from isolates with primers specific for the bla (KPC) produced an amplicon confirmed to be bla (KPC-2) by sequence analysis. Pulsed-field gel electrophoresis (PFGE) typing revealed that three isolates belonged to two closely related types. Plasmids electrophoresis and restriction analysis revealed that the bla (KPC-2) was located on different plasmids. The transfer of carbapenem resistance from the three original isolates to Escherichia coli EC600 was successful by conjugation. An examination of the outer membrane proteins showed a lack of a 38-kDa outer membrane protein (OMP) compared with M. morganii susceptible to carbapenems. The production of KPC-2 and ESBLs, combined with OMP deficiency, resulted in high-level carbapenem resistance in the M. morganii strains. The genetic environment around bla (KPC-2) analysis revealed that this β-lactamase was located on the same mobile genetic elements which could transfer between different plasmids.

  13. Construction of a recombinant bacterial plasmid containing DNA sequences for a mouse embryonic globin chain.

    Science.gov (United States)

    Fantoni, A; Bozzoni, I; Ullu, E; Farace, M G

    1979-08-10

    Messenger RNAs for mouse embryonic globins were purified from yolk sac derived eyrthroid cells in mouse fetuses. Double stranded DNAs complementary to these messengers were synthesized and blunt end ligated to a EcoRI digested and DNA polymerase I repaired pBR322 plasmid. Of the ampicillin resistant transformants, one contained a plasmid with globin-specific cDNA. The inserted sequence is about 350 base pairs long. It contains one restriction site for EcoRI and one restriction site for HinfI about 170 and 80 base pairs from one end. The insert is not cleaved by HindIII, HindII, BamHI, PstI, SalI, AvaI, TaqI, HpaII, BglI. A mixture of purified messengers coding for alpha chains and for x, y and z embryonic chains was incubated with the recombinant plasmid and the hybridized messenger was translated in a mRNA depleted reticulocyte lysate protein synthesizing system. The product of translation was identified as a z chain by carboxymethylcellulose cromatography. The recombinant plasmid is named "pBR322-egz" after embryonic globin z.

  14. Development of new plasmid DNA vaccine vectors with R1-based replicons

    Directory of Open Access Journals (Sweden)

    Bower Diana M

    2012-08-01

    Full Text Available Abstract Background There has been renewed interest in biopharmaceuticals based on plasmid DNA (pDNA in recent years due to the approval of several veterinary DNA vaccines, on-going clinical trials of human pDNA-based therapies, and significant advances in adjuvants and delivery vehicles that have helped overcome earlier efficacy deficits. With this interest comes the need for high-yield, cost-effective manufacturing processes. To this end, vector engineering is one promising strategy to improve plasmid production. Results In this work, we have constructed a new DNA vaccine vector, pDMB02-GFP, containing the runaway R1 origin of replication. The runaway replication phenotype should result in plasmid copy number amplification after a temperature shift from 30°C to 42°C. However, using Escherichia coli DH5α as a host, we observed that the highest yields of pDMB02-GFP were achieved during constant-temperature culture at 30°C, with a maximum yield of approximately 19 mg pDNA/g DCW being observed. By measuring mRNA and protein levels of the R1 replication initiator protein, RepA, we determined that RepA may be limiting pDMB02-GFP yield at 42°C. A mutant plasmid, pDMB-ATG, was constructed by changing the repA start codon from the sub-optimal GTG to ATG. In cultures of DH5α[pDMB-ATG], temperature-induced plasmid amplification was more dramatic than that observed with pDMB02-GFP, and RepA protein was detectable for several hours longer than in cultures of pDMB02-GFP at 42°C. Conclusions Overall, we have demonstrated that R1-based plasmids can produce high yields of high-quality pDNA without the need for a temperature shift, and have laid the groundwork for further investigation of this class of vectors in the context of plasmid DNA production.

  15. The heat-shock DnaK protein is required for plasmid R1 replication and it is dispensable for plasmid ColE1 replication.

    Science.gov (United States)

    Giraldo-Suárez, R; Fernández-Tresguerres, E; Díaz-Orejas, R; Malki, A; Kohiyama, M

    1993-01-01

    Plasmid R1 replication in vitro is inactive in extracts prepared from a dnaK756 strain but is restored to normal levels upon addition of purified DnaK protein. Replication of R1 in extracts of a dnaKwt strain can be specifically inhibited with polyclonal antibodies against DnaK. RepA-dependent replication of R1 in dnaK756 extracts supplemented with DnaKwt protein at maximum concentration is partially inhibited by rifampicin and it is severely inhibited at sub-optimal concentrations of DnaK protein. The copy number of a run-away R1 vector is reduced in a dnaK756 background at 30 degrees C and at 42 degrees C the amplification of the run-away R1 vector is prevented. However a runaway R1 vector containing dnaK gene allows the amplification of the plasmid at high temperature. These data indicate that DnaK is required for both in vitro and in vivo replication of plasmid R1 and show a partial compensation for the low level of DnaK by RNA polymerase. In contrast ColE1 replication is not affected by DnaK as indicated by the fact that ColE1 replicates with the same efficiency in extracts from dnaKwt and dnaK756 strains. Images PMID:8265367

  16. Genotypic and phenotypic diversity does not affect productivity and drought response in competitive stands of Trifolium repens

    Directory of Open Access Journals (Sweden)

    Heidrun eHuber

    2016-03-01

    Full Text Available Clonal plants can form dense canopies in which plants of different genetic origin are competing for the uptake of essential resources. The competitive relationships among these clones are likely to be affected by extreme environmental conditions, such as prolonged drought spells, which are predicted to occur more frequently due to global climate change. This, in turn, may alter characteristics of the ecological system and its associated functioning.We hypothesized that the relative success of individual clones will depend on the size of the ramets as ramets with larger leaves and longer petioles (large ramets were predicted to have a competitive advantage in terms of increased light interception over smaller-sized ramets. Under drier conditions the relative performances of genotypes were expected to change leading to a change in genotype ranking. We also hypothesized that increased genotypic and phenotypic diversity will increase stand performance and resistance to drought. These hypotheses and the mechanisms responsible for shifts in competitive relationships were investigated by subjecting genotypes of the important pasture legume Trifolium repens to competition with either genetically identical clones, genetically different but similarly sized clones, or genetically as well as morphologically different clones under well-watered and dry conditions.Competitive relationships were affected by ramet size with large genotypes outperforming small genotypes in diverse stands in terms of biomass production. However, large genotypes also produced relatively fewer ramets than small genotypes and could not benefit in terms of clonal reproduction from competing with smaller genotypes, indicating that evolutionary shifts in genotype composition will depend on whether ramet size or ramet number is under selection. In contrast to our hypotheses, diversity did not increase stand performance under different selection regimes and genotype ranking was hardly

  17. Deletion mutants of region E1 a of AD12 E1 plasmids: Effect on oncogenic transformation

    NARCIS (Netherlands)

    Bos, J.L.; Jochemsen, A.G.; Bernards, R.A.; Schrier, P.I.; Ormondt, H. van; Eb, A.J. van der

    1983-01-01

    Plasmids containing the El region of Ad12 DNA can transform certain rodent cells into oncogenic cells. To study the role of the Ela subregion in the process of oncogenic transformation, Ad12 region El mutants carrying deletions in the Ela region were constructed. Deletion mutants pR7 and pR8 affect

  18. Properties of the arsenate reductase of plasmid R773.

    Science.gov (United States)

    Gladysheva, T B; Oden, K L; Rosen, B P

    1994-06-14

    Resistance to toxic oxyanions in Escherichia coli is conferred by the ars operon carried on plasmid R773. The gene products of this operon catalyze extrusion of antimonials and arsenicals from cells of E. coli, thus providing resistance to those toxic oxyanions. In addition, resistance to arsenate is conferred by the product of the arsC gene. In this report, purified ArsC protein was shown to catalyze reduction of arsenate to arsenite. The enzymatic activity of the ArsC protein required glutaredoxin as a source of reducing equivalents. Other reductants, including glutathione and thioredoxin, were not effective electron donors. A spectrophotometric assay was devised in which arsenate reduction was coupled to NADPH oxidation. The results obtained with the coupled assay corresponded to those found by direct reduction of radioactive arsenate to arsenite. The only substrate of the reaction was arsenate (Km = 8 mM); other oxyanions including phosphate, sulfate, and antimonate were not reduced. Phosphate and sulfate were weak inhibitors, while the product, arsenite, was a stronger inhibitor (Ki = 0.1 mM). Arsenate reductase activity exhibited a pH optimum of 6.3-6.8. These results indicate that the ArsC protein is a novel reductase, and elucidation of its enzymatic mechanism should be of interest.

  19. Homology of plasmids in strains of unicellular cyanobacteria

    NARCIS (Netherlands)

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

    1979-01-01

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

  20. Multilocus sequence typing of IncN plasmids

    DEFF Research Database (Denmark)

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

    2011-01-01

    in different countries from both animals and humans belonged to ST1, suggesting dissemination of an epidemic plasmid through the food chain. Fifteen of 17 plasmids carrying blaVIM-1 from Klebsiella pneumoniae and Escherichia coli, isolated during a 5year period in Greece were assigned to ST10, suggesting...

  1. 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.

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

    NARCIS (Netherlands)

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

    1992-01-01

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

  3. Plasmid cloning vehicle for Haemophilus influenzae and Escherichia coli

    Energy Technology Data Exchange (ETDEWEB)

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

    1982-09-01

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

  4. Functional analysis of three plasmids from Lactobacillus plantarum

    NARCIS (Netherlands)

    Kranenburg, R. van; Golic, N.; Bongers, R.; Leer, R.J.; Vos, W.M. de; Siezen, R.J.; Kleerebezem, M.

    2005-01-01

    Lactobacillus plantarum WCFS1 harbors three plasmids, pWCFS101, pWCFS102, and pWCFS103, with sizes of 1,917, 2,365, and 36,069 bp, respectively. The two smaller plasmids are of unknown function and contain replication genes that are likely to function via the rolling-circle replication mechanism. Th

  5. Identification of IncA/C Plasmid Replication and Maintenance Genes and Development of a Plasmid Multilocus Sequence Typing Scheme.

    Science.gov (United States)

    Hancock, Steven J; Phan, Minh-Duy; Peters, Kate M; Forde, Brian M; Chong, Teik Min; Yin, Wai-Fong; Chan, Kok-Gan; Paterson, David L; Walsh, Timothy R; Beatson, Scott A; Schembri, Mark A

    2017-02-01

    Plasmids of incompatibility group A/C (IncA/C) are becoming increasingly prevalent within pathogenic Enterobacteriaceae They are associated with the dissemination of multiple clinically relevant resistance genes, including blaCMY and blaNDM Current typing methods for IncA/C plasmids offer limited resolution. In this study, we present the complete sequence of a blaNDM-1-positive IncA/C plasmid, pMS6198A, isolated from a multidrug-resistant uropathogenic Escherichia coli strain. Hypersaturated transposon mutagenesis, coupled with transposon-directed insertion site sequencing (TraDIS), was employed to identify conserved genetic elements required for replication and maintenance of pMS6198A. Our analysis of TraDIS data identified roles for the replicon, including repA, a toxin-antitoxin system; two putative partitioning genes, parAB; and a putative gene, 053 Construction of mini-IncA/C plasmids and examination of their stability within E. coli confirmed that the region encompassing 053 contributes to the stable maintenance of IncA/C plasmids. Subsequently, the four major maintenance genes (repA, parAB, and 053) were used to construct a new plasmid multilocus sequence typing (PMLST) scheme for IncA/C plasmids. Application of this scheme to a database of 82 IncA/C plasmids identified 11 unique sequence types (STs), with two dominant STs. The majority of blaNDM-positive plasmids examined (15/17; 88%) fall into ST1, suggesting acquisition and subsequent expansion of this blaNDM-containing plasmid lineage. The IncA/C PMLST scheme represents a standardized tool to identify, track, and analyze the dissemination of important IncA/C plasmid lineages, particularly in the context of epidemiological studies.

  6. Expansion of the IncX plasmid family for improved identification and typing of novel plasmids in drug-resistant Enterobacteriaceae

    DEFF Research Database (Denmark)

    Johnson, Timothy J.; Bielak, Eliza Maria; Fortini, Daniela;

    2012-01-01

    and biofilm formation. Previous plasmid-based replicon typing procedures have indicated that the prevalence of IncX plasmids is low among members of the Enterobacteriaceae. However, examination of a number of IncX-like plasmid sequences and their occurrence in various organisms suggests that IncX plasmid...

  7. Environmental Factors Affecting Production, Release, and Field Populations of Conidia of Alternaria alternata, the Cause of Brown Spot of Citrus.

    Science.gov (United States)

    Timmer, L W; Solel, Z; Gottwald, T R; Ibañez, A M; Zitko, S E

    1998-11-01

    ABSTRACT Alternaria brown spot, caused by Alternaria alternata pv. citri, affects many tangerines and their hybrids, causing loss of immature leaves and fruit and reducing the marketability of the remaining fruit. Conidial production of A. alternata was greatest on mature leaves moistened and maintained at near 100% relative humidity (RH) for 24 h, whereas leaves that had been soaked or maintained at moderate RH produced few conidia. Conidial release from filter paper cultures and infected leaves was studied in a computer-controlled environmental chamber. Release of large numbers of conidia was triggered from both substrates by sudden drops in RH or by simulated rainfall events. Vibration induced release of low numbers of conidia, but red/infrared irradiation had no effect. In field studies from 1994 to 1996, air sampling with a 7-day recording volumetric spore trap indicated that conidia were present throughout the year with periodic large peaks. The number of conidia captured was not closely related to rainfall amounts or average wind speed, but was weakly related to the duration of leaf wetness. Likewise, disease severity on trap plants placed in the field weekly during 1995 to 1996 was not closely related to conidial numbers or rainfall amounts, but was weakly related to leaf wetness duration. Sufficient inoculum appears to be available to allow infection to occur throughout the year whenever susceptible host tissue and moisture are available.

  8. N-acetyl-L-cysteine affects growth, extracellular polysaccharide production, and bacterial biofilm formation on solid surfaces.

    Science.gov (United States)

    Olofsson, Ann-Cathrin; Hermansson, Malte; Elwing, Hans

    2003-08-01

    N-Acetyl-L-cysteine (NAC) is used in medical treatment of patients with chronic bronchitis. The positive effects of NAC treatment have primarily been attributed to the mucus-dissolving properties of NAC, as well as its ability to decrease biofilm formation, which reduces bacterial infections. Our results suggest that NAC also may be an interesting candidate for use as an agent to reduce and prevent biofilm formation on stainless steel surfaces in environments typical of paper mill plants. Using 10 different bacterial strains isolated from a paper mill, we found that the mode of action of NAC is chemical, as well as biological, in the case of bacterial adhesion to stainless steel surfaces. The initial adhesion of bacteria is dependent on the wettability of the substratum. NAC was shown to bind to stainless steel, increasing the wettability of the surface. Moreover, NAC decreased bacterial adhesion and even detached bacteria that were adhering to stainless steel surfaces. Growth of various bacteria, as monocultures or in a multispecies community, was inhibited at different concentrations of NAC. We also found that there was no detectable degradation of extracellular polysaccharides (EPS) by NAC, indicating that NAC reduced the production of EPS, in most bacteria tested, even at concentrations at which growth was not affected. Altogether, the presence of NAC changes the texture of the biofilm formed and makes NAC an interesting candidate for use as a general inhibitor of formation of bacterial biofilms on stainless steel surfaces.

