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Sample records for vfr rpos pvds

  1. Role of GacA, LasI, RhlI, Ppk, PsrA, Vfr and ClpXP in the regulation of the stationary-phase sigma factor rpoS/RpoS in Pseudomonas.

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    Bertani, Iris; Sevo, Milica; Kojic, Milan; Venturi, Vittorio

    2003-10-01

    RpoS is the stationary phase sigma factor responsible for increased transcription of a set of genes when bacterial cells enter stationary phase and under stress conditions. In Escherichia coli, RpoS expression is modulated at the level of transcription, translation, and post-translational stability whereas in Pseudomonas, previous studies have implicated four genetic loci ( psrA, gacA, lasI and rhlI) involved in rpoS transcription. In this report, the transcription, translation and proteins profiles of rpoS/RpoS were analyzed in response to growth phase of knockout genomic mutants in the P. aeruginosa transcriptional regulatory loci psrA, gacA, vfr, and in the las and rhl quorum-sensing systems. Gene expression and protein profiles were also analyzed in the ppk genomic mutant. This gene is responsible for the biosynthesis of polyphosphate, an alarmone involved in the regulation of RpoS accumulation in E. coli. Finally, the role of the ClpXP protease in RpoS regulation was also studied; in E. coli, this protease has been shown to rapidly degrade RpoS during exponential growth. These studies confirm the significant role of PsrA in rpoS transcription during the late-exponential and stationary growth phases, the probable role of Vfr in transcriptional repression during exponential phase, and the function of the ClpXP protease in RpoS degradation during exponential phase. GacA/GacS, the quorum-sensing systems, and the polyphosphate alarmone molecule were not significant in rpoS/RpoS regulation. These results demonstrate important similarities and differences with the regulation of this sigma factor in E. coli and in Pseudomonas.

  2. 14 CFR 125.359 - Flight release under VFR.

    Science.gov (United States)

    2010-01-01

    ... 14 Aeronautics and Space 3 2010-01-01 2010-01-01 false Flight release under VFR. 125.359 Section...,000 POUNDS OR MORE; AND RULES GOVERNING PERSONS ON BOARD SUCH AIRCRAFT Flight Release Rules § 125.359 Flight release under VFR. No person may release an airplane for VFR operation unless the ceiling and...

  3. Further studies on RpoS in enterobacteria: identification of rpoS in Enterobacter cloacae and Kluyvera cryocrescens.

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    Martínez-García, E; Tormo, A; Navarro-Llorens, J M

    2001-06-01

    RpoS, the alternative sigma factor sigma(s), is important for bacterial survival under extreme conditions. Many enterobacteria are opportunistic human pathogens and their ability to survive in a changing environment could be an essential step for their virulence. To determine the presence of this gene in enteric bacteria, an Escherichia coli rpoS probe was constructed and used to detect the presence of this gene in different species. A gene homologous to rpoS was found in Citrobacter amalonaticus, Enterobacter cloacae, Klebsiella planticola, Kluyvera cryocrescens, Serratia rubidaea, Shigella sonnei, and Yersinia ruckeri. Providencia stuartii and Proteus vulgaris were the only tested enterobacteria that did not show any signal with the E. coli rpoS probe or that did not lead to amplification of an rpoS fragment using specific primers. The rpoS gene from E. cloacae and from K. cryocrescens was cloned and sequenced and a mutant allele was constructed in E. cloacae. Survival rates under different harsh conditions were followed in order to determine the effect of rpoS inactivation in exponential- and stationary-phase cells of both strains. E. cloacae rpoS mutants were more sensitive to extreme pH, high osmolarity, and high temperature than the wild-type.

  4. RpoS controls the Vibrio cholerae mucosal escape response.

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    Alex Toftgaard Nielsen

    2006-10-01

    Full Text Available Vibrio cholerae causes a severe diarrhoeal disease by secreting a toxin during colonization of the epithelium in the small intestine. Whereas the initial steps of the infectious process have been intensively studied, the last phases have received little attention. Confocal microscopy of V. cholerae O1-infected rabbit ileal loops captured a distinctive stage in the infectious process: 12 h post-inoculation, bacteria detach from the epithelial surface and move into the fluid-filled lumen. Designated the "mucosal escape response," this phenomenon requires RpoS, the stationary phase alternative sigma factor. Quantitative in vivo localization assays corroborated the rpoS phenotype and showed that it also requires HapR. Expression profiling of bacteria isolated from ileal loop fluid and mucus demonstrated a significant RpoS-dependent upregulation of many chemotaxis and motility genes coincident with the emigration of bacteria from the epithelial surface. In stationary phase cultures, RpoS was also required for upregulation of chemotaxis and motility genes, for production of flagella, and for movement of bacteria across low nutrient swarm plates. The hapR mutant produced near-normal numbers of flagellated cells, but was significantly less motile than the wild-type parent. During in vitro growth under virulence-inducing conditions, the rpoS mutant produced 10- to 100-fold more cholera toxin than the wild-type parent. Although the rpoS mutant caused only a small over-expression of the genes encoding cholera toxin in the ileal loop, it resulted in a 30% increase in fluid accumulation compared to the wild-type. Together, these results show that the mucosal escape response is orchestrated by an RpoS-dependent genetic program that activates chemotaxis and motility functions. This may furthermore coincide with reduced virulence gene expression, thus preparing the organism for the next stage in its life cycle.

  5. Final Environmental Assessment (EA) for Modification of Airspace Units R-3008A/B/C from Visual Flight Rules (VFR) to VFR-Instrument Flight Rules (IFR) at Moody Air Force Base, Georgia

    Science.gov (United States)

    2015-09-30

    RULES ( IFR ) AT MOODY AIR FORCE BASE, GEORGIA September 2015 Finding of No Significant Impact Modification of Airspace Units R-3008A/B/C from...Vlsual Flight Rules (VFR) to VFR-lnstrument Flight Rules ( IFR ) Moody Air Force Base, Georgia Pursuant to provisions of the National Environmental...rules (VFR) to VFR-lnstrument flight rules (VFR- IFR ). This action would minimize the number of training hours lost by allowing full utilization of Grand

  6. Characterization of the Pseudomonas aeruginosa metalloendopeptidase, Mep72, a member of the Vfr regulon.

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    Balyimez, Aysegul; Colmer-Hamood, Jane A; San Francisco, Michael; Hamood, Abdul N

    2013-11-27

    Pseudomonas aeruginosa Vfr (the virulence factor regulator) enhances P. aeruginosa virulence by positively regulating the expression of numerous virulence genes. A previous microarray analysis identified numerous genes positively regulated by Vfr in strain PAK, including the yet uncharacterized PA2782 and PA2783. In this study, we report the detailed characterization of PA2783 in the P. aeruginosa strain PAO1. RT-PCR analysis confirmed that PA2782-PA2783 constitute an operon. A mutation in vfr significantly reduced the expression of both genes. The predicted protein encoded by PA2783 contains a typical leader peptide at its amino terminus end as well as metalloendopeptidase and carbohydrate binding motifs at its amino terminus and carboxy terminus regions, respectively. An in-frame PA2783::phoA fusion encoded a hybrid protein that was exported to the periplasmic space of Escherichia coli and P. aeruginosa. In PAO1, the proteolytic activity of the PA2783-encoded protein was masked by other P. aeruginosa extracellular proteases but an E. coli strain carrying a PA2783 recombinant plasmid produced considerable proteolytic activity. The outer membrane fraction of an E. coli strain in which PA2783 was overexpressed contained specific endopeptidase activity. In the presence of cAMP, purified recombinant Vfr (rVfr) bound to a 98-bp fragment within the PA2782-PA2783 upstream region that carries a putative Vfr consensus sequence. Through a series of electrophoretic mobility shift assays, we localized rVfr binding to a 33-bp fragment that contains part of the Vfr consensus sequence and a 5-bp imperfect (3/5) inverted repeat at its 3' and 5' ends (TGGCG-N22-CGCTG). Deletion of either repeat eliminated Vfr binding. PA2782 and PA2783 constitute an operon whose transcription is positively regulated by Vfr. The expression of PA2783 throughout the growth cycle of P. aeruginosa follows a unique pattern. PA2783 codes for a secreted metalloendopeptidase, which we named Mep72. Mep72

  7. Increasing RpoS expression causes cell death in Borrelia burgdorferi.

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    Linxu Chen

    Full Text Available RpoS, one of the two alternative σ factors in Borrelia burgdorferi, is tightly controlled by multiple regulators and, in turn, determines expression of many critical virulence factors. Here we show that increasing RpoS expression causes cell death. The immediate effect of increasing RpoS expression was to promote bacterial division and as a consequence result in a rapid increase in cell number before causing bacterial death. No DNA fragmentation or degradation was observed during this induced cell death. Cryo-electron microscopy showed induced cells first formed blebs, which were eventually released from dying cells. Apparently blebbing initiated cell disintegration leading to cell death. These findings led us to hypothesize that increasing RpoS expression triggers intracellular programs and/or pathways that cause spirochete death. The potential biological significance of induced cell death may help B. burgdorferi regulate its population to maintain its life cycle in nature.

  8. Compensatory evolution of gene regulation in response to stress by Escherichia coli lacking RpoS.

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    Daniel M Stoebel

    2009-10-01

    Full Text Available The RpoS sigma factor protein of Escherichia coli RNA polymerase is the master transcriptional regulator of physiological responses to a variety of stresses. This stress response comes at the expense of scavenging for scarce resources, causing a trade-off between stress tolerance and nutrient acquisition. This trade-off favors non-functional rpoS alleles in nutrient-poor environments. We used experimental evolution to explore how natural selection modifies the regulatory network of strains lacking RpoS when they evolve in an osmotically stressful environment. We found that strains lacking RpoS adapt less variably, in terms of both fitness increase and changes in patterns of transcription, than strains with functional RpoS. This phenotypic uniformity was caused by the same adaptive mutation in every independent population: the insertion of IS10 into the promoter of the otsBA operon. OtsA and OtsB are required to synthesize the osmoprotectant trehalose, and transcription of otsBA requires RpoS in the wild-type genetic background. The evolved IS10 insertion rewires expression of otsBA from RpoS-dependent to RpoS-independent, allowing for partial restoration of wild-type response to osmotic stress. Our results show that the regulatory networks of bacteria can evolve new structures in ways that are both rapid and repeatable.

  9. RpoS regulates a novel type of plasmid DNA transfer in Escherichia coli.

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    Zhang, Yanmei; Shi, Chunyu; Yu, Jiafei; Ren, Jingjing; Sun, Dongchang

    2012-01-01

    Spontaneous plasmid transformation of Escherichia coli is independent of the DNA uptake machinery for single-stranded DNA (ssDNA) entry. The one-hit kinetic pattern of plasmid transformation indicates that double-stranded DNA (dsDNA) enters E. coli cells on agar plates. However, DNA uptake and transformation regulation remain unclear in this new type of plasmid transformation. In this study, we developed our previous plasmid transformation system and induced competence at early stationary phase. Despite of inoculum size, the development of competence was determined by optical cell density. DNase I interruption experiment showed that DNA was taken up exponentially within the initial 2 minutes and most transforming DNA entered E. coli cells within 10 minutes on LB-agar plates. A half-order kinetics between recipient cells and transformants was identified when cell density was high on plates. To determine whether the stationary phase master regulator RpoS plays roles in plasmid transformation, we investigated the effects of inactivating and over-expressing its encoding gene rpoS on plasmid transformation. The inactivation of rpoS systematically reduced transformation frequency, while over-expressing rpoS increased plasmid transformation. Normally, RpoS recognizes promoters by its lysine 173 (K173). We found that the K173E mutation caused RpoS unable to promote plasmid transformation, further confirming a role of RpoS in regulating plasmid transformation. In classical transformation, DNA was transferred across membranes by DNA uptake proteins and integrated by DNA processing proteins. At stationary growth phase, RpoS regulates some genes encoding membrane/periplasmic proteins and DNA processing proteins. We quantified transcription of 22 of them and found that transcription of only 4 genes (osmC, yqjC, ygiW and ugpC) encoding membrane/periplasmic proteins showed significant differential expression when wildtype RpoS and RpoS(K173E) mutant were expressed. Further

  10. RpoS regulates a novel type of plasmid DNA transfer in Escherichia coli.

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    Yanmei Zhang

    Full Text Available Spontaneous plasmid transformation of Escherichia coli is independent of the DNA uptake machinery for single-stranded DNA (ssDNA entry. The one-hit kinetic pattern of plasmid transformation indicates that double-stranded DNA (dsDNA enters E. coli cells on agar plates. However, DNA uptake and transformation regulation remain unclear in this new type of plasmid transformation. In this study, we developed our previous plasmid transformation system and induced competence at early stationary phase. Despite of inoculum size, the development of competence was determined by optical cell density. DNase I interruption experiment showed that DNA was taken up exponentially within the initial 2 minutes and most transforming DNA entered E. coli cells within 10 minutes on LB-agar plates. A half-order kinetics between recipient cells and transformants was identified when cell density was high on plates. To determine whether the stationary phase master regulator RpoS plays roles in plasmid transformation, we investigated the effects of inactivating and over-expressing its encoding gene rpoS on plasmid transformation. The inactivation of rpoS systematically reduced transformation frequency, while over-expressing rpoS increased plasmid transformation. Normally, RpoS recognizes promoters by its lysine 173 (K173. We found that the K173E mutation caused RpoS unable to promote plasmid transformation, further confirming a role of RpoS in regulating plasmid transformation. In classical transformation, DNA was transferred across membranes by DNA uptake proteins and integrated by DNA processing proteins. At stationary growth phase, RpoS regulates some genes encoding membrane/periplasmic proteins and DNA processing proteins. We quantified transcription of 22 of them and found that transcription of only 4 genes (osmC, yqjC, ygiW and ugpC encoding membrane/periplasmic proteins showed significant differential expression when wildtype RpoS and RpoS(K173E mutant were expressed

  11. rpoS Mutations and Loss of General Stress Resistance in Escherichia coli Populations as a Consequence of Conflict between Competing Stress Responses

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    Notley-McRobb, Lucinda; King, Thea; Ferenci, Thomas

    2002-01-01

    The general stress resistance of Escherichia coli is controlled by the RpoS sigma factor (ϕS), but mutations in rpoS are surprisingly common in natural and laboratory populations. Evidence for the selective advantage of losing rpoS was obtained from experiments with nutrient-limited bacteria at different growth rates. Wild-type bacteria were rapidly displaced by rpoS mutants in both glucose- and nitrogen-limited chemostat populations. Nutrient limitation led to selection and sweeps of rpoS nu...

  12. In silico Analysis and Molecular Modeling of RNA Polymerase, Sigma S (RpoS Protein in Pseudomonas aeruginosa PAO1

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    Mansour Sedighi

    2015-10-01

    Full Text Available Background: Sigma factors are proteins that regulate transcription in bacteria. Sigma factors can be activated in response to different environmental conditions. The rpoS (RNA polymerase, sigma S gene encodes sigma-38 (σ38, or RpoS, a 37.8 kDa protein in Pseudomonas aeruginosa (P. aeruginosa strains. RpoS is a central regulator of the general stress response and operates in both retroactive and proactive manners; not only does it allow the cell to survive environmental challenges; it also prepares the cell for subsequent stresses (cross-protection. Methods: The significance of RpoS for stress resistance and protein expression in stationary-phase P. aeruginosa cells was assessed. The goal of the current study was to characterize RpoS of P. aeruginosa PAO1 using bioinformatics tools. Results: The results showed that RpoS is an unstable protein that belongs to the sigma-70 factor family. Secondary structure analysis predicted that random coil is the predominant structure followed by extended alpha helix. The three-dimensional (3D structure was modeled using SWISS-MODEL Workspace. Conclusion: Determination of sequence, function, structure, and predicted epitopes of RpoS is important for modeling of inhibitors that will help in the design of new drugs to combat multi-drug-resistant (MDR strains. Such information may aid in the development of new diagnostic tools, drugs, and vaccines for treatment in endemic regions.

  13. Interactions of ribosomal protein S1 with DsrA and rpoS mRNA.

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    Koleva, Rositsa I; Austin, Christina A; Kowaleski, Jeffrey M; Neems, Daniel S; Wang, Leyi; Vary, Calvin P H; Schlax, Paula Jean

    2006-09-22

    Ribosomal protein S1 is shown to interact with the non-coding RNA DsrA and with rpoS mRNA. DsrA is a non-coding RNA that is important in controlling expression of the rpoS gene product in Escherichia coli. Photochemical crosslinking, quadrupole-time of flight tandem mass spectrometry, and peptide sequencing have identified an interaction between DsrA and S1 in the 30S ribosomal subunit. Purified S1 binds both DsrA (K(obs) approximately 6 x 10(6) M(-1)) and rpoS mRNA (K(obs) approximately 3 x 10(7) M(-1)). Ribonuclease probing experiments indicate that S1 binding has a weak but detectable effect on the secondary structure of DsrA or rpoS mRNA.

  14. Perspective Tools of the Strategic Management of VFR Tourism Development at the Regional Level

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    Gorbunov, Aleksandr P.; Efimova, Ekaterina V.; Kobets, Margarita V.; Kilinkarova, Sofiya G.

    2016-01-01

    This study is aimed at identifying the perspective tools of strategic management in general and strategic planning of VFR tourism (for the purpose of visiting friends and relatives) at the regional level in particular. It is based on dialectical and logical methods, analysis and synthesis, induction and deduction, the concrete historical and…

  15. The Polymorphic Aggregative Phenotype of Shiga Toxin-Producing Escherichia coli O111 Depends on RpoS and Curli.

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    Diodati, M E; Bates, A H; Miller, W G; Carter, M Q; Zhou, Y; Brandl, M T

    2015-12-28

    Escherichia coli O111 is an emerging non-O157:H7 serotype of Shiga toxin-producing E. coli (STEC). We previously reported that outbreak and environmental, but not sporadic-case, strains of STEC O111 share a distinct aggregation phenotype (M. E. Diodati, A. H. Bates, M. B. Cooley, S. Walker, R. E. Mandrell, and M. T. Brandl, Foodborne Pathog Dis 12:235-243, 2015, http://dx.doi.org/10.1089/fpd.2014.1887). We show here the natural occurrence of nonaggregative variants in single STEC O111 strains. These variants do not produce curli fimbriae and lack RpoS function but synthesize cellulose. The deletion of csgBAC or rpoS in an aggregative outbreak strain abolished aggregate formation, which was rescued when curli biogenesis or RpoS function, respectively, was restored. Complementation of a nonaggregative variant with RpoS also conferred curli production and aggregation. These observations were supported by Western blotting with an anti-CsgA antibody. Immunomicroscopy revealed that curli were undetectable on the cells of the nonaggregative variant and the RpoS mutant but were present in large quantities in the intercellular matrix of the assemblages formed by aggregative strains. Sequence analysis of rpoS in the aggregative strain and its variant showed a single substitution of threonine for asparagine at amino acid 124. Our results indicate that the multicellular behavior of STEC O111 is RpoS dependent via positive regulation of curli production. Aggregation may confer a fitness advantage in O111 outbreak strains under stressful conditions in hydrodynamic environments along the food production chain and in the host, while the occurrence of nonaggregative variants may allow the cell population to adapt to conditions benefiting a planktonic lifestyle. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  16. Impact of the rpoS genotype for acid resistance patterns of pathogenic and probiotic Escherichia coli

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    Schmidt Dorothea S

    2007-03-01

    Full Text Available Abstract Background Enterohemorrhagic E. coli (EHEC, a subgroup of Shiga toxin (Stx producing E. coli (STEC, may cause severe enteritis and hemolytic uremic syndrome (HUS and is transmitted orally via contaminated foods or from person to person. The infectious dose is known to be very low, which requires most of the bacteria to survive the gastric acid barrier. Acid resistance therefore is an important mechanism of EHEC virulence. It should also be a relevant characteristic of E. coli strains used for therapeutic purposes such as the probiotic E. coli Nissle 1917 (EcN. In E. coli and related enteric bacteria it has been extensively demonstrated, that the alternative sigma factor σS, encoded by the rpoS gene, acts as a master regulator mediating resistance to various environmental stress factors. Methods Using rpoS deletion mutants of a highly virulent EHEC O26:H11 patient isolate and the sequenced prototype EHEC EDL933 (ATCC 700927 of serotype O157:H7 we investigated the impact of a functional rpoS gene for orchestrating a satisfactory response to acid stress in these strains. We then functionally characterized rpoS of probiotic EcN and five rpoS genes selected from STEC isolates pre-investigated for acid resistance. Results First, we found out that ATCC isolate 700927 of EHEC EDL933 has a point mutation in rpoS, not present in the published sequence, leading to a premature stop codon. Moreover, to our surprise, one STEC strain as well as EcN was acid sensitive in our test environment, although their cloned rpoS genes could effectively complement acid sensitivity of an rpoS deletion mutant. Conclusion The attenuation of sequenced EHEC EDL933 might be of importance for anyone planning to do either in vitro or in vivo studies with this prototype strain. Furthermore our data supports recently published observations, that individual E. coli isolates are able to significantly modulate their acid resistance phenotype independent of their rpoS

  17. The LuxR-type regulator VpsT negatively controls the transcription of rpoS, encoding the general stress response regulator, in Vibrio cholerae biofilms.

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    Wang, Hongxia; Ayala, Julio C; Benitez, Jorge A; Silva, Anisia J

    2014-03-01

    Cholera is a waterborne diarrheal disease caused by Vibrio cholerae strains of serogroups O1 and O139. Expression of the general stress response regulator RpoS and formation of biofilm communities enhance the capacity of V. cholerae to persist in aquatic environments. The transition of V. cholerae between free-swimming (planktonic) and biofilm life-styles is regulated by the second messenger cyclic di-GMP (c-di-GMP). We previously reported that increasing the c-di-GMP pool by overexpression of a diguanylate cyclase diminished RpoS expression. Here we show that c-di-GMP repression of RpoS expression is eliminated by deletion of the genes vpsR and vpsT, encoding positive regulators of biofilm development. To determine the mechanism of this regulation, we constructed a strain expressing a vpsT-FLAG allele from native transcription and translation signals. Increasing the c-di-GMP pool induced vpsT-FLAG expression. The interaction between VpsT-FLAG and the rpoS promoter was demonstrated by chromatin immunoprecipitation. Furthermore, purified VpsT interacted with the rpoS promoter in a c-di-GMP-dependent manner. Primer extension analysis identified two rpoS transcription initiation sites located 43 bp (P1) and 63 bp (P2) upstream of the rpoS start codon. DNase I footprinting showed that the VpsT binding site at the rpoS promoter overlaps the primary P1 transcriptional start site. Deletion of vpsT significantly enhanced rpoS expression in V. cholerae biofilms that do not make HapR. This result suggests that VpsT and c-di-GMP contribute to the transcriptional silencing of rpoS in biofilms prior to cells entering the quorum-sensing mode.

  18. Epidemiologic and biogeographic analysis of 542 VFR traveling children in Catalonia (Spain). A rising new population with specific needs.

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    Valerio, Lluís; Roure, Sílvia; Sabrià, Miquel; de Balanzó, Xavier; Moreno, Nemesio; Martinez-Cuevas, Octavio; Peguero, Carme

    2011-01-01

    Imported diseases recorded in the European Union (EU) increasingly involve traveling immigrants returning from visits to their relatives and friends (VFR). Children of these immigrant families can represent a population of extreme vulnerability. A randomized cross-sectional study of 698 traveling children under the age of 15 was performed. VFR traveling children and non-VFR (or tourist) children groups were compared. A total of 698 individuals were analyzed: 354 males (50.7%) and 344 females (49.3%), with a median age (interquartile range) of 4 (2-9) years. Of these, 578 (82.8%) had been born in the EU with 542 (77.7%) being considered as VFR, whereas 156 (22.3%) were considered tourists. VFR children were younger (4.7 vs 8.2 yr; p travel time interval was shorter (21.8 vs 32.2 d; p traveling children showed a greater risk of exposure to infectious diseases compared with tourists. Immigrant families may represent a target group to prioritize international preventive activities. © 2011 International Society of Travel Medicine.

  19. RpoS contributes to phagocyte oxidase-mediated stress resistance during urinary tract infection by Escherichia coli CFT073.