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

    Science.gov (United States)

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

    2016-01-01

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

  10. 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...

  11. Engineering Complex Microbial Phenotypes with Continuous Genetic Integration and Plasmid Based Multi-gene Library

    Science.gov (United States)

    2013-10-09

    various fragments from L plant arum. Cells were grown in LB medium with 5% (V/V) ethanol and starting optical density of 0.3. Survival comparison...in final titer is observed. Expression of the HSP plasmid appears to slow both growth and production throughout the entire fermentation . However...this may be an effect of overexpression of the HSPs during the early portions of the fermentation where there are low concentrations of ethanol and

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

    Science.gov (United States)

    Kuwahara, S

    1978-09-01

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

  13. Complex nature of enterococcal pheromone-responsive plasmids.

    Science.gov (United States)

    Wardal, Ewa; Sadowy, Ewa; Hryniewicz, Waleria

    2010-01-01

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

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

    DEFF Research Database (Denmark)

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

    2003-01-01

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

  15. 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

  16. Differential S1P Receptor Profiles on M1- and M2-Polarized Macrophages Affect Macrophage Cytokine Production and Migration

    Science.gov (United States)

    Müller, Jan; von Bernstorff, Wolfram; Heidecke, Claus-Dieter

    2017-01-01

    Introduction. Macrophages are key players in complex biological processes. In response to environmental signals, macrophages undergo polarization towards a proinflammatory (M1) or anti-inflammatory (M2) phenotype. Sphingosine 1-phosphate (S1P) is a bioactive lysophospholipid that acts via 5 G-protein coupled receptors (S1P1–5) in order to influence a broad spectrum of biological processes. This study assesses S1P receptor expression on macrophages before and after M1 and M2 polarization and performs a comparative analysis of S1P signalling in the two activational states of macrophages. Methods. Bone marrow derived macrophages (BMDM) from C57 BL/6 mice were cultured under either M1- or M2-polarizing conditions. S1P-receptor expression was determined by quantitative RT-PCR. Influence of S1P on macrophage activation, migration, phagocytosis, and cytokine secretion was assessed in vitro. Results. All 5 S1P receptor subclasses were expressed in macrophages. Culture under both M1- and M2-polarizing conditions led to significant downregulation of S1P1. In contrast, M1-polarized macrophages significantly downregulated S1P4. The expression of the remaining three S1P receptors did not change. S1P increased expression of iNOS under M2-polarizing conditions. Furthermore, S1P induced chemotaxis in M1 macrophages and changed cytokine production in M2 macrophages. Phagocytosis was not affected by S1P-signalling. Discussion. The expression of different specific S1P receptor profiles may provide a possibility to selectively influence M1- or M2-polarized macrophages.

  17. Strawberry processing does not affect the production and urinary excretion of urolithins, ellagic acid metabolites, in humans.

    Science.gov (United States)

    Truchado, Pilar; Larrosa, Mar; García-Conesa, María Teresa; Cerdá, Begoña; Vidal-Guevara, María Luisa; Tomás-Barberán, Francisco A; Espín, Juan Carlos

    2012-06-13

    The study of fruit and vegetable processing and its effects on the levels of health-promoting constituents and their bioavailability and metabolism is very relevant to understanding the role of these constituents in human health. Strawberry polyphenols, and particularly ellagitannins and ellagic acid, have been associated with the health benefits of this berry for humans. These compounds are transformed into urolithins by the gut microbiota, and these metabolites exert several biological activities that could be responsible for the health effects of strawberries. Processing potentially increases the extraction of ellagitannins from the strawberry achenes and the release of ellagic acid from ellagitannins. It is of interest to evaluate the effect of processing on strawberry ellagitannin microbial metabolism compared with fresh strawberries. This study shows that no significant differences in the production and excretion of urolithins were found between the intake of fresh strawberries and that of a thermally processed strawberry puree containing the same amount of strawberries. Processing increases the amount of free ellagic acid 2.5-fold, but this had no effect on the transformation in urolithins by the gut microbiota or in the excretion of urolithin metabolites (urolithin glucuronides) in urine, showing that the release of ellagic acid from ellagitannins is not a relevant factor affecting the microbial metabolism. All of the volunteers produced urolithin A, but only 3 of 20 volunteers produced and excreted urolithin B. It is confirmed that some volunteers were efficient producers of urolithins, whereas other produced much lower amounts. These results show that processing does not modify the potential health effects of strawberry polyphenols.

  18. Advanced glycation end-products reduce collagen molecular sliding to affect collagen fibril damage mechanisms but not stiffness.

    Directory of Open Access Journals (Sweden)

    Gion Fessel

    Full Text Available Advanced glycation end-products (AGE contribute to age-related connective tissue damage and functional deficit. The documented association between AGE formation on collagens and the correlated progressive stiffening of tissues has widely been presumed causative, despite the lack of mechanistic understanding. The present study investigates precisely how AGEs affect mechanical function of the collagen fibril--the supramolecular functional load-bearing unit within most tissues. We employed synchrotron small-angle X-ray scattering (SAXS and carefully controlled mechanical testing after introducing AGEs in explants of rat-tail tendon using the metabolite methylglyoxal (MGO. Mass spectrometry and collagen fluorescence verified substantial formation of AGEs by the treatment. Associated mechanical changes of the tissue (increased stiffness and failure strength, decreased stress relaxation were consistent with reports from the literature. SAXS analysis revealed clear changes in molecular deformation within MGO treated fibrils. Underlying the associated increase in tissue strength, we infer from the data that MGO modified collagen fibrils supported higher loads to failure by maintaining an intact quarter-staggered conformation to nearly twice the level of fibril strain in controls. This apparent increase in fibril failure resistance was characterized by reduced side-by-side sliding of collagen molecules within fibrils, reflecting lateral molecular interconnectivity by AGEs. Surprisingly, no change in maximum fibril modulus (2.5 GPa accompanied the changes in fibril failure behavior, strongly contradicting the widespread assumption that tissue stiffening in ageing and diabetes is directly related to AGE increased fibril stiffness. We conclude that AGEs can alter physiologically relevant failure behavior of collagen fibrils, but that tissue level changes in stiffness likely occur at higher levels of tissue architecture.

  19. Impact of plasmids, including those encodingVirB4/D4 type IV secretion systems, on Salmonella enterica serovar Heidelberg virulence in macrophages and epithelial cells.

    Directory of Open Access Journals (Sweden)

    Kuppan Gokulan

    Full Text Available Salmonella enterica serovar Heidelberg (S. Heidelberg can cause foodborne illness in humans following the consumption of contaminated meat and poultry products. Recent studies from our laboratory have demonstrated that certain S. Heidelberg isolated from food-animal sources harbor multiple transmissible plasmids with genes that encode antimicrobial resistance, virulence and a VirB4/D4 type-IV secretion system. This study examines the potential role of these transmissible plasmids in bacterial uptake and survival in intestinal epithelial cells and macrophages, and the molecular basis of host immune system modulation that may be associated with disease progression. A series of transconjugant and transformant strains were developed with different combinations of the plasmids to determine the roles of the individual and combinations of plasmids on virulence. Overall the Salmonella strains containing the VirB/D4 T4SS plasmids entered and survived in epithelial cells and macrophages to a greater degree than those without the plasmid, even though they carried other plasmid types. During entry in macrophages, the VirB/D4 T4SS encoding genes are up-regulated in a time-dependent fashion. When the potential mechanisms for increased virulence were examined using an antibacterial Response PCR Array, the strain containing the T4SS down regulated several host innate immune response genes which likely contributed to the increased uptake and survival within macrophages and epithelial cells.

  20. Impact of plasmids, including those encodingVirB4/D4 type IV secretion systems, on Salmonella enterica serovar Heidelberg virulence in macrophages and epithelial cells.

    Science.gov (United States)

    Gokulan, Kuppan; Khare, Sangeeta; Rooney, Anthony W; Han, Jing; Lynne, Aaron M; Foley, Steven L

    2013-01-01

    Salmonella enterica serovar Heidelberg (S. Heidelberg) can cause foodborne illness in humans following the consumption of contaminated meat and poultry products. Recent studies from our laboratory have demonstrated that certain S. Heidelberg isolated from food-animal sources harbor multiple transmissible plasmids with genes that encode antimicrobial resistance, virulence and a VirB4/D4 type-IV secretion system. This study examines the potential role of these transmissible plasmids in bacterial uptake and survival in intestinal epithelial cells and macrophages, and the molecular basis of host immune system modulation that may be associated with disease progression. A series of transconjugant and transformant strains were developed with different combinations of the plasmids to determine the roles of the individual and combinations of plasmids on virulence. Overall the Salmonella strains containing the VirB/D4 T4SS plasmids entered and survived in epithelial cells and macrophages to a greater degree than those without the plasmid, even though they carried other plasmid types. During entry in macrophages, the VirB/D4 T4SS encoding genes are up-regulated in a time-dependent fashion. When the potential mechanisms for increased virulence were examined using an antibacterial Response PCR Array, the strain containing the T4SS down regulated several host innate immune response genes which likely contributed to the increased uptake and survival within macrophages and epithelial cells.

  1. 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.

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

    Directory of Open Access Journals (Sweden)

    Lauren C Frazer

    Full Text Available Loss of the conserved "cryptic" plasmid from C. trachomatis and C. muridarum is pleiotropic, resulting in reduced innate inflammatory activation via TLR2, glycogen accumulation and infectivity. The more genetically distant C. caviae GPIC is a natural pathogen of guinea pigs and induces upper genital tract pathology when inoculated intravaginally, modeling human disease. To examine the contribution of pCpGP1 to C. caviae pathogenesis, a cured derivative of GPIC, strain CC13, was derived and evaluated in vitro and in vivo. Transcriptional profiling of CC13 revealed only partial conservation of previously identified plasmid-responsive chromosomal loci (PRCL in C. caviae. However, 2-deoxyglucose (2DG treatment of GPIC and CC13 resulted in reduced transcription of all identified PRCL, including glgA, indicating the presence of a plasmid-independent glucose response in this species. In contrast to plasmid-cured C. muridarum and C. trachomatis, plasmid-cured C. caviae strain CC13 signaled via TLR2 in vitro and elicited cytokine production in vivo similar to wild-type C. caviae. Furthermore, inflammatory pathology induced by infection of guinea pigs with CC13 was similar to that induced by GPIC, although we observed more rapid resolution of CC13 infection in estrogen-treated guinea pigs. These data indicate that either the plasmid is not involved in expression or regulation of virulence in C. caviae or that redundant effectors prevent these phenotypic changes from being observed in C. caviae plasmid-cured strains.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    1990-08-01

    Spores and vegetative cells of Bacillus thuringiensis were more sensitive to UV light than were spores or cells of plasmid-cured B. thuringiensis strains or of the closely related Bacillus cereus. Introduction of B. thuringiensis plasmids into B. cereus by cell mating increased the UV sensitivity of the cells and spores. Protoxins encoded by one or more B. thuringiensis plasmids were not involved in spore sensitivity, since a B. thuringiensis strain conditional for protoxin accumulation was equally sensitive at the permissive and nonpermissive temperatures. In addition, introduction of either a cloned protoxin gene, the cloning vector, or another plasmid not containing a protoxin gene into a plasmid-cured strain of B. thuringiensis all increased the UV sensitivity of the spores. Although the variety of small, acid-soluble proteins was the same in the spores of all strains examined, the quantity of dipicolinic acid was about twice as high in the plasmid-containing strains, and this may account for the differences in UV sensitivity of the spores. The cells of some strains harboring only B. thuringiensis plasmids were much more sensitive than cells of any of the other strains, and the differences were much greater than observed with spores.

  4. Construction of plasmids with tunable copy numbers in Saccharomyces cerevisiae and their applications in pathway optimization and multiplex genome integration.

    Science.gov (United States)

    Lian, Jiazhang; Jin, Run; Zhao, Huimin

    2016-11-01

    The CEN/ARS-based low-copy plasmids and 2 μ-based high-copy plasmids have been broadly used for both fundamental studies and practical applications in Saccharomyces cerevisiae. However, the relative low copy numbers and narrow dynamic range limit their applications in many cases. In this study, the expression level of the selection marker proteins was engineered to increase the plasmid copy numbers. A series of plasmids with step-wise increased copy numbers were constructed. The copy number of the plasmids with engineered dominant markers (5-100 copies per cell) showed a positive correlation with the concentration of antibiotics supplemented to the growth media. Based on this finding, we developed a simple yet highly efficient strategy, named Pathway Optimization by Tuning Antibiotic Concentrations (POTAC) to rapidly balance the flux of multi-gene pathways at the DNA level in S. cerevisiae. As proof of concept, POTAC was used to optimize the lycopene and n-butanol biosynthetic pathways, increasing the production of lycopene and n-butanol by 10- and 100-fold, respectively. Additionally, multiplex genome integration with controllable copy numbers was attempted by combining the engineered dominant markers with the CRISPR/Cas9 system. Biotechnol. Bioeng. 2016;113: 2462-2473. © 2016 Wiley Periodicals, Inc.

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

    Science.gov (United States)

    Baumgartner, A; Breer, C; Schopfer, K

    1989-04-05

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

  7. 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.

  8. Characterization of a Novel Plasmid-Borne Thiopeptide Gene Cluster in Staphylococcus epidermidis Strain 115

    Science.gov (United States)

    Bennallack, Philip R.; Burt, Scott R.; Heder, Michael J.

    2014-01-01

    Thiopeptides are small (12- to 17-amino-acid), heavily modified peptides of bacterial origin. This antibiotic family, with more than 100 known members, is characterized by the presence of sulfur-containing heterocyclic rings and dehydrated residues within a macrocyclic peptide structure. Thiopeptides, including micrococcin P1, have garnered significant attention in recent years for their potent antimicrobial activity against bacteria, fungi, and even protozoa. Micrococcin P1 is known to target the ribosome; however, like those of other thiopeptides, its biosynthesis and mechanisms of self-immunity are poorly characterized. We have discovered an isolate of Staphylococcus epidermidis harboring the genes for thiopeptide production and self-protection on a 24-kb plasmid. Here we report the characterization of this plasmid, identify the antimicrobial peptide that it encodes, and provide evidence of a target replacement-mediated mechanism of self-immunity. PMID:25313391

  9. Diversity and epidemiology of plasmids from Enterobacteriaceae from human and non-human reservoirs

    DEFF Research Database (Denmark)

    Bielak, Eliza Maria

    The family of Enterobacteriaceae is comprised of Gram negative bacteria found in a variety of natural environments as well as in the gastrointestinal (GI) tracts of humans and many animals including diverse mammals, birds and reptiles. Three species of the enteric bacteria are largely responsible....... It is believed that these practices lead to the generation of reservoirs of antimicrobial resistance genes in the GI tracts of intensively reared food - production animals like pigs, poultry and cattle. Moreover, it has been previously shown that the bla genes (e.g. genes encoding resistance to ß-lactams) could...... be transmitted between different bacteria on mobile genetic elements (MGEs) like plasmids and variety of transposons. Evidences were also published indicating that zoonotic bacteria like E. coli or S. enterica resistant to diverse antimicrobials and harbouring plasmids might have been transmitted from farm...

  10. Dissemination of NDM-1-Producing Enterobacteriaceae Mediated by the IncX3-Type Plasmid.

    Directory of Open Access Journals (Sweden)

    Qing Yang

    Full Text Available The emergence and spread of NDM-1-producing Enterobacteriaceae have resulted in a worldwide public health risk that has affected some provinces of China. China is an exceptionally large country, and there is a crucial need to investigate the epidemic of blaNDM-1-positive Enterobacteriaceae in our province. A total of 186 carbapenem-resistant Enterobacteriaceae isolates (CRE were collected in a grade-3 hospital in Zhejiang province. Carbapenem-resistant genes, including blaKPC, blaIMP, blaVIM, blaOXA-48 and blaNDM-1 were screened and sequenced. Ninety isolates were identified as harboring the blaKPC-2 genes, and five blaNDM-1-positive isolates were uncovered. XbaI-PFGE revealed that three blaNDM-1-positive K. pneumoniae isolates belonged to two different clones. S1-PFGE and southern blot suggested that the blaNDM-1 genes were located on IncX3-type plasmids with two different sizes ranging from 33.3 to 54.7 kb (n=4 and 104.5 to 138.9 kb (n=1, respectively, all of which could easily transfer to Escherichia coli by conjugation and electrotransformation. The high-throughput sequencing of two plasmids was performed leading to the identification of a smaller 54-kb plasmid, which had high sequence similarity with a previously reported pCFNDM-CN, and a larger plasmid in which only a 7.8-kb sequence of a common gene environment around blaNDM-1 (blaNDM-1-trpF- dsbC-cutA1-groEL-ΔInsE, was detected. PCR mapping and sequencing demonstrated that four smaller blaNDM-1 plasmids contained a common gene environment around blaNDM-1 (IS5-blaNDM-1-trpF- dsbC-cutA1-groEL. We monitored the CRE epidemic in our hospital and determined that KPC-2 carbapenemase was a major risk to patient health and the IncX3-type plasmid played a vital role in the spread of the blaNDM-1 gene among the CRE.

  11. Partition of the Linear Plasmid N15: Interactions of N15 Partition Functions with the sop Locus of the F Plasmid

    OpenAIRE

    1999-01-01

    A locus close to one end of the linear N15 prophage closely resembles the sop operon which governs partition of the F plasmid; the promoter region contains similar operator sites, and the two putative gene products have extensive amino acid identity with the SopA and -B proteins of F. Our aim was to ascertain whether the N15 sop homologue functions in partition, to identify the centromere site, and to examine possible interchangeability of function with the F Sop system. When expressed at a m...

  12. Improved determination of plasmid copy number using quantitative real-time PCR for monitoring fermentation processes

    Directory of Open Access Journals (Sweden)

    Štrukelj Borut

    2008-03-01

    Full Text Available Abstract Background Recombinant protein production in Escherichia coli cells is a complex process, where among other parameters, plasmid copy number, structural and segregational stability of plasmid have an important impact on the success of productivity. It was recognised that a method for accurate and rapid quantification of plasmid copy number is necessary for optimization and better understanding of this process. Lately, qPCR is becoming the method of choice for this purpose. In the presented work, an improved qPCR method adopted for PCN determination in various fermentation processes was developed. Results To avoid experimental errors arising from irreproducible DNA isolation, whole cells, treated by heating at 95°C for 10 minutes prior to storage at -20°C, were used as a template source. Relative quantification, taking into account different amplification efficiencies of amplicons for chromosome and plasmid, was used in the PCN calculation. The best reproducibility was achieved when the efficiency estimated for specific amplicon, obtained within one run, was averaged. It was demonstrated that the quantification range of 2 log units (100 to 10000 bacteria per well enable quantification in each time point during fermentation. The method was applied to study PCN variation in fermentation at 25°C and the correlation between PCN and protein accumulation was established. Conclusion Using whole cells as a template source and relative quantification considering different PCR amplification efficiencies are significant improvements of the qPCR method for PCN determination. Due to the approaches used, the method is suitable for PCN determination in fermentation processes using various media and conditions.

  13. The good, the bad, and the expert: How consumer expertise affects review valence effects on purchase intentions in online product reviews

    NARCIS (Netherlands)

    Ketelaar, P.E.; Willemsen, L.M.; Sleven, L.; Kerkhof, P.

    2015-01-01

    This study aims to shed more light on the question whether, and under what circumstances, valence affects consumers' intention to buy a product after reading an online review. We hypothesize that receiver expertise could possibly moderate (a) the impact of review valence on consumers' purchase inten

  14. Mechanisms of Evolution in High-Consequence Drug Resistance Plasmids

    Directory of Open Access Journals (Sweden)

    Susu He

    2016-12-01

    Full Text Available The dissemination of resistance among bacteria has been facilitated by the fact that resistance genes are usually located on a diverse and evolving set of transmissible plasmids. However, the mechanisms generating diversity and enabling adaptation within highly successful resistance plasmids have remained obscure, despite their profound clinical significance. To understand these mechanisms, we have performed a detailed analysis of the mobilome (the entire mobile genetic element content of a set of previously sequenced carbapenemase-producing Enterobacteriaceae (CPE from the National Institutes of Health Clinical Center. This analysis revealed that plasmid reorganizations occurring in the natural context of colonization of human hosts were overwhelmingly driven by genetic rearrangements carried out by replicative transposons working in concert with the process of homologous recombination. A more complete understanding of the molecular mechanisms and evolutionary forces driving rearrangements in resistance plasmids may lead to fundamentally new strategies to address the problem of antibiotic resistance.

  15. 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...

  16. 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.

  17. Hypermutable ligation of plasmid DNA ends in cells from patients with Werner syndrome.

    Science.gov (United States)

    Rünger, T M; Bauer, C; Dekant, B; Möller, K; Sobotta, P; Czerny, C; Poot, M; Martin, G M

    1994-01-01

    Werner Syndrome is a rare autosomal recessive disorder characterized by an increased cancer risk and by symptoms suggestive of premature aging. Cells from these patients demonstrate a typical pattern of chromosomal instability and a spontaneous hypermutability with a high rate of unusually large deletions. We have studied the in vivo DNA ligation in three lymphoblast cell lines from Werner syndrome patients and three from normal donors. In our host cell ligation assay we transfected linearized plasmid pZ189 and measured the amount of plasmid DNA ends rejoined by these host cells as the ability of the recovered plasmid to transform bacteria. A mutagenesis marker gene close to the ligation site allowed screening for mutations. Subsequent mutation analysis provided information about the accuracy of the ligation process. The cells from Werner syndrome patients were as effective as normal cells in ligating DNA ends. However, mutation analysis revealed that the three Werner syndrome cell lines introduced 2.4-4.6 times more mutations (p < 0.001) than the normal cell lines during ligation of the DNA ends: the mutation rates were 69.4, 97.2, and 58.7%, as compared to 23.6, 21.7, and 24.4% in the normal cell lines. These increased mutation frequencies in plasmids ligated during passage through Werner syndrome cells were mainly due to a significant (p < 0.001) increase in deletions. This error-prone DNA ligation might be responsible for the spontaneous hypermutability and the genomic instability in Werner syndrome cells and related to the apparently accelerated aging and high cancer risk in affected patients.

  18. 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.

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

    DEFF Research Database (Denmark)

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

    2009-01-01

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

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

    Science.gov (United States)

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

    2012-01-01

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

  1. Construtcion of Neisseria Gonorrhoeae Porin B Plasmid Recombinant and Its Expression in E.coli

    Institute of Scientific and Technical Information of China (English)

    SONG Qifa; LIAO Fang; YE Siying; CUI Bing; XIONG Ping

    2005-01-01

    Summary: A prokaryotic expression recombinant plasmid pET-PIB to express porin B (PIB) of Neisseria gonorrhoeae in E.coli DE3 was constructed in order to provide a basis of research in detection, prophylactic and therapeutic vaccine against the pathogen infection. The gene encoding PIB was amplified by PCR from Neisseria gonorrhoeae and cloned into prokaryotic expression plasmid pET-28a(+) to construct a pET-PIB recombinant, which was verified by restriction endonuclease and DNA sequencing. Protein PIB was expressed in E.coli DE3 induced with IPTG. The antigenicity of the expressed protein was evaluated by indirect ELISA. Rabbits were immunized with the protein and serum was collected after immunization. To assess the immunogenicity of the protein, the titer of serum to protein PIB was determined by ELISA. DNA sequence analysis showed that the nucleic acid sequence of PIB gene was 99.28 % of homology compared with that (NGPIB18) published in GenBank. A 41 kD fused protein was detected by SDS-PAGE and was proven to have reactivity with anti-PIB polyclonal antibody from mouse. A polyclonal antibody to PIB of 1:4000 titer determined by indirect ELISA was obtained from rabbit immunized with the purified product. Recombinant plasmid encoding PIB of Neisseria gonorrhoeae was constructed. Protein PIB with antigenicity and immunogenicity was successfully expressed.

  2. How does co-product handling affect the carbon footprint of milk? Case study of milk production in New Zealand and Sweden

    DEFF Research Database (Denmark)

    Flysjö, Anna Maria; Cederberg, Christel; Henriksson, Maria;

    2011-01-01

    Purpose This paper investigates different methodologies of handling co-products in life cycle assessment (LCA) or carbon footprint (CF) studies. Co-product handling can have a significant effect on final LCA/CF results, and although there are guidelines on the preferred order for different methods...... for handling co-products, no agreed understanding on applicable methods is available. In the present study, the greenhouse gases (GHG) associated with the production of 1 kg of energy-corrected milk (ECM) at farm gate is investigated considering co-product handling. Materials and methods Two different milk...... (when slaughtered), calves, manure, hides, etc., the environmental burden (here GHG emissions) must be distributed between these outputs (in the present study no emissions are attributed to hides specifically, or to manure which is recycled on-farm). Different methodologically approaches, (1) system...

  3. Do blood components affect the production of reactive oxygen species (ROS by equine synovial cells in vitro?

    Directory of Open Access Journals (Sweden)

    Patrícia M. Brossi

    2012-12-01

    Full Text Available Blood-derived products are commonly administered to horses and humans to treat many musculoskeletal diseases, due to their potential antioxidant and anti-inflammatory effects. Nevertheless, antioxidant effects have never been shown upon horse synovial fluid cells in vitro. If proved, this could give a new perspective to justify the clinical application of blood-derived products. The aim of the present study was to investigate the antioxidant effects of two blood-derived products - plasma (unconditioned blood product - UBP and a commercial blood preparation (conditioned blood product - CBP¹ - upon stimulated equine synovial fluid cells. Healthy tarsocrural joints (60 were tapped to obtain synovial fluid cells; these cells were pooled, processed, stimulated with lipopolysaccharide (LPS or phorbol 12-myristate 13-acetate (PMA, and evaluated by flow cytometry for the production of reactive oxygen species (ROS. Upon addition of any blood-derived product here used - UBP and CBP - there was a significant decrease in the oxidative burst of synovial fluid cells (P<0.05. There was no difference between UBP and CBP effects. In conclusion, treatment of stimulated equine synovial cells with either UBP or CBP efficiently restored their redox equilibrium.

  4. Genetic islands in pome fruit pathogenic and nonpathogenic Erwinia species and related plasmids

    Directory of Open Access Journals (Sweden)

    Pablo eLlop

    2015-08-01

    Full Text Available New pathogenic bacteria species belonging to the genus Erwinia associated with pome fruit trees (Erwinia pyrifoliae, E. piriflorinigrans, E. uzenensis have been increasingly described in the last years, and comparative analyses have found that all these species share several genetic characteristics. Studies at different level (whole genome comparison, virulence genes, plasmid content, etc. show a high intraspecies homogeneity (i.e. among E. amylovora strains and also abundant similarities appear between the different Erwinia species: presence of plasmids of similar size in the pathogenic species; high similarity in several genes associated with exopolysaccharide production and hence, with virulence, as well as in some other genes, in the chromosomes. Many genetic similarities have been observed also among some of the plasmids (and genomes from the pathogenic species and E. tasmaniensis or E. billingiae, two epiphytic species on the same hosts. The amount of genetic material shared in this genus varies from individual genes to clusters, genomic islands and genetic material that even may constitute a whole plasmid. Recent research on evolution of erwinias point out the horizontal transfer acquisition of some genomic islands that were subsequently lost in some species and several pathogenic traits that are still present. How this common material has been obtained and is efficiently maintained in different species belonging to the same genus sharing a common ecological niche provides an idea of the origin and evolution of the pathogenic Erwinia and the interaction with nonpathogenic species present in the same niche, and the role of the genes that are conserved in all of them.

  5. Genetic islands in pome fruit pathogenic and non-pathogenic Erwinia species and related plasmids.

    Science.gov (United States)

    Llop, Pablo

    2015-01-01

    New pathogenic bacteria belonging to the genus Erwinia associated with pome fruit trees (Erwinia, E. piriflorinigrans, E. uzenensis) have been increasingly described in the last years, and comparative analyses have found that all these species share several genetic characteristics. Studies at different level (whole genome comparison, virulence genes, plasmid content, etc.) show a high intraspecies homogeneity (i.e., among E. amylovora strains) and also abundant similarities appear between the different Erwinia species: presence of plasmids of similar size in the pathogenic species; high similarity in several genes associated with exopolysaccharide production and hence, with virulence, as well as in some other genes, in the chromosomes. Many genetic similarities have been observed also among some of the plasmids (and genomes) from the pathogenic species and E. tasmaniensis or E. billingiae, two epiphytic species on the same hosts. The amount of genetic material shared in this genus varies from individual genes to clusters, genomic islands and genetic material that even may constitute a whole plasmid. Recent research on evolution of erwinias point out the horizontal transfer acquisition of some genomic islands that were subsequently lost in some species and several pathogenic traits that are still present. How this common material has been obtained and is efficiently maintained in different species belonging to the same genus sharing a common ecological niche provides an idea of the origin and evolution of the pathogenic Erwinia and the interaction with non-pathogenic species present in the same niche, and the role of the genes that are conserved in all of them.