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    Hryckowian, Andrew J; Welch, Rodney A

    2013-02-12

    Uropathogenic Escherichia coli (UPEC) is the most common causative agent of community-acquired urinary tract infection (UTI). In order to cause UTI, UPEC must endure stresses ranging from nutrient limitation to host immune components. RpoS (σ(S)), the general stress response sigma factor, directs gene expression under a variety of inhibitory conditions. Our study of rpoS in UPEC strain CFT073 began after we discovered an rpoS-frameshift mutation in one of our laboratory stocks of "wild-type" CFT073. We demonstrate that an rpoS-deletion mutation in CFT073 leads to a colonization defect during UTI of CBA/J mice at 48 hours postinfection (hpi). There is no difference between the growth rates of CFT073 and CFT073 rpoS in urine. This indicates that rpoS is needed for replication and survival in the host rather than being needed to address limitations imposed by urine nutrients. Consistent with previous observations in E. coli K-12, CFT073 rpoS is more sensitive to oxidative stress than the wild type. We demonstrate that peroxide levels are elevated in voided urine from CFT073-infected mice compared to urine from mock-infected mice, which supports the notion that oxidative stress is generated by the host in response to UPEC. In mice that lack phagocyte oxidase, the enzyme complex expressed by phagocytes that produces superoxide, the competitive defect of CFT073 rpoS in bladder colonization is lost. These results demonstrate that σ(S) is important for UPEC survival under conditions of phagocyte oxidase-generated stress during UTI. Though σ(S) affects the pathogenesis of other bacterial species, this is the first work that directly implicates σ(S) as important for UPEC pathogenesis. UPEC must cope with a variety of stressful conditions in the urinary tract during infection. RpoS (σ(S)), the general stress response sigma factor, is known to direct the expression of many genes under a variety of stressful conditions in laboratory-adapted E. coli K-12. Here, we show that

  20. The Cyclic AMP-Vfr Signaling Pathway in Pseudomonas aeruginosa Is Inhibited by Cyclic Di-GMP

    DEFF Research Database (Denmark)

    Almblad, Henrik; Harrison, Joe J; Rybtke, Morten

    2015-01-01

    as a direct result of elevated c-di-GMP content. Overproduction of c-di-GMP causes a decrease in the transcription of virulence factor genes that are regulated by the global virulence regulator Vfr. The low level of Vfr-dependent transcription is caused by a low level of its coactivator, cyclic AMP (cAMP...... infection give rise to rugose small colony variants (RSCVs), which are hyper-biofilm-forming mutants that commonly possess mutations that increase production of the biofilm-promoting secondary messenger cyclic di-GMP (c-di-GMP). We show that RSCVs display a decreased production of acute virulence factors......), which is decreased in response to a high level of c-di-GMP. Mutations that cause reversion of the RSCV phenotype concomitantly reactivate Vfr-cAMP signaling. Attempts to uncover the mechanism underlying the observed c-di-GMP-mediated lowering of cAMP content provided evidence that it is not caused...

  1. Roles for RpoS in survival of Escherichia coli during protozoan predation and in reduced moisture conditions highlight its importance in soil environments.

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    Somorin, Yinka; Bouchard, Guillaume; Gallagher, Joseph; Abram, Florence; Brennan, Fiona; O'Byrne, Conor

    2017-10-16

    The soil is a complex ecosystem where interactions between biotic and abiotic factors determine the survival and fate of microbial inhabitants of the system. Having previously shown that Escherichia coli requires the general stress response regulator, RpoS, to survive long term in soil, it was important to determine what specific conditions in this environment necessitate a functional RpoS. This study investigated the susceptibility of soil-persistent E. coli to predation by the single-celled eukaryotes Acanthamoeba polyphaga and Tetrahymena pyriformis, and the role RpoS plays in resisting this predation. Strain-specific differences were observed in the predation of E. coli strains, with soil-persistent strain COB583 being the most resistant to predation by both protozoans. RpoS and curli, proteinaceous fibres used for attachment to biotic and abiotic surfaces, increased the ability of E. coli to resist predation by A. polyphaga and T. pyriformis. Furthermore, soil moisture content impacted the survival of E. coli BW25113 but wild-type COB583 had similar survival irrespective of soil moisture content. Overall, this study confirmed that RpoS contributes to the resistance of E. coli to protozoan predation and that RpoS is crucial for the increased fitness of soil-persistent E. coli against predation and reduced moisture in soil. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  2. The SOS and RpoS Regulons Contribute to Bacterial Cell Robustness to Genotoxic Stress by Synergistically Regulating DNA Polymerase Pol II.

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    Dapa, Tanja; Fleurier, Sébastien; Bredeche, Marie-Florence; Matic, Ivan

    2017-07-01

    Mitomycin C (MMC) is a genotoxic agent that induces DNA cross-links, DNA alkylation, and the production of reactive oxygen species (ROS). MMC induces the SOS response and RpoS regulons in Escherichia coli SOS-encoded functions are required for DNA repair, whereas the RpoS regulon is typically induced by metabolic stresses that slow growth. Thus, induction of the RpoS regulon by MMC may be coincidental, because DNA damage slows growth; alternatively, the RpoS regulon may be an adaptive response contributing to cell survival. In this study, we show that the RpoS regulon is primarily induced by MMC-induced ROS production. We also show that RpoS regulon induction is required for the survival of MMC-treated growing cells. The major contributor to RpoS-dependent resistance to MMC treatment is DNA polymerase Pol II, which is encoded by the polB gene belonging to the SOS regulon. The observation that polB gene expression is controlled by the two major stress response regulons that are required to maximize survival and fitness further emphasizes the key role of this DNA polymerase as an important factor in genome stability. Copyright © 2017 by the Genetics Society of America.

  3. Visible laser and UV-A radiation impact on a PNP degrading Moraxella strain and its rpoS mutant.

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    Nandakumar, Kanavillil; Keeler, Werden; Schraft, Heidi; Leung, Kam T

    2006-07-05

    The role of stationary phase sigma factor gene (rpoS) in the stress response of Moraxella strain when exposed to radiation was determined by comparing the stress responses of the wild-type (WT) and its rpoS knockout (KO) mutant. The rpoS was turned on by starving the WT cultures for 24 h in minimal salt medium. Under non-starved condition, both WT and KO planktonic Moraxella cells showed an increase in mortality with the increase in duration of irradiation. In the planktonic non-starved Moraxella, for the power intensity tested, UV radiation caused a substantially higher mortality rate than did by the visible laser light (the mortality rate observed for 15-min laser radiation was 53.4 +/- 10.5 and 48.7 +/- 8.9 for WT and KO, respectively, and 97.6 +/- 0 and 98.5 +/- 0 for 25 s of UV irradiation in WT and KO, respectively). However, the mortality rate decreased significantly in the starved WT when exposed to these two radiations. In comparison, rpoS protected the WT against the visible laser light more effectively than it did for the UV radiation. The WT and KO strains of Moraxella formed distinctly different types of biofilms on stainless steel coupons. The KO strain formed a denser biofilm than did the WT. Visible laser light removed biofilms from the surfaces more effectively than did the UV. This was true when comparing the mortality of bacteria in the biofilms as well. The inability of UV radiation to penetrate biofilms due to greater rates of surface absorption is considered to be the major reason for the weaker removal of biofilms in comparison to that of the visible laser light. This result suggests that high power visible laser light might be an effective tool for the removal of biofilms. (c) 2006 Wiley Periodicals, Inc.

  4. Characterization of a stress-induced alternate sigma factor, RpoS, of Coxiella burnetii and its expression during the development cycle.

    Science.gov (United States)

    Seshadri, R; Samuel, J E

    2001-08-01

    Coxiella burnetii is an obligate intracellular bacterium that resides in an acidified phagolysosome and has a remarkable ability to persist in the extracellular environment. C. burnetii has evolved a developmental cycle that includes at least two morphologic forms, designated large cell variants (LCV) and small cell variants (SCV). Based on differential protein expression, distinct ultrastructures, and different metabolic activities, we speculated that LCV and SCV are similar to typical logarithmic- and stationary-phase growth stages. We hypothesized that the alternate sigma factor, RpoS, a global regulator of genes expressed under stationary-phase, starvation, and stress conditions in many bacteria, regulates differential expression in life cycle variants of C. burnetii. To test this hypothesis, we cloned and characterized the major sigma factor, encoded by an rpoD homologue, and the stress response sigma factor, encoded by an rpoS homologue. The rpoS gene was cloned by complementation of an Escherichia coli rpoS null mutant containing an RpoS-dependent lacZ fusion (osmY::lacZ). Expression of C. burnetii rpoS was regulated by growth phase in E. coli (induced upon entry into stationary phase). A glutathione S-transferase-RpoS fusion protein was used to develop polyclonal antiserum against C. burnetii RpoS. Western blot analysis detected abundant RpoS in LCV but not in SCV. These results suggest that LCV and SCV are not comparable to logarithmic and stationary phases of growth and may represent a novel adaptation for survival in both the phagolysosome and the extracellular environment.

  5. Increase in imported malaria in the Netherlands in asylum seekers and VFR travellers.

    Science.gov (United States)

    de Gier, Brechje; Suryapranata, Franciska S T; Croughs, Mieke; van Genderen, Perry J J; Keuter, Monique; Visser, Leo G; van Vugt, Michele; Sonder, Gerard J B

    2017-02-02

    Malaria is a notifiable disease in the Netherlands, a non-endemic country. Imported malaria infections occur regularly among travellers, migrants and visitors. Surveillance data were analysed from 2008 to 2015. Trends in amounts of notifications among risk groups were analysed using Poisson regression. For asylum seekers, yearly incidence was calculated per region of origin, using national asylum request statistics as denominator data. For tourists, denominator data were used from travel statistics to estimate incidence per travel region up to 2012. A modest increase in overall imported malaria notifications occurred in 2008-2015 (from 222 in 2008 to 344 in 2015). Notably, in 2014 and 2015 sharp increases were seen in malaria among travellers visiting friends and relatives (VFR), and in asylum seekers. Of all Plasmodium falciparum infections, most (1254/1337; 93.8%) were imported from Africa; 1037/1337 (77.6%) were imported from Central and West Africa. Malaria in VFR was mostly caused by P. falciparum infection after visiting Ghana (22%) or Nigeria (19%). Malaria in asylum seekers was mostly caused by Plasmodium vivax infection from the Horn of Africa. The large number of notifications in asylum seekers resulted from both an increase in number of asylum seekers and a striking increase of malaria incidence in this group. Incidence of malaria in asylum seekers from the Horn of Africa ranged between 0.02 and 0.3% in 2008-2013, but rose to 1.6% in 2014 and 1.3% in 2015. In 2008-2012, incidence in tourists visiting Central and West Africa dropped markedly. Imported malaria is on the rise again in the Netherlands, most notably since 2013. This is mostly due to immigration of asylum seekers from the Horn of Africa. The predominance of P. vivax infection among asylum seekers warrants vigilance in health workers when a migrant presents with fever, as relapses of this type of malaria can occur long after arrival in the Netherlands.

  6. Transcript accumulation from the rpoS gene encoding a stationary-phase sigma factor in Pseudomonas chlororaphis strain O6 is regulated by the polyphosphate kinase gene.

    Science.gov (United States)

    Kim, H J; Yang, K Y; Cho, B H; Kim, K Y; Lee, M C; Kim, Y H; Anderson, A J; Kim, Y C

    2007-03-01

    Polyphosphate levels are modulated by the actions of polyphosphate kinase, encoded by ppk, and exopolyphosphatase, encoded by ppx. The genes ppk and ppx are adjacent to each other in the genome of the root colonizer, Pseudomonas chlororaphis O6. A ppk-deficient mutant was more sensitive to oxidative stress than the wild-type and the ppx mutant. Transcripts from ppx increased as cultures matured from mid- to late-logarithmic and stationary phases, whereas abundance was greater for ppk in the late-logarithmic phase than in the stationary phase. Transcript accumulation from the rpoS gene, encoding the stationary-phase sigma factor RpoS, was decreased in the mid- and late-logarithmic and stationary phases in the ppk mutant. Thus, ppk regulates rpoS transcript accumulation in P. chlororaphis 06. However, mutations in either the ppk or ppx genes had no effect on induction of systemic resistance in plants colonized by P. chlororaphis O6.

  7. The role of RpoS, H-NS and AcP on the pH-dependent OmpC and ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-05-04

    May 4, 2009 ... mutations in envZ, ompR, hns, rpoS and pta on the expression of porin proteins were also investigated. In all cases expression of the relevant porin was followed using a lacZ reporter system. MATERIALS AND METHODS. Bacterial strains and culture conditions. Bacterial strains used in this study are listed ...

  8. Crl binds to domain 2 of σ(S) and confers a competitive advantage on a natural rpoS mutant of Salmonella enterica serovar Typhi.

    Science.gov (United States)

    Monteil, Véronique; Kolb, Annie; Mayer, Claudine; Hoos, Sylviane; England, Patrick; Norel, Françoise

    2010-12-01

    The RpoS sigma factor (σ(S)) is the master regulator of the bacterial response to a variety of stresses. Mutants in rpoS arise in bacterial populations in the absence of stress, probably as a consequence of a subtle balance between self-preservation and nutritional competence. We characterized here one natural rpoS mutant of Salmonella enterica serovar Typhi (Ty19). We show that the rpoS allele of Ty19 (rpoS(Ty19)) led to the synthesis of a σ(S)(Ty19) protein carrying a single glycine-to-valine substitution at position 282 in σ(S) domain 4, which was much more dependent than the wild-type σ(S) protein on activation by Crl, a chaperone-like protein that increases the affinity of σ(S) for the RNA polymerase core enzyme (E). We used the bacterial adenylate cyclase two-hybrid system to demonstrate that Crl bound to residues 72 to 167 of σ(S) domain 2 and that G282V substitution did not directly affect Crl binding. However, this substitution drastically reduced the ability of σ(S)(Ty19) to bind E in a surface plasmon resonance assay, a defect partially rescued by Crl. The modeled structure of the Eσ(S) holoenzyme suggested that substitution G282V could directly disrupt a favorable interaction between σ(S) and E. The rpoS(Ty19) allele conferred a competitive fitness when the bacterial population was wild type for crl but was outcompeted in Δcrl populations. Thus, these results indicate that the competitive advantage of the rpoS(Ty19) mutant is dependent on Crl and suggest that crl plays a role in the appearance of rpoS mutants in bacterial populations.

  9. Crl Binds to Domain 2 of σS and Confers a Competitive Advantage on a Natural rpoS Mutant of Salmonella enterica Serovar Typhi▿

    Science.gov (United States)

    Monteil, Véronique; Kolb, Annie; Mayer, Claudine; Hoos, Sylviane; England, Patrick; Norel, Françoise

    2010-01-01

    The RpoS sigma factor (σS) is the master regulator of the bacterial response to a variety of stresses. Mutants in rpoS arise in bacterial populations in the absence of stress, probably as a consequence of a subtle balance between self-preservation and nutritional competence. We characterized here one natural rpoS mutant of Salmonella enterica serovar Typhi (Ty19). We show that the rpoS allele of Ty19 (rpoSTy19) led to the synthesis of a σSTy19 protein carrying a single glycine-to-valine substitution at position 282 in σS domain 4, which was much more dependent than the wild-type σS protein on activation by Crl, a chaperone-like protein that increases the affinity of σS for the RNA polymerase core enzyme (E). We used the bacterial adenylate cyclase two-hybrid system to demonstrate that Crl bound to residues 72 to 167 of σS domain 2 and that G282V substitution did not directly affect Crl binding. However, this substitution drastically reduced the ability of σSTy19 to bind E in a surface plasmon resonance assay, a defect partially rescued by Crl. The modeled structure of the EσS holoenzyme suggested that substitution G282V could directly disrupt a favorable interaction between σS and E. The rpoSTy19 allele conferred a competitive fitness when the bacterial population was wild type for crl but was outcompeted in Δcrl populations. Thus, these results indicate that the competitive advantage of the rpoSTy19 mutant is dependent on Crl and suggest that crl plays a role in the appearance of rpoS mutants in bacterial populations. PMID:20935100

  10. Ribosomal protein S1 specifically binds to the 5' untranslated region of the Pseudomonas aeruginosa stationary-phase sigma factor rpoS mRNA in the logarithmic phase of growth.

    Science.gov (United States)

    Sevo, Milica; Buratti, Emanuele; Venturi, Vittorio

    2004-08-01

    The rpoS gene encodes the stationary-phase sigma factor (RpoS or sigma(s)), which was identified in several gram-negative bacteria as a central regulator controlling the expression of genes involved in cell survival in response to cessation of growth (stationary phase) and providing cross-protection against various stresses. In Pseudomonas aeruginosa, the levels of sigma(s) increase dramatically at the onset of the stationary phase and are regulated at the transcriptional and posttranscriptional levels. The P. aeruginosa rpoS gene is transcribed as a monocistronic rpoS mRNA transcript comprised of an unusually long 373-bp 5' untranslated region (5' UTR). In this study, the 5' UTR and total protein extracts from P. aeruginosa logarithmic and stationary phases of growth were used in order to investigate the protein-RNA interactions that may modulate the translational process. It was observed that a 69-kDa protein, which corresponded to ribosomal protein S1, preferentially binds the 5' UTR of the rpoS mRNA in the logarithmic phase and not in the stationary phase. This is the first report of a protein-rpoS mRNA 5' UTR interaction in P. aeruginosa, and the possible involvement of protein S1 in translation regulation of rpoS is discussed.

  11. The role of RpoS, H-NS and AcP on the pH-dependent OmpC and ...

    African Journals Online (AJOL)

    This work was carried out to find out the effect of envZ, ompR, rpoS, pta and hns mutations on the pH-dependent ompC and ompF expression in E. coli in minimal medium at different pH. The expression of ompF was higher in cells grown in an alkaline pH, and the expression of ompC increased at the acidic pH value.

  12. The role of ClpP, RpoS and CsrA in growth and filament formation of Salmonella enterica serovar Typhimurium at low temperature

    DEFF Research Database (Denmark)

    Knudsen, Gitte Maegaard; Nielsen, Maj-Britt; Thomsen, Line Elnif

    2014-01-01

    Background: Salmonellae are food-borne pathogens of great health and economic importance. To pose a threat to humans, Salmonellae normally have to cope with a series of stressful conditions in the food chain, including low temperature. In the current study, we evaluated the importance of the Clp...... proteolytic complex and the carbon starvation protein, CsrA, for the ability of Salmonella Typhimurium to grow at low temperature. Results: A clpP mutant was severely affected in growth and formed pin point colonies at 10 degrees C. Contrary to this, rpoS and clpP/rpoS mutants were only slightly affected....... The clpP mutant formed cold resistant suppressor mutants at a frequency of 2.5 x 10(-3) and these were found not to express RpoS. Together these results indicated that the impaired growth of the clpP mutant was caused by high level of RpoS. Evaluation by microscopy of the clpP mutant revealed...

  13. ClpP deletion causes attenuation of Salmonella Typhimurium virulence through mis-regulation of RpoS and indirect control of CsrA and the SPI genes

    DEFF Research Database (Denmark)

    Knudsen, Gitte Maegaard; Olsen, John E.; Aabo, Søren

    2013-01-01

    the proteolytic component ClpP, the stationary phase regulator RpoS and the carbon-storage regulator CsrA. However, the mechanism behind the ClpP regulation is not fully understood. To elucidate this we examined differentially expressed genes in a ΔclpP mutant compared with WT using global transcriptomic analysis......, suggesting the repression of invasion was directed through RpoS. The expression of the csrA virulence regulator was increased in the ΔclpP mutant and decreased in the rpoS : : amp and ΔclpP/rpoS : : amp mutants, indicating that ClpP affects the csrA expression level as well. Thus, this study suggests...... that ClpP affects SPI1 expression and thereby virulence indirectly through its regulation of both RpoS and CsrA....

  14. Escherichia coli K-12 survives anaerobic exposure at pH 2 without RpoS, Gad, or hydrogenases, but shows sensitivity to autoclaved broth products.

    Directory of Open Access Journals (Sweden)

    Daniel P Riggins

    Full Text Available Escherichia coli and other enteric bacteria survive exposure to extreme acid (pH 2 or lower in gastric fluid. Aerated cultures survive via regulons expressing glutamate decarboxylase (Gad, activated by RpoS, cyclopropane fatty acid synthase (Cfa and others. But extreme-acid survival is rarely tested under low oxygen, a condition found in the stomach and the intestinal tract. We observed survival of E. coli K-12 W3110 at pH 1.2-pH 2.0, conducting all manipulations (overnight culture at pH 5.5, extreme-acid exposure, dilution and plating in a glove box excluding oxygen (10% H2, 5% CO2, balance N2. With dissolved O2 concentrations maintained below 6 µM, survival at pH 2 required Cfa but did not require GadC, RpoS, or hydrogenases. Extreme-acid survival in broth (containing tryptone and yeast extract was diminished in media that had been autoclaved compared to media that had been filtered. The effect of autoclaved media on extreme-acid survival was most pronounced when oxygen was excluded. Exposure to H2O2 during extreme-acid treatment increased the death rate slightly for W3110 and to a greater extent for the rpoS deletion strain. Survival at pH 2 was increased in strains lacking the anaerobic regulator fnr. During anaerobic growth at pH 5.5, strains deleted for fnr showed enhanced transcription of acid-survival genes gadB, cfa, and hdeA, as well as catalase (katE. We show that E. coli cultured under oxygen exclusion (<6 µM O2 requires mechanisms different from those of aerated cultures. Extreme acid survival is more sensitive to autoclave products under oxygen exclusion.

  15. Effect of Global Regulators RpoS and Cyclic-AMP/CRP on the Catabolome and Transcriptome of Escherichia coli K12 during Carbon- and Energy-Limited Growth.

    Directory of Open Access Journals (Sweden)

    Alessandro G Franchini

    Full Text Available For heterotrophic microbes, limited availability of carbon and energy sources is one of the major nutritional factors restricting the rate of growth in most ecosystems. Physiological adaptation to this hunger state requires metabolic versatility which usually involves expression of a wide range of different catabolic pathways and of high-affinity carbon transporters; together, this allows for simultaneous utilization of mixtures of carbonaceous compounds at low concentrations. In Escherichia coli the stationary phase sigma factor RpoS and the signal molecule cAMP are the major players in the regulation of transcription under such conditions; however, their interaction is still not fully understood. Therefore, during growth of E. coli in carbon-limited chemostat culture at different dilution rates, the transcriptomes, expression of periplasmic proteins and catabolomes of strains lacking one of these global regulators, either rpoS or adenylate cyclase (cya, were compared to those of the wild-type strain. The inability to synthesize cAMP exerted a strong negative influence on the expression of alternative carbon source uptake and degradation systems. In contrast, absence of RpoS increased the transcription of genes belonging to high-affinity uptake systems and central metabolism, presumably due to reduced competition of σ(D with σ(S. Phenotypical analysis confirmed this observation: The ability to respire alternative carbon substrates and to express periplasmic high-affinity binding proteins was eliminated in cya and crp mutants, while these properties were not affected in the rpoS mutant. As expected, transcription of numerous stress defence genes was negatively affected by the rpoS knock-out mutation. Interestingly, several genes of the RpoS stress response regulon were also down-regulated in the cAMP-negative strain indicating a coordinated global regulation. The results demonstrate that cAMP is crucial for catabolic flexibility during slow

  16. Expression of the Pseudomonas aeruginosa biofilm antibiotic resistance gene ndvB requires the RpoS stationary phase sigma factor.

    Science.gov (United States)

    Hall, Clayton W; Hinz, Aaron J; Gagnon, Luke B-P; Zhang, Li; Nadeau, Jean-Paul; Copeland, Sarah; Saha, Bratati; Mah, Thien-Fah

    2018-01-19

    Chronic, biofilm-based bacterial infections are exceptionally difficult to eradicate due to the high degree of antibiotic recalcitrance exhibited by cells in biofilm communities. In the opportunistic pathogen Pseudomonas aeruginosa, biofilm recalcitrance is multifactorial and arises in part from the preferential expression of resistance genes in biofilms compared to exponential phase planktonic cells. One such mechanism involves ndvB, which we have previously shown to be expressed specifically in biofilms. In this study, we investigated the regulatory basis of this lifestyle-specific expression by developing an unstable GFP transcriptional reporter to observe the expression pattern of ndvB We found that, in addition to its expression in biofilms, ndvB was also upregulated in planktonic cells as they enter stationary phase. The transcription of ndvB in both growth phases was shown to be dependent on the stationary phase sigma factor RpoS, and mutation of a putative RpoS binding site in the ndvB promoter abolished the activity of the promoter in stationary phase cells. Overall, we have expanded our understanding of the temporal expression of ndvB in P. aeruginosa and have uncovered a regulatory basis for its growth phase-dependent expression.Importance Bacterial biofilms are more resistant to antibiotics than free-living planktonic cells, and understanding the mechanistic basis of this resistance can inform treatments of biofilm-based infections. In addition to chemical and structural barriers that can inhibit antibiotic entry, the upregulation of specific genes in biofilms contributes to the resistance. We investigated this biofilm-specific gene induction by examining expression patterns of ndvB, a gene involved in biofilm resistance of the opportunistic pathogen Pseudomonas aeruginosa We characterized ndvB expression in planktonic and biofilm growth conditions with an unstable green fluorescent protein (GFP) reporter and found that the expression of ndvB in

  17. Phenotype and specificity of T cells in primary human cytomegalovirus infection during pregnancy: IL-7Rpos long-term memory phenotype is associated with protection from vertical transmission.

    Science.gov (United States)

    Mele, Federico; Fornara, Chiara; Jarrossay, David; Furione, Milena; Arossa, Alessia; Spinillo, Arsenio; Lanzavecchia, Antonio; Gerna, Giuseppe; Sallusto, Federica; Lilleri, Daniele

    2017-01-01

    Congenital human cytomegalovirus (HCMV) infection is the major cause of birth defects and a precise definition of the HCMV-specific T-cell response in primary infection may help define reliable correlates of immune protection during pregnancy. In this study, a high throughput method was used to define the frequency of CD4+ and CD8+ T cells specific for four HCMV proteins in the naïve compartment of seronegative subjects and the effector/memory compartments of subjects with primary/remote HCMV infection. The naïve repertoire displayed comparable frequencies of T cells that were reactive with HCMV structural (pp65, gB and the pentamer gHgLpUL128L) and non-structural (IE-1) proteins. Whereas, following natural infection, the majority of effector/memory CD4+ and CD8+ T cells recognized either gB or IE-1, respectively, and pp65. The pattern of T cell reactivity was comparable at early and late stages of infection and in pregnant women with primary HCMV infection transmitting or not transmitting the virus to the fetus. At an early stage of primary infection, about 50% of HCMV-reactive CD4+ T cells were long-term IL-7Rpos memory cells, while 6-12 months later, the frequency of these cells increased to 70%, approaching 100% in remote infections. In contrast, only 10-20% of HCMV-specific CD8+ T cells were long-term memory cells up to 12 months after infection onset, thereafter increasing to 70% in remote infections. Interestingly, a significantly higher frequency of HCMV-specific CD4+ T cells with a long-term IL-7Rpos memory phenotype was observed in non-transmitting compared to transmitting women. These findings indicate that immunodominance in HCMV infection is not predetermined in the naïve compartment, but is the result of virus-host interactions and suggest that prompt control of HCMV infection in pregnancy is associated with the rapid development of long-term IL-7Rpos memory HCMV-specific CD4+ T cells and a low risk of virus transmission to the fetus.

  18. Positive Effect of Carbon Sources on Natural Transformation in Escherichia coli: Role of Low-Level Cyclic AMP (cAMP)-cAMP Receptor Protein in the Derepression of rpoS.

    Science.gov (United States)

    Guo, Mengyue; Wang, Huanyu; Xie, Nengbin; Xie, Zhixiong

    2015-10-01

    Natural plasmid transformation of Escherichia coli is a complex process that occurs strictly on agar plates and requires the global stress response factor σ(S). Here, we showed that additional carbon sources could significantly enhance the transformability of E. coli. Inactivation of phosphotransferase system genes (ptsH, ptsG, and crr) caused an increase in the transformation frequency, and the addition of cyclic AMP (cAMP) neutralized the promotional effect of carbon sources. This implies a negative role of cAMP in natural transformation. Further study showed that crp and cyaA mutations conferred a higher transformation frequency, suggesting that the cAMP-cAMP receptor protein (CRP) complex has an inhibitory effect on transformation. Moreover, we observed that rpoS is negatively regulated by cAMP-CRP in early log phase and that both crp and cyaA mutants show no transformation superiority when rpoS is knocked out. Therefore, it can be concluded that both the crp and cyaA mutations derepress rpoS expression in early log phase, whereby they aid in the promotion of natural transformation ability. We also showed that the accumulation of RpoS during early log phase can account for the enhanced transformation aroused by additional carbon sources. Our results thus demonstrated that the presence of additional carbon sources promotes competence development and natural transformation by reducing cAMP-CRP and, thus, derepressing rpoS expression during log phase. This finding could contribute to a better understanding of the relationship between nutrition state and competence, as well as the mechanism of natural plasmid transformation in E. coli. Escherichia coli, which is not usually considered to be naturally transformable, was found to spontaneously take up plasmid DNA on agar plates. Researching the mechanism of natural transformation is important for understanding the role of transformation in evolution, as well as in the transfer of pathogenicity and antibiotic

  19. IraL is an RssB anti-adaptor that stabilizes RpoS during logarithmic phase growth in Escherichia coli and Shigella.

    Science.gov (United States)

    Hryckowian, Andrew J; Battesti, Aurelia; Lemke, Justin J; Meyer, Zachary C; Welch, Rodney A

    2014-05-27

    RpoS (σ(S)), the general stress response sigma factor, directs the expression of genes under a variety of stressful conditions. Control of the cellular σ(S) concentration is critical for appropriately scaled σ(S)-dependent gene expression. One way to maintain appropriate levels of σ(S) is to regulate its stability. Indeed, σ(S) degradation is catalyzed by the ClpXP protease and the recognition of σ(S) by ClpXP depends on the adaptor protein RssB. Three anti-adaptors (IraD, IraM, and IraP) exist in Escherichia coli K-12; each interacts with RssB and inhibits RssB activity under different stress conditions, thereby stabilizing σ(S). Unlike K-12, some E. coli isolates, including uropathogenic E. coli strain CFT073, show comparable cellular levels of σ(S) during the logarithmic and stationary growth phases, suggesting that there are differences in the regulation of σ(S) levels among E. coli strains. Here, we describe IraL, an RssB anti-adaptor that stabilizes σ(S) during logarithmic phase growth in CFT073 and other E. coli and Shigella strains. By immunoblot analyses, we show that IraL affects the levels and stability of σ(S) during logarithmic phase growth. By computational and PCR-based analyses, we reveal that iraL is found in many E. coli pathotypes but not in laboratory-adapted strains. Finally, by bacterial two-hybrid and copurification analyses, we demonstrate that IraL interacts with RssB by a mechanism distinct from that used by other characterized anti-adaptors. We introduce a fourth RssB anti-adaptor found in E. coli species and suggest that differences in the regulation of σ(S) levels may contribute to host and niche specificity in pathogenic and nonpathogenic E. coli strains. Bacteria must cope with a variety of environmental conditions in order to survive. RpoS (σ(S)), the general stress response sigma factor, directs the expression of many genes under stressful conditions in both pathogenic and nonpathogenic Escherichia coli strains

  20. Role of bolA and rpoS genes in biofilm formation and adherence pattern by Escherichia coli K-12 MG1655 on polypropylene, stainless steel, and silicone surfaces.

    Science.gov (United States)

    Adnan, Mohd; Sousa, Ana Margarida; Machado, Idalina; Pereira, Maria Olivia; Khan, Saif; Morton, Glyn; Hadi, Sibte

    2017-06-01

    Escherichia coli has developed sophisticated means to sense, respond, and adapt in stressed environment. It has served as a model organism for studies in molecular genetics and physiology since the 1960s. Stress response genes are induced whenever a cell needs to adapt and survive under unfavorable growth conditions. Two of the possible important genes are rpoS and bolA. The rpoS gene has been known as the alternative sigma (σ) factor, which controls the expression of a large number of genes, which are involved in responses to various stress factors as well as transition to stationary phase from exponential form of growth. Morphogene bolA response to stressed environment leads to round morphology of E. coli cells, but little is known about its involvement in biofilms and its development or maintenance. This study has been undertaken to address the adherence pattern and formation of biofilms by E. coli on stainless steel, polypropylene, and silicone surfaces after 24 h of growth at 37 °C. Scanning electron microscopy was used for direct examination of the cell attachment and biofilm formation on various surfaces and it was found that, in the presence of bolA, E. coli cells were able to attach to the stainless steel and silicone very well. By contrast, polypropylene surface was not found to be attractive for E. coli cells. This indicates that bolA responded and can play a major role in the presence and absence of rpoS in cell attachment.

  1. Sigma factor selectivity in Borrelia burgdorferi: RpoS recognition of the ospE/ospF/elp promoters is dependent on the sequence of the -10 region.

    Science.gov (United States)

    Eggers, Christian H; Caimano, Melissa J; Radolf, Justin D

    2006-03-01

    Members of the ospE/ospF/elp lipoprotein gene families of Borrelia burgdorferi, the Lyme disease agent, are transcriptionally upregulated in response to the influx of blood into the midgut of an infected tick. We recently have demonstrated that despite the high degree of similarity between the promoters of the ospF (P(ospF)) and ospE (P(ospE)) genes of B. burgdorferi strain 297, the differential expression of ospF is RpoS-dependent, while ospE is controlled by sigma(70). Herein we used wild-type and RpoS-deficient strains of B. burgdorferi and Escherichia coli to analyse transcriptional reporters consisting of a green fluorescent protein (gfp) gene fused to P(ospF), P(ospE), or two hybrid promoters in which the -10 regions of P(ospF) and P(ospE) were switched [P(ospF ) ((E - 10)) and P(ospE) ((F - 10)) respectively]. We found that the P(ospF)-10 region is both necessary and sufficient for RpoS-dependent recognition in B. burgdorferi, while sigma(70) specificity for P(ospE) is dependent on elements outside of the -10 region. In E. coli, sigma factor selectivity for these promoters was much more permissive, with expression of each being primarily due to sigma(70). Alignment of the sequences upstream of each of the ospE/ospF/elp genes from B. burgdorferi strains 297 and B31 revealed that two B31 ospF paralogues [erpK (BBM38) and erpL (BBO39)] have -10 regions virtually identical to that of P(ospF). Correspondingly, expression of gfp reporters based on the erpK and erpL promoters was RpoS-dependent. Thus, the sequence of the P(ospF)-10 region appears to serve as a motif for RpoS recognition, the first described for any B. burgdorferi promoter. Taken together, our data support the notion that B. burgdorferi utilizes sequence differences at the -10 region as one mechanism for maintaining the transcriptional integrity of RpoS-dependent and -independent genes activated at the onset of tick feeding.

  2. Identification of IbeR as a Stationary-Phase Regulator in Meningitic Escherichia coli K1 that Carries a Loss-of-Function Mutation in rpoS

    Directory of Open Access Journals (Sweden)

    Feng Chi

    2009-01-01

    Full Text Available IbeR is a regulator present in meningitic Escherichia coli strain E44 that carries a loss-of-function mutation in the stationary-phase (SP regulatory gene rpoS. In order to determine whether IbeR is an SP regulator in E44, two-dimensional gel electrophoresis and LC-MS were used to compare the proteomes of a noninvasive ibeR deletion mutant BR2 and its parent strain E44 in the SP. Four up-regulated (TufB, GapA, OmpA, AhpC and three down-regulated (LpdA, TnaA, OpmC proteins in BR2 were identified when compared to E44. All these proteins contribute to energy metabolism or stress resistance, which is related to SP regulation. One of the down-regulated proteins, tryptophanase (TnaA, which is regulated by RpoS in other E. coli strains, is associated with SP regulation via production of a signal molecule indole. Our studies demonstrated that TnaA was required for E44 invasion, and that indole was able to restore the noninvasive phenotype of the tnaA mutant. The production of indole was significantly reduced in BR2, indicating that ibeR is required for the indole production via tnaA. Survival studies under different stress conditions indicated that IbeR contributed to bacteria stress resistance in the SP. Taken together, IbeR is a novel regulator contributing to the SP regulation.

  3. Methodology of VFR night flying

    Directory of Open Access Journals (Sweden)

    Ľubomír Stanko

    2017-01-01

    Full Text Available Night is defined, for aviation purposes, as the period of darkness from the end of evening civil twilight to the beginning of morning civil twilight. Night flying is risky and more dangerous, comparing with flying during daylight, so it is essential to seek training with a flight instructor specifically for night flying. This article looks briefly at some underlying principles and practices, including: illusions, planning considerations, and handling emergencies.

  4. 75 FR 31288 - Plant-Verified Drop Shipment (PVDS)-Nonpostal Documentation

    Science.gov (United States)

    2010-06-03

    ... documents such as delivery receipts or bills of lading presented by a mailer's transportation provider are... document such as a delivery receipt or a bill of lading furnished by a USPS customer's carrier as proof of...

  5. Influence of general stress-response alternative sigma factors σ(S) (RpoS) and σ(B) (SigB) on bacterial tolerance to the essential oils from Origanum vulgare L. and Rosmarinus officinalis L. and pulsed electric fields.

    Science.gov (United States)

    Gomes Neto, Nelson Justino; Magnani, Marciane; Chueca, Beatriz; García-Gonzalo, Diego; Pagán, Rafael; de Souza, Evandro Leite

    2015-10-15

    This study assessed the influence of general stress-response alternative sigma factors RpoS (σ(S)) and SigB (σ(B)) on tolerance of Escherichia coli (E. coli MG1655 and its isogenic mutant E. coli MG1655 ΔrpoS) and Listeria monocytogenes (L. monocytogenes EGD-e and its isogenic mutant L. monocytogenes EGD-e ΔsigB) to the essential oils (EOs) from Origanum vulgare L.-oregano (OVEO) and Rosmarinus officinalis L.-rosemary (ROEO), as well as the changes in tolerance of parental and ΔrpoS and ΔsigB mutant strains to OVEO, ROEO and pulsed electric fields (PEF) following overnight exposure to subinhibitory concentrations (1/2×minimum inhibitory concentration-MIC) of each tested EO. MIC values of OVEO and ROEO against the mutant cells were usually lower than those found against the parental cells. Survivor curves showed that mutant cells were more sensitive to these EOs than parental cells. The recovery of survivors in selective media showed a greater proportion of cells sublethally injured at their cell envelopes in the mutant strains compared with the parental strains. Induction of increased direct-tolerance to OVEO and ROEO or cross-tolerance to PEF was not observed after pre-exposure of parental and mutant cells to EOs. Otherwise, parental and mutant cells of E. coli and L. monocytogenes pre-exposed to OVEO or ROEO showed decreased tolerance when further treated with the homologous stressing agent at 2×MIC. Still, mutant cells pre-exposed to OVEO or ROEO showed lower tolerance to PEF than parental strains. These results showed the influence of σ(S) and σ(B) in tolerance of single strains of E. coli and L. monocytogenes, respectively, to OVEO and ROEO. Moreover, the deletion of σ(S) and σ(B) resulted in decreased tolerance to OVEO, ROEO or PEF in tested strains following exposure to OVEO or ROEO at a subinhibitory concentration. Copyright © 2015 Elsevier B.V. All rights reserved.

  6. Grand Canyon VFR Chart - Aeronautical Information Services Digital Products

    Data.gov (United States)

    Department of Transportation — The Federal Aviation Administration (FAA) digital-Visual Chart series is designed to meet the needs of users who require georeferenced raster images of a FAA Visual...

  7. Increase in imported malaria in the Netherlands in asylum seekers and VFR travellers

    NARCIS (Netherlands)

    Gier, B. de; Suryapranata, F.S.; Croughs, M.; Genderen, P.J. van; Keuter, M.; Visser, L.G.; Vugt, M. De; Sonder, G.J.

    2017-01-01

    BACKGROUND: Malaria is a notifiable disease in the Netherlands, a non-endemic country. Imported malaria infections occur regularly among travellers, migrants and visitors. Surveillance data were analysed from 2008 to 2015. Trends in amounts of notifications among risk groups were analysed using

  8. Increase in imported malaria in the Netherlands in asylum seekers and VFR travellers

    NARCIS (Netherlands)

    de Gier, Brechje; Suryapranata, Franciska S. T.; Croughs, Mieke; van Genderen, Perry J. J.; Keuter, Monique; Visser, Leo G.; van Vugt, Michele; Sonder, Gerard J. B.

    2017-01-01

    Malaria is a notifiable disease in the Netherlands, a non-endemic country. Imported malaria infections occur regularly among travellers, migrants and visitors. Surveillance data were analysed from 2008 to 2015. Trends in amounts of notifications among risk groups were analysed using Poisson

  9. 78 FR 50313 - Final Additional Airworthiness Design Standards: Night Visual Flight Rules (VFR) Under the...

    Science.gov (United States)

    2013-08-19

    ... control toward lean or shut-off position. (b) Each manual engine mixture control must be designed so that... Federal Aviation Administration 14 CFR Part 21 Final Additional Airworthiness Design Standards: Night... design standards. SUMMARY: This document is an issuance of Final Airworthiness design criteria for night...

  10. Vertebral fracture risk (VFR) score for fracture prediction in postmenopausal Women

    DEFF Research Database (Denmark)

    Lillholm, Martin; Ghosh, A.; Pettersen, P. C.

    2011-01-01

    Early prognosis of osteoporosis risk is not only important to individual patients but is also a key factor when screening for osteoporosis drug trial populations. We present an osteoporosis fracture risk score based on vertebral heights. The score separated individuals who sustained fractures (by...

  11. Natural rpoS mutations contribute to the phenotypic heterogeneity of clonal populations in Escherichia coli O157:H7

    Science.gov (United States)

    We previously reported the distinct acid resistance between the curli-producing (C+) and curli-deficient (C-) variants of E. coli O157:H7, although the curli fimbriae were not associated with this intra-strain phenotypic divergence. Here we investigated the underlying molecular mechanism by examinin...

  12. Educating international students about tuberculosis and infections associated with travel to visit friends and relatives (VFR-travel).

    Science.gov (United States)

    Gibney, Katherine B; Brass, Amanda; Hume, Sam C; Leder, Karin

    2014-01-01

    International students in Victoria, Australia, originate from over 140 different countries. They are over-represented in disease notifications for tuberculosis and travel-associated infections, including enteric fever, hepatitis A, and malaria. We describe a public health initiative aimed to increase awareness of these illnesses among international students and their support staff. We identified key agencies including student support advisors, medical practitioners, health insurers, and government and professional organisations. We developed health education materials targeting international students regarding tuberculosis and travel-related infections to be disseminated via a number of different media, including electronic and printed materials. We sought informal feedback from personnel in all interested agencies regarding the materials developed, their willingness to deliver these materials to international students, and their preferred media for disseminating these materials. Education institutions with dedicated international student support staff and on-campus health clinics were more easily engaged to provide feedback and disseminate the health education materials than institutions without such dedicated personnel. Response to contacting off-campus medical practices was poor. Delivery of educational materials via electronic and social media was preferred over face-to-face education. It is feasible to provide health education messages targeting international students for dissemination via appropriately-staffed educational institutions. This initiative could be expanded in terms of age-group, geographic range, and health issues to be targeted. Copyright © 2013 Elsevier Ltd. All rights reserved.

  13. The role of RpoS, H-NS and AcP on the pH-dependent OmpC and ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-05-04

    2008) showed the rearrange- ment of outer membrane proteins in Escherichia coli entering the Viable But Non Culturable (VBNC) state. The mechanisms of OmpC and OmpF porin regulation are particularly complex.

  14. Deployment support leading to implementation

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1999-11-01

    This quarterly report discusses progress on the following tasks and subtasks: (Task 1)--Field demonstration of prefabricated vertical drains (PVD) technology: Subtask 1.1--Pilot scale demonstration of TCE flushing through PVDs at the DOE/RMI Extrusion Plant; Subtask 1.2--Technical support on surfactants and QA/QC for field operations employing PVDs at the RMI Ashtabula, Ohio, site; Subtask 1.3--Operational support of the PVDs and engineering/design/construction/operations of a wastewater treatment plant for extracted fluids; Subtask 1.4--Installation of PVDs at the RMI Ashtabulah, Ohio, site; and Subtask 1.5--Removal of surfactants and trichloroethylene (TCE) from extracted groundwater at the RMI Ashtabula, Ohio, site; (Task 2)--Watershed economics and overview studies; and (Task 3)--Project management.

  15. Transcriptional profiling of ParA and ParB mutants in actively dividing cells of an opportunistic human pathogen Pseudomonas aeruginosa.

    Directory of Open Access Journals (Sweden)

    Aneta A Bartosik

    Full Text Available Accurate chromosome segregation to progeny cells is a fundamental process ensuring proper inheritance of genetic material. In bacteria with simple cell cycle, chromosome segregation follows replication initiation since duplicated oriC domains start segregating to opposite halves of the cell soon after they are made. ParA and ParB proteins together with specific DNA sequences are parts of the segregation machinery. ParA and ParB proteins in Pseudomonas aeruginosa are important for optimal growth, nucleoid segregation, cell division and motility. Comparative transcriptome analysis of parA null and parB null mutants versus parental P. aeruginosa PAO1161 strain demonstrated global changes in gene expression pattern in logarithmically growing planktonic cultures. The set of genes similarly affected in both mutant strains is designated Par regulon and comprises 536 genes. The Par regulon includes genes controlled by two sigma factors (RpoN and PvdS as well as known and putative transcriptional regulators. In the absence of Par proteins, a large number of genes from RpoS regulon is induced, reflecting the need for slowing down the cell growth rate and decelerating the metabolic processes. Changes in the expression profiles of genes involved in c-di-GMP turnover point out the role of this effector in such signal transmission. Microarray data for chosen genes were confirmed by RT-qPCR analysis. The promoter regions of selected genes were cloned upstream of the promoter-less lacZ gene and analyzed in the heterologous host E. coliΔlac. Regulation by ParA and ParB of P. aeruginosa was confirmed for some of the tested promoters. Our data demonstrate that ParA and ParB besides their role in accurate chromosome segregation may act as modulators of genes expression. Directly or indirectly, Par proteins are part of the wider regulatory network in P. aeruginosa linking the process of chromosome segregation with the cell growth, division and motility.

  16. Shopping for Diasporic Belonging: Being ‘Local’ or Being ‘Mobile’ as a VFR Visitor in the Ancestral Homeland

    NARCIS (Netherlands)

    Wagner, L.B.

    2015-01-01

    Visits to friends and relatives are often part of circulatory migration mobilities between resident and ancestral homelands. These journeys are characteristic of the diasporic lives of post-migrant generations of Moroccans from Europe, where close proximity and relatively unimpeded mobility between

  17. Architecture of Burkholderia cepacia complex σ70 gene family: evidence of alternative primary and clade-specific factors, and genomic instability

    Directory of Open Access Journals (Sweden)

    Menard Aymeric

    2007-09-01

    Full Text Available Abstract Background The Burkholderia cepacia complex (Bcc groups bacterial species with beneficial properties that can improve crop yields or remediate polluted sites but can also lead to dramatic human clinical outcomes among cystic fibrosis (CF or immuno-compromised individuals. Genome-wide regulatory processes of gene expression could explain parts of this bacterial duality. Transcriptional σ70 factors are components of these processes. They allow the reversible binding of the DNA-dependent RNA polymerase to form the holoenzyme that will lead to mRNA synthesis from a DNA promoter region. Bcc genome-wide analyses were performed to investigate the major evolutionary trends taking place in the σ70 family of these bacteria. Results Twenty σ70 paralogous genes were detected in the Burkholderia cenocepacia strain J2315 (Bcen-J2315 genome, of which 14 were of the ECF (extracytoplasmic function group. Non-ECF paralogs were related to primary (rpoD, alternative primary, stationary phase (rpoS, flagellin biosynthesis (fliA, and heat shock (rpoH factors. The number of σ70 genetic determinants among this genome was of 2,86 per Mb. This number is lower than the one of Pseudomonas aeruginosa, a species found in similar habitats including CF lungs. These two bacterial groups showed strikingly different σ70 family architectures, with only three ECF paralogs in common (fecI-like, pvdS and algU. Bcen-J2315 σ70 paralogs showed clade-specific distributions. Some paralogs appeared limited to the ET12 epidemic clone (ecfA2, particular Bcc species (sigI, the Burkholderia genus (ecfJ, ecfF, and sigJ, certain proteobacterial groups (ecfA1, ecfC, ecfD, ecfE, ecfG, ecfL, ecfM and rpoS, or were broadly distributed in the eubacteria (ecfI, ecfK, ecfH, ecfB, and rpoD-, rpoH-, fliA-like genes. Genomic instability of this gene family was driven by chromosomal inversion (ecfA2, recent duplication events (ecfA and RpoD, localized (ecfG and large scale deletions (sig

  18. Sigma factor selectivity in Borrelia burgdorferi: RpoS recognition of the ospE/ospF/elp promoters is dependent on the sequence of the −10 region

    National Research Council Canada - National Science Library

    Eggers, Christian H; Caimano, Melissa J; Radolf, Justin D

    2006-01-01

    Members of the ospE/ospF/elp lipoprotein gene families of Borrelia burgdorferi , the Lyme disease agent, are transcriptionally upregulated in response to the influx of blood into the midgut of an infected tick...

  19. The role of each compartment in a two-compartment vertical flow reactor for ferruginous mine water treatment.

    Science.gov (United States)

    Yim, G J; Cheong, Y W; Hong, J H; Hur, W

    2014-10-01

    A vertical flow reactor (VFR) has been suggested for remediation of ferruginous mine drainage that passes down through an accreting bed of ochre. However, a VFR has a limited operation time until the system begins to overflow. In this study, a mathematical model was developed as a part of the effort to explore the operation of a VFR, showing dynamic changes in the head differences, ochre depths, and Fe(II)/Fe(III) concentrations in the effluent flow. The analysis showed that VFR operation time extended from 148.5 days to 163 days in an equally divided and to 168.4 days in asymmetrically (0.72:0.28) divided two-compartment VFR, suggesting that an optimum compartment ratio exists that maximizes the VFR operation time. A constant head filtration in the first compartment maximized filtration efficiency and thus prolonged VFR longevity in the two-compartment VFR. Fe(II) oxidation and ochre formation should be balanced with the permeability of the ochre bed to maximize the VFR operation time and minimize the residual Fe(II) in the effluent. Accelerated Fe(II) oxidation affected the optimum ratio of the compartment area and reduced the residual Fe(II) in the effluent. The VFR operation time can be prolonged significantly from 764 days to 3620 days by increasing the rate of ochre formation, much more than by accelerating the Fe(II) oxidation. During the prolonged VFR operation, ochre formed largely in the first compartment, while overflowing mine water with reduced iron content was effectively filtered in the second compartment. These results not only provide a better understanding of VFR operation but also suggest the direction of evolution of two-compartment VFR toward a compact and highly efficient facility integrated with an aerated cascade and with automatic coagulant feeding. Copyright © 2014 Elsevier Ltd. All rights reserved.