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

    OpenAIRE

    Casselli, Timothy; Tourand, Yvonne; Bankhead, Troy

    2012-01-01

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

  7. Study of factors affecting the productivity of nurses based on the ACHIEVE model and prioritizing them using analytic hierarchy process technique, 2012

    Directory of Open Access Journals (Sweden)

    Payam Farhadi

    2013-01-01

    Full Text Available Objective: Improving productivity is one of the most important strategies for social-economic development. Human resources are known as the most important resources in the organizations′ survival and success. Aims: To determine the factors affecting the human resource productivity using the ACHIEVEa model from the nurses′ perspective and then prioritize them from the perspective of head nurses using Analytic Hierarchy Process (AHP technique. Settings and Design: Iran, Shiraz University of Medical Sciences teaching hospitals in 2012. Materials and Methods: This was an applied, cross-sectional and analytical-descriptive study conducted in two phases. In the first phase, to determine the factors affecting the human resource productivity from nurses′ perspective, 110 nurses were selected using a two-stage cluster sampling method. Required data were collected using the Persian version of Hersey and Goldsmith′s Human Resource Productivity Questionnaire. In the second phase, in order to prioritize the factors affecting human resource productivity based on the ACHIEVE model using AHP technique, pairwise comparisons matrices were given to the 19 randomly selected head nurses to express their opinions about those factors relative priorities or importance. Statistical Analysis Used: Collected data and matrices in two mentioned phases were analyzed using SPSS 15.0 and some statistical tests including Independent-Samples T-Test and Pearson Correlation coefficient, as well as, Super Decisions software (Latest Beta. Results: The human resource productivity had significant relationships with nurses′ sex (P = 0.008, marital status (P < 0.001, education level (P < 0.001, and all questionnaire factors (P < 0.05. Nurses′ productivity from their perspective was below average (44.97 ΁ 7.43. Also, the priorities of factors affecting the productivity of nurses based on the ACHIEVE model from the head nurses′ perspective using AHP technique, from the

  8. Estimating benthic secondary production from aquatic insect emergence in streams affected by mountaintop removal coal mining, West Virginia USA

    Science.gov (United States)

    Mountaintop removal and valley fill (MTR/VF) coal mining recountours the Appalachian landscape, buries headwater stream channels, and degrades downstream water quality. The goal of this study was to compare benthic community production estimates, based on seasonal insect emergen...

  9. Factors affecting Import Shares of Powdered Milk and other Milk Products and their Implications in Sri Lanka

    OpenAIRE

    Bogahawatte, C.; Herath, Janaranjana

    2006-01-01

    Import shares of liquid milk, powdered milk, condensed milk and other milk products were estimated to determine their relative competitiveness. The change of import shares with changes of exchange rate and world price of milk. The analysis based on yearly data between 1975-2006 showed that relative CIF prices and incomes were important factors influencing the market shares of milk and milk products. The results also showed that imported milk powder is price inelastic and a weak substitute for...

  10. Mechanisms driving diversity-productivity relationships differ between exotic and native communities and are affected by gastropod herbivory.

    Science.gov (United States)

    Korell, Lotte; Schmidt, Robin; Bruelheide, Helge; Hensen, Isabell; Auge, Harald

    2016-04-01

    Biodiversity experiments have shown that productivity usually increases with plant species richness. However, most of those studies disregarded the importance of trophic interactions to the diversity-productivity relationship, and focused on the loss of native species while ignoring invasions by exotic species. Yet, as functional complementarity and the impact of plant antagonists are likely to differ between native and exotic communities, the diversity-productivity relationship may change when native communities are invaded by exotic species. We conducted a mesocosm experiment to test how diversity effects, evenness, and productivity differed between exotic and native assemblages of grassland plants, and how these communities were influenced by slug herbivory. In line with other experiments, we found higher productivity in exotic than in native communities. However, different mechanisms (complementarity vs. selection effect) contributed to the positive diversity-productivity relationships in exotic vs. native communities. Against expectations, native communities showed much lower evenness and a greater selection effect, suggesting that competitive dominance among native species may be even stronger than among exotic species. Slug herbivory decreased productivity independently of species origin and species diversity. However, exotic communities showed a threefold higher complementarity effect than native communities in the absence of slugs, which was mainly driven by differences in the responses of native and exotic legumes and nonleguminous herbs. Our results imply that underlying mechanisms for the positive diversity-productivity relationship differ between native and exotic communities in the early stages of community development, and that differential responses of plant functional groups to generalist herbivory can contribute to this pattern.

  11. Susceptibility of Candida albicans biofilms to caspofungin and anidulafungin is not affected by metabolic activity or biomass production.

    Science.gov (United States)

    Marcos-Zambrano, Laura Judith; Escribano, Pilar; Bouza, Emilio; Guinea, Jesús

    2016-02-01

    Micafungin is more active against biofilms with high metabolic activity; however, it is unknown whether this observation applies to caspofungin and anidulafungin and whether it is also dependent on the biomass production. We compare the antifungal activity of anidulafungin, caspofungin, and micafungin against preformed Candida albicans biofilms with different degrees of metabolic activity and biomass production from 301 isolates causing fungemia in patients admitted to Gregorio Marañon Hospital (January 2007 to September 2014). Biofilms were classified as having low, moderate, or high metabolic activity according XTT reduction assay or having low, moderate, or high biomass according to crystal violet assay. Echinocandin MICs for planktonic and sessile cells were measured using the EUCAST E.Def 7.2 procedure and XTT reduction assay, respectively. Micafungin showed the highest activity against biofilms classified according to the metabolic activity and biomass production (P caspofungin and anidulafungin was not dependent on the metabolic activity of the biofilm or the biomass production. These observations were confirmed by scanning electron microscopy. None of the echinocandins produced major changes in the structure of biofilms with low metabolic activity and biomass production when compared with the untreated biofilms. However, biofilm with high metabolic activity or high biomass production was considerably more susceptible to micafungin; this effect was not shown by caspofungin or anidulafungin.

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

    Directory of Open Access Journals (Sweden)

    Joseph J Gillespie

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

  13. Performance and energy metabolism in restrictively fed weanling pigs are not affected by feeding either fermented cereals or their end-products.

    Science.gov (United States)

    Bruininx, E M A M; Binnendijk, G P; Zandstra, T; Heetkamp, M J W; Van Der Peet-Schwering, C M C; Gerrits, W J J

    2010-12-01

    To study the effects of feeding fermented cereals or just fermentation end-products on performance and energy metabolism, 18 restrictedly fed groups of eight pigs each were assigned to one of three dietary treatments: (i) a liquid control diet (C) containing 40% of a mixture of barley and wheat; or (ii) a liquid diet (F) containing 40% fermented barley and wheat; or (iii) a liquid diet as C with the addition of some important fermentation end-products (FP; organic acids and ethanol) in concentrations similar to those in the fermented F-diet. Energy and nitrogen balances, heat production, and performance traits were measured during two consecutive periods (days 1-5 and days 6-14). There was a considerable increase in average dry matter intake that tended (p = 0.06) to be higher in the FP-group than in the other groups. Apparent fecal digestibility of dry matter, ash, nitrogen and energy during period 2 were not affected (p > 0.1). Averaged over both periods, none of the energy metabolism parameters were affected by the diets (p > 0.1). However, there were diet × period interactions for metabolizable energy-intake (p = 0.07), energy retention (p feeding of either 40% fermented cereals nor their fermentation end-products affected performance and energy metabolism traits in weanling pigs. Nevertheless, lower postprandial activity-related heat production by pigs given the fermented cereals suggest a stimulating effect of fermented cereals on short term satiety that was not seen in pigs given fermentation end-products only.

  14. Factors affecting the production of a single-chain antibody fragment by Aspergillus awamori in a stirred tank reactor.

    Science.gov (United States)

    Sotiriadis, A; Keshavarz, T; Keshavarz-Moore, E

    2001-01-01

    A recombinant strain of Aspergillus awamori expressing anti-lysozyme single chain antibody fragments (scFv), under the control of a xylanase promoter, was studied in order to investigate the impact of medium, induction regime and protease production on the expression of the product. Experiments with the time of induction showed that the optimum results are achieved when induction is started in the late exponential phase (21 h after inoculation) improving the titer of the product from 14.5 mg L(-1), obtained in the early exponential phase (7 h after inoculation), to 16.2 mg L(-1). A 100% increase of the carbon (fructose) and nitrogen (ammonium sulfate) sources in the growth medium resulted in an increase in product concentration from 16.2 to 108.9 mg L(-1) and an increase in maximum dry cell weight from 7.5 to 11.5 g L(-1). A 50% reduction in the concentration of the inducer resulted in an increase in the product yield from 10 mg g(-1) dry cell weight to 12 mg g(-1). Proteolytic enzymes were produced during the fermentation up to concentrations equivalent to 1.4 g L(-1) trypsin, but they had no detrimental effect on the concentration of the antibody fragment.

  15. Factors affecting the production of seeds in fully fertile tomatoes (Lycopersicon esculentum L. Mill. and those showing a tendency to parthenocarpy

    Directory of Open Access Journals (Sweden)

    Barbara Gabara

    2014-01-01

    Full Text Available Comparative studies on the development of the female gametophyte, pollination and fertilization in two lines of Lycopersicon esculentum, Kholodostoykye (Kh, fertile and A33 (with a tendency to parthenocarpy have revealed that seed production is affected by disturbances in embryo sac formation but mainly by its degeneration after anthesis, which is especially visible in line A33. Moreover, delayed development of some embryo sacs and incomplete pollination due to various stigma levels seem to be responsible for the diminution of seed number in line A33. Deep fluorescence of numerous pollen grains as well as whole pollen tubes in 83.3 per cent of A33 stigmas and only 24.1 per cent in the Kh line points to the heterogeneity of pollen. This could be one more reason for reduced fertility. The results of application of plant growth regulators (auxin, PCIB which affect seed production in tomato of line A33 remain inconclusive.

  16. In vitro fermentation profiles, gas production rates, and microbiota modulation as affected by certain fructans, galactooligosaccharides, and polydextrose.

    Science.gov (United States)

    Hernot, David C; Boileau, Thomas W; Bauer, Laura L; Middelbos, Ingmar S; Murphy, Michael R; Swanson, Kelly S; Fahey, George C

    2009-02-25

    It is of interest to benefit from the positive intestinal health outcomes of prebiotic consumption but with minimal gas production. This study examined gas production potential, fermentation profile, and microbial modulation properties of several types of oligosaccharides. Substrates studied included short-chain, medium-chain, and long-chain fructooligosaccharides, oligofructose-enriched inulin, galactooligosaccharide, and polydextrose. Each substrate was fermented in vitro using human fecal inoculum, and fermentation characteristics were quantified at 0, 4, 8, and 12 h. Gas and short-chain fatty acid (SCFA) production data showed that short-chain oligosaccharides were more rapidly fermented and produced more SCFA and gas than substrates with greater degrees of polymerization. Lactobacilli increased similarly among substrates. Short-chain oligosaccharides fermentation resulted in the greatest increase in bifidobacteria concentrations. Mixing short- and long-chain oligosaccharides attenuated short-chain oligosaccharide fermentation rate and extent. This study provides new information on the fermentation characteristics of some oligosaccharides used in human nutrition.

  17. Long-term affected energy production of waste to energy technologies identified by use of energy system analysis

    DEFF Research Database (Denmark)

    Münster, Marie; Meibom, Peter

    2010-01-01

    incineration continues to treat the largest amount of waste. Investments in new waste incineration capacity may, however, be superseded by investments in new Waste-to-Energy technologies, particularly those utilising sorted fractions such as organic waste and refuse derived fuel. The changed use of waste...... production is identified by use of energy system analysis. The focus is on different uses of waste for energy production. The Waste-to-Energy technologies analysed include co-combustion of coal and waste, anaerobic digestion and thermal gasification. The analysis is based on optimization of both investments...... and production of electricity, district heating and bio-fuel in a future possible energy system in 2025 in the countries of the Northern European electricity market (Denmark, Norway, Sweden, Finland and Germany). Scenarios with different CO2 quota costs are analysed. It is demonstrated that the waste...

  18. Pollen production in sunflower (Helianthus annuus L.) is affected by air temperature and relative humidity during early reproductive growth

    OpenAIRE

    Astiz, V.; Hernández, Luis Francisco

    2013-01-01

    During microsporogenesis, sunflower florets might be exposed to episodes of day- and night-time temperatures exceeding 30 and 20 °C, respectively, that can affect pollen availability and consequently plant’s yield. The relationship between air temperature and relative humidity during the meiotic phase of microsporogenesis, and the quantity and quality of pollen produced by the flowers, were studied in two sunflower hybrids during two years. The hybrids were grown on irrigated plots in Bahía B...

  19. The Ratio of Blood Products Transfused Affects Mortality in Patients Receiving Massive Transfusions at a Combat Support Hospital

    Science.gov (United States)

    2007-10-01

    therapy resuscitation, and exacer- bated by hemorrhagic shock, metabolic acidosis , hypother- mia, hyperfibrinolysis, hypocalcemia, and anemia.11,14–19...persistent hypothermia and progressive metabolic acidosis .10 Several other retrospec- tive studies have confirmed the presence of coagulopathy in patients...Affects Mortality in Trauma Volume 63 • Number 4 811 metabolic acidosis , and hypocalcemia need continued study to determine whether they can improve

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

    Science.gov (United States)

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

    1992-01-01

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

  1. Field emissions of N2O during biomass production may affect the sustainability of agro-biofuels

    DEFF Research Database (Denmark)

    Carter, Mette Sustmann; Hauggaard-Nielsen, Henrik; Heiske, Stefan

    emissions is calculated as the avoided fossil fuel‐derived CO2, where the N2O emission has been subtracted. This value does not account for CO2 emissions from farm machinery and during biofuel production. We obtained the greatest net reduction in greenhouse gas emissions by co‐production of bioethanol......One way of reducing the emissions of fossil fuel‐derived CO2 is to replace fossil fuels with biofuels produced from agricultural biomasses or residuals. However, cultivation of soils results in emission of other greenhouse gasses, especially nitrous oxide (N2O). Previous studies have shown...