  20. Relative Lyapunov Center Bifurcations

    DEFF Research Database (Denmark)

    Wulff, Claudia; Schilder, Frank

    2014-01-01

    Relative equilibria (REs) and relative periodic orbits (RPOs) are ubiquitous in symmetric Hamiltonian systems and occur, for example, in celestial mechanics, molecular dynamics, and rigid body motion. REs are equilibria, and RPOs are periodic orbits of the symmetry reduced system. Relative Lyapunov...... center bifurcations are bifurcations of RPOs from REs corresponding to Lyapunov center bifurcations of the symmetry reduced dynamics. In this paper we first prove a relative Lyapunov center theorem by combining recent results on the persistence of RPOs in Hamiltonian systems with a symmetric Lyapunov...

  1. Which Air Force Civil Engineer Capabilities Can Complement USNORTHCOM’s Role in Defense Support to Civil Authorities (DSCA)?

    Science.gov (United States)

    2014-05-21

    fixed wing or rotary wing aircraft, airfield/landing zone under Visual Flight Rules (VFR) and Instrument Flight Rules ( IFR ) conditions for multiple...wing aircraft, airfield/landing zone under Visual Flight Rules (VFR) and Instrument Flight Rules ( IFR ) conditions for multiple aircraft types on small...under Visual Flight Rules (VFR) and Instrument Flight Rules ( IFR ) conditions for multiple aircraft types on small to large temporary/ permanent

  2. Due Regard Encounter Model Version 1.0

    Science.gov (United States)

    2013-08-19

    Note that no existing model covers encoun- ters between two IFR aircraft in oceanic airspace. The reason for this is that one cannot observe encounters...encounters between instrument flight rules ( IFR ) and non- IFR traffic beyond 12NM. 2 TABLE 1 Encounter model categories. Aircraft of Interest Intruder...Aircraft Location Flight Rule IFR VFR Noncooperative Noncooperative Conventional Unconventional CONUS IFR C C U X VFR C U U X Offshore IFR C C U X VFR C U

  3. Preliminary Uncorrelated Encounter Model of the National Airspace System

    National Research Council Canada - National Science Library

    Kochenderfer, M. J; Kuchar, J. K; Espindle, L. P; Gertz, J. L

    2008-01-01

    ... procedures and collision avoidance systems. Of particular relevance to Unmanned Aircraft Systems is the potential for encountering general aviation aircraft that are flying under Visual Flight Rules (VFR...

  4. Numerical bifurcation of Hamiltonian relative periodic orbits

    DEFF Research Database (Denmark)

    Wulff, Claudia; Schilder, Frank

    2009-01-01

    Relative periodic orbits (RPOs) are ubiquitous in symmetric Hamiltonian systems and occur, for example, in celestial mechanics, molecular dynamics, and the motion of rigid bodies. RPOs are solutions which are periodic orbits of the symmetry-reduced system. In this paper we analyze certain symmetry...

  5. Variable frame rate analysis for automatic speech recognition

    Science.gov (United States)

    Tan, Zheng-Hua

    2007-09-01

    In this paper we investigate the use of variable frame rate (VFR) analysis in automatic speech recognition (ASR). First, we review VFR technique and analyze its behavior. It is experimentally shown that VFR improves ASR performance for signals with low signal-to-noise ratios since it generates improved acoustic models and substantially reduces insertion and substitution errors although it may increase deletion errors. It is also underlined that the match between the average frame rate and the number of hidden Markov model states is critical in implementing VFR. Secondly, we analyze an effective VFR method that uses a cumulative, weighted cepstral-distance criterion for frame selection and present a revision for it. Lastly, the revised VFR method is combined with spectral- and cepstral-domain enhancement methods including the minimum statistics noise estimation (MSNE) based spectral subtraction and the cepstral mean subtraction, variance normalization and ARMA filtering (MVA) process. Experiments on the Aurora 2 database justify that VFR is highly complementary to the enhancement methods. Enhancement of speech both facilitates the frame selection in VFR and provides de-noised speech for recognition.

  6. Perceived Visual Distortions in Juvenile Amblyopes During/Following Routine Amblyopia Treatment.

    Science.gov (United States)

    Piano, Marianne E F; Bex, Peter J; Simmers, Anita J

    2016-08-01

    To establish the point prevalence of perceived visual distortions (PVDs) in amblyopic children; the association between severity of PVDs and clinical parameters of amblyopia; and the relationship between PVDs and amblyopia treatment outcomes. Perceived visual distortions were measured using a 16-point dichoptic alignment paradigm in 148 visually normal children (aged, 9.18 ± 2.51 years), and 82 amblyopic children (aged, 6.33 ± 1.48 years) receiving or following amblyopia treatment. Global distortion (GD; vector sum of mean-centered individual alignment error between physical and perceived target location) and Global uncertainty (GU; SD of GD over two experiment runs) were compared to age-matched control data, and correlated against clinical parameters of amblyopia (type, monocular visual acuity, pretreatment interocular acuity difference, refractive error, age at diagnosis, motor fusion, stereopsis, near angle of deviation) and amblyopia treatment outcomes (refractive adaption duration, treatment duration, occlusion dosage, posttreatment interocular acuity difference, number of lines improvement). Point prevalence of PVDs in amblyopes was 56.1%. Strabismic amblyopes experienced more severe distortions than anisometropic or microtropic amblyopes (GD Kruskal Wallis H = 16.89, P amblyopia treatment outcomes, or the amblyopic visual acuity deficit. Perceived visual distortions persisted in more than one-half of treated amblyopic cases whose treatment was deemed successful. Perceived visual distortions are common symptoms of amblyopia and are correlated with binocular (stereoacuity, angle of deviation), but not monocular (visual acuity) clinical outcomes. This adds to evidence demonstrating the role of decorrelated binocular single vision in many aspects of amblyopia, and emphasizes the importance of restoring and improving binocular single vision in amblyopic individuals.

  7. Expression dynamics of RpoS/Crl-dependent genes in Escherichia coli.

    Science.gov (United States)

    Dudin, Omaya; Lacour, Stéphan; Geiselmann, Johannes

    2013-10-01

    The alternative sigma factor RpoS is a central regulator of the stress response in many Proteobacteria, acting both during exponential growth and in stationary phase. The small protein Crl increases the interaction between RpoS and RNA polymerase and thereby activates certain RpoS-dependent promoters. However, the growth-phase dependence of the interaction of Crl with different forms of polymerase remains unknown. We use 41 GFP transcriptional fusions to study the dynamics of gene regulation by RpoS and Crl during growth transition from exponential to stationary phase in Escherichia coli. We confirm that RpoS can regulate gene expression in exponential phase, both positively and negatively. Crl slightly stimulates transcription by RpoS in exponential phase and controls a subset of RpoS-dependent genes in stationary phase. Growth temperature strongly affects induction of specific promoters by RpoS, whereas its impact on gene regulation by Crl is much less significant. In addition, we identify five new genes regulated by Crl (ada, cbpA, glgS, sodC and flgM) and demonstrate that Crl improves promoter binding and opening by RpoS-containing RNA polymerase at the hdeA promoter. Our study also shows that Crl is a cognate enhancer of RpoS activity under different growth conditions, since its deletion has no effect on genes transcribed by other sigma factors. Copyright © 2013 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  8. Cell-mediated contraction of vitreous explants from chicken embryo: Possibility of screening for therapeutic agents against proliferative vitreoretinal diseases

    Science.gov (United States)

    Oki, Keitaro; Shimada, Arata; Nagase, Terumasa; Katsura, Yoshiya; Kosano, Hiroshi

    2013-01-01

    Purpose We aimed to establish a novel screening system for identifying potential therapeutic agents for treating proliferative vitreoretinal diseases (PVDs). In this study, we focused on vitreous explants from chicken embryos and evaluated the usefulness of quantitatively analyzing the effects of potential candidates on cell-mediated vitreous contraction, which leads to blindness in PVDs. Methods Vitreous explants were extracted from 19-day-old embryonic chickens and then incubated with retinal Müller cells or endothelial cells to permit cell adhesion. After cell adhesion occurred, we examined the effect of the attached cells on the wet weight of vitreous explants as an index of vitreous contraction. We also performed hematoxylin and eosin staining to characterize the cell morphology on the vitreous surface. Results Contraction of the vitreous explants was observed after cell adhesion of not only retinal Müller cells but also endothelial cells. We confirmed the adhesion of these cells on vitreous explants and estimated the number of adherent cells with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) analysis. The cells on the vitreous surface presented an elongated fibroblast-like phenotype. Integrin was found to be a receptor involved in cell adhesion on the vitreous surface. Discussion Our results suggest that vitreous explants from chicken embryos may be novel useful tools for screening antiadhesion therapeutic agents in PVDs. This preliminary study must be validated with human vitreous and human retinal pigment epithelial cells. PMID:24319331

  9. Chinese travellers visiting friends and relatives--A review of infectious risks.

    Science.gov (United States)

    Ma, Tara; Heywood, Anita; MacIntyre, C Raina

    2015-01-01

    Travellers are potential vectors in the transmission of infectious diseases across international borders. Travellers visiting friends and relatives (VFR) have a particularly high risk of acquiring certain infections during travel. Chinese VFR travellers account for a substantial proportion of all travel in Western countries with high migrant populations. A literature review was undertaken regarding major infectious disease risks for VFR travellers visiting China. This included an examination of the previous pandemics arising in China, the likelihood of future outbreaks in China from H5N1 and H7N9 avian influenza viruses, the potential role of travellers in disease transmission, and the special risks for VFR travellers. China has been the origin of several influenza pandemics in past few decades, and the origin of several emerging infectious diseases with pandemic potential, including SARS. Travel to and from China has the potential for global spread of emergent infectious diseases, as seen in the SARS outbreak in 2003. For VFR travellers, the risk of other infectious diseases may also be greater in China compared to their countries of migration, including hepatitis A and B, dengue fever, typhoid, and other diseases. VFR travel to China may be associated with increased risk of acquiring a range of infectious diseases, and also poses a potential risk for importation of future pandemics to other countries. Chinese VFR travellers need to be cognisant of these risks and health professionals should consider educational interventions to minimise these risks. Copyright © 2015 Elsevier Ltd. All rights reserved.

  10. Texas RPO workshop implementation project summary.

    Science.gov (United States)

    2012-05-01

    This report documents rural planning organization (RPO) workshops conducted throughout Texas. An RPO is a voluntary organization created and governed by locally elected officials responsible for transportation decisions at the local level. RPOs addre...

  11. Development of performance measures for the assessment of rural planning organizations.

    Science.gov (United States)

    2011-04-27

    In order for the Transportation Planning Board to provide oversight and assistance to the 20 RPOs in the state, they : need effective evaluation criteria and performance measures. The existing measures, including the annual : performance report, do n...

  12. Two mechanisms for putrescine-dependent transcriptional expression of the putrescine aminotransferase gene, ygjG, in Escherichia coli.

    Science.gov (United States)

    Kim, Young-Sik; Shin, Hyun-Chul; Lee, Jong-Ho

    2014-09-01

    In this study, on evaluating the physiological function and mechanism of putrescine, we found that putrescine supplementation (1 mM) increases transcription of the putrescine aminotransferase gene, ygjG. Putrescine-dependent expression was confirmed by measuring β-galactosidase activity and with reverse transcription-polymerase chain reaction. To understand the role of putrescine in ygjG expression, we genetically characterized and found that a knockout mutation in an alternative sigma factor, rpoS, abolished putrescine-dependent ygjG-lacZ expression. In the rpoS mutant, RpoS overexpression complemented the mutant phenotype. However, RpoS overexpression induced ygjG-lacZ expression with putrescine supplementation but not without supplementation. We also found that the loss of putrescine-dependent ygjG-lacZ expression induced by rpoS was completely restored under nitrogen-starvation conditions. The putrescine-dependent expression of ygjG-lacZ under this condition was clearly dependent on another alternative sigma factor, rpoN, and its cognate activator ntrC. These results show that rpoS is required for putrescine-dependent ygjG-lacZ expression, but the effect of putrescine on this expression is not caused by simple modulation of RpoS synthesis. Putrescine-dependent expression of ygjG-lacZ was controlled by at least two sigma factors: rpoS under excess nitrogen conditions and rpoN under nitrogen-starvation conditions. These results suggest that putrescine plays an important role in the nitrogen regulation system.

  13. FA

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — An Area Forecast (FA) is a forecast of Visual Flight Rules (VFR) clouds and weather conditions over an area as large as the size of several states.

  14. SigWx-Low

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — The low-level graphics product is a forecast of aviation weather hazards, primarily intended to be used as a guidance product for briefing the VFR pilot. The...

  15. 78 FR 63861 - Modification of Class B Airspace; Minneapolis, MN

    Science.gov (United States)

    2013-10-25

    ... Class B airspace as required by FAA directives to enhance safety and efficient management of aircraft... airspace with an expected minimal impact on non- participatory VFR operations. The FAA anticipates that...

  16. Chart Supplements - Aeronautical Information Services Digital Products

    Data.gov (United States)

    Department of Transportation — The Chart Supplements are searchable by individual airport in PDF format. They contain data on public and joint use airports, seaplane bases, heliports, VFR airport...

  17. 77 FR 6663 - Airworthiness Directives; CPAC, Inc. Airplanes

    Science.gov (United States)

    2012-02-09

    ... is a Rockwell ``marketing name'' for the Model 112. The Model 115 is a Rockwell ``marketing name... permitted for daytime visual flight rules (VFR) only, restricted to crew, calm weather, reduced speed not to...

  18. 78 FR 34608 - Proposed Establishment of Class D Airspace; Bryant AAF, Anchorage, AK

    Science.gov (United States)

    2013-06-10

    ... Association (AOPA), Alaska Airmen's Association (AAA), and the Alaskan Aviation Safety Foundation (AASF). One... AAF. AOPA identified three issues that affect pilots using the Eastside VFR corridor they would like...

  19. Exploiting the Automatic Dependent Surveillance-Broadcast System via False Target Injection

    Science.gov (United States)

    2012-03-01

    IEEE Xplore . 8. Hal Stoen. (2001) VFR Flight. Retrieved February 15, 2012 from http://stoenworks.com/vfr%20flight.html 9. Cohen, B., & Smith, A...7. Gilbert, G. (1973). Historical development of the air traffic control system. IEEE Transactions on Communications, 364-375. Retrieved from...1998. Proceedings., 17th DASC. the AIAA/ IEEE /SAE,2 F44/1- F44/8 vol.2. 10. Livack, G. S., McDaniel, J. I., Battiste, V., & Johnson, W. W. (2000

  20. Impact of traveling to visit friends and relatives on chronic disease management.

    Science.gov (United States)

    Gurgle, Holly E; Roesel, David J; Erickson, Tiffany N; Devine, Emily Beth

    2013-01-01

    Travelers visiting friends and relatives (VFR) are known to be at high risk of acquiring infectious diseases during travel. However, little is known about the impact of VFR travel on chronic diseases. This was a nonrandomized, retrospective observational study. Patients were adult VFR travelers who received care from an internal medical clinic serving immigrants and refugees. The primary objective was to determine the impact of VFR travel on markers of chronic disease management including: blood pressure, glycosylated hemoglobin, body mass index, serum creatinine, and anticoagulation. Of the 110 VFR travelers in our study, N = 48 traveled to Africa and N = 62 traveled to Asia for a mean duration of 59 (range 21-303) days. Of the 433 counseling points discussed at pre-travel visits, 71% were infectious disease prevention, 16% chronic disease related, and 13% travel safety. A total of 63 patients (57%) reported one or more health problems while traveling. Of these, 35 patients (32%) experienced a problem related to a chronic condition. In comparison, 24 (22%) patients experienced an acute infection. Sixty percent of patients were nonadherent to medications during travel. An average increase in diastolic blood pressure of 3.6 mmHg among patients with hypertension was the only statistically significant change in a chronic disease marker when values before and after travel were compared. Subgroup analysis revealed that travel to Africa and nonadherence to medications were also associated with worsening blood pressure control, and patients traveling to Africa experienced a decrease in body mass index. This study identified a high proportion of problems related to chronic conditions experienced during VFR travel, while pre-travel appointments tended to focus on infectious disease prevention. A greater emphasis on medication adherence and chronic disease management during VFR travel is also needed during pre-travel preparations. © 2013 International Society of

  1. Correlation of volumetric flow rate and skin blood flow with cold intolerance in digital replantation.

    Science.gov (United States)

    Zhao, Gang; Mi, Jingyi; Rui, Yongjun; Pan, Xiaoyun; Yao, Qun; Qiu, Yang

    2017-12-01

    Cold intolerance is a common complication of digital replantation. The exact etiology is unclear, but it is considered to be multifactorial, including nonsurgical characteristics, vascular, and neurologic conditions. Blood flow may play a significant role in cold intolerance. This study was designed to evaluate the correlation of digital blood flow, including volumetric flow rate (VFR) and skin blood flow (SkBF), with cold intolerance in replanted fingers.A retrospective study was conducted among patients who underwent digital replantation between 2010 and 2013. Patients were selected into study cohort based on the inclusion criteria. Surgical data was collected on each patient, including age, sex, injury mechanism, amputation level, ischemia time, number of arteries repaired, and whether or not vascular crisis occurred. Patients were included as study cohort with both nerves repaired and without chronic disease. Cold intolerance was defined as a Cold Intolerance Symptom Severity (CISS) score over 30. The arterial flow velocity and caliber were measured by Color Doppler Ultrasound and the digital VFR was calculated. The SkBF was measured by Laser Speckle Imager. Both VFR and SkBF were calculated as a percentage of the contralateral fingers. Comparative study of surgical data and blood flow was performed between the patient with and without cold intolerance. Correlation between VFR and SkBF was also analyzed.A total of 93 patients met inclusion criteria for the study. Approximately, 42 patients were identified as having cold intolerance. Fingers that survived vascular crisis had a higher incidence of cold intolerance with a lower VFR and SkBF. The VFR was higher in 2-artery replantation, but the SkBF and incidence of cold intolerance did not differ significantly. No differences were found in age, sex, injury mechanism, amputation level, or ischemia time. Furthermore, no correlation was found between VFR and SkBF.Cold intolerance of digital replantation is associated

  2. 77 FR 12770 - Approval, Disapproval and Promulgation of Implementation Plans; Nebraska; Regional Haze State...

    Science.gov (United States)

    2012-03-02

    ... and related issues. The RPOs first evaluated technical information to better understand how their... referred to as the ``regional haze program''). States are required to ensure reasonable progress toward the... of sources until monitoring, modeling and scientific knowledge about the relationships between...

  3. Statistical Analysis of Pseudomonas aeruginosa Biofilm Development: Impact of Mutations in Genes Involved in Twitching Motility, Cell-to-Cell Signaling, and Stationary-Phase Sigma Factor Expression

    DEFF Research Database (Denmark)

    Heydorn, Arne; Ersbøll, Bjarne Kjær; Kato, Junichi

    2002-01-01

    Four strains of Pseudomonas aeruginosa (wild type, DeltapilHIJK mutant, lasI mutant, and rpoS mutant) were genetically tagged with the green fluorescent protein, and the development of flow chamber-grown biofilms by each of them was investigated by confocal laser scanning microscopy. The structur...

  4. Lysophosphatidic acid inhibition of the accumulation of Pseudomonas aeruginosa PAO1 alginate, pyoverdin, elastase and LasA

    DEFF Research Database (Denmark)

    Laux, D.C.; Corson, J.M.; Givskov, Michael Christian

    2002-01-01

    pyoverdin. MPPA also inhibited biofilm formation. The inhibitory effects of MPPA occur independently of rpoS expression and without affecting the accumulation of the autoinducers N-(3-oxododecanoyl) homoserine lactone and N-butyryl-(L)-homoserine lactone, and may be due, at least in part, to the ability...

  5. Comparative survival of Escherichia coli O157:H7, Salmonella Typhimurium, and Murine Norovirus on spinach plants

    Science.gov (United States)

    Introduction: Outbreaks resulting from the consumption of leafy greens contaminated with E. coli O157:H7, Salmonella spp., and norovirus have occurred. It is unclear how the stress response factor rpoS in E. coli O157:H7 and Salmonella spp. affects their survival on spinach. Purpose: A comparison ...

  6. Accumulation of hns mutations specifically in stationary phase in an ...

    Indian Academy of Sciences (India)

    Home; Journals; Journal of Genetics; Volume 85; Issue 3. Accumulation of hns mutations specifically in stationary phase in an E. coli strain carrying an impaired rpoS locus. Stuti K. Desai S. Mahadevan. Research Note Volume 85 Issue 3 December 2006 pp 221-224 ...

  7. Initial characterization of a bolA homologue from Pseudomonas fluorescens indicates different roles for BolA-like proteins in em>P. fluorescens and Escherichia coli

    DEFF Research Database (Denmark)

    Koch, Birgit; Nybroe, Ole

    2006-01-01

    The RpoS-regulated bolA gene in Escherichia coli is important for the decrease in cell size during stationary phase or sudden carbon starvation. A Pseudomonas fluorescens strain mutated in a gene with homology to bolA reduced its cell size upon carbon starvation, and RpoS had little effect on bol...

  8. Accumulation of hns mutations specifically in stationary phase in an ...

    Indian Academy of Sciences (India)

    ditions for growth such as nutrient deprivation experienced during stationary ... taneous mutants that preexisted in the exponential-phase cul- ture used for plating. The same plates on further incubation. (96 h) gave rise to additional Bgl. + mutants that ... Landini 2004) we made use of an additional rpoS. + con- trol strain that ...

  9. Publications | Page 590 | IDRC - International Development ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    Results 5891 - 5900 of 6371 ... Chilean Management Centers are companies specialized in developing the management and competitiveness capacities of individual family farms and of market-oriented Rural Producer Organizations (RPOs; known in Chile as associative peasant businesses firms). In order to develop their ...

  10. Pyoverdine synthesis by the Mn(II-oxidizing bacterium Pseudomonas putida GB-1

    Directory of Open Access Journals (Sweden)

    Dorothy Lundquist Parker

    2014-05-01

    Full Text Available When iron-starved, the Mn(II-oxidizing bacteria Pseudomonas putida strains GB-1 and MnB1 produce pyoverdines (PVDGB-1 and PVDMnB1, siderophores that both influence iron uptake and inhibit manganese(II oxidation by these strains. To explore the properties and genetics of a PVD that can affect manganese oxidation, LC-MS/MS and various siderotyping techniques were used to identify the peptides of PVDGB-1 and PVDMnB1 as being (for both PVDs: chromophore-Asp-Lys-OHAsp-Ser-Gly-aThr-Lys-cOHOrn, resembling a structure previously reported for P. putida CFML 90-51, which does not oxidize Mn. All three strains also produced an azotobactin and a sulfonated PVD, each with the peptide sequence above, but with unknown regulatory or metabolic effects. Bioinformatic analysis of the sequenced genome of P. putida GB-1 suggested that a particular non-ribosomal peptide synthetase, coded by the operon PputGB1_4083-4086, could produce the peptide backbone of PVDGB-1. To verify this prediction, plasmid integration disruption of PputGB1_4083 was performed and the resulting mutant failed to produce detectable PVD. In silico analysis of the modules in PputGB1_4083-4086 predicted a peptide sequence of Asp-Lys-Asp-Ser-Ala-Thr-Lsy-Orn, which closely matches the peptide determined by MS/MS. To extend these studies to other organisms, various Mn(II-oxidizing and non-oxidizing isolates of P. putida, P. fluorescens, P. marincola, P. fluorescens-syringae group, P. mendocina-resinovorans group and P. stutzerii group were screened for PVD synthesis. The PVD producers (12 out of 16 tested strains were siderotyped and placed into four sets of differing PVD structures, some corresponding to previously characterized PVDs and some to novel PVDs. These results combined with previous studies suggested that the presence of OHAsp or the flexibility of the pyoverdine polypeptide may enable efficient binding of Mn(III.