  2. Functional amyloids as inhibitors of plasmid DNA replication

    Science.gov (United States)

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

    2016-01-01

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

  3. Identification of Genetic Associations and Functional Polymorphisms of SAA1 Gene Affecting Milk Production Traits in Dairy Cattle

    Science.gov (United States)

    Zhang, Shengli; Zhang, Qin; Sun, Dongxiao

    2016-01-01

    Our initial RNA sequencing (RNA-seq) revealed that the Serum amyloid A1 (SAA1) gene was differentially expressed in the mammary glands of lactating Holstein cows with extremely high versus low phenotypic values of milk protein and fat percentage. To further validate the genetic effect and potential molecular mechanisms of SAA1 gene involved in regulating milk production traits in dairy cattle, we herein performed a study through genotype-phenotype associations. Six identified SNPs were significantly associated with one or more milk production traits (0.00002milk production traits in dairy cows. Subsequently, both luciferase assay and electrophoretic mobility shift assay (EMSA) clearly demonstrated that the allele A of g.-963C>A increased the promoter activity by binding the PARP factor while allele C did not. Bioinformatics analysis indicated that the secondary structure of SAA protein changed by the substitution A/G in the locus c. +2510A>G. Our findings were the first to reveal the significant associations of the SAA1 gene with milk production traits, providing basis for further biological function validation, and two identified SNPs, g.-963C>A and c. +2510A>G, may be considered as genetic markers for breeding in dairy cattle. PMID:27610623

  4. A 10-day heatwave at flowering superimposed on climate change conditions strongly affects production of 22 barley accessions

    DEFF Research Database (Denmark)

    Ingvordsen, Cathrine Heinz; Lyngkjær, Michael F.; Peltonen-Sainio, Pirjo

    Extreme climate events are projected to be among the future most challenging constraints to plant development. Heatwaves as well as floods and droughts cause acute changes in the growth environment determining our primary production (Collins et al., 2013). Europe experienced extreme heatwaves...

  5. Experimental investigation and optimization of process variables affecting the production of extracellular lipase by Kluyveromyces marxianus IFO 0288.

    Science.gov (United States)

    Stergiou, Panagiota-Yiolanda; Foukis, Athanasios; Sklivaniti, Helen; Zacharaki, Paraskevi; Papagianni, Maria; Papamichael, Emmanuel M

    2012-10-01

    In this study, the production and optimization of extracellular lipase from Kluyveromyces marxianus IFO 0288 was investigated by using optimized nutritional and cultural conditions in a yeast medium containing glucose as the carbon source in fully aerobic batch fermentation (150 rpm). The influence of four fermentation parameters (type of lipidic source, initial culture pH, temperature, and length of fermentation) on growth and lipase production was investigated and evaluated using the conventional "one variable at a time" approach and response surface methodology. An 18-fold increase in lipase production during 65 h of fermentation was obtained with optimized nutritional (0.5 % olive oil) and cultivation (pH 6.5, 35 °C) conditions by employing the conventional optimization method. By applying the response surface methodology technique the initial pH value of 6.4 and temperature of 32.5 °C were identified as optimal and led to further improvements (up to 18-fold) of extracellular lipase production. The results provide, for the first time, evidence that K. marxianus has the potential to be used as an efficient producer of extracellular lipase with prospective application in a variety of industrial and biotechnological areas.

  6. Light affects fumonisin production in strains of Fusarium fujikuroi, Fusarium proliferatum, and Fusarium verticillioides isolated from rice.

    Science.gov (United States)

    Matić, Slavica; Spadaro, Davide; Prelle, Ambra; Gullino, Maria Lodovica; Garibaldi, Angelo

    2013-09-16

    Three Fusarium species associated with bakanae disease of rice (Fusarium fujikuroi, Fusarium proliferatum, and Fusarium verticillioides) were investigated for their ability to produce fumonisins (FB1 and FB2) under different light conditions, and for pathogenicity. Compared to darkness, the conditions that highly stimulated fumonisin production were yellow and green light in F. verticillioides strains; white and blue light, and light/dark alternation in F. fujikuroi and F. proliferatum strains. In general, all light conditions positively influenced fumonisin production with respect to the dark. Expression of the FUM1 gene, which is necessary for the initiation of fumonisin production, was in accordance with the fumonisin biosynthetic profile. High and low fumonisin-producing F. fujikuroi strains showed typical symptoms of bakanae disease, abundant fumonisin-producing F. verticillioides strains exhibited chlorosis and stunting of rice plants, while fumonisin-producing F. proliferatum strains were asymptomatic on rice. We report that F. fujikuroi might be an abundant fumonisin producer with levels comparable to that of F. verticillioides and F. proliferatum, highlighting the need of deeper mycotoxicological analyses on rice isolates of F. fujikuroi. Our results showed for the first time the influence of light on fumonisin production in isolates of F. fujikuroi, F. proliferatum, and F. verticillioides from rice.

  7. Parameters affecting diacylglycerol formation during the production of specific-structured lipids by lipase-catalyzed interesterification

    DEFF Research Database (Denmark)

    Xu, Xuebing; Mu, Huiling; Skands, Anja;

    1999-01-01

    Diacylglycerols (DAGs) are important intermediates in lipase-catalyzed interesterification, but a high DAG concentration in the reaction mixture results in a high DAG content in the final product. We have previously shown that a high DAG concentration in the reaction mixture increases the degree ...

  8. Temperature Effects on Microbial CH4 and CO2 Production in Permafrost-Affected Soils From the Barrow Environmental Observatory

    Science.gov (United States)

    Graham, D. E.; Roy Chowdhury, T.; Zheng, J.; Moon, J. W.; Yang, Z.; Gu, B.; Wullschleger, S. D.

    2015-12-01

    Warmer Arctic temperatures are increasing the annual soil thaw depth and prolonging the thaw season in Alaskan permafrost zones. This change exposes organic matter buried in the soils and permafrost to microbial degradation and mineralization to form CO2 and CH4. The proportion and fluxes of these greenhouse gases released into the atmosphere control the global feedback on warming. To improve representations of these biogeochemical processes in terrestrial ecosystem models we compared soil properties and microbial activities in core samples of polygonal tundra from the Barrow Environmental Observatory. Measurements of soil water potential through the soil column characterized water binding to the organic and mineral components. This suction combines with temperature to control freezing, gas diffusion and microbial activity. The temperature-dependence of CO2 and CH4 production from anoxic soil incubations at -2, +4 or +8 °C identified a significant lag in methanogenesis relative to CO2 production by anaerobic respiration and fermentation. Changes in the abundance of methanogen signature genes during incubations indicate that microbial population shifts caused by thawing and warmer temperatures drive changes in the mixtures of soil carbon degradation products. Comparisons of samples collected across the microtopographic features of ice-wedge polygons address the impacts of water saturation, iron reduction and organic matter content on CH4 production and oxidation. These combined measurements build process understanding that can be applied across scales to constrain key response factors in models that address Arctic soil warming.

  9. Factors affecting the outcome of in vitro bovine embryo production using ovum pick-up-derived cumulus oocyte complexes

    NARCIS (Netherlands)

    Merton, J.S.

    2013-01-01

    Optimization of bovine ovum pick up (OPU) followed by in vitro embryo production (IVP) has been driven by the desire of both beef and dairy cattle breeders to enhance genetic improvement. The work presented in this thesis focuses on optimizing the efficiency and efficacy of the OPU-IVP program. Atte

  10. Do Proficiency and Study-Abroad Experience Affect Speech Act Production? Analysis of Appropriateness, Accuracy, and Fluency

    Science.gov (United States)

    Taguchi, Naoko

    2011-01-01

    This cross-sectional study examined the effect of general proficiency and study-abroad experience in production of speech acts among learners of L2 English. Participants were 25 native speakers of English and 64 Japanese college students of English divided into three groups. Group 1 (n = 22) had lower proficiency and no study-abroad experience.…

  11. Genetic and biochemical analyses of chromosome and plasmid gene homologues encoding ICL and ArCP domains in Vibrio anguillarum strain 775.

    Science.gov (United States)

    Di Lorenzo, Manuela; Stork, Michiel; Crosa, Jorge H

    2011-08-01

    Anguibactin, the siderophore produced by Vibrio anguillarum 775 is synthesized from 2,3-dihydroxybenzoic acid (DHBA), cysteine and hydroxyhistamine via a nonribosomal peptide synthetase (NRPS) mechanism. Most of the genes encoding anguibactin biosynthetic proteins are harbored by the pJM1 plasmid. In this work we report the identification of a homologue of the plasmid-encoded angB on the chromosome of strain 775. The product of both genes harbor an isochorismate lyase (ICL) domain that converts isochorismic acid to 2,3-dihydro-2,3-dihydroxybenzoic acid, one of the steps of DHBA synthesis. We show in this work that both ICL domains are functional in the production of DHBA in V. anguillarum as well as in E. coli. Substitution by alanine of the aspartic acid residue in the active site of both ICL domains completely abolishes their isochorismate lyase activity in vivo. The two proteins also carry an aryl carrier protein (ArCP) domain. In contrast with the ICL domains only the plasmid encoded ArCP can participate in anguibactin production as determined by complementation analyses and site-directed mutagenesis in the active site of the plasmid encoded protein, S248A. The site-directed mutants, D37A in the ICL domain and S248A in the ArCP domain of the plasmid encoded AngB were also tested in vitro and clearly show the importance of each residue for the domain function and that each domain operates independently.

  12. Short communication: A missense mutation in the PROP1 (prophet of Pit 1) gene affects male fertility and milk production traits in the US Holstein population.

    Science.gov (United States)

    Lan, X Y; Peñagaricano, F; DeJung, L; Weigel, K A; Khatib, H

    2013-02-01

    In previous studies, we reported significant associations of the POU1F1 pathway genes with reproduction and production traits in several dairy cattle populations. Polymorphisms in genes of this pathway were found to be associated with both female and male fertility traits in dairy cattle. The POU1F1 gene is a direct downstream target for the regulation of the prophet of Pit1 (PROP1) gene, also known as PROP paired-like homeobox 1. Interestingly, the position of PROP1 coincides with a quantitative trait locus affecting ovulation rate in cattle. Therefore, the objective of this study was to investigate whether PROP1 affects fertility and milk production traits in Holstein cattle. Using the DNA pooling sequencing approach, a missense single nucleotide polymorphism that replaces a histidine amino acid with an arginine was detected in exon 3 of PROP1. The arginine allele was found to be associated with a decrease in sire conception rate and an increase in productive life, protein yield, and net merit index in a population of 1,951 Holstein bulls. The transcription factors produced from the histidine and arginine isoforms are known to have different transcription, DNA binding, and regulation activities. As such, we propose that the association of the arginine isoform with decreased bull fertility is likely caused by reduced activity of this allele in male functions. The findings of this study suggest PROP1 polymorphisms as candidates in selection programs for fertility, health, and milk production traits in dairy cattle.

  13. Acute central neuropeptide Y administration increases food intake but does not affect hepatic very low-density lipoprotein (VLDL production in mice.

    Directory of Open Access Journals (Sweden)

    Janine J Geerling

    Full Text Available OBJECTIVE: Central neuropeptide Y (NPY administration stimulates food intake in rodents. In addition, acute modulation of central NPY signaling increases hepatic production of very low-density lipoprotein (VLDL-triglyceride (TG in rats. As hypertriglyceridemia is an important risk factor for atherosclerosis, for which well-established mouse models are available, we set out to validate the effect of NPY on hepatic VLDL-TG production in mice, to ultimately investigate whether NPY, by increasing VLDL production, contributes to the development of atherosclerosis. RESEARCH DESIGN AND METHODS: Male C57Bl/6J mice received an intracerebroventricular (i.c.v. cannula into the lateral (LV or third (3V ventricle of the brain. One week later, after a 4 h fast, the animals received an intravenous (i.v. injection of Tran(35S (100 µCi followed by tyloxapol (500 mg/kg body weight; BW, enabling the study of hepatic VLDL-apoB and VLDL-TG production, respectively. Immediately after the i.v. injection of tyloxapol, the animals received either an i.c.v. injection of NPY (0.2 mg/kg BW in artificial cerebrospinal fluid; aCSF, synthetic Y1 receptor antagonist GR231118 (0.5 mg/kg BW in aCSF or vehicle (aCSF, or an i.v. injection of PYY3-36 (0.5 mg/kg BW in PBS or vehicle (PBS. RESULTS: Administration of NPY into both the LV and 3V increased food intake within one hour after injection (+164%, p<0.001 and +367%, p<0.001, respectively. NPY administration neither in the LV nor in the 3V affected hepatic VLDL-TG or VLDL-apoB production. Likewise, antagonizing central NPY signaling by either PYY3-36 or GR231118 administration did not affect hepatic VLDL production. CONCLUSION: In mice, as opposed to rats, acute central administration of NPY increases food intake without affecting hepatic VLDL production. These results are of great significance when extrapolating findings on the central regulation of hepatic VLDL production between species.

  14. Agrobacterium tumefaciens pTAR parA promoter region involved in autoregulation, incompatibility and plasmid partitioning.

    Science.gov (United States)

    Gallie, D R; Kado, C I

    1987-02-05

    The locus responsible for directing proper plasmid partitioning of Agrobacterium tumefaciens pTAR is contained within a 1259 base-pair region. Insertions or deletions within this locus can result in the loss of the plasmid's ability to partition properly. One protein product (parA), approximately 25,000 Mr, is expressed from the par locus in Escherichia coli and A. tumefaciens protein analysis systems in vitro. DNA sequence analysis of the locus revealed a single 23,500 Mr open reading frame, confirming the protein data. A 248 base-pair region immediately upstream from the 23,500 Mr open reading frame, containing an array of 12 seven-base-pair palindromic repeats each of which are separated by exactly ten base-pairs of A + T-rich (75%) sequence, not only serves to provide the promoter but is also involved in parA autoregulation. In addition, this region containing a set of 12 seven-base-pair palindromic repeats, is responsible for plasmid-associated incompatibility within Inc Ag-1 and also functions as the cis-acting recognition site at which parA interacts to bring about partitioning. Transcriptional analysis indicated that only the DNA strand responsible for parA is actively transcribed, and that active transcription of the opposite strand of par can inhibit the production of parA, resulting in plasmid destabilization. The presence of the par locus in a plasmid results in stable inheritance within a wide range of members of Rhizobiaceae. Segregation rates of par-defective derivatives can be influenced by the host.

  15. Market-oriented new product development: How can a means-end chain approach affect the process?

    DEFF Research Database (Denmark)

    Søndergaard, Helle Alsted

    2005-01-01

    Purpose - Aims to ascertain whether a well-known theory within consumer research - a means-end chain (MEC) - can be used as a catalyst to achieve market oriented product development. Design/methodology/approach - Describes a case study, involving a Danish food manufacturer, where a MEC approach...... was introduced to a cross-functional development team at two different stages of the development process. Findings - Results show that MEC data is perceived as a good way of gaining knowledge about consumers; that the information serves well as the basis of discussions and for keeping project goals fixed....... The resukts also indicate that MEC data are most valuable to the team in the early stages of the development process and that lack of a learning orientation may inhibit the effects of a MEC approach. Originality/value - The MEC approach shows clear advantages for market oriented product development....

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

    Science.gov (United States)

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

    2014-10-01

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

  17. Water availability and calcium propionate affect fungal population and aflatoxins production in broiler finisher feed during storage.