  11. Eye Tracking in the Cockpit: a Review of the Relationships between Eye Movements and the Aviators Cognitive State

    Science.gov (United States)

    2014-12-01

    frequency. In particular, Spady (1978) examined eye movements during simulated landing approach under instrument flight rules ( IFR ). Simulated turbulence...al. (2007) found that NNI varied across phases of simulated IFR flight, showing the least random (most clustered) distribution of fixations during...decreased for visually demanding flight segments ( IFR vs. VFR, landing vs. cruise). This general pattern has been replicated across several studies

  12. Supplemental Environmental Impact Statement for F-35 Beddown at Eglin Air Force Base, Florida

    Science.gov (United States)

    2014-06-26

    associated with placing additional instrument flight rules ( IFR ) operations at other airfields within the region. These flight restrictions degrade... IFR instrument flight rules IJTS Initial Joint Training Site ILLUM Illuminating ILS Instrument Landing System IMPLAN an economic impact modeling...primary runway for instrument flight rules ( IFR ) activities, while RW 12/30 is the primary runway for visual flight rules (VFR) activities. Although

  13. A comparison between the order and the volume fill rate for a base-stock inventory control system under a compound renewal demand process

    DEFF Research Database (Denmark)

    Larsen, Christian; Thorstenson, Anders

    2008-01-01

    The order fill rate (OFR) is sometimes suggested as an alternative to the volume fill rate (VFR) (most often just denoted fill rate) as a performance measure for inventory control systems. We consider a continuous review, base-stock policy, where replenishment orders have a constant lead time...

  14. Derivation of confidence intervals of service measures in a base-stock inventory control system with low-frequent demand

    DEFF Research Database (Denmark)

    Larsen, Christian

    We explore a base-stock system with backlogging where the demand process is a compound renewal process and the compound element is a delayed geometric distribution. For this setting it is proven in [4] that the long-run average service measures order fill rate (OFR) and volume fill rate (VFR) are...

  15. Derivation of confidence intervals of service measures in a base-stock inventory control system with low-frequent demand

    DEFF Research Database (Denmark)

    Larsen, Christian

    2011-01-01

    We explore a base-stock system with backlogging where the demand process is a compound renewal process and the compound element is a delayed geometric distribution. For this setting it holds that the long-run average service measures order fill rate (OFR) and volume fill rate (VFR) are equal in v...

  16. 14 CFR 135.227 - Icing conditions: Operating limitations.

    Science.gov (United States)

    2010-01-01

    ... 14 Aeronautics and Space 3 2010-01-01 2010-01-01 false Icing conditions: Operating limitations... VFR/IFR Operating Limitations and Weather Requirements § 135.227 Icing conditions: Operating... to be free of frost, ice, or snow. (3) The certificate holder has an approved deicing/anti-icing...

  17. 14 CFR 91.527 - Operating in icing conditions.

    Science.gov (United States)

    2010-01-01

    ... 14 Aeronautics and Space 2 2010-01-01 2010-01-01 false Operating in icing conditions. 91.527...-Powered Multiengine Airplanes and Fractional Ownership Program Aircraft § 91.527 Operating in icing...— (1) Under IFR into known or forecast moderate icing conditions; or (2) Under VFR into known light or...

  18. 14 CFR 1.1 - General definitions.

    Science.gov (United States)

    2010-01-01

    ... gear of an aircraft and their accessories and controls. Airplane means an engine-driven fixed-wing... rules. IFR over-the-top, with respect to the operation of aircraft, means the operation of an aircraft over-the-top on an IFR flight plan when cleared by air traffic control to maintain “VFR conditions” or...

  19. 78 FR 37103 - Amendment of VOR Federal Airways V-55 and V-169 in Eastern North Dakota

    Science.gov (United States)

    2013-06-20

    ... contained within the associated MOA lateral boundary and altitudes from the affected airway. In 1980, the FAA circularized a proposal to change the boundary between the Devils Lake East and Devils Lake West... pilot awareness, respectively. Additionally, although pilots operating under visual flight rules (VFR...

  20. 78 FR 10564 - Proposed Modification of Class B Airspace; Minneapolis, MN

    Science.gov (United States)

    2013-02-14

    ... and Airspace Docket No. 09-AWA-1) and be submitted in triplicate to the Docket Management Facility... efficient management of aircraft operations in the Minneapolis, MN, terminal area. Class B airspace areas... traffic within Class B airspace with an expected minimal impact on non-participatory VFR operations. The...

  1. 14 CFR 121.689 - Flight release form: Supplemental operations.

    Science.gov (United States)

    2010-01-01

    ...) Company or organization name. (2) Make, model, and registration number of the aircraft being used. (3....g., IFR, VFR). (8) For each flight released as an ETOPS flight, the ETOPS diversion time for which the flight is released. (b) The aircraft flight release must contain, or have attached to it, weather...

  2. 76 FR 64038 - Airworthiness Directives; CPAC, Inc. Airplanes

    Science.gov (United States)

    2011-10-17

    ... does not include Models 112A and 115. The Model 112A is a Rockwell ``marketing name'' for the Model 112. The Model 115 is a Rockwell ``marketing name'' for the Model 114. Since they are type-certificated as... visual flight rules (VFR) only, restricted to crew, calm weather, reduced speed not to exceed 111 knots...

  3. 77 FR 30585 - Operating Limitations at New York LaGuardia Airport; Technical Amendment

    Science.gov (United States)

    2012-05-23

    ... January 1, 2007; to the primary marketing air carrier in the case of AIR-21 small hub/nonhub airport slot... marketing control up to 5 business days after the actual operation. This post-transfer approval is limited... requirement. Reservations for unscheduled flights operating under visual flight rules (VFR) are granted when...

  4. 14 CFR 61.195 - Flight instructor limitations and qualifications.

    Science.gov (United States)

    2010-01-01

    ... Instructors Other than Flight Instructors With a Sport Pilot Rating § 61.195 Flight instructor limitations and... instructor certificate with the applicable category and class rating; and (2) If appropriate, a type rating... instrument rating, a type rating not limited to VFR, or the instrument training required for commercial pilot...

  5. 77 FR 48476 - Proposed Amendment to Class B Airspace; Detroit, MI

    Science.gov (United States)

    2012-08-14

    ... July 20, 2010, at the Troy, MI, Holiday Inn; the second on July 21, 2010, at the Ypsilanti, MI, campus... Class B airspace area. Non- participating VFR aircraft would continue to have their choice of flying... at 8,000 feet MSL. Canadian regulations do not have an equivalent requirement to the FAA's Mode C...

  6. Efficacy of Low-Cost PC-Based Aviation Training Devices

    Directory of Open Access Journals (Sweden)

    Savern l Reweti

    2017-03-01

    Full Text Available Aim/Purpose: The aim of this study was to explore whether a full cost flight training device (FTD was significantly better for simulator training than a low cost PC-Based Aviation Training Device (PCATD. Background: A quasi-transfer study was undertaken to ascertain whether a Civil Aviation Authority certified Flight Training Device (FTD was more effective at improving pilot proficiency in the performance of a standard VFR traffic pattern (Overhead Rejoin Procedure than a customised low cost PCATD. Methodology: In this quasi-transfer study, a high fidelity FTD rather than an aircraft was used to test both training and transfer tasks. Ninety-three pilots were recruited to participate in the study. Contribution: The use of PCATDs is now well established for pilot training, especially for Instrument Flight Rules (IFR skills training. However, little substantive research has been undertaken to examine their efficacy for VFR training. Findings: There was no evidence of a pre-test/post-test difference in VFR task perfor-mance between participants trained on the PCATD and the FTD, when post tested on the FTD. The use of both PCATD and FTD demonstrated signifi-cant improvements in VFR task performance compared to a control group that received no PCATD or FTD training. Recommendations for Practitioners\t: We discuss the possibility that low cost PCATDs may be a viable alternative for flight schools wishing to use a flight simulator but not able to afford a FTD. Recommendation for Researchers: We discuss the introduction of improved low cost technologies that allow PCATDs to be used more effectively for training in VFR procedures. The development and testing of new technologies requires more research. Impact on Society: Flight training schools operate in a difficult economic environment with continued increases in the cost of aircraft maintenance, compliance costs, and aviation fuel. The increased utilisation of low cost PCATD’s especially for VFR

  7. Characteristics of nasal resonance and perceptual rating in prelingual hearing impaired adults.

    Science.gov (United States)

    Kim, Eun Yeon; Yoon, Mi Sun; Kim, Hyang Hee; Nam, Chung Mo; Park, Eun Sook; Hong, Sung Hwa

    2012-03-01

    Resonance problems in hearing impaired (HI) individuals have been described as aspects of nasality. However, there are limitations in being able to explain the range of resonance problems. Therefore, this study suggests a perceptual rating that will effectively explain the characteristics of resonance problems in HI individuals. Nasalance scores were obtained from 32 subjects in each of HI and normal hearing (NH) groups using a nasometer. The subjects were categorized into groups based on normal and abnormal nasalance ranges. The abnormal nasalance range group was further divided into hyper-, hypo-, and mixed-nasal groups. Nasalance scores were based the individuals performance in a series of passage and syllable tasks. The perceptual rating was evaluated using a newly introduced tool, 'vertical focus of resonance' (VFR), which focuses on the resonance energy in the frontal, throat, pharyngeal and nasal locations. The NH group demonstrated a significantly lower nasalance score in the oral coupling and passage tasks than the HI group. Based on the results of nasalance correlation analysis, the HI group showed highly significant correlations between syllable and passage tasks, as contrasted with the NH group. There were significant differences in VFR between the nasalance types in both the NH and the HI groups. The HI hyper-nasal group showed tendencies of velopharyngeal opening, as opposed to the HI hypo-nasal group which showed tendencies of velopharyngeal closure. The HI mixed-nasal group showed inappropriate coordination of velopharyngeal function. In the HI group, the results of VFR showed that the air flow and the resonance energy were not released from the cavity of resonance. The suggested VFR tool explains the focusing characteristics of resonance energy within a continuation of speech sound regardless of the phonetic environment. Therefore, VFR may be a useful tool in explaining the deviant resonance patterns of HI individuals.

  8. Decontamination Systems Information and Research Program

    Energy Technology Data Exchange (ETDEWEB)

    Cook, Echol E; Beatty, Tia Maria

    1998-07-01

    The following paragraphs comprise the research efforts during the second quarter of 1998 (April 1 - June 30.) These tasks have been granted a continuation until the end of August 1998. This report represents the last technical quarterly report deliverable for the WVU Cooperative Agreement - Decontamination Systems Information and Research Program. Final draft technical reports will be the next submission. During this period, work was completed on the Injection and Circulation of Potable Water Through PVDs on Task 1.6 - Pilot Scale Demonstration of TCE Flushing Through PVDs at the DOE/RMI Extrusion Plant. The data has been evaluated and representative graphs are presented. The plot of Cumulative Injected Volume vs. Cumulative Week Time show the ability to consistently inject through the two center PVDs at a rate of approximately ten (10) gallons per hour. This injection rate was achieved under a static head that varied from five (5) feet to three (3) feet. The plot of Extracted Flow Rate vs. Cumulative Week Time compares the extraction rate with and without the injection of water. The injection operation was continuous for eight hour periods while the extraction operation was executed over a pulsing schedule. Extraction rates as high as forty-five (45) gallons per hour were achieved in conjunction with injection (a 350% increase over no injection.) The retrieved TCE in the liquid phase varied to a considerable degree depending on the pulsing scheme, indicating a significant amount of stripping (volatilization) took place during the extraction process. A field experiment was conducted to confirm this. A liquid sample was obtained using the same vacuum system used in the pad operation and a second liquid sample was taken by a bailer. Analyzation of TCE concentration showed 99.5% volatilization when the vacuum system was used for extraction. This was also confirmed by data from the air monitoring program which indicated that 92%-99% of the retrieved TCE was being

  9. Decontamination Systems Information and Reseach Program

    Energy Technology Data Exchange (ETDEWEB)

    Cook, Echol E

    1998-04-01

    The following paragraphs comprise the research efforts during the first quarter of 1998 (January 1 - March 31). These tasks have been granted a continuation from the 1997 work and will all end in June 1998. This report represents the last technical quarterly report deliverable for the WVU Cooperative Agreement - Decontamination Systems Information and Research Program. Final reports for all of the 1997 projects will be submitted afterwards as one document. During this period, groundwater extraction operations were completed on Task 1.6 - Pilot Scale Demonstration of TCE Flushing Through PVDs at the DOE/RMI Extrusion Plant. The data have been evaluated and graphs are presented. The plot of TCE Concentration versus Time shows that the up-gradient groundwater monitoring well produced consistent levels of TCE contamination. A similar trend was observed for the down-gradient wells via grab samples tested. Groundwater samples from the PVD test pad Zone of Influence showed consistent reductions in TCE concentrations with respect to time. In addition, a natural pulse frequency is evident which will have a significant impact on the efficiency of the contaminant removal under natural groundwater advection/diffusion processes. The relationships between the PVD Extraction Flow Rate versus Cumulative Time shows a clear trend in flow rate. Consistent values between 20 to 30 g.p.m. at the beginning of the extraction duration, to less than 10 g.p.m. by the end of the extraction cycle are observed. As evidenced by the aquifer's diminishing recharge levels, the PVD extraction is affecting the response of the aquifer's natural attenuation capability. Progress was also marked on the Injection and Circulation of Potable Water Through PVDs task. Data reduction from this sequence of testing is ongoing. Work planned for next quarter includes completing the Injection / Extraction of potable water task and beginning the Surfactant Injection and removal task.

  10. Analysis of promoter elements involved in the transcriptional initiation of RpoS-dependent Borrelia burgdorferi genes.

    Science.gov (United States)

    Eggers, Christian H; Caimano, Melissa J; Radolf, Justin D

    2004-11-01

    Borrelia burgdorferi, the causative agent of Lyme disease, encodes an RpoS ortholog (RpoS(Bb)) that controls the temperature-inducible differential expression of at least some of the spirochete's lipoprotein genes, including ospC and dbpBA. To begin to dissect the determinants of RpoS(Bb) recognition of, and selectivity for, its dependent promoters, we linked a green fluorescent protein reporter to the promoter regions of several B. burgdorferi genes with well-characterized expression patterns. Consistent with the expression patterns of the native genes/proteins in B. burgdorferi strain 297, we found that expression of the ospC, dbpBA, and ospF reporters in the spirochete was RpoS(Bb) dependent, while the ospE and flaB reporters were RpoS(Bb) independent. To compare promoter recognition by RpoS(Bb) with that of the prototype RpoS (RpoS(Ec)), we also introduced our panel of constructs into Escherichia coli. In this surrogate, maximal expression from the ospC, dbpBA, and ospF promoters clearly required RpoS, although in the absence of RpoS(Ec) the ospF promoter was weakly recognized by another E. coli sigma factor. Furthermore, RpoS(Bb) under the control of an inducible promoter was able to complement an E. coli rpoS mutant, although RpoS(Ec) and RpoS(Bb) each initiated greater activity from their own dependent promoters than they did from those of the heterologous sigma factor. Genetic analysis of the ospC promoter demonstrated that (i) the T(-14) in the presumptive -10 region plays an important role in sigma factor recognition in both organisms but is not as critical for transcriptional initiation by RpoS(Bb) as it is for RpoS(Ec); (ii) the nucleotide at the -15 position determines RpoS or sigma(70) selectivity in E. coli but does not serve the same function in B. burgdorferi; and (iii) the 110-bp region upstream of the core promoter is not required for RpoS(Ec)- or RpoS(Bb)-dependent activity in E. coli but is required for maximal expression from this promoter in

  11. Subalkaline andesite from Valu Fa Ridge, a back-arc spreading center in southern Lau Basin: petrogenesis, comparative chemistry, and tectonic implications

    Science.gov (United States)

    Vallier, T.L.; Jenner, G.A.; Frey, F.A.; Gill, J.B.; Davis, A.S.; Volpe, A.M.; Hawkins, J.W.; Morris, J.D.; Cawood, Peter A.; Morton, J.L.; Scholl, D. W.; Rautenschlein, M.; White, W.M.; Williams, Ross W.; Stevenson, A.J.; White, L.D.

    1991-01-01

    Tholeiitic andesite was dredged from two sites on Valu Fa Ridge (VFR), a back-arc spreading center in Lau Basin. Valu Fa Ridge, at least 200 km long, is located 40-50 km west of the active Tofua Volcanic Arc (TVA) axis and lies about 150 km above the subducted oceanic plate. One or more magma chambers, traced discontinuously for about 100 km along the ridge axis, lie 3-4 km beneath the ridge. The mostly aphyric and glassy lavas had high volatile contents, as shown by the abundance and large sizes of vesicles. An extensive fractionation history is inferred from the high SiO2 contents and FeO* MgO ratios. Chemical data show that the VFR lavas have both volcanic arc and back-arc basin affinities. The volcanic arc characteristics are: (1) relatively high abundances of most alkali and alkaline earth elements; (2) low abundances of high field strength elements Nb and Ta; (3) high U/Th ratios; (4) similar radiogenic isotope ratios in VFR and TVA lavas, in particular the enrichment of 87Sr 86Sr relative to 206Pb 204Pb; (5) high 238U 230Th, 230Th 232Th, and 226Ra 230Th activity ratios; and (6) high ratios of Rb/Cs, Ba/Nb, and Ba/La. Other chemical characteristics suggest that the VFR lavas are related to MORB-type back-arc basin lavas. For example, VFR lavas have (1) lower 87Sr 86Sr ratios and higher 143Nd 144Nd ratios than most lavas from the TVA, except samples from Ata Island, and are similar to many Lau Basin lavas; (2) lower Sr/REE, Rb/Zr, and Ba/Zr ratios than in arc lavas; and (3) higher Ti, Fe, and V, and higher Ti/V ratios than arc lavas generally and TVA lavas specifically. Most characteristics of VFR lavas can be explained by mixing depleted mantle with either small amounts of sediment and fluids from the subducting slab and/or an older fragment of volcanic arc lithosphere. The eruption of subalkaline andesite with some arc affinities along a back-arc spreading ridge is not unique. Collision of the Louisville and Tonga ridges probably activated back-arc extension

  12. [Studies on modulation and control sigma 38 and RMF for the expression of some genes].

    Science.gov (United States)

    Ding, Q; Akira, I

    1996-10-01

    The E. coli mutants and wild type strains of rpoS and rmf were cultured in rich medium LB and limited component medium EP respectively. During the stationary phase, the viable cells of mutants were less than wild type strains's. The change of the product of serial proteins was quantified with Western blot. Sigma 38 has not effects on the product of rpoA, rpoB, rpoC, groE and tu gene, depress the transcription of crp and promote the expression of rmf. RMF can promote expression of rpoA, rpoD, groEl, rho, ompA and tufA gene in rich medium, but not in limited medium, and then depress and promote the expression of crp and rpoS respectively.

  13. Toward a better understanding of resonant annihilation on molecules

    Science.gov (United States)

    Surko, C. M.; Danielson, J. R.; Jones, A. C. L.

    2011-05-01

    For many if not most molecules, annihilation at positron energies ɛ in the range of the vibrational modes proceeds via vibrational Feshbach resonances (VFR) in which positrons attach to these targets., In small molecules, the theory of Gribakin and Lee provides a quantitative description of the annihilation rates, Zeff. However other effects are currently less well understood. Described here are some open questions and experiments using deuterium substitution that are designed to address them. They include the effect of intramolecular vibrational energy redistribution (IVR) on Zeff in large molecules (e.g., alkanes), the role of molecular rotations on VFR in very small molecules (e.g., ammonia), and the observation of combination and overtone modes in molecules of small to intermediate size (e.g., acetylene and ethylene). Work supported by NSF grant PHY 07-55809.

  14. Characterising stationary and translating vortex flow using magnetic resonance

    Science.gov (United States)

    Vallatos, Antoine; Wilson, Mark C. T.; Taylor, Annette F.; Britton, Melanie M.

    2012-09-01

    We report magnetic resonance (MR) velocity and diffusion maps in three directions for stationary vortices (Taylor vortex flow —TVF) and velocity maps for translating vortices (vortex flow reactor —VFR) produced in a Couette cell. Motion artefacts in the translating vortex flow are removed by synchronising data acquisition with the translation period of the vortices. MR propagator experiments, which measure the conditional probability density for displacement, were performed to characterise molecular displacements in these systems. Simulations were performed using the experimental velocity and diffusion maps to aid interpretation of experimentally measured propagators and enable characterisation of the macro-mixing and transport properties within TVF and VFR systems. These simulations enabled molecular transport and mixing to be assessed over longer-time scales than are accessible experimentally, allowing plug flow, by-pass flow and inter-vortex mixing to be quantified.

  15. Use of satellite navigation for General Aviation (GA) and Aerial Work (AW)

    Energy Technology Data Exchange (ETDEWEB)

    Pade, D. [AOPA-Germany, Egelsbach (Germany)

    1999-07-01

    The European airspace strategy strongly relies on the new navigation strategy, which predominantly considers airline type of operation and equipment. Since this type of traffic does only limit the cases operated under visual flight rules, the airspace is shaped and organised without regard to VFR. As I said before, the bulk of our operations are conducted under visual flight rules (VFR), in other words, we are left out, if not other cost beneficial systems are offered for General Aviation, to keep the same safety level for all airspace user, while traffic will double till 2015. What worries us are the airline driven CNS/ATM strategies under development taking into consideration only the needs of the big boys. Their solutions at the moment are unaffordable for use and if nothing changes strategically, will close a large portion of the airspace and will deny access to large airports. (orig.)

  16. Cases of typhoid fever in Copenhagen region

    DEFF Research Database (Denmark)

    Barrett, Freja Cecille; Knudsen, Jenny Dahl; Johansen, Isik Somuncu

    2013-01-01

    Typhoid fever is a systemic illness which in high-income countries mainly affects travellers. The incidence is particularly high on the Indian subcontinent. Travellers who visit friends and relatives (VFR) have been shown to have a different risk profile than others. We wished to identify main ch...... characteristics for travellers infected with S. Typhi considering both clinical and laboratory findings in order to provide for faster and better diagnostics in the future. The outcome of treatment, especially concerning relapse, was evaluated as well.......Typhoid fever is a systemic illness which in high-income countries mainly affects travellers. The incidence is particularly high on the Indian subcontinent. Travellers who visit friends and relatives (VFR) have been shown to have a different risk profile than others. We wished to identify main...

  17. Stationary-phase genes upregulated by polyamines are responsible for the formation of Escherichia coli persister cells tolerant to netilmicin.

    Science.gov (United States)

    Tkachenko, Alexander G; Kashevarova, Natalya M; Tyuleneva, Elena A; Shumkov, Mikhail S

    2017-05-01

    Persisters are rare phenotypic variants of regular bacterial cells that survive lethal antibiotics or stresses owing to slowing down of their metabolism. Recently, we have shown that polyamine putrescine can upregulate persister cell formation in Escherichia coli via the stimulation of rpoS expression, encoding a master regulator of general stress response. We hypothesized that rmf and yqjD, the stationary-phase genes responsible for ribosome inactivation, might be good candidates for the similar role owing to their involvement in translational arrest and the ability to be affected by polyamines. Using reporter gene fusions or single and multiple knockout mutations in rpoS, rmf and yqjD genes, we show in this work that (i) E. coli polyamines spermidine and cadaverine can upregulate persistence, like putrescine; (ii) polyamine effects on persister cell formation are mediated through stimulation of expression of rpoS, rmf and yqjD genes; (iii) these genes are involved in persister cell formation sequentially in a dynamic fashion as cells enter the stationary phase. The data obtained in this work can be used to develop novel tools relying on a suppression of polyamine metabolism in bacteria to combat persister cells as an important cause of infections refractory to antibiotics. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  18. Uncorrelated Encounter Model of the National Airspace System, Version 2.0

    Science.gov (United States)

    2013-08-19

    between two IFR aircraft in oceanic airspace. The reason for this is that one cannot observe encounters of sufficient fidelity in the available data...does not observe a sufficient number of encounters between instrument flight rules ( IFR ) and non- IFR traffic beyond 12 NM from the shore. 4 TABLE 1...Encounter model categories. Aircraft of Interest Intruder Aircraft Location Flight Rule IFR VFR Noncooperative Noncooperative Conventional

  19. Study of the Light Utility Helicopter (LUH) Acquisition Program as a Model for Defense Acquisition of Nondevelopmental Items

    Science.gov (United States)

    2014-12-01

    Headquarters, Department of the Army IFR instrument flight rules IOTE initial operational test and evaluation IR infrared KO contracting officer kt...instrument flight rules ( IFR ) and visual flight rules (VFR) capabilities, thereby allowing flight at night and under low visibility weather...entire United States, as well as nautical charts for all coastal areas, and IFR and aeronautical charts. It can house topographical maps for the

  20. Value of Forecaster in the Loop

    Science.gov (United States)

    2014-09-01

    forecast system IFR instrument flight rules IMC instrument meteorological conditions LAMP Localized Aviation Model Output Statistics Program METOC...but less than 3000 ft and/or visibility is greater than or equal to 3 miles but less than 5 miles. The terms Instrument Flight Rules ( IFR ) and Visual...dictated by the meteorological conditions. IMC puts IFR into effect while VMC and MVMC put VFR into effect. Outside of lightening, IMC conditions

  1. Leveraging DMO’s Hi-Tech Simulation Against the F-16 Flying Training Gap

    Science.gov (United States)

    2005-04-01

    technologies such as JHMCS , LINK 16, HTS, TGP, AAI, and all the associated "smart precision" weapons into one platform. This exponential leap in capability on...implementation of the Common Configuration Implementation Program (CCIP). Taking a basic day-VFR designed fighter, CCIP now merges technologies such as JHMCS ...F-16 as a machine the ability to; elevate its established air-to-air prowess through the addition of the JHMCS and an AAI, enhance SEAD mission

  2. Defense Small Business Innovation Research Program (SBIR). Volume 3. Air Force Abstracts of Phase 1 Awards 1991

    Science.gov (United States)

    1991-01-01

    COMMUNICATIONS. COMMERCIAL BENEFITS INCLUDE PREDICTING VISUAL FLIGHT RULE (VFR) CONDITIONS AND SOLAR INSOLATION FOR ALTERNATIVE ENERGY SITES. SHORT-TERM...PARAMETRIC DATA FOR SCALE-UP ACTIVITIES. M. L. ENERGIA , INC. TopicS: 91-057 l .91CES-224 P.O. BOX 1468, Office: AFCESA PRINCETON, NJ 08542 Contract 0...pbolochromic dyes previously developed for thermal protection Systems to achieve passive solar control charactersics in aircraft cockpit transparncics. Tbe

  3. Elements of quantum probability

    OpenAIRE

    Kummerer, B.; Maassen, Hans

    1996-01-01

    This is an introductory article presenting some basic ideas of quantum probability. From a discussion of simple experiments with polarized light and a card game we deduce the necessity of extending the body of classical probability theory. For a class of systems, containing classical systems with finitely many states, a probabilistic model is developed. It can describe, in particular, the polarization experiments. Some examples of ‘quantum coin tosses’ are discussed, closely related to V.F.R....

  4. The competing effects of sulfide saturation versus degassing on the behavior of the chalcophile elements during the differentiation of hydrous melts

    Science.gov (United States)

    Jenner, Frances E.; Hauri, Erik H.; Bullock, Emma S.; König, Stephan; Arculus, Richard J.; Mavrogenes, John A.; Mikkelson, Nicole; Goddard, Charlotte

    2015-05-01

    There is a lack of consensus regarding the roles of sulfide saturation versus volatile degassing on the partitioning of Cu and Ag during differentiation and eruption of convergent margin magmas. Because of their oxidized character, volatile-rich magmas from the Eastern Manus Back-arc Basin (EMBB) only reach sulfide saturation following magnetite-driven reduction of the melt: the so-called "magnetite crisis." If sulfide saturation typically precedes volatile saturation, the magnetite crisis will limit the proportion of Cu and Ag that can partition from the melt into an exsolving volatile-rich phase, which may contribute to the sporadic occurrence of magmatic-hydrothermal ore deposits at convergent margins. However, it is unclear whether the magnetite crisis is a common or rare event during differentiation of volatile-rich magmas. We report major and trace element data for submarine volcanic glasses from the Tonga arc-proximal Valu Fa Ridge (VFR; SW Pacific). Cu-Se-Ag systematics of samples erupting at the southern VFR suggest magnetite fractionation-triggered sulfide saturation. The similarity in chalcophile element systematics of the southern VFR and EMBB samples is unlikely to be coincidental, and may indicate that the magnetite crisis is a common event during differentiation of hydrous melts. However, unlike many convergent margin magmas, it is unlikely that the evolving VFR and EMBB were saturated in a S-bearing volatile phase prior to magnetite fractionation. Hence, the metal-depleting magnetite crisis may be restricted to back-arc basin magmas that do not degas volatiles prior to magnetite fractionation and potentially convergent margin magmas fractionating at high pressures in the continental crust.

  5. U.S. Army Unmanned Aircraft Systems Roadmap 2010-2035: Eyes of the Army

    Science.gov (United States)

    2010-04-09

    synchronizing this transformation. In 1915, Nicola Tesla introduced the concept of unmanned flight in his dissertation that described an armed, pilotless...Background As stated earlier, the genesis of unmanned flight began in 1915 when Tesla believed an armed, pilotless-aircraft could be used to defend...to 2500 feet AGL) that have an operating control tower . Flights in Class D airspace must establish and maintain communications with ATC, but VFR

  6. Oceanic Area System Improvement Study (OASIS). Volume V. North Atlantic, Central East Pacific, and Caribbean Regions Communication Systems Description.

    Science.gov (United States)

    1981-09-01

    system SENEAM Servicies a la Navagacion en el Espacio Aero Mexicano SSB Single side band USB Upper side band VFR Visual flight rules VHF Very-high...items 2 and 7 in Table 18) may be somewhat arbitrary because lines are often purchased from government institutions that may choose to charge more or...Transport Association, International User Charges (September 1978). 19. Servicies a la Navegacion en el Espacio Aero Mexicano (SENZAM), "Informal note

  7. An increase in visceral fat is associated with a decrease in the taste and olfactory capacity.

    Directory of Open Access Journals (Sweden)

    Jose Carlos Fernandez-Garcia

    Full Text Available Sensory factors may play an important role in the determination of appetite and food choices. Also, some adipokines may alter or predict the perception and pleasantness of specific odors. We aimed to analyze differences in smell-taste capacity between females with different weights and relate them with fat and fat-free mass, visceral fat, and several adipokines.179 females with different weights (from low weight to morbid obesity were studied. We analyzed the relation between fat, fat-free mass, visceral fat (indirectly estimated by bioelectrical impedance analysis with visceral fat rating (VFR, leptin, adiponectin and visfatin. The smell and taste assessments were performed through the "Sniffin' Sticks" and "Taste Strips" respectively.We found a lower score in the measurement of smell (TDI-score (Threshold, Discrimination and Identification in obese subjects. All the olfactory functions measured, such as threshold, discrimination, identification and the TDI-score, correlated negatively with age, body mass index (BMI, leptin, fat mass, fat-free mass and VFR. In a multiple linear regression model, VFR mainly predicted the TDI-score. With regard to the taste function measurements, the normal weight subjects showed a higher score of taste functions. However a tendency to decrease was observed in the groups with greater or lesser BMI. In a multiple linear regression model VFR and age mainly predicted the total taste scores.We show for the first time that a reverse relationship exists between visceral fat and sensory signals, such as smell and taste, across a population with different body weight conditions.

  8. Mode coupling and multiquantum vibrational excitations in Feshbach-resonant positron annihilation in molecules

    Science.gov (United States)

    Gribakin, G. F.; Stanton, J. F.; Danielson, J. R.; Natisin, M. R.; Surko, C. M.

    2017-12-01

    The dominant mechanism of low-energy positron annihilation in polyatomic molecules is through positron capture in vibrational Feshbach resonances (VFR). In this paper, we investigate theoretically the effect of anharmonic terms in the vibrational Hamiltonian on positron annihilation rates. Such interactions enable positron capture in VFRs associated with multiquantum vibrational excitations, leading to enhanced annihilation. Mode coupling can also lead to faster depopulation of VFRs, thereby reducing their contribution to the annihilation rates. To analyze this complex picture, we use coupled-cluster methods to calculate the anharmonic vibrational spectra and dipole transition amplitudes for chloroform, chloroform-d1, 1,1-dichloroethylene, and methanol, and use these data to compute positron resonant annihilation rates for these molecules. Theoretical predictions are compared with the annihilation rates measured as a function of incident positron energy. The results demonstrate the importance of mode coupling in both enhancement and suppression of the VFR. There is also experimental evidence for the direct excitation of multimode VFR. Their contribution is analyzed using a statistical approach, with an outlook towards more accurate treatment of this phenomenon.

  9. Health challenges of young travelers visiting friends and relatives compared with those traveling for other purposes.

    Science.gov (United States)

    Han, Pauline; Yanni, Emad; Jentes, Emily S; Hamer, Davidson H; Chen, Lin H; Wilson, Mary E; Macleod, William B; Ooi, Winnie W; Kogelman, Laura; Karchmer, Adolf W; Barnett, Elizabeth D

    2012-09-01

    The study objective was to assess differences in demographics and travel health challenges between youths ≤18 years old traveling internationally to visit friends and relatives (VFRs) compared with those traveling for other purposes (non-VFR). The Boston Area Travel Medicine Network consists of 5 clinics collecting anonymous data from international pretravel consultations. Data on all travelers ≤18 years of age seen between January 2008 and July 2010 were used. VFRs were compared with non-VFRs on demographics, primary language, trip characteristics, travel vaccinations administered, malaria prophylaxis and antidiarrheal medications prescribed. Thirty-five percent (610/1731) listed VFR as their purpose of travel. Almost half of VFRs were travel to countries that were yellow fever holoendemic, had malaria risk and were high-risk for typhoid (44% versus 20%, 39% versus 12%, 25% versus 15%, P travel-related morbidity, healthcare providers should be prepared to give travel advice to parents of VFR infants and children, particularly those US-born VFRs with foreign-born parents, regarding antimalarial and antidiarrheal medications and preventing yellow fever, malaria and typhoid.

  10. Associations of Maternal Dietary Patterns during Pregnancy with Offspring Adiposity from Birth Until 54 Months of Age

    Directory of Open Access Journals (Sweden)

    Ling-Wei Chen

    2016-12-01

    Full Text Available Most studies linking maternal diet with offspring adiposity have focused on single nutrients or foods, but a dietary pattern approach is more representative of the overall diet. We thus aimed to investigate the relations between maternal dietary patterns and offspring adiposity in a multi-ethnic Asian mother–offspring cohort in Singapore. We derived maternal dietary patterns using maternal dietary intake information at 26–28 weeks of gestation, of which associations with offspring body mass index (BMI, abdominal circumference (AC, subscapular skinfold (SS, and triceps skinfold (TS were assessed using longitudinal data analysis (linear mixed effects (LME and multiple linear regression at ages 0, 3, 6, 9, 12, 15, 18, 24, 36, 48, and 54 months. Three dietary patterns were derived: (1 vegetables-fruit-and-white rice (VFR; (2 seafood-and-noodles (SfN; and (3 pasta-cheese-and-bread (PCB. In the LME model adjusting for potential confounders, each standard deviation (SD increase in maternal VFR pattern score was associated with 0.09 mm lower offspring TS. Individual time-point analysis additionally revealed that higher VFR score was generally associated with lower postnatal offspring BMI z-score, TS, SS, and sum of skinfolds (SS + TS at ages 18 months and older. Maternal adherence to a dietary pattern characterized by higher intakes of fruit and vegetables and lower intakes of fast food was associated with lower offspring adiposity.

  11. [Prior health advice to immigrants who travel to visit family and friends].

    Science.gov (United States)

    Roca, Carme; Aguilar, Anna; Valerio, Lluís; Solsona, Lluís; Carrasco, Clara; Gimeno-Feliu, Luis Andrés

    2014-04-01

    Immigrants who make or plan journeys to visit their families in their countries of origin (immigrants -visiting friends and relatives, I-VFR) have a higher risk of acquiring travel-associated diseases than other travellers. The main aim of this study is to analyse the knowledge of the immigrant population on the need to receive health advice (HA) before making international journeys in general and in particular before travelling to their country of origin. Observational, multicentre study. Ten Family Doctors from 10 Health Centres in Catalonia and Aragon participated A total of 555 immigrants ≥ 15 years of age, who consulted their Family Doctor and agreed to answer a questionnaire. Opportunity sampling was used. A total of 389 (70.1%) of those surveyed considered it necessary to receive HA before making an international journey, 406 (73.2%) were I-VFR and 145 (35.7%) had requested HA prior to the journey, mostly from their Family Doctor (n=60; 41.1%). Almost two-thirds (261, 65.2%) of the subjects did not seek HA, with the most common reason being that they did not consider it necessary (173, 42.6%). I-VFR do not usually request HA prior to travelling, basic due to considering it unnecessary. When they do request it, they are very often initially directed to their Family Doctor. Copyright © 2013 Elsevier España, S.L. All rights reserved.

  12. Composite Blastoid Variant of Mantle Cell Lymphoma and Classical Hodgkin Lymphoma.

    Science.gov (United States)

    Murray, Ciara; Quinn, Fiona; Illyes, Gyorgy; Walker, Jan; Castriciano, Giussepa; O'Sullivan, Paul; Grant, Cliona; Vandenberghe, Elisabeth; Bird, Brian; Flavin, Richard

    2017-05-01

    Composite lymphoma (CL) describes the rare occurrence of 2 or more distinct types of lymphoma in a single anatomical location. We present the case of a 78-year-old man presenting with a 3-month history of weakness, malaise, and increasing dyspnea. A lymph node excised from the posterior triangle of the neck revealed the coexistence of 2 morphologically and phenotypically distinct lymphoid neoplasms consistent with a blastoid variant of mantle cell lymphoma (MCL) occurring in composite with classical Hodgkin lymphoma (cHL), mixed cellularity subtype. A t(11;14)(q13;q32) translocation was demonstrated by fluorescence in situ hybridization in the MCL and Hodgkin Reed-Sternberg cells of the cHL. Multiplex polymerase chain reaction detected clonal Immunoglobulin heavy chain (VFR1-J, VFR2-J, and VFR3-J), clonal immunoglobulin light chain kappa (V-J and V/JC intron-kde) and clonal immunoglobulin light chain lambda (V-J) gene rearrangements in the MCL. This report represents the first case of a blastoid variant of MCL occurring in composite with cHL.

  13. Wear resistance of four types of vacuum-formed retainer materials: a laboratory study.

    Science.gov (United States)

    Raja, Taiyub A; Littlewood, Simon J; Munyombwe, Theresa; Bubb, Nigel L

    2014-07-01

    To investigate the resistance to wear of four different vacuum-formed retainer (VFR) materials: Essix C+, Essix ACE, Duran, and Tru-Tain. Essix C+ is a polypropylene polymer; the other materials are polyethylene co-polymers. The study was undertaken at the Leeds Dental Institute, Leeds, UK, with 26 samples in each group. The specimens were vacuum-formed according to the manufacturers' guidelines, and a custom-made wear-simulation machine was used to conduct the test. Each specimen was subjected to 1000 cycles of the wear simulation, with steatite balls as the antagonist material. The resistance to wear of the VFR materials was evaluated by measuring the maximum wear depth using noncontact, three-dimensional surface profilometry. The wear depth was given in micrometers. The median wear depth was 63.20 µm for the Essix C+ group, 7.88 µm for the Essix ACE group, 9.75 µm for the Duran group, and 12.08 µm for the Tru-Tain group. The Kruskal-Wallis test to compare the four VFR materials detected a statistically significant difference between the groups (P materials-Essix ACE, Duran, and Tru-Tain-exhibited significantly less wear than the polypropylene material, Essix C+.

  14. Performance of clear vacuum-formed thermoplastic retainers depending on retention protocol: a systematic review.

    Science.gov (United States)

    Kaklamanos, Eleftherios G; Kourakou, Maria; Kloukos, Dimitrios; Doulis, Ioannis; Kavvadia, Smaragda

    2017-04-01

    We aimed at comparing the performance of vacuum-formed thermoplastic retainers (VFR) worn either full-time or part-time, in maintaining orthodontic treatment results in terms of tooth alignment, arch form and occlusion. We reviewed randomized and prospective controlled clinical trials comparing VFR wearing protocols and searched databases, without restrictions, for published and unpublished literature. The risk of bias was assessed using the Cochrane Risk of Bias tool and the overall level of certainty in the evidence following ADA methodology. 184 studies were initially identified and reduced to the 3 randomized controlled trials included in the systematic review by means of specific criteria. One study followed patients 1 year into retention, and the other two for 6 months. Little's Irregularity Index, intermolar and intercanine width, arch length, overjet and PAR score did not differ significantly between the patients wearing their retainers part time or full time. We observed a slight increase in the overbite in the part-time group in only one trial. With a moderate level of certainty, we found that during the observation period, full-time VFR wear is not superior to part-time, bearing in mind the potential implications for health burden, retainer longevity and cost-effectiveness, as well as patient satisfaction and compliance.

  15. [Evaluation of pulmonary venous flow pattern in hypertrophied and dilated hearts: a study with transesophageal pulsed Doppler echocardiography].

    Science.gov (United States)

    Iuchi, A; Oki, T; Ogawa, S; Kawano, T; Hayashi, M; Aoyama, Y; Emi, S; Hosoi, K; Fukuda, N; Mori, H

    1991-01-01

    In order to evaluate the clinical significance of pulmonary venous flow (PVF) pattern, transesophageal pulsed Doppler echocardiography (TEE) was performed in 25 patients with hypertrophied heart (all with hypertrophic cardiomyopathy), 15 patients with dilated heart (10 with old myocardial infarction and 5 with dilated cardiomyopathy) and 10 normal controls. Parameters obtained from the PVF pattern were compared with those of transmitral flow (MVF) pattern, % fractional shortening (%FS) of left ventricle (LV) and amplitude of mitral anular motion (MAM) during a cardiac cycle. Results were as follows: 1. PVF pattern in cases of sinus rhythm was divided into four components, atrial systolic backward flow (PVA), ventricular systolic (PVS1, PVS2) and diastolic (PVD) forward flows. 2. In patients with dilated heart, peak velocities of PVS1 and PVS2 were markedly decreased compared with those of hypertrophied and normal hearts. 3. Peak velocity of PVD in hypertrophied and dilated hearts was significantly decreased compared with that of normal controls, and PV-D/S (ratio of peak velocity of PVD to PVS2) was significantly lower in hypertrophied heart than in normal controls. 4. Time interval from the first heart sound to the peak of PVS2 (TS) was significantly longer in dilated heart, and time interval from the second heart sound to the peak of PVD (TD) was longer in hypertrophied heart than in the other two groups. 5. MAM and %FS of dilated heart were significantly lower than those in normal and hypertrophied hearts, and peak velocity of PVS2 in dilated heart group correlated well with MAM or %FS. 6. There were significant correlations among the diastolic parameters from PVD of PVF (peak velocity of PVD, PV-D/S) and early diastolic wave (D) of MVF (peak velocity, deceleration time and deceleration of rapid filling). 7. In a case of hypertrophic cardiomyopathy with mid-diastolic wave of MVF, distinct forward wave was observed after PVD of PVF, and this wave coincided in

  16. Impact of physical deconditioning on ventricular tachyarrhythmias in trained athletes.

    Science.gov (United States)

    Biffi, Alessandro; Maron, Barry J; Verdile, Luisa; Fernando, Fredrick; Spataro, Antonio; Marcello, Giuseppe; Ciardo, Roberto; Ammirati, Fabrizio; Colivicchi, Furio; Pelliccia, Antonio

    2004-09-01

    The purpose of this research was to evaluate the impact of athletic training and, in particular, physical deconditioning, on frequent and/or complex ventricular tachyarrhythmias assessed by 24-h ambulatory (Holter) electrocardiogram (ECG). Sudden deaths in athletes are usually mediated by ventricular tachyarrhythmias. Twenty-four hour ambulatory ECGs were recorded at peak training and after a deconditioning period of 19 +/- 6 weeks (range, 12 to 24 weeks) in a population of 70 trained athletes selected on the basis of frequent and/or complex ventricular tachyarrhythmias (i.e., > or =2,000 premature ventricular depolarization [PVD] and/or > or =1 burst of non-sustained ventricular tachycardia [NSVT]/24 h). A significant decrease in the frequency and complexity of ventricular arrhythmias was evident after deconditioning: PVDs/24 h: 10,611 +/- 10,078 to 2,165 +/- 4,877 (80% reduction; p deconditioning. In athletes with heart disease, the resolution of such arrhythmias with detraining may represent a mechanism by which risk for sudden death is reduced. Conversely, in athletes without cardiovascular abnormalities, reduction in frequency of ventricular tachyarrhythmias and the absence of cardiac events in the follow-up support the benign clinical nature of these rhythm disturbances as another expression of athlete's heart.

  17. Regional vascular density-visual field sensitivity relationship in glaucoma according to disease severity.

    Science.gov (United States)

    Shin, Joong Won; Lee, Jiyun; Kwon, Junki; Choi, Jaewan; Kook, Michael S

    2017-12-01

    To study whether there are global and regional relationships between peripapillary vascular density (pVD) assessed by optical coherence tomography angiography (OCT-A) and visual field (VF) mean sensitivity at different glaucoma stages. Microvascular images and peripapillary retinal nerve fibre layer (pRNFL) thicknesses were obtained using a Cirrus OCT-A device in 91 glaucoma subjects. The pVD was measured at various spatial locations according to the Garway-Heath map, using a MATLAB software (The MathWorks, Natick, Massachusetts). VF mean sensitivity (VFMS) was recorded in the 1/L scale. Global and regional vasculature-function (pVD vs VFMS) relationships were assessed in separate patient groups at mild and moderate-to-advanced stages of glaucoma. The pVDs at superotemporal and inferotemporal regions were significantly associated with corresponding VFMS in mild glaucoma (pglaucoma, there were significant associations between pVD and VFMS, regardless of location. The association between global pVD and VFMS was significantly stronger than that between global pRNFL thickness and VFMS in moderate-to-advanced stage glaucoma (p glaucoma. OCT-A may be useful in monitoring glaucoma at various stages. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2017. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

  18. Carbon and clay nanoparticles induce minimal stress responses in gram negative bacteria and eukaryotic fish cells.

    Science.gov (United States)

    Taylor, Alicia A; Aron, Gary M; Beall, Gary W; Dharmasiri, Nihal; Zhang, Yixin; McLean, Robert J C

    2014-08-01

    We investigated in vitro the potential mutagenic and toxic effects of two clay-based nanoparticles, Cloisite® Na(+) (Cloisite) and halloysite; and multi-walled carbon nanotubes (MWCNT), commonly used in the polymer composite industry. Using the Ames test, the three nanoparticles did not have a true mutagenic effect, although growth of Salmonella enterica var. Typhimurium (S.typhimurium) was diminished at higher nanoparticle concentrations. We investigated the impact of nanoparticles on Escherichia coli and S. typhimurium including oxyR and rpoS mutants, which are susceptible to oxidative stress. The oxyR mutants were inhibited in the presence of nanoparticles, when grown aerobically with light. Toxicity was not observed in the absence of light or during anaerobic growth. E. coli rpoS mutants exhibited some toxicity when cultured with Cloisite and MWCNT only when grown aerobically with light. There was no effect with other nanoparticles, or with S. typhimurium rpoS mutants. MWCNT exhibited a slight toxic effect against Epithelioma papulosum cyprini (EPC) cells only at the highest concentration tested. There was no discernable toxicity to EPC cells caused by the clay nanoparticles. We conclude that clay-based nanoparticles and MWCNT do not exert a mutagenic effect and do not have a general toxic effect across all bacterial species or between prokaryotic and eukaryotic cells. Modest toxicity was only observed in eukaryotic EPC cells against MWCNT at the highest concentration tested. Limited species-specific toxicity to clay based and MWCNT nanoparticles was seen in bacterial strains primarily due to culture conditions and mutations that exacerbate oxidative stress. Copyright © 2012 Wiley Periodicals, Inc., a Wiley company.

  19. Regulation of RNA polymerase sigma subunit synthesis in Escherichia coli: intracellular levels of four species of sigma subunit under various growth conditions.

    OpenAIRE

    Jishage, M; Iwata, A.; Ueda, S.; Ishihama, A

    1996-01-01

    By a quantitative Western immunoblot analysis, the intracellular levels of two principal sigma subunits, sigma 70 (sigma D, the rpoD gene product) and sigma 38 (sigma S, the rpoS gene product), and of two minor sigma subunits, sigma 54 (sigma N, the rpoN gene product) and sigma 28 (sigma F, the rpoF gene product), were determined in two Escherichia coli strains, W3110 and MC4100. The results indicated that the levels of sigma 54 and sigma 28 are maintained at 10 and 50%, respectively, of the ...

  20. Acetyl-Phosphate Is Not a Global Regulatory Bridge between Virulence and Central Metabolism in Borrelia burgdorferi.

    Directory of Open Access Journals (Sweden)

    Crystal L Richards

    Full Text Available In B. burgdorferi, the Rrp2-RpoN-RpoS signaling cascade is a distinctive system that coordinates the expression of virulence factors required for successful transition between its arthropod vector and mammalian hosts. Rrp2 (BB0763, an RpoN specific response regulator, is essential to activate this regulatory pathway. Previous investigations have attempted to identify the phosphate donor of Rrp2, including the cognate histidine kinase, Hk2 (BB0764, non-cognate histidine kinases such as Hk1, CheA1, and CheA2, and small molecular weight P-donors such as carbamoyl-phosphate and acetyl-phosphate (AcP. In a report by Xu et al., exogenous sodium acetate led to increased expression of RpoS and OspC and it was hypothesized this effect was due to increased levels of AcP via the enzyme AckA (BB0622. Genome analyses identified only one pathway that could generate AcP in B. burgdorferi: the acetate/mevalonate pathway that synthesizes the lipid, undecaprenyl phosphate (C55-P, lipid I, which is essential for cell wall biogenesis. To assess the role of AcP in Rrp2-dependent regulation of RpoS and OspC, we used a unique selection strategy to generate mutants that lacked ackA (bb0622: acetate to AcP or pta (bb0589: AcP to acetyl-CoA. These mutants have an absolute requirement for mevalonate and demonstrate that ackA and pta are required for cell viability. When the ΔackA or Δpta mutant was exposed to conditions (i.e., increased temperature or cell density that up-regulate the expression of RpoS and OspC, normal induction of those proteins was observed. In addition, adding 20mM acetate or 20mM benzoate to the growth media of B. burgdorferi strain B31 ΔackA induced the expression of RpoS and OspC. These data suggest that AcP (generated by AckA is not directly involved in modulating the Rrp2-RpoN-RpoS regulatory pathway and that exogenous acetate or benzoate are triggering an acid stress response in B. burgdorferi.