    Science.gov (United States)

    Alam, Sahib; Shah, Hamid Ullah; Khan, Nazir Ahmad; Zeb, Alam; Shah, Abdul Sattar; Magan, Naresh

    2014-01-01

    The aim of this study was to investigate the effects of calcium propionate, water activity (aw) and incubation time on the total fungal count and aflatoxins B₁ (AFB₁), B₂ (AFB₂), G₁ (AFG₁) and G₂ (AFG₂) production in the broiler finisher feed. The feed was added with calcium propionate (5 g kg(-1)), adjusted to 0.85, 0.90 and 0.95 aw and stored for 28 days at 25°C, analysing for mould growth and aflatoxins production every 7 days. Analysis of variance indicated that all the factors (preservative, aw and storage time) alone and in combination significantly (p aflatoxins production in the feed. Minimum total fungal counts (1.99 × 10(2) CFU g(-1)) were observed in calcium propionate feed at 0.85 aw on day 1 and the highest (4.36 × 10(9) CFUs g(-1)) in control sample at 0.95 aw on day 28 of storage. During the storage period, AFB₁ content in control samples increased from 11.35 to 73.44, from 11.58 to 81.81 and from 11.54 to 102.68 ng g(-1), whereas in preserved feed the content of B₁ increased from 11.47 to 37.83, from 11.54 to 49.07 and from 11.20 to 53.14 ng g(-1) at 0.85, 0.90 and 0.95 aw, respectively. Similar patterns were noted for AFB2, AFG₁ and AFG₂ contents. All the aflatoxins readily increased over storage time; however, the increase was much slower in preserved feed that contained a lower amount of available water. This study reveals that calcium propionate addition to poultry litter along with water activity amelioration is an effective tool for controlling mould incidence and aflatoxin production in poultry feed.

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

    Science.gov (United States)

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

    2014-12-01

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

  19. Lipogems Product Treatment Increases the Proliferation Rate of Human Tendon Stem Cells without Affecting Their Stemness and Differentiation Capability.

    Science.gov (United States)

    Randelli, Pietro; Menon, Alessandra; Ragone, Vincenza; Creo, Pasquale; Bergante, Sonia; Randelli, Filippo; De Girolamo, Laura; Alfieri Montrasio, Umberto; Banfi, Giuseppe; Cabitza, Paolo; Tettamanti, Guido; Anastasia, Luigi

    2016-01-01

    Increasing the success rate of rotator cuff healing remains tremendous challenge. Among many approaches, the possibility of activating resident stem cells in situ, without the need to isolate them from biopsies, could represent valuable therapeutic strategy. Along this line, it has been recently demonstrated that lipoaspirate product, Lipogems, contains and produces growth-factors that may activate resident stem cells. In this study, human tendon stem cells (hTSCs) from the rotator cuff were cocultured in a transwell system with the Lipogems lipoaspirate product and compared to control untreated cells in terms of cell proliferation, morphology, stem cell marker and VEGF expression, and differentiation and migration capabilities. Results showed that the Lipogems product significantly increases the proliferation rate of hTSCs without altering their stemness and differentiation capability. Moreover, treated cells increase the expression of VEGF, which is crucial for the neovascularization of the tissue during the healing process. Overall, this study supports that directly activating hTSCs with the Lipogems lipoaspirate could represent a new practical therapeutic approach. In fact, obtaining a lipoaspirate is easier, safer, and more cost-effective than harvesting cells from tendon or bone marrow biopsies, expanding them in GMP facility and then reinjecting them in the patient.

  20. Overall Quality of Fruits and Vegetables Products Affected by the Drying Processes with the Assistance of Vacuum-Microwaves.

    Science.gov (United States)

    Figiel, Adam; Michalska, Anna

    2016-12-30

    The seasonality of fruits and vegetables makes it impossible to consume and use them throughout the year, thus numerous processing efforts have been made to offer an alternative to their fresh consumption and application. To prolong their availability on the market, drying has received special attention as currently this method is considered one of the most common ways for obtaining food and pharmaceutical products from natural sources. This paper demonstrates the weakness of common drying methods applied for fruits and vegetables and the possible ways to improve the quality using different drying techniques or their combination with an emphasis on the microwave energy. Particular attention has been drawn to the combined drying with the assistance of vacuum-microwaves. The quality of the dried products was ascribed by chemical properties including the content of polyphenols, antioxidant capacity and volatiles as well as physical parameters such as color, shrinkage, porosity and texture. Both these fields of quality classification were considered taking into account sensory attributes and energy aspects in the perspective of possible industrial applications. In conclusion, the most promising way for improving the quality of dried fruit and vegetable products is hybrid drying consisting of osmotic dehydration in concentrated fruit juices followed by heat pump drying and vacuum-microwave finish drying.

  1. Sodium houttuyfonate affects production of N-acyl homoserine lactone and quorum sensing-regulated genes expression in Pseudomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    Daqiang eWu

    2014-11-01

    Full Text Available Quorum sensing (QS is a means of cell-to-cell communication that uses diffusible signaling molecules that are sensed by the population to determine population density, thus allowing co-ordinate gene regulation in response to population density. In Pseudomonas aeruginosa, production of the QS signaling molecule, N-acyl homoserine lactone (AHL, co-ordinates expression of key factors of pathogenesis, including biofilm formation and toxin secretion. It is predicted that the inhibition of AHL sensing would provide an effective clinical treatment to reduce the expression of virulence factors and increase the effectiveness of antimicrobial agents. We previously demonstrated that sodium houttuyfonate (SH, commonly used in traditional Chinese medicine to treat infectious diseases, can effectively inhibit QS-regulated processes, including biofilm formation. Here, using a model system, we demonstrate that SH causes the dose-dependent inhibition of AHL production, through down-regulation of the AHL biosynthesis gene, lasI. Addition of SH also resulted in down-regulation of expression of the AHL sensor and transcriptional regulator, LasR, and inhibited the production of the QS-regulated virulence factors, pyocyanin and LasA. These results suggest that the antimicrobial activity of SH may be due to its ability to disrupt QS in P. aeruginosa.

  2. Mediating Effects of Intention On The Factors Affecting Organic Food Products Consumption Among Chinese Generation Y In Malaysia

    Directory of Open Access Journals (Sweden)

    Tan Poh Leong

    2015-02-01

    Full Text Available This study aimed to measure the effect of intention as a mediator in the relationship between internal factors and external factors on consumption. The factors were a internal factors: knowledge, awareness, health consciousness, product attributes, environmental concern, and b external factors: subjective norms, perceived behaviour control, and media and advertisement. From the review of literature, there were limited studies done on the mediating effect of intention on the relationship between (internal factor and external factors and organic food product consumption among CGY in Malaysia. A total of 410 respondents completed the online questionnaires that evaluated the factors. Structural Equation Modelling was used as the main practical approach for data analysis. The results showed that intention was a full or a complete mediator between the six exogenous constructs; knowledge, health consciousness, product attributes, environmental concern, perceived behaviour, and media and advertisement, while it only acted as a partial mediator between subjective norm and consumption. However, the intention was not a full or a complete mediator between awareness and consumption. Eventually, the implications for theory improvement and practices were discussed.

  3. Lipogems Product Treatment Increases the Proliferation Rate of Human Tendon Stem Cells without Affecting Their Stemness and Differentiation Capability

    Directory of Open Access Journals (Sweden)

    Pietro Randelli

    2016-01-01

    Full Text Available Increasing the success rate of rotator cuff healing remains tremendous challenge. Among many approaches, the possibility of activating resident stem cells in situ, without the need to isolate them from biopsies, could represent valuable therapeutic strategy. Along this line, it has been recently demonstrated that lipoaspirate product, Lipogems, contains and produces growth-factors that may activate resident stem cells. In this study, human tendon stem cells (hTSCs from the rotator cuff were cocultured in a transwell system with the Lipogems lipoaspirate product and compared to control untreated cells in terms of cell proliferation, morphology, stem cell marker and VEGF expression, and differentiation and migration capabilities. Results showed that the Lipogems product significantly increases the proliferation rate of hTSCs without altering their stemness and differentiation capability. Moreover, treated cells increase the expression of VEGF, which is crucial for the neovascularization of the tissue during the healing process. Overall, this study supports that directly activating hTSCs with the Lipogems lipoaspirate could represent a new practical therapeutic approach. In fact, obtaining a lipoaspirate is easier, safer, and more cost-effective than harvesting cells from tendon or bone marrow biopsies, expanding them in GMP facility and then reinjecting them in the patient.

  4. Lipogems Product Treatment Increases the Proliferation Rate of Human Tendon Stem Cells without Affecting Their Stemness and Differentiation Capability

    Science.gov (United States)

    Randelli, Pietro; Menon, Alessandra; Ragone, Vincenza; Creo, Pasquale; Bergante, Sonia; Randelli, Filippo; De Girolamo, Laura; Alfieri Montrasio, Umberto; Banfi, Giuseppe; Cabitza, Paolo; Tettamanti, Guido; Anastasia, Luigi

    2016-01-01

    Increasing the success rate of rotator cuff healing remains tremendous challenge. Among many approaches, the possibility of activating resident stem cells in situ, without the need to isolate them from biopsies, could represent valuable therapeutic strategy. Along this line, it has been recently demonstrated that lipoaspirate product, Lipogems, contains and produces growth-factors that may activate resident stem cells. In this study, human tendon stem cells (hTSCs) from the rotator cuff were cocultured in a transwell system with the Lipogems lipoaspirate product and compared to control untreated cells in terms of cell proliferation, morphology, stem cell marker and VEGF expression, and differentiation and migration capabilities. Results showed that the Lipogems product significantly increases the proliferation rate of hTSCs without altering their stemness and differentiation capability. Moreover, treated cells increase the expression of VEGF, which is crucial for the neovascularization of the tissue during the healing process. Overall, this study supports that directly activating hTSCs with the Lipogems lipoaspirate could represent a new practical therapeutic approach. In fact, obtaining a lipoaspirate is easier, safer, and more cost-effective than harvesting cells from tendon or bone marrow biopsies, expanding them in GMP facility and then reinjecting them in the patient. PMID:27057170

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

    Science.gov (United States)

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

    2000-01-01

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

  6. Production Conditions Affect the In Vitro Anti-Tumoral Effects of a High Concentration Multi-Strain Probiotic Preparation

    Science.gov (United States)

    Cinque, Benedetta; La Torre, Cristina; Lombardi, Francesca; Palumbo, Paola; Van der Rest, Michel

    2016-01-01

    A careful selection of the probiotic agent, standardization of the dose and detailed characterization of the beneficial effects are essential when considering use of a probiotic for the dietary management of serious diseases. However, changes in the manufacturing processes, equipment or facilities can result in differences in the product itself due to the live nature of probiotics. The need to reconfirm safety and/or efficacy for any probiotic product made at a different factory is therefore mandatory. Recently, under the brand VSL#3®, a formulation produced by a manufacturer different from the previous one, has been commercialized in some European countries (the UK and Holland). VSL#3 is a high concentration multi-strain preparation which has been recognized by the main Gastroenterology Associations for the dietary management of pouchitis as well as ulcerative colitis. We have compared the “original” VSL#3 produced in USA with the “newfound” VSL#3 produced in Italy. According to our results, the “newfound” VSL#3 has 130–150% more “dead bacteria” compared to the “original” product, raising concerns for the well-known association between dead microbes with adverse effects. The abilities of bacterial lysates from the two formulations to influence in vitro viability and proliferation of different tumor cell lines also resulted different. The repair of previously scratched monolayers of various adherent tumor cell lines (i.e. HT1080, and Caco-2 cells) was inhibited more significantly by the “original” VSL#3 when compared to the “newfound” VSL#3. Tumor cell cycle profile, in particular cell cycle arrest and apoptotic death of the cancer cells, further confirms that the “original” VSL#3 has a better functional profile than the “newfound” VSL#3, at least in in vitro. Our data stress the importance of the production conditions for the “newfound” VSL#3 considering that this product is intended to be used for the dietary management

  7. 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.

  8. Establishment of a novel anabolism-based addiction system with an artificially introduced mevalonate pathway: complete stabilization of plasmids as universal application in white biotechnology.

    Science.gov (United States)

    Kroll, Jens; Steinle, Anna; Reichelt, Rudolf; Ewering, Christian; Steinbüchel, Alexander

    2009-05-01

    Plasmid stability in recombinant microorganisms is a very important requirement for highly efficient plasmid-based production processes in biotechnology. To stably maintain plasmids, we developed in this study an efficient and stringent novel anabolism-based addiction system, which can be widely used. This novel addiction system is based on two components: (i) an Escherichia coli HMS174(DE3) knockout mutant of the ispH gene coding for 4-hydroxy-3-methylbut-2-enyl diphosphate reductase (EC 1.17.1.2) of the deoxyxylulose 5-phosphate (DXP) pathway, impairing the synthesis of isopentenyl pyrophosphate (IPP) and (ii) a completely synthetic and episomal mevalonate (MVA) pathway as an alternative supplier of essential IPP. The latter is encoded by a plasmid that contains the genes for HMG-CoA reductases from Lactococcus lactis and Staphylococcus aureus plus HMG-CoA-synthase, MVA kinase, MVP kinase and MVPP decarboxylase from S. aureus. This plasmid should then also harbor the genes for the protein or for the pathway that will be produced or that will be utilized for production of a chemical. To demonstrate the functionality of this addiction system, a mutated cyanophycin synthetase gene (cphA(6308)C595S) was used. To determine plasmid stabilities, flasks experiments in media supplied or not supplied with antibiotics were carried out with the knockout mutant and two control strains, one harboring plasmid pCOLADuet-1::MVA1-5::cphA(6308) and the other harboring a conventional expression plasmid pET-23a::cphA(6308). As revealed by measuring the colony-forming units of aliquots spread on solid media with or without antibiotics, the knockout mutant revealed a plasmid stability of 100% whereas the control strains exhibited plasmid stabilities of only 64% and 2%, respectively. Radiometric enzyme activity measurements for CphA revealed only 95% and 12.5% of the activity in the control strains harboring pCOLADuet-1::MVA1-5::cphA(6308) and pET-23a::cphA(6308), respectively, in

  9. Parameters affecting incorporation and by-product formation during the production of structured phospholipids by lipase-catalyzed acidolysis in solvent free system

    DEFF Research Database (Denmark)

    Vikbjerg, Anders Falk; Mu, Huiling; Xu, Xuebing

    2005-01-01

    to minimize by-products during reaction. In the present study we examined the Lipozyme RM IM-catalyzed acidolysis for the production of structured phospholipids between phosphatidylcholine (PC) and caprylic acid in the solvent free system. A five-factor response surface design was used to evaluate...... at three levels with two star points. All parameters besides water addition had an effect on the incorporation of caprylic acid into PC and lysophosphatidylcholine (LPC). Reaction time and enzyme dosage showed increased effect on incorporation into PC, while substrate ratio and reaction temperature showed...... opposite effect. The PC content decreased with increase of all parameters except for substrate ratio. Optimal conditions are recommended as enzyme dosage 40%, reaction temperature 55°C, water addition 1%, reaction time 70h, and substrate ratio 6 mol/mol caprylic acid/PC. Under these conditions...

  10. Mitochondrial pAL2-1 plasmid homologs are senescence factors in Podospora anserina independent of intrinsic senescence

    NARCIS (Netherlands)

    Diepeningen, van A.D.; Debets, A.J.M.; Slakhorst-Wandel, S.M.; Hoekstra, R.F.