  1. Variable Persister Gene Interactions with (pppGpp for Persister Formation in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Shuang Liu

    2017-09-01

    Full Text Available Persisters comprise a group of phenotypically heterogeneous metabolically quiescent bacteria with multidrug tolerance and contribute to the recalcitrance of chronic infections. Although recent work has shown that toxin-antitoxin (TA system HipAB depends on stringent response effector (pppGppin persister formation, whether other persister pathways are also dependent on stringent response has not been explored. Here we examined the relationship of (pppGpp with 15 common persister genes (dnaK, clpB, rpoS, pspF, tnaA, sucB, ssrA, smpB, recA, umuD, uvrA, hipA, mqsR, relE, dinJ using Escherichia coli as a model. By comparing the persister levels of wild type with their single gene knockout and double knockout mutants with relA, we divided their interactions into five types, namely A “dependent” (dnaK, recA, B “positive reinforcement” (rpoS, pspF, ssrA, recA, C “antagonistic” (clpB, sucB, umuD, uvrA, hipA, mqsR, relE, dinJ, D “epistasis” (clpB, rpoS, tnaA, ssrA, smpB, hipA, and E “irrelevant” (dnaK, clpB, rpoS, tnaA, sucB, smpB, umuD, uvrA, hipA, mqsR, relE, dinJ. We found that the persister gene interactions are intimately dependent on bacterial culture age, cell concentrations (diluted versus undiluted culture, and drug classifications, where the same gene may belong to different groups with varying antibiotics, culture age or cell concentrations. Together, this study represents the first attempt to systematically characterize the intricate relationships among the different mechanisms of persistence and as such provide new insights into the complexity of the persistence phenomenon at the level of persister gene network interactions.

  2. [Travelers attending an international vaccine center. Is the risk for the pediatric traveler increasing?].

    Science.gov (United States)

    Calavia Garsaball, O; Otero Romero, S; Campins Martí, M; Martínez-Gómez, X; Rodrigo Pendas, J A; Armadans Gil, L

    2013-09-01

    There is a perception that the number of pediatric travelers is increasing, mainly due to the so-called visiting friends and relatives (VFRs) group. Both the demographic and trips characteristics in this group may lead to an increased risk and a greater complexity in vaccinations and other preventive recommendations. To analyze the outcomes of different groups of pediatric travelers. To describe the demographic and travel characteristics within the pediatric population, and to analyze whether the VFRs differ from non-VFRs with regards to the factors that may contribute to a greater risk. A cross sectional descriptive study of the characteristics of pediatric travelers treated in the International Pre-travel Consultation Unit of the University Hospital Vall d'Hebron, from July 2002 to January 2009. Of the 692 children analyzed, with a mean age of 8 years (SD 5.4), an increase in the overall number of travelers was identified, along with an initial increase in the number of VFR children in the early years of the study, although later on, the numbers of this group stabilized. The mean age of the VFR travelers was also found to be lower. A lack of planning prior to the start of the travel was also noted in the VFRs group, as well as longer trip durations. A routine vaccine was administered to 29.2% of children, and malaria prophylaxis was recommended for 52% of travelers. Despite the progressive increase in international travel and the initial increase in VFR travelers, the percentage of this group has remained stable in recent years. However, the perception of a low risk among the immigrant population suggests the need to encourage an adequate pre-travel consultation within this group. Copyright © 2012 Asociación Española de Pediatría. Published by Elsevier Espana. All rights reserved.

  3. Thermo-Economic Performance Analysis of a Regenerative Superheating Organic Rankine Cycle for Waste Heat Recovery

    Directory of Open Access Journals (Sweden)

    Zhonghe Han

    2017-10-01

    Full Text Available The Organic Rankine Cycle (ORC is a promising form of technology for recovering low-grade waste heat. In this study, a regenerative ORC system is established to recover the waste flue gas of 160 °C. Focusing on thermodynamic and economic performance while simultaneously considering the limitations of volume flow ratio (VFR and the effect of superheat, working fluid selection and parameter optimization have been investigated. The optimization of the evaporation temperature is carried out by analyzing the variation of net power output and specific investment cost (SIC. Then, the net power output, specific net power output, total exergy destruction rate, VFR, total capital cost, and levelized electricity cost (LEC are selected as criteria, and a fuzzy multi-criteria evaluation method is adopted to select a more suitable working fluid and determine the optimal degree of superheat. In addition, the preheating coefficient, latent heat coefficient, superheating coefficient, and internal heat coefficient were proposed to explore the effect of working fluid critical temperature on thermal efficiency. Research studies demonstrate that there is an optimal evaporation temperature, maximizing net power output and minimizing the SIC. Isohexane and butane have greater specific net power output due to greater latent heat. A suitable degree of superheat is not only conducive to improving the working capacity of working fluids, but also reduces the VFR, total capital cost, SIC, and LEC for different working fluids. Thus, the system’s thermodynamic and economic performance—as well as the operational stability—are improved. Among the six working fluids, butane exhibits the best comprehensive performance, and its optimal evaporation temperature and degree of superheat are 100 °C and 5 °C, respectively.

  4. FimL regulates cAMP synthesis in Pseudomonas aeruginosa.

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    Yuki F Inclan

    2011-01-01

    Full Text Available Pseudomonas aeruginosa, a ubiquitous bacteria found in diverse ecological niches, is an important cause of acute infections in immunocompromised individuals and chronic infections in patients with Cystic Fibrosis. One signaling molecule required for the coordinate regulation of virulence factors associated with acute infections is 3', 5'-cyclic adenosine monophosphate, (cAMP, which binds to and activates a catabolite repressor homolog, Vfr. Vfr controls the transcription of many virulence factors, including those associated with Type IV pili (TFP, the Type III secretion system (T3SS, the Type II secretion system, flagellar-mediated motility, and quorum sensing systems. We previously identified FimL, a protein with histidine phosphotransfer-like domains, as a regulator of Vfr-dependent processes, including TFP-dependent motility and T3SS function. In this study, we carried out genetic and physiologic studies to further define the mechanism of action of FimL. Through a genetic screen designed to identify suppressors of FimL, we found a putative cAMP-specific phosphodiesterase (CpdA, suggesting that FimL regulates cAMP levels. Inactivation of CpdA increases cAMP levels and restores TFP-dependent motility and T3SS function to fimL mutants, consistent with in vivo phosphodiesterase activity. By constructing combinations of double and triple mutants in the two adenylate cyclase genes (cyaA and cyaB, fimL, and cpdA, we show that ΔfimL mutants resemble ΔcyaB mutants in TM defects, decreased T3SS transcription, and decreased cAMP levels. Similar to some of the virulence factors that they regulate, we demonstrate that CyaB and FimL are polarly localized. These results reveal new complexities in the regulation of diverse virulence pathways associated with acute P. aeruginosa infections.

  5. Sistem Informasi Geografi untuk Simulasi Rute Penerbangan Virtual

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    Fahrul Agus

    2017-02-01

    Full Text Available Pilot penerbang vitual menggunakan rute penerbangan berbasis peta spasial untuk mengoperasikan pesawat virtualnya. Untuk itu diperlukan sistem yang mampu melakukan analisis rute yang mesti diambil oleh seorang penerbang virtual.  Studi ini bertujuan membuat Sistem Informasi Geografi untuk simulasi rute penerbangan virtual menggunakan teknik pemrograman dinamis mundur (Backward Dynamic Programming-BDP.  Teknik BDB dipilih karena kemampuannya melakukan analisis untuk optimasi rute terutama pada jenis pernerbangan VFR (Visual Flight Rules dengan rute berbentuk multistage graph.  Pada bagian hasil dideskripsikan sistem  simulasi rute penerbangan virtual berupa antar muka program, pencarian ruter penerbanga dan hasil pengujian sistem.

  6. Some thoughts on the implementation of pilot night vision devices for helicopters

    Science.gov (United States)

    Tucker, G. E.

    1984-01-01

    Night vision enhancement devices greatly expand the range and quality of services by extending night operational capabilities. Evolving military tactical concepts for helicopters survivability and battlefield effectiveness necessitate nap-of-the-earth (NOE) flying under both day and night conditions. From a pilot workload standpoint, flying a helicopter NOE in day VFR conditions with minimum clearance between rotors and obstacles is quite demanding. Doing the same job at night is several times more difficult. There are two general categories of night vision devices in operation in helicopter aviation: the Night Vision Goggles (NVG) and forward looking infrared (FLIR) system. The capabilities and limitations of those two devices are discussed.

  7. Telerobotics with whole arm collision avoidance

    Energy Technology Data Exchange (ETDEWEB)

    Wilhelmsen, K.; Strenn, S.

    1993-09-01

    The complexity of teleorbotic operations in a cluttered environment is exacerbated by the need to present collision information to the operator in an understandable fashion. In addition to preventing movements which will cause collisions, a system providing some form of virtual force reflection (VFR) is desirable. With this goal in mind, Lawrence Livermore National Laboratory (LLNL) has installed a kinematically master/slave system and developed a whole arm collision avoidance system which interacts directly with the telerobotic controller. LLNL has also provided a structure to allow for automated upgrades of workcell models and provide collision avoidance even in a dynamically changing workcell.

  8. Air Traffic Management: Civil/Military Systems and Technologies.

    Science.gov (United States)

    1980-02-01

    for cruise cC = f4 (w,v,l) (In this case c is the specific (unit distance) consur.ption). - for descent Id = f5 (D) td = f6 (DI Moreover the...ce value, using f4 and f7; d)assume a temptative take-off weight Pt (o) and put i = 1; e)compute, by means of f2 and fl relationships he distance Ds (i...collision at a low rate of closure. 3.3 Midair collision 4 F-4 Phantoms / powered glider In 1976, a formation of 4 ]F-4 Phantoms were flying VFR in a

  9. Volatility and hygroscopicity of aging secondary organic aerosol in a smog chamber

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    T. Tritscher

    2011-11-01

    Full Text Available The evolution of secondary organic aerosols (SOA during (photo-chemical aging processes was investigated in a smog chamber. Fresh SOA from ozonolysis of 10 to 40 ppb α-pinene was formed followed by aging with OH radicals. The particles' volatility and hygroscopicity (expressed as volume fraction remaining (VFR and hygroscopicity parameter κ were measured in parallel with a volatility and hygroscopicity tandem differential mobility analyzer (V/H-TDMA. An aerosol mass spectrometer (AMS was used for the chemical characterization of the aerosol. These measurements were used as sensitive parameters to reveal the mechanisms possibly responsible for the changes in the SOA composition during aging. A change of VFR and/or κ during processing of atmospheric aerosols may occur either by addition of SOA mass (by condensation or by a change of SOA composition leading to different aerosol properties. The latter may occur either by heterogeneous reactions on the surface of the SOA particles, by condensed phase reactions like oligomerization or by an evaporation – gas-phase oxidation – recondensation cycle. The condensation mechanism showed to be dominant when there is a substantial change in the aerosol mass by addition of new molecules to the aerosol phase with time. Experiments could be divided into four periods based on the temporal evolution (qualitative changes of VFR, κ and organic mass: O3 induced condensation, ripening, and OH induced chemical aging first with substantial mass gain and then without significant mass gain.

    During the O3 induced condensation the particles' volatility decreased (increasing VFR while the hygroscopicity increased. Thereafter, in the course of ripening volatility continued to decrease, but hygroscopicity stayed roughly constant. After exposing the SOA to OH radicals an OH induced chemical aging with substantial mass gain started resulting in the production of at least 50 % more SOA mass

  10. IbeR facilitates stress-resistance, invasion and pathogenicity of avian pathogenic Escherichia coli.

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    Shaohui Wang

    Full Text Available Systemic infections by avian pathogenic Escherichia coli (APEC are economically devastating to poultry industries worldwide. IbeR, located on genomic island GimA, was shown to serve as an RpoS-like regulator in rpoS gene mutation neonatal meningitis E. coli (NMEC RS218. However, the role of IbeR in pathogenicity of APEC carrying active RpoS has not yet been investigated. We showed that the APEC IbeR could elicit antibodies in infected ducks, suggesting that IbeR might be involved in APEC pathogenicity. To investigate the function of IbeR in APEC pathogenesis, mutant and complementation strains were constructed and characterized. Inactivation of ibeR led to attenuated virulence and reduced invasion capacity towards DF-1 cells, brains and cerebrospinal fluid (CSF in vitro and in vivo. Bactericidal assays demonstrated that the mutant strain had impaired resistance to environmental stress and specific pathogen-free (SPF chicken serum. These virulence-related phenotypes were restored by genetic complementation. Quantitative real-time reverse transcription PCR revealed that IbeR controlled expression of stress-resistance genes and virulence genes, which might led to the associated virulence phenotype.

  11. Decrease in cell viability in an RMF, sigma(38), and OmpC triple mutant of Escherichia coli.

    Science.gov (United States)

    Samuel Raj, V; Füll, Christine; Yoshida, Madoka; Sakata, Kaori; Kashiwagi, Keiko; Ishihama, Akira; Igarashi, Kazuei

    2002-11-29

    In a speG-disrupted Escherichia coli mutant, which cannot metabolize spermidine to acetylspermidine, addition of spermidine to the medium caused a decrease in cell viability at the late stationary phase of growth. There were parallel decreases in the levels of ribosome modulation factor (RMF), the sigma(38) subunit of RNA polymerase, and the outer membrane protein C (OmpC). To clarify that these three proteins are strongly involved in cell viability, the rmf, rpoS (encoding sigma(38)), and ompC genes were disrupted. Viability of the triple mutant decreased to less than 1% of normal cells. The triple mutant had a reduced cell viability compared to any combination of double mutants, which also had a reduced cell viability. The single rmf and rpoS, but not ompC, mutant only slightly reduced cell viability. The results indicate that cooperative functions of these three proteins are necessary for cell viability at the late stationary phase. The triple mutant had a reduced level of ribosomes and of intracellular cations.

  12. Putrescine controls the formation of Escherichia coli persister cells tolerant to aminoglycoside netilmicin.

    Science.gov (United States)

    Tkachenko, Alexander G; Kashevarova, Natalya M; Karavaeva, Elena A; Shumkov, Mikhail S

    2014-12-01

    Persisters are suggested to be the products of a phenotypic variability that are quasi-dormant forms of regular bacterial cells highly tolerant to antibiotics. Our previous investigations revealed that a decrease in antibiotic tolerance of Escherichia coli cells could be reached through the inhibition of key enzymes of polyamine synthesis (putrescine, spermidine). We therefore assumed that polyamines could be involved in persister cell formation. Data obtained in our experiments with the polyamine-deficient E. coli strain demonstrate that the formation of persisters tolerant to netilmicin is highly upregulated by putrescine in a concentration-dependent manner when cells enter the stationary phase. This period is also accompanied by dissociation of initially homogenous subpopulation of persister cells to some fractions differing in their levels of tolerance to netilmicin. With three independent experimental approaches, we demonstrate that putrescine-dependent upregulation of persister cell formation is mediated by stimulation of rpoS expression. Complementary activity of putrescine and RpoS results in ~ 1000-fold positive effect on persister cell formation. © 2014 Federation of European Microbiological Societies.

  13. [Advances in molecular mechanisms of bacterial resistance caused by stress-induced transfer of resistance genes--a review].

    Science.gov (United States)

    Sun, Dongchang; Wang, Bing; Zhu, Lihong

    2013-07-04

    The transfer of resistance gene is one of the most important causes of bacterial resistance. Recent studies reveal that stresses induce the transfer of antibiotic resistance gene through multiple mechanisms. DNA damage stresses trigger bacterial SOS response and induce the transfer of resistance gene mediated by conjugative DNA. Antibiotic stresses induce natural bacterial competence for transformation in some bacteria which lack the SOS system. In addition, our latest studies show that the general stress response regulator RpoS regulates a novel type of resistance gene transfer which is mediated by double-stranded plasmid DNA and occurs exclusively on the solid surface. In this review, we summarized recent advances in SOS dependent and independent stress-induced DNA transfer which is mediated by conjugation and transformation respectively, and the transfer of double-stranded plasmid DNA on the solid surface which is regulated by RpoS. We propose that future work should address how stresses activate the key regulators and how these regulators control the expression of gene transfer related genes. Answers to the above questions would pave the way for searching for candidate targets for controlling bacterial resistance resulted from the transfer of antibiotic genes.

  14. Fitness of Outbreak and Environmental Strains of Escherichia coli O157:H7 in Aerosolizable Soil and Association of Clonal Variation in Stress Gene Regulation

    Directory of Open Access Journals (Sweden)

    Subbarao V. Ravva

    2014-06-01

    Full Text Available Airborne dust from feedlots is a potential mechanism of contamination of nearby vegetable crops with Escherichia coli O157:H7 (EcO157. We compared the fitness of clinical and environmental strains of EcO157 in <45 µm soil from a spinach farm. Differences in survival were observed among the 35 strains with D-values (days for 90% decreases ranging from 1–12 days. Strains that survived longer, generally, were from environmental sources and lacked expression of curli, a protein associated with attachment and virulence. Furthermore, the proportion of curli-positive (C+ variants of EcO157 strains decreased with repeated soil exposure and the strains that were curli-negative (C− remained C− post-soil exposure. Soil exposure altered expression of stress-response genes linked to fitness of EcO157, but significant clonal variation in expression was measured. Mutations were detected in the stress-related sigma factor, rpoS, with a greater percentage occurring in parental strains of clinical origin prior to soil exposure. We speculate that these mutations in rpoS may confer a differential expression of genes, associated with mechanisms of survival and/or virulence, and thus may influence the fitness of EcO157.

  15. Antimicrobials, stress and mutagenesis.

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    Alexandro Rodríguez-Rojas

    2014-10-01

    Full Text Available Cationic antimicrobial peptides are ancient and ubiquitous immune effectors that multicellular organisms use to kill and police microbes whereas antibiotics are mostly employed by microorganisms. As antimicrobial peptides (AMPs mostly target the cell wall, a microbial 'Achilles heel', it has been proposed that bacterial resistance evolution is very unlikely and hence AMPs are ancient 'weapons' of multicellular organisms. Here we provide a new hypothesis to explain the widespread distribution of AMPs amongst multicellular organism. Studying five antimicrobial peptides from vertebrates and insects, we show, using a classic Luria-Delbrück fluctuation assay, that cationic antimicrobial peptides (AMPs do not increase bacterial mutation rates. Moreover, using rtPCR and disc diffusion assays we find that AMPs do not elicit SOS or rpoS bacterial stress pathways. This is in contrast to the main classes of antibiotics that elevate mutagenesis via eliciting the SOS and rpoS pathways. The notion of the 'Achilles heel' has been challenged by experimental selection for AMP-resistance, but our findings offer a new perspective on the evolutionary success of AMPs. Employing AMPs seems advantageous for multicellular organisms, as it does not fuel the adaptation of bacteria to their immune defenses. This has important consequences for our understanding of host-microbe interactions, the evolution of innate immune defenses, and also sheds new light on antimicrobial resistance evolution and the use of AMPs as drugs.

  16. The ftsQ1p gearbox promoter of Escherichia coli is a major sigma S-dependent promoter in the ddlB-ftsA region.

    Science.gov (United States)

    Ballesteros, M; Kusano, S; Ishihama, A; Vicente, M

    1998-10-01

    The most potent promoters in the ddlB-ftsA region of the dcw cluster have been analysed for sigmaS-dependent transcription. Only the gearbox promoter ftsQ1p was found to be transcribed in vitro by RNA polymerase holoenzyme coupled to sigmaS (EsigmaS). This dependency on sigmaS was also found in vivo when single-copy fusions to a reporter gene were analysed in rpoS and rpoS+ backgrounds. Although ftsQ1p can be transcribed by RNA polymerase containing either sigmaD or sigmaS, there is a preference for EsigmaS when the assay conditions include potassium glutamate and supercoiled templates, a property shared with the bolA1p gearbox promoter. The rest of the promoters assayed, ftsQ2p and ftsZ2p3p4p, similarly to the control bolA2p promoter, were preferentially transcribed by EsigmaD, the housekeeper polymerase. The ftsQ1p and the bolA1p promoters also share the presence of AT-rich sequences upstream of the - 35 region and the requirement for an intact wild-type alpha-subunit for a proficient transcription, allowing their joint classification as gearboxes.

  17. Efeitos das restrições pré e pós-natal sobre o crescimento e o desempenho de cordeiros Santa Inês do nascimento ao desmame Effects of pre and postnatal feed restriction on growth and production of Santa Inês lambs from birth to weaning

    Directory of Open Access Journals (Sweden)

    Luciana Castro Geraseev

    2006-02-01

    Full Text Available Objetivou-se com este estudo avaliar os efeitos das restrições pré e pós-natal sobre o crescimento e o desempenho de cordeiros Santa Inês do nascimento ao desmame. Foram utilizados 68 cordeiros machos, divididos em três grupos: restrição pré-natal, restrição pós-natal e nenhum tipo de restrição (controle. O peso ao nascer dos animais dos grupos controle (4,628 kg e RPOS (4,421 kg foram semelhantes e superiores ao do grupo RPRE (3,024 kg. Os consumos médios de sucedâneo foram de 1,308; 0,957 e 0,952 litros/dia para os grupos controle, RPOS e RPRE, respectivamente. Os cordeiros sob restrição alimentar apresentaram consumo inferior aos do grupo controle, em razão do controle imposto aos animais RPOS e do menor tamanho do trato gastrintestinal dos animais RPRE. Os ganhos de peso médios dos cordeiros durante a fase de aleitamento foram de 200 g/dia (controle, 162 g/dia (RPRE e 153 g/dia (RPOS. O maior ganho de peso dos cordeiros do grupo controle resultou em maior peso ao desmame (17,12 kg em comparação aos cordeiros RPOS (14,15 kg e RPRE (13,00 kg. O desempenho inferior dos cordeiros RPRE em relação aos do grupo controle indica que estes animais não compensaram a restrição imposta durante a fase pré-natal, o que resultou em comprometimento do crescimento pós-natal e, conseqüentemente, em menor peso ao desmame.This trial was conducted to study the effects of pre and postnatal feed restriction on growth and production of Santa Inês lambs from birth to weaning. Sixty-eight lambs were assigned to three treatments as follows: prenatal feed restriction (PRER, postnatal feed restriction (POSTR, and no feed restriction (Control. Average birth body weight of lambs on Control (4.628 kg and POSTR (4.421 kg treatments were similar but higher than that of animals on PRER (3.024 kg. Milk replacer intake averaged 1.308, 0.957, and 0.952 L/d on Control, POSTR, and PRER treatments, respectively. The lower intake of feed

  18. On the trends of Fukui potential and hardness potential derivatives in isolated atoms vs. atoms in molecules.

    Science.gov (United States)

    Bhattacharjee, Rituparna; Roy, Ram Kinkar

    2014-10-28

    In the present study, trends of electronic contribution to molecular electrostatic potential [Vel(r¯)(r=0)], Fukui potential [v(+)f|(r=0) and v(-)f|(r=0)] and hardness potential derivatives [Δ(+)h(k) and Δ(-)h(k)] for isolated atoms as well as atoms in molecules are investigated. The generated numerical values of these three reactivity descriptors in these two electronically different situations are critically analyzed through the relevant formalism. Values of Vel(r¯) (when r → 0, i.e., on the nucleus) are higher for atoms in molecules than that of isolated atoms. In contrast, higher values of v(+)|(r=0) and v(-)|(r=0) are observed for isolated atoms compared to the values for atoms in a molecule. However, no such regular trend is observed for the Δ(+)h(k) and Δ(-)h(k) values, which is attributed to the uncertainty in the Fukui function values of atoms in molecules. The sum of Fukui potential and the sum of hardness potential derivatives in molecules are also critically analyzed, which shows the efficacy of orbital relaxation effects in quantifying the values of these parameters. The chemical consequence of the observed trends of these descriptors in interpreting electron delocalization, electronic relaxation and non-negativity of atomic Fukui function indices is also touched upon. Several commonly used molecules containing carbon as well as heteroatoms are chosen to make the investigation more insightful.