    2008-01-01

    Since the first description of a linear mitochondrial plasmid in Podospora anserina, pAL2-1, and homologous plasmids have gone from being considered beneficial longevity plasmids, via neutral genetic elements, toward mutator plasmids causing senescence. The plasmid has an invertron structure, with t

  11. Recombinogenic engineering of conjugative plasmids with fluorescent marker cassettes

    DEFF Research Database (Denmark)

    Reisner, A.; Molin, Søren; Zechner, E.L.

    2002-01-01

    An efficient approach for the insertion of fluorescent marker genes with sequence specificity into conjugative plasmids in Escherichia coli is described. For this purpose, homologous recombination of linear double-stranded targeting DNA was mediated by the bacteriophage lambda recombination...... resistance genes and fluorescent markers. The choice of 5' non-homologous extensions in primer pairs used for amplifying the marker cassettes determines the site specificity of the targeting DNA. This methodology is applicable to the modification of all plasmids that replicate in E coli and is not restricted...

  12. 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.

  13. 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...... of costly regions from the plasmid backbone, effectively expanding the host-range of the plasmid. Although these adaptations were also beneficial to plasmid persistence in a naïve K. pneumoniae host, they were never observed in this species, indicating that differential evolvability can limit opportunities...

  14. Storage of parbaked bread affects shelf life of fully baked end product: a ¹H NMR study.

    Science.gov (United States)

    Bosmans, Geertrui M; Lagrain, Bert; Ooms, Nand; Fierens, Ellen; Delcour, Jan A

    2014-12-15

    Full baking of earlier partially baked (parbaked) bread can supply fresh bread to the consumer at any time of the day. When parbaked bread loaves were stored at -25, 4 or 23°C, the extent of crumb to crust moisture migration and amylopectin retrogradation differed with storage temperature, and the firming rate was evidently lowest during frozen storage. The extent of crumb to crust moisture migration during parbaked bread storage largely determined the mass of the fresh finished bread, and its crumb and crust moisture contents. Initial NMR proton mobility, initial resilience, the extent of amylopectin retrogradation and changes in firmness and resilience during storage of fully baked bread were affected by its crumb moisture content. The lowest firming rate was observed for finished bread resulting from parbaked bread stored at -25°C, while the highest firming rate was observed for finished bread from parbaked bread stored at 23°C.

  15. Breed of cow and herd productivity affect milk composition and modeling of coagulation, curd firming, and syneresis.

    Science.gov (United States)

    Stocco, G; Cipolat-Gotet, C; Bobbo, T; Cecchinato, A; Bittante, G

    2017-01-01

    Milk coagulation properties (MCP) have been widely investigated in the past using milk collected from different cattle breeds and herds. However, to our knowledge, no previous studies have assessed MCP in individual milk samples from several multi-breed herds characterized by either high or low milk productivity, thereby allowing the effects of herd and cow breed to be evaluated independently. Multi-breed herds (n=41) were classified into 2 categories based on milk productivity (high vs. low), defined according to the average milk net energy yielded daily by lactating cows. Milk samples were taken from 1,508 cows of 6 different breeds: 3 specialized dairy (Holstein-Friesian, Brown Swiss, Jersey) and 3 dual-purpose (Simmental, Rendena, Alpine Grey) breeds, and analyzed in duplicate (3,016 tests) using 2 lactodynamographs to obtain 240 curd firming (CF) measurements over 60min (1 every 15 s) for each duplicate. The 5 traditional single-point MCP (RCT, k20, a30, a45, and a60) were yielded directly by the instrument from the available CF measures. All 240 CF measures of each replicate were also used to estimate 4 individual equation parameters: RCT estimated according to curd firm change over time modeling (RCTeq), asymptotic potential curd firmness (CFP), curd firming instant rate constant (kCF), and syneresis instant rate constant (kSR) and 2 derived traits: maximum curd firmness achieved within 45min (CFmax) and time at achievement of CFmax (tmax) by curvilinear regression using a nonlinear procedure. Results showed that the effect of herd-date on traditional and modeled MCP was modest, ranging from 6.1% of total variance for k20 to 10.7% for RCT, whereas individual animal variance was the highest, ranging from 32.0% for tmax to 82.5% for RCTeq. The repeatability of MCP was high (>80%) for all traits except those associated with the last part of the lactodynamographic curve (i.e., a60, kSR, kCF, and tmax: 57 to 71%). Reproducibility, taking into account the effect

  16. Nitrate reductase-mediated NO production enhances Cd accumulation in Panax notoginseng roots by affecting root cell wall properties.

    Science.gov (United States)

    Kan, Qi; Wu, Wenwei; Yu, Wenqian; Zhang, Jiarong; Xu, Jin; Rengel, Zed; Chen, Limei; Cui, Xiuming; Chen, Qi

    2016-04-01

    Panax notoginseng (Burk) F. H. Chen is a traditional medicinal herb in China. However, the high capacity of its roots to accumulate cadmium (Cd) poses a potential risk to human health. Although there is some evidence for the involvement of nitric oxide (NO) in mediating Cd toxicity, the origin of Cd-induced NO and its function in plant responses to Cd remain unknown. In this study, we examined NO synthesis and its role in Cd accumulation in P. notoginseng roots. Cd-induced NO production was significantly decreased by application of the nitrate reductase inhibitor tungstate but not the nitric oxide synthase inhibitor L-NAME (N(G)-methyl-l-arginine acetate), indicating that nitrate reductase is the major contributor to Cd-induced NO production in P. notoginseng roots. Under conditions of Cd stress, sodium nitroprusside (SNP, an NO donor) increased Cd accumulation in root cell walls but decreased Cd translocation to the shoot. In contrast, the NO scavenger cPTIO (2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide) and tungstate both significantly decreased NO-increased Cd retention in root cell walls. The amounts of hemicellulose 1 and pectin, together with pectin methylesterase activity, were increased with the addition of SNP but were decreased by cPTIO and tungstate. Furthermore, increases or decreases in hemicellulose 1 and pectin contents as well as pectin methylesterase activity fit well with the increased or decreased retention of Cd in the cell walls of P. notoginseng roots. The results suggest that nitrate reductase-mediated NO production enhances Cd retention in P. notoginseng roots by modulating the properties of the cell wall.

  17. Feedbacks between earlywood anatomy and non-structural carbohydrates affect spring phenology and wood production in ring-porous oaks

    Science.gov (United States)

    Pérez-de-Lis, Gonzalo; García-González, Ignacio; Rozas, Vicente; Olano, José Miguel

    2016-10-01

    Non-structural carbohydrates (NSC) play a central role in the construction and maintenance of a tree's vascular system, but feedbacks between the NSC status of trees and wood formation are not fully understood. We aimed to evaluate multiple dependencies among wood anatomy, winter NSC, and phenology for coexisting temperate (Quercus robur) and sub-Mediterranean (Q. pyrenaica) oaks along a water-availability gradient in the NW Iberian Peninsula. Sapwood NSC concentrations were quantified at three sites in December 2012 (N = 240). Leaf phenology and wood anatomy were surveyed in 2013. Structural equation modelling was used to analyse the interplay among hydraulic diameter (Dh), winter NSC, budburst date, and earlywood vessel production (EVP), while the effect of Dh and EVP on latewood width was assessed by using a mixed-effects model. NSC and wood production increased under drier conditions for both species. Q. robur showed a narrower Dh and lower soluble sugar (SS) concentration (3.88-5.08 % dry matter) than Q. pyrenaica (4.06-5.57 % dry matter), but Q. robur exhibited larger EVP and wider latewood (1403 µm) than Q. pyrenaica (667 µm). Stem diameter and Dh had a positive effect on SS concentrations, which were related to an earlier leaf flushing in both species. Sapwood sugar content appeared to limit EVP exclusively in Q. pyrenaica. In turn, Dh and EVP were found to be key predictors of latewood growth. Our results confirm that sapwood SS concentrations are involved in modulating growth resumption and xylem production in spring. Q. pyrenaica exhibited a tighter control of carbohydrate allocation to wood formation than Q. robur, which would play a role in protecting against environmental stress in the sub-Mediterranean area.

  18. A critical appraisal of the factors affecting energy production from amorphous silicon photovoltaic arrays in a maritime climate

    Energy Technology Data Exchange (ETDEWEB)

    Gottschalg, R.; Betts, T.R.; Williams, S.R.; Sauter, D.; Infield, D.G. [Loughborough University (United Kingdom). Department of Electronic and Electrical Engineering, Centre for Renewable Energy Systems Technology; Kearney, M.J. [University of Surrey, Guildford (United Kingdom). School of Electronics and Physical Sciences, Advanced Technology Institute

    2004-12-01

    Contradictory reports exist in the literature regarding the energy production from amorphous silicon photovoltaic arrays. The majority claims high-energy output compared to crystalline silicon arrays of the same power rating (i.e. high kW h/kW{sub p}), but some reports point to less favourable comparisons. The reasons for these conflicting reports are investigated using long-term measurements of the I-V characteristics of a number of amorphous silicon devices, in conjunction with in situ measurements of the solar spectrum and other relevant environmental parameters. It is shown that the variation in the performance of devices produced by different manufacturers is so significant that one cannot speak of the performance of amorphous silicon devices in general; one has to investigate each type of amorphous silicon panel separately. The causes of differences in energy production are investigated in detail. The major factor impacting on the seasonal performance in the UK is identified to be variations in the solar spectrum. Single junction devices exhibit some seasonal thermal annealing but multi-junctions do not show this effect at a significant level. Scope for further improvement is identified, largely in the photon absorption. The response to different spectra can be modified to some extent, which would bridge the gap between the best and the worst performers in the field. It is also shown that in the case of multi-junction devices an optimised current matching might bring a 5% increase in energy production for this location. Differences in the magnitude of the fill factor have been identified to be the second most significant cause for performance variation between the different samples in the test, suggesting additional scope for improvement. (author)

  19. SUBTASK 1.7 EVALUATION OF KEY FACTORS AFFECTING SUCCESSFUL OIL PRODUCTION IN THE BAKKEN FORMATION, NORTH DAKOTA PHASE II

    Energy Technology Data Exchange (ETDEWEB)

    Darren D. Schmidt; Steven A. Smith; James A. Sorensen; Damion J. Knudsen; John A. Harju; Edward N. Steadman

    2011-10-31

    Production from the Bakken and Three Forks Formations continues to trend upward as forecasts predict significant production of oil from unconventional resources nationwide. As the U.S. Geological Survey reevaluates the 3.65 billion bbl technically recoverable estimate of 2008, technological advancements continue to unlock greater unconventional oil resources, and new discoveries continue within North Dakota. It is expected that the play will continue to expand to the southwest, newly develop in the northeastern and northwestern corners of the basin in North Dakota, and fully develop in between. Although not all wells are economical, the economic success rate has been near 75% with more than 90% of wells finding oil. Currently, only about 15% of the play has been drilled, and recovery rates are less than 5%, providing a significant future of wells to be drilled and untouched hydrocarbons to be pursued through improved stimulation practices or enhanced oil recovery. This study provides the technical characterizations that are necessary to improve knowledge, provide characterization, validate generalizations, and provide insight relative to hydrocarbon recovery in the Bakken and Three Forks Formations. Oil-saturated rock charged from the Bakken shales and prospective Three Forks can be produced given appropriate stimulation treatments. Highly concentrated fracture stimulations with ceramic- and sand-based proppants appear to be providing the best success for areas outside the Parshall and Sanish Fields. Targeting of specific lithologies can influence production from both natural and induced fracture conductivity. Porosity and permeability are low, but various lithofacies units within the formation are highly saturated and, when targeted with appropriate technology, release highly economical quantities of hydrocarbons.

  20. Amino acid and glucose metabolism in fed-batch CHO cell culture affects antibody production and glycosylation

    DEFF Research Database (Denmark)

    Fan, Yuzhou; Jimenez Del Val, Ioscani; Müller, Christian

    2015-01-01

    and feeds were studied using two IgG-producing cell lines. Our results indicate that the balance of glucose and amino acid concentration in the culture is important for cell growth, IgG titer and N-glycosylation. Accordingly, the ideal fate of glucose and amino acids in the culture could be mainly towards...... optimization, especially media optimization. Gaining knowledge on their interrelations could provide insight for obtaining higher immunoglobulin G (IgG) titer and better controlling glycosylationrelated product quality. In this work, different fed-batch processes with two chemically defined proprietary media...

  1. Determination of some significant batch culture conditions affecting acetyl-xylan esterase production by Penicillium notatum NRRL-1249

    Directory of Open Access Journals (Sweden)

    Akhtar MN

    2011-05-01

    Full Text Available Abstract Background Acetyl-xylan esterase (AXE, EC 3.1.1.72 hydrolyses acetate group from the linear chain of xylopyranose residues bound by β-1,4-linkage. The enzyme finds commercial applications in bio-bleaching of wood pulp, treating animal feed to increase digestibility, processing food to increase clarification and converting lignocellulosics to feedstock and fuel. In the present study, we report on the production of an extracellular AXE from Penicillium notatum NRRL-1249 by solid state fermentation (SSF. Results Wheat bran at a level of 10 g (with 4 cm bed height was optimized as the basal substrate for AXE production. An increase in enzyme activity was observed when 7.5 ml of mineral salt solution (MSS containing 0.1% KH2PO4, 0.05% KCl, 0.05% MgSO4.7H2O, 0.3% NaNO3, 0.001% FeSO4.2H2O and 0.1% (v/w Tween-80 as an initial moisture content was used. Various nitrogen sources including ammonium sulphate, urea, peptone and yeast extract were compared for enzyme production. Maximal enzyme activity of 760 U/g was accomplished which was found to be highly significant (p ≤ 0.05. A noticeable enhancement in enzyme activity was observed when the process parameters including incubation period (48 h, initial pH (5, 0.2% (w/w urea as nitrogen source and 0.5% (v/w Tween-80 as a stimulator were further optimized using a 2-factorial Plackett-Burman design. Conclusion From the results it is clear that an overall improvement of more than 35% in terms of net enzyme activity was achieved compared to previously reported studies. This is perhaps the first report dealing with the use of P. notatum for AXE production under batch culture SSF. The Plackett-Burman model terms were found highly significant (HS, suggesting the potential commercial utility of the culture used (df = 3, LSD = 0.126.

  2. How does the entering of copper nanoparticles into biological wastewater treatment system affect sludge treatment for VFA production.

    Science.gov (United States)

    Chen, Hong; Chen, Yinguang; Zheng, Xiong; Li, Xiang; Luo, Jingyang

    2014-10-15

    Usually the studies regarding the effect of engineered nanoparticles (NPs), which are released to wastewater treatment plant, on sludge anaerobic treatment in the literature have been conducted by directly adding NPs to sludge treatment system. Actually, NPs must enter into the wastewater treatment facility from influent before sludge being treated. Thus, the documented results can not reflect the real situations. During sludge anaerobic treatment for producing volatile fatty acids (VFA, the preferred carbon source for wastewater biological nutrient removal), it was found in this study that the entering of CuNPs to biological wastewater treatment system had no significant effect on sludge-derived VFA generation, while direct addition of CuNPs to sludge fermentation reactor caused a much lower VFA production, when compared to the control test. Further investigation revealed that the entering of CuNPs into wastewater biological treatment system improved sludge solubilization due to the decline of sludge particle size and the increase of sludge microorganism cells breakage. In addition, there was no obvious influence on hydrolysis, while significant inhibition was observed on acidification, resulting in the final VFA production similar to the control. When CuNPs were directly added to the fermentation system, the solubilization was little influenced, however the hydrolysis and acidification were seriously inhibited, causing the ultimate VFA generation decreased. Therefore, selecting proper method close to the real situation is vital to accurately assess the toxicity of nanoparticles on sludge anaerobic fermentation.

  3. Do Water Rights Affect Technical Efficiency and Social Disparities of Crop Production in the Mediterranean? The Spanish Ebro Basin Evidence

    Directory of Open Access Journals (Sweden)

    Sonia Quiroga

    2014-11-01

    Full Text Available The coming agenda for the European Common Agricultural Policy includes more incentives for the environmental compliance of farmer’s activities. This will be particularly important in the case of water risk management in Mediterranean countries. Among the new challenges is the need to evaluate some of the instruments necessary to comply with the Water Framework Directive requirements that emphasize the management of water demand to achieve the environmental targets. Here we analyze the implications of changing water rights as a policy response to these challenges. We analyze two important aspects of the decision: (i the effects on the crop productivity and efficiency and (ii the effects on the rural income distribution. We provide the empirical estimations for the marginal effects on the two considered aspects. First, we calculate a stochastic frontier production function for five representative crops using historical data to estimate technical efficiency. Second, we use a decomposition of the Gini coefficient to estimate the impact of irrigation rights changes on yield disparity. In our estimates, we consider both bio-physical and socio-economic aspects to conclude that there are long term implications on both efficiency and social disparities. We find disparities in the adaptation strategies depending on the crop and the region analyzed.

  4. Management options for food production systems affected by a nuclear accident. Task 2 options for minimising the production of contaminated milk

    CERN Document Server

    Smith, J G; Mercer, J A; Nisbet, A F; Wilkins, B T

    2002-01-01

    This report describes an evaluation of three possible means by which the production of waste milk could be reduced following a nuclear accident. The three options studied are the reduction of contaminated pasture in the diet, the drying off of lactating dairy cattle and the slaughter of dairy cattle. The practicability of each of these is considered using criteria such as technical feasibility, capacity, cost, impact and acceptability, where appropriate. In theory reductions in waste milk arisings can be achieved with each option, however, there are a number of limitations associated with their practical application.

  5. Dynamics of a Class 1 integron located on plasmid or chromosome in two Aeromonas spp. strains

    Directory of Open Access Journals (Sweden)

    Abigail Perez-Valdespino

    2016-09-01

    Full Text Available Integrons are non-mobile bacterial genetic elements that carry different cassettes conferring antibiotic resistance. Cassettes can excise or integrate by action of an integron-encoded integrase, enabling bacteria to face environmental challenges. In this work the functionality and dynamics of two integrons carrying the same cassette arrangement (dfrA12-orfF-aadA2, but located on plasmid or chromosome in two different strains were studied. In order to demonstrate the functionality of the Class 1 integrase, circular cassette integration intermediaries were PCR amplified by PCR using extrachromosomal DNA extracted from bacteria grown in the presence or absence of cassette-encoded antibiotics. Circular aadA2 and dfrA12-orfF-aadA2 cassettes were detected in cultures grown either in the presence or absence of antibiotics in both strains. No dfrA12-orfF circular intermediates could be detected under any culture conditions. These results show that both integrons are functional. However, these elements show different dynamics and functionality since the presence of streptomycin led to detectable gene rearrangements in the variable region only in the strain with the plasmid-born integron. In addition, complete integration products were demonstrated using a receptor molecule carrying an empty integron. In this case integration products were observed in both strains even in the absence of antibiotics, but they were more evident in the strain with the plasmid-located integron when streptomycin was present in the culture medium. This suggests that integrons in the two strains respond differently to streptomycin even though DNA sequences upstream the intI1 gene, including the lexA boxes of both integrons are identical.

  6. Dynamics of a Class 1 Integron Located on Plasmid or Chromosome in Two Aeromonas spp. Strains

    Science.gov (United States)

    Pérez-Valdespino, Abigail; Lazarini-Martínez, Alfredo; Rivera-González, Alejandro X.; García-Hernández, Normand; Curiel-Quesada, Everardo

    2016-01-01

    Integrons are non-mobile bacterial genetic elements that carry different cassettes conferring antibiotic resistance. Cassettes can excise or integrate by action of an integron-encoded integrase, enabling bacteria to face environmental challenges. In this work, the functionality and dynamics of two integrons carrying the same cassette arrangement (dfrA12–orfF–aadA2), but located on plasmid or chromosome in two different strains were studied. In order to demonstrate the functionality of the Class 1 integrase, circular cassette integration intermediaries were PCR amplified by PCR using extrachromosomal DNA extracted from bacteria grown in the presence or absence of cassette-encoded antibiotics. Circular aadA2 and dfrA12–orfF–aadA2 cassettes were detected in cultures grown either in the presence or absence of antibiotics in both strains. No dfrA12–orfF circular intermediates could be detected under any culture conditions. These results show that both integrons are functional. However, these elements show different dynamics and functionality since the presence of streptomycin led to detectable gene rearrangements in the variable region only in the strain with the plasmid-born integron. In addition, complete integration products were demonstrated using a receptor molecule carrying an empty integron. In this case, integration products were observed in both strains even in the absence of antibiotics, but they were more evident in the strain with the plasmid-located integron when streptomycin was present in the culture medium. This suggests that integrons in the two strains respond differently to streptomycin even though DNA sequences upstream the intI1 gene, including the lexA boxes of both integrons are identical. PMID:27733851

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

    Directory of Open Access Journals (Sweden)

    Bryon A Nicholson

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

  9. 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.

  10. Measuring New Product Portfolio Innovativeness: How Differences in Scale Width and Evaluator Perspectives Affect its Relationship with Performance

    DEFF Research Database (Denmark)

    Schultz, Carsten; Salomo, Søren; Talke, Katrin

    2013-01-01

    Portfolio innovativeness is a central variable in innovation management. However, the impact of portfolio innovativeness on new product development (NPD) performance is unclear, which may partly be due to the construct’s multifaceted nature. Different facets may reflect different degrees...... of innovativeness and may have different relationships with performance. In addition, firm members with different functional backgrounds may perceive and thus assess these facets differently, which again may influence the performance effect of portfolio innovativeness. Based on a sample of 746 CEOs and marketing...... as well as technology professionals from 117 firms and using Item Response Theory (IRT), a multifaceted scale of portfolio innovativeness, whose facets are able to cover the entire innovativeness spectrum, is developed. In addition, it is shown that the performance impact of portfolio innovativeness...

  11. [Hemipteran diversity (Cicadellidae and Clastopteridae) in three coffee production zones affected by Xylella fastidiosa (Wells et al.) in Costa Rica].

    Science.gov (United States)

    Garita-Cambronero, Jerson; Villalobos, William; Godoy, Carolina; Rivera, Carmen

    2008-01-01

    A survey was conducted during 2002, 2003 and 2004 to determine the leafhopper species composition, abundance, richness, diversity, evenness, occurrence and flight activity among three coffee production zones of Costa Rica. Yellow sticky traps were used to qualify and quantify the number of aerial leafhoppers during the sampling period. A total of 82,500 individuals, belonging to 139 species within nine leafhopper subfamilies, were trapped. San Isidro de León Cortés site presented the highest diversity from the three surveyed sites. Twenty five species were frequently trapped at least in one of the studied zones, and only Coelidiana sp.1, Osbornellus sp.1, Scaphytopius sp.1 and Empoasca sp. were trapped throughout the sampling period. The flight activity of the taxa that contain the main vectors of Xylella fastidiosa Wells et al. showed differences among the sampling zones.

  12. Elevated atmospheric CO2 affected photosynthetic products in wheat seedlings and biological activity in rhizosphere soil under cadmium stress.

    Science.gov (United States)

    Jia, Xia; Liu, Tuo; Zhao, Yonghua; He, Yunhua; Yang, Mingyan

    2016-01-01

    The objective of this study was to investigate the effects of elevated CO2 (700 ± 23 μmol mol(-1)) on photosynthetic products in wheat seedlings and on organic compounds and biological activity in rhizosphere soil under cadmium (Cd) stress. Elevated CO2 was associated with decreased quantities of reducing sugars, starch, and soluble amino acids, and with increased quantities of soluble sugars, total sugars, and soluble proteins in wheat seedlings under Cd stress. The contents of total soluble sugars, total free amino acids, total soluble phenolic acids, and total organic acids in the rhizosphere soil under Cd stress were improved by elevated CO2. Compared to Cd stress alone, the activity of amylase, phenol oxidase, urease, L-asparaginase, β-glucosidase, neutral phosphatase, and fluorescein diacetate increased under elevated CO2 in combination with Cd stress; only cellulase activity decreased. Bacterial abundance in rhizosphere soil was stimulated by elevated CO2 at low Cd concentrations (1.31-5.31 mg Cd kg(-1) dry soil). Actinomycetes, total microbial abundance, and fungi decreased under the combined conditions at 5.31-10.31 mg Cd kg(-1) dry soil. In conclusion, increased production of soluble sugars, total sugars, and proteins in wheat seedlings under elevated CO2 + Cd stress led to greater quantities of organic compounds in the rhizosphere soil relative to seedlings grown under Cd stress only. Elevated CO2 concentrations could moderate the effects of heavy metal pollution on enzyme activity and microorganism abundance in rhizosphere soils, thus improving soil fertility and the microecological rhizosphere environment of wheat under Cd stress.

  13. Regulatory T cells negatively affect IL-2 production of effector T cells through CD39/adenosine pathway in HIV infection.

    Directory of Open Access Journals (Sweden)

    Mohammad-Ali Jenabian

    Full Text Available The mechanisms by which Regulatory T cells suppress IL-2 production of effector CD4+ T cells in pathological conditions are unclear. A subpopulation of human Treg expresses the ectoenzyme CD39, which in association with CD73 converts ATP/ADP/AMP to adenosine. We show here that Treg/CD39+ suppress IL-2 expression of activated CD4+ T-cells more efficiently than Treg/CD39-. This inhibition is due to the demethylation of an essential CpG site of the il-2 gene promoter, which was reversed by an anti-CD39 mAb. By recapitulating the events downstream CD39/adenosine receptor (A2AR axis, we show that A2AR agonist and soluble cAMP inhibit CpG site demethylation of the il-2 gene promoter. A high frequency of Treg/CD39+ is associated with a low clinical outcome in HIV infection. We show here that CD4+ T-cells from HIV-1 infected individuals express high levels of A2AR and intracellular cAMP. Following in vitro stimulation, these cells exhibit a lower degree of demethylation of il-2 gene promoter associated with a lower expression of IL-2, compared to healthy individuals. These results extend previous data on the role of Treg in HIV infection by filling the gap between expansion of Treg/CD39+ in HIV infection and the suppression of CD4+ T-cell function through inhibition of IL-2 production.

  14. Survey Probability and Factors affecting Farmers Participation in Future and Option Markets Case Study: Cotton product in Gonbad kavos city

    Directory of Open Access Journals (Sweden)

    F. sakhi

    2016-03-01

    Full Text Available Introduction: Farmers are facing with a variety of natural and unnatural risks in agricultural activities, and thus their income is unstable. A wide range of risks such as risks of production, price risk, financial and human risks, influence the income of agricultural products. One of the major risks that farmers faced is the risk of price volatility of agricultural products. Cotton is one of the agricultural products with high real price volatility. Numerous tools for marketing and risk management for agricultural products in the face of price risks are available. Futures and options contracts may be the most important available tools (to reduce price volatility in agricultural products. The purpose of the current study was to look at the possibility of farmers participations in the future and option markets that presented as a means to reduce the cotton prices volatility. The dependent variable for this purpose had four categories and these included: participate in both the market, participation in the future market, participation in the option market and participation in both future and option markets. Materials and Methods: data gathered with interview and completing 200 questionnaires of cotton growers using simple random sampling. Multinomial Logit Regression Model was used for data analysis. Results and Discussion: To measure content validity of the preliminary study the validity of confirmatory factor analysis were used. For calculating reliability, the pre-test done with 30 questionnaires and reliability, coefficient Cronbach alpha was 0.79. The independence of dependent variables categories was confirmed by Hausman test results. The Likelihood ratio and Wald showed these categories are not combinable. Results indicated into period 2014 -2015 and the sample under study, 35% of cotton growers unwilling to participate in future and option markets. Farmers willingness to participate in future and option market was 19% and %21

  15. Monitoring bacterial processes by Fourier transform infrared spectroscopy : Helicobacter pylori drug inactivation and plasmid bioproduction in recombinant Escherichia coli cultures

    OpenAIRE

    Scholz, Teresa; Lopes, Vitor V.; Calado, Cecília R. C.

    2011-01-01

    Fourier transform infrared (FTIR) spectroscopy is evaluated as a tool to monitor two bacterial processes: strain discrimination and drug inactivation studies with the gastric pathogen Helicobacter pylori and the plasmid production process based on high-density cultures of recombinant Escherichia coli. Results show, that after evaluation of different incubation conditions of H.pylori with the drug model, the application of principal component analysis to the FTIR spectra assembles the samples ...

  16. Homology between nucleotide sequences of promoter regions of nah and sal operons of NAH7 plasmid of Pseudomonas putida.

    OpenAIRE

    1986-01-01

    The in vivo transcription start sites of the nah and sal operons of the NAH7 plasmid were determined by S1 nuclease mapping and the nucleotide sequence surrounding these transcription start sites was determined. Since expression of both of these operons is coordinately controlled by the product of the transcriptional activator gene nahR, the sequences were compared to locate potential sites involved in common regulation. In the 100-base-pair region preceding transcription start sites of both ...

  17. Effects of maternal plasmid GHRH treatment on offspring growth

    Science.gov (United States)

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

  18. Pharmaceutical development of the plasmid DNA vaccine pDERMATT

    NARCIS (Netherlands)

    Quaak, S.G.L.

    2009-01-01

    The discovery of tumor specific antigens and self tolerance mechanisms against these antigens led to the assumption that antigens circulating at sufficient concentration levels could break this self tolerance mechanism and evoke immunological antitumor effects. pDERMATT (plasmid DNA encoding recombi

  19. Use of plasmid DNA for induction of protective immunity

    DEFF Research Database (Denmark)

    Lorenzen, Niels

    2004-01-01

    Vaccines