  19. Calculation of the Runway Capacity of Ljubljana- Brnik Airport

    Directory of Open Access Journals (Sweden)

    Andrej Grebenšek

    2004-01-01

    Full Text Available LjubljanaAirport is the main and central airport of the Republicof Slovenia that serves the needs of the major part of thecountry as well as the needs of the eminent regions across theborder. Due to its proximity to the capital of Slovenia, it is ofspecial importance for the country. The traffic at Ljubljana Airportconsists of IFR and VFR traffic. In calculations, however,only IFR traffic is considered. Although the VFR traffic participateswith a significant share in the total amount of traffic, it isso flexible that in the end, it only contributes to the overall capacityof the runway.LjubljanaAirport is specific because there is no taxiway tillthe holding position of runway 31. Aircraft generally land onrunway 31. This is the only available instrumental runway forlanding. Take-offs can be executed in the directions 31 or 13.Both directions are considered in the calculations. The basiccalculations are done on the assumption that take-offs andlandings are executed in the same direction, that is, on runway31. This presents a normal procedure at traffic peaks since sucha situation is clearer, easier and safer for an Air Traffic Controllerto handle. The present traffic load at the airport is not soheavy, therefore in quieter periods of the day aircraft land onrunway 31 and take off from runway 13. This procedure reducesthe capacity of the runway and the calculations for ithave been carried out at the end.

  20. Autonomous Flight Rules - A Concept for Self-Separation in U.S. Domestic Airspace

    Science.gov (United States)

    Wing, David J.; Cotton, William B.

    2011-01-01

    Autonomous Flight Rules (AFR) are proposed as a new set of operating regulations in which aircraft navigate on tracks of their choice while self-separating from traffic and weather. AFR would exist alongside Instrument and Visual Flight Rules (IFR and VFR) as one of three available flight options for any appropriately trained and qualified operator with the necessary certified equipment. Historically, ground-based separation services evolved by necessity as aircraft began operating in the clouds and were unable to see each other. Today, technologies for global navigation, airborne surveillance, and onboard computing enable the functions of traffic conflict management to be fully integrated with navigation procedures onboard the aircraft. By self-separating, aircraft can operate with more flexibility and fewer restrictions than are required when using ground-based separation. The AFR concept is described in detail and provides practical means by which self-separating aircraft could share the same airspace as IFR and VFR aircraft without disrupting the ongoing processes of Air Traffic Control.

  1. For Spacious Skies: Self-Separation with "Autonomous Flight Rules" in US Domestic Airspace

    Science.gov (United States)

    Wing, David J.; Cotton, William B.

    2011-01-01

    Autonomous Flight Rules (AFR) are proposed as a new set of operating regulations in which aircraft navigate on tracks of their choice while self-separating from traffic and weather. AFR would exist alongside Instrument and Visual Flight Rules (IFR and VFR) as one of three available flight options for any appropriately trained and qualified operator with the necessary certified equipment. Historically, ground-based separation services evolved by necessity as aircraft began operating in the clouds and were unable to see each other. Today, technologies for global precision navigation, emerging airborne surveillance, and onboard computing enable traffic conflict management to be fully integrated with navigation procedures onboard the aircraft. By self-separating, aircraft can operate with more flexibility and fewer flight restrictions than are required when using ground-based separation. The AFR concept proposes a practical means in which self-separating aircraft could share the same airspace as IFR and VFR aircraft without disrupting the ongoing processes of Air Traffic Control. The paper discusses the context and motivation for implementing self-separation in US domestic airspace. It presents a historical perspective on separation, the proposed way forward in AFR, the rationale behind mixed operations, and the expected benefits of AFR for the airspace user community.

  2. Onion Peel Ethylacetate Fraction and Its Derived Constituent Quercetin 4′-O-β-D Glucopyranoside Attenuates Quorum Sensing Regulated Virulence and Biofilm Formation

    Directory of Open Access Journals (Sweden)

    Hanan M. Al-Yousef

    2017-09-01

    Full Text Available The resistance and pathogenesis of bacteria could be related to their ability to sense and respond to population density, termed quorum sensing (QS. Inhibition of the QS system is considered as a novel strategy for the development of antipathogenic agents, especially for combating drug-resistant bacterial infections. In the present study, the anti-QS activity of Onion peel ethylacetate fraction (ONE was tested against Chromobacterium violaceum CV12472 and Pseudomonas aeruginosa PAO1. ONE inhibit the QS-mediated virulence factors production such as violacein in C. violaceum and elastase, pyocyanin in P. aeruginosa. Further, the treatment with sub-MICs of ONE significantly inhibited the QS-mediated biofilm formation, EPS (Extracellular polymeric substances production and swarming motility. Further, quercetin 4′-O-β-D glucopyranoside (QGP was isolated from ONE and its anti-QS potential was confirmed after observing significant inhibition of QS-controlled virulence factors such as violacein, elastase, pyocyanin and biofilm formation in test pathogens. Molecular docking analysis predicted that QGP should be able to bind at the active sites of Vfr and LasR, and if so blocks the entry of active sites in Vfr and LasR.

  3. Ferric uptake regulator mutants of Pseudomonas aeruginosa with distinct alterations in the iron-dependent repression of exotoxin A and siderophores in aerobic and microaerobic environments.

    Science.gov (United States)

    Barton, H A; Johnson, Z; Cox, C D; Vasil, A I; Vasil, M L

    1996-09-01

    assessed under aerobic and microaerobic conditions. Iron-dependent repression of ETA synthesis in both parental strains and A2 and A4 mutants was found to be 50-100-fold under aerobic and microaerobic conditions, as assayed by quantitative Western dot-blot assays. By contrast in the CS and C6 mutants, while iron-dependent repression os ETA synthesis was similar to both parental strains under aerobic conditions, ETA production in these mutants was constitutive in a microaerobic environment. RNase protection analysis of toxA and regAB transcription in PAO1, PAO6261 and the C6 mutant corroborated the results of quantitative dot-blot assays of ETA. The mutant Fur proteins were purified and examined for their ability to bind to the promoter of a gene (pvdS) that positively regulates the expression of siderophores and ETA. Fur from the A2 and A4 mutants and from the C6Rv revertant was able to bind to the target DNA, but with reduced affinity by comparison to wild-type Fur. Fur from the C6 mutant in DNase I footprint experiments failed to protect the promoter region of the pvdS gene, but it retained some weak binding activity in gel mobility shift assays. The data presented in this study not only furnish some additional insights into the structure-function relationships of Fur, but also afford novel perspectives of virulence factors in P. aeruginosa under environmental conditions that have not previously been considered.

  4. Iron-Regulated Expression of Alginate Production, Mucoid Phenotype, and Biofilm Formation by Pseudomonas aeruginosa

    Science.gov (United States)

    Wiens, Jacinta R.; Vasil, Adriana I.; Schurr, Michael J.; Vasil, Michael L.

    2014-01-01

    ABSTRACT Pseudomonas aeruginosa strains of non-cystic fibrosis (non-CF) origin do not produce significant amounts of extracellular alginate and are nonmucoid. In CF, such isolates can become mucoid through mutation of one of the genes (mucA, mucB, mucC, or mucD) that produce regulatory factors that sequester AlgU, required for increased expression of alginate genes. Mutation of the muc genes in the nonmucoid PAO1, PA14, PAKS-1, and Ps388 strains led to increased levels of extracellular alginate and an obvious mucoid phenotype, but only under iron-limiting growth conditions (≤5 µM), not under iron-replete conditions (≥10 µM). In contrast, >50% of P. aeruginosa isolates from chronic CF pulmonary infections expressed increased levels of alginate and mucoidy both under iron-limiting and iron-replete conditions (i.e., iron-constitutive phenotype). No single iron regulatory factor (e.g., Fur, PvdS) was associated with this loss of iron-regulated alginate expression and mucoidy in these CF isolates. However, the loss of only pyoverdine production, or its uptake, abrogated the ability of P. aeruginosa to produce a robust biofilm that represents the Psl-type of biofilm. In contrast, we show that mutation of the pyoverdine and pyochelin biosynthesis genes and the pyoverdine receptor (FpvA) lead to iron-constitutive expression of the key alginate biosynthesis gene, algD, and an explicitly mucoid phenotype in both iron-limiting and iron-replete conditions. These data indicate that alginate production and mucoidy, in contrast to other types of biofilms produced by P. aeruginosa, are substantially enhanced under iron limitation. These results also have compelling implications in relation to the use of iron chelators in the treatment of P. aeruginosa CF infections. PMID:24496793

  5. Block and boost DNA transfer: opposite roles of OmpA in natural and artificial transformation of Escherichia coli.

    Science.gov (United States)

    Sun, Dongchang; Wang, Bing; Zhu, Lihong; Chen, Mengyao; Zhan, Linlin

    2013-01-01

    Our previous work established that DNA is naturally transferable on agar plates through a new transformation system which is regulated by the stationary phase master regulator RpoS in Escherichia coli. In this transformation system, neither additional Ca(2+) nor heat shock is required. Instead, transformation is stimulated by agar. The membrane protein OmpA, a gated pore permeable to ions and larger solutes, serves as a receptor for DNA transfer during bacteriophage infection and conjugation. However, it remains unknown how DNA transfers across membranes and whether OmpA is involved in transformation of E. coli. Here, we explored potential roles of OmpA in natural and chemical transformation of E. coli. We observed that ompA inactivation significantly improved natural transformation on agar plates, indicating that OmpA blocks DNA transfer. Transformation promotion by ompA inactivation also occurred on soft plates, indicating that OmpA blocks DNA transfer independent of agar. By contrast, compared with the wild-type strain, chemical transformation of the ompA mutant was lower, indicating that OmpA has a role in DNA transfer. Inactivation of ompA also reduced chemical transformation in solution containing less Ca(2+) or with a shortened time for heat shock, suggesting that the promotion effect of OmpA on DNA transfer does not solely rely on Ca(2+) or heat shock. We conclude that OmpA plays opposite roles in natural and chemical transformation: it blocks DNA uptake on agar plates but promotes DNA transfer in the liquid Ca(2+) solution. Considering that no single factor was identified to reverse the function of OmpA, we propose that multiple factors may cooperate in the functional reversal of OmpA during natural and artificial transformation of E. coli. Finally, we observed that ompA transcription was not affected by the expression of RpoS, excluding the possibility that RpoS regulates DNA transfer by suppressing ompA transcription.

  6. Block and boost DNA transfer: opposite roles of OmpA in natural and artificial transformation of Escherichia coli.

    Directory of Open Access Journals (Sweden)

    Dongchang Sun

    Full Text Available Our previous work established that DNA is naturally transferable on agar plates through a new transformation system which is regulated by the stationary phase master regulator RpoS in Escherichia coli. In this transformation system, neither additional Ca(2+ nor heat shock is required. Instead, transformation is stimulated by agar. The membrane protein OmpA, a gated pore permeable to ions and larger solutes, serves as a receptor for DNA transfer during bacteriophage infection and conjugation. However, it remains unknown how DNA transfers across membranes and whether OmpA is involved in transformation of E. coli. Here, we explored potential roles of OmpA in natural and chemical transformation of E. coli. We observed that ompA inactivation significantly improved natural transformation on agar plates, indicating that OmpA blocks DNA transfer. Transformation promotion by ompA inactivation also occurred on soft plates, indicating that OmpA blocks DNA transfer independent of agar. By contrast, compared with the wild-type strain, chemical transformation of the ompA mutant was lower, indicating that OmpA has a role in DNA transfer. Inactivation of ompA also reduced chemical transformation in solution containing less Ca(2+ or with a shortened time for heat shock, suggesting that the promotion effect of OmpA on DNA transfer does not solely rely on Ca(2+ or heat shock. We conclude that OmpA plays opposite roles in natural and chemical transformation: it blocks DNA uptake on agar plates but promotes DNA transfer in the liquid Ca(2+ solution. Considering that no single factor was identified to reverse the function of OmpA, we propose that multiple factors may cooperate in the functional reversal of OmpA during natural and artificial transformation of E. coli. Finally, we observed that ompA transcription was not affected by the expression of RpoS, excluding the possibility that RpoS regulates DNA transfer by suppressing ompA transcription.

  7. Profile of travel-associated illness in children, Zürich, Switzerland.

    Science.gov (United States)

    Hunziker, Thomas; Berger, Christoph; Staubli, Georg; Tschopp, Alois; Weber, Rainer; Nadal, David; Hatz, Christoph; Schlagenhauf, Patricia

    2012-01-01

    The number of families traveling with their children to their country of origin and/or to tropical destinations has increased in Switzerland and includes a changing profile and multinational range of patients. Defining the profile of reported travel-associated illnesses will help to improve the prevention and treatment of such illnesses in children. This study includes children aged up to 16 years who sought medical advice for a presumed travel-related illness at the emergency room of the University of Zürich Children's Hospital during the period July 2007 to December 2008. We analyzed data on 328 children (58.8% male, mean age: 4.62 y) who presented with travel-associated illness. Our analysis included 155 traditional (mainly tourist) travelers, 162 children who were visiting friends and relatives (VFR), and 11 immigrants. Some 11% were hospitalized. No deaths occurred. The main conditions recorded were diarrheal illness (39%), respiratory (28.7%) and febrile/systemic illness (13.4%). With increasing age, the proportion of children with diarrheal disease increased, while the proportion with respiratory illness declined. There were significant associations between geographic area of exposure and the profile of travel-related disease (p < 0.001). Among 36 children with more serious diseases requiring hospitalization, 12 (3.7% overall) presented with potentially serious diseases: malaria (n = 2), Salmonella typhi (n = 3), Salmonella paratyphi (n = 2), meningococcal meningitis (n = 1), tuberculosis (n = 2), visceral leishmania (n = 1), and hepatitis A (n = 1). Eleven of the 12 children presenting with these potentially serious illnesses were VFR or immigrant children. The main diagnoses for ill-returned Zürich children who presented for emergency care were diarrhea, respiratory, and febrile/systemic illness. A broad spectrum of morbidity was seen including meningococcal meningitis, malaria, tuberculosis, typhoid fever, leishmania, and hepatitis A. Diagnoses varied

  8. Imported malaria in an area in southern Madrid, 2005-2008.

    Science.gov (United States)

    Rey, Sonia; Zuza, Inés; Martínez-Mondéjar, Belén; Rubio, José M; Merino, Francisco J

    2010-10-20

    In Spain, malaria cases are mostly due to migrants and travellers returning from endemic areas. The objective of this work was to describe the malaria cases diagnosed at the Severo Ochoa University Hospital (HUSO) in Leganés in the south of the Madrid Region from 2005 to 2008. Descriptive retrospective study performed at HUSO. Data sources are registries from the Microbiology Department and malaria cases notified to the Preventive Medicine Department. Analysed parameters were: administrative, demographical, related to the stay at the endemic country, clinical, microbiological diagnosis method, pregnancy, treatment and prophylaxis, co-infections, and days of hospital stay. Fifty-seven patients diagnosed with malaria were studied. Case distribution per year was 13 in 2005, 15 in 2006, 15 in 2007 and 14 in 2008. Thirty-three patients were female (57.9%) and 24 male (42.1%). Mean age was 27.8 years. Most of the malaria cases were acquired in Nigeria (49.1%) and Equatorial Guinea (32.7%). 29.1% of the patients were immigrants who had arrived recently, and 61.8% acquired malaria when travelling to their countries of origin to visit friends and relatives (VFR). Majority of cases were diagnosed between June and September. Microscopy was positive in 39 cases (68.4%) immunochromatography in 42 (73.7%) and PCR in the 55 cases where performed. Plasmodium falciparum was responsible for 94.7% of the cases. The more frequent symptoms were fever (77.2%), followed by headache and gastrointestinal symptoms (33.3%). Nine cases needed hospital admittance, a pregnant woman, three children, four VFR and an African tourist, but all evolved favourably. Chemoprophylaxis data was known from 55 patients. It was taken correctly in one case (1.8%), in five (9.1%) the prophylaxis was improper while the others 49 (89.1%) cases had not followed any anti-malarial prophylaxis. Children, pregnant women and the VFR have the highest risk to present severe malaria and to need hospital admittance

  9. Malaria knowledge and utilization of chemoprophylaxis in the UK population and in UK passengers departing to malaria-endemic areas.

    Science.gov (United States)

    Behrens, Ron H; Alexander, Neal

    2013-12-21

    The burden of imported malaria is predominantly in travellers visiting friends and relatives (VFR) in sub-Saharan Africa. The failure of this group to use chemoprophylaxis is recognized as the most important risk factor for the high incidence of disease. Understanding the reasons for failure to follow national recommendations may relate to knowledge, risk perception, cost, and peer pressure. Research into these variables is critical to understand and change practices in this group and this study was designed to explore whether knowledge, risk perception and prophylaxis use differs between travellers' to various destinations and the rest of the UK population. Two face-to-face questionnaire surveys were conducted to collect information on demographics, malaria knowledge, source, and quality of pre-travel advice, past travel experience and perceived malaria threat. One was an IPSOS survey of individuals representative of the UK population. The other was a departure lounge survey (Civil Aviation Authority (CAA)) of passengers departing to malarious regions detailing destinations and use of chemoprophylaxis. Around a quarter of the 1,991 UK population surveyed had previously travelled to a malarious area. Five-hundred departing passengers were interviewed, of which 80% travelled for leisure (56% VFR's) and 42% were travelling to West Africa. Malaria knowledge among the UK population (score 58.6) was significantly lower than that of individuals who had previously travelled or were travelling (63.8 and 70.7 respectively). Malaria knowledge was similar in individuals who had and had not sought pre-travel advice and travellers using and not using chemoprophylaxis for their journey. Leisure travellers to Ghana and Nigeria were predominantly VFRs (74%), whilst 66% of travellers to Kenya were tourists. Despite similar high knowledge scores and perceived (>90%) threat of the lethality of malaria in the three groups, chemoprophylaxis use in Nigerians (50%) was substantially

  10. Imported malaria in an area in southern Madrid, 2005-2008

    Directory of Open Access Journals (Sweden)

    Rubio José M

    2010-10-01

    Full Text Available Abstract Background In Spain, malaria cases are mostly due to migrants and travellers returning from endemic areas. The objective of this work was to describe the malaria cases diagnosed at the Severo Ochoa University Hospital (HUSO in Leganés in the south of the Madrid Region from 2005 to 2008. Methods Descriptive retrospective study performed at HUSO. Data sources are registries from the Microbiology Department and malaria cases notified to the Preventive Medicine Department. Analysed parameters were: administrative, demographical, related to the stay at the endemic country, clinical, microbiological diagnosis method, pregnancy, treatment and prophylaxis, co-infections, and days of hospital stay. Results Fifty-seven patients diagnosed with malaria were studied. Case distribution per year was 13 in 2005, 15 in 2006, 15 in 2007 and 14 in 2008. Thirty-three patients were female (57.9% and 24 male (42.1%. Mean age was 27.8 years. Most of the malaria cases were acquired in Nigeria (49.1% and Equatorial Guinea (32.7%. 29.1% of the patients were immigrants who had arrived recently, and 61.8% acquired malaria when travelling to their countries of origin to visit friends and relatives (VFR. Majority of cases were diagnosed between June and September. Microscopy was positive in 39 cases (68.4% immunochromatography in 42 (73.7% and PCR in the 55 cases where performed. Plasmodium falciparum was responsible for 94.7% of the cases. The more frequent symptoms were fever (77.2%, followed by headache and gastrointestinal symptoms (33.3%. Nine cases needed hospital admittance, a pregnant woman, three children, four VFR and an African tourist, but all evolved favourably. Chemoprophylaxis data was known from 55 patients. It was taken correctly in one case (1.8%, in five (9.1% the prophylaxis was improper while the others 49 (89.1% cases had not followed any anti-malarial prophylaxis. Conclusions Children, pregnant women and the VFR have the highest risk to

  11. Buffer capacity of food components influences the acid tolerance response in Salmonella Typhimurium during simulated gastric passage

    DEFF Research Database (Denmark)

    Henriksen, Sidsel; Buschhardt, Tasja; Hansen, Tina Beck

    2014-01-01

    Food composition, buffer capacity, and fat and protein content have been shown to effect the gastric acid survival of pathogens (Waterman & Small 1998). In this study, simple food-model substances with different buffer capacities were investigated for their ability to support survival of stationary...... major stationary phase ATR regulators, we found an approx. four-fold increase in expression of ompR and an approx. three-fold increase of rpoS in saline and buffered saline, respectively, after 15 min of gastric acid challenge. The relative expression of these genes, were significantly lower in Brain...... Heart Infusion Broth having a higher buffer capacity. We suggest this to be associated with a varying ability of Salmonella Typhimurium to mount a stationary phase acid tolerance response (ATR) depending on the buffer capacity of the food vehicle....

  12. Survival of Salmonella serovars on beef carcasses and molecular mechanisms to survive low temperature stress and desiccation

    DEFF Research Database (Denmark)

    Knudsen, Gitte Maegaard; Thomsen, Line Elnif; Aabo, Søren

    2006-01-01

    . Infantis and S. Newport. This experiment indicates that storage at low temperature of beef carcasses can be used as a reduction strategy for Salmonella spp. in beef. In addition molecular mechanisms to resist low temperature stress and desiccation have been investigated. Mutants in the otsA, rpoS and clp...... at 10°C and 15°C, the ¿clpP mutant was severely affected in ability to form colonies compared to the wildtype. Neither the ¿otsA nor the ¿rpoS were affected in growth or survival at low temperature. In a desiccation model, both the ¿rpoS and ¿clpP mutant were more sensitive than the C5 wildtype...

  13. Efeitos das restrições pré e pós-natal sobre o crescimento e desempenho de cordeiros Santa Inês do desmame ao abate Effects of pre and postnatal feed restriction on growth and production of Santa Inês lambs from weaning to slaughter

    Directory of Open Access Journals (Sweden)

    Luciana Castro Geraseev

    2006-02-01

    Full Text Available Objetivou-se com este estudo avaliar os efeitos das restrições pré e pós-natal sobre o crescimento e desempenho de cordeiros Santa Inês do desmame ao abate. Foram utilizados 48 cordeiros machos, divididos em três grupos: restrição pré-natal (RPRE, restrição pós-natal (RPOS e nenhum tipo de restrição (controle. Após o desmame, o consumo médio de ração dos cordeiros da RPOS foi inferior aos dos demais, em razão do controle no fornecimento do alimento, sendo que as médias para os grupo controle, RPRE e RPOS foram, respectivamente, de 757, 765 e 624g/dia. Os animais do grupo controle apresentaram os menores consumos totais de ração, em decorrência da menor idade de abate, em comparação aos cordeiros com alimentação restrita, acarretando menor tempo de confinamento. A análise da curva de crescimento comprovou que o ganho médio dos cordeiros do grupo controle (257 g/dia foi superior ao dos cordeiros RPOS (155g/dia e RPRE (198 g/dia. O desempenho inferior dos cordeiros com alimentação restrita em relação aos do grupo controle após o desmame indica que os animais não compensaram a restrição imposta e, portanto, apresentaram valores mais altos para idade de abate, consumo total de ração e tempo de ocupação da instalação.This experiment was conducted to evaluate the effects of pre and postnatal feed restriction on growth and production of Santa Inês lambs from weaning to slaughter. Forty-eight lambs were assigned to three treatments as follows: lambs with prenatal feed restriction (PRER, lambs with postnatal feed restriction (POSTR, and lambs with no feed restriction (Control. Average daily feed intake (as fed basis of lambs on POSTR treatment (624 g/d was significantly lower (P<0.05 than that of lambs on control (757 g/d and PRER (765 g/d treatments possibly because of the feed restriction after birth. However, total feed intake was lowest on control, which may be explained by the earlier slaughter age of lambs

  14. Competitive growth advantage of nontoxigenic mutants in the stationary phase in archival cultures of pathogenic Vibrio cholerae strains.

    Science.gov (United States)

    Paul, Kalidas; Ghosh, Amalendu; Sengupta, Nilanjan; Chowdhury, Rukhsana

    2004-09-01

    Spontaneous nontoxigenic mutants of highly pathogenic Vibrio cholerae O1 strains accumulate in large numbers during long-term storage of the cultures in agar stabs. In these mutants, production of the transcriptional regulator ToxR was reduced due to the presence of a mutation in the ribosome-binding site immediately upstream of the toxR open reading frame. Consequently, the ToxR-dependent virulence regulon was turned off, with concomitant reduction in the expression of cholera toxin and toxin-coregulated pilus. An intriguing feature of these mutants is that they have a competitive fitness advantage when grown in competition with the parent strains in stationary-phase cocultures which is independent of RpoS, the only locus known to be primarily associated with acquisition of a growth advantage phenotype in bacteria.

  15. Carbon and nitrogen substrate utilization by archival Salmonella typhimurium LT2 cells

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    Edwards Kelly K

    2002-09-01

    Full Text Available Abstract Background A collection of over 20,000 Salmonella typhimurium LT2 mutants, sealed for four decades in agar stabs, is a unique resource for study of genetic and evolutionary changes. Previously, we reported extensive diversity among descendants including diversity in RpoS and catalase synthesis, diversity in genome size, protein content, and reversion from auxotrophy to prototrophy. Results Extensive and variable losses and a few gains of catabolic functions were observed by this standardized method. Thus, 95 catabolic reactions were scored in each of three plates in wells containing specific carbon and nitrogen substrates. Conclusion While the phenotype microarray did not reveal a distinct pattern of mutation among the archival isolates, the data did confirm that various isolates have used multiple strategies to survive in the archival environment. Data from the MacConkey plates verified the changes in carbohydrate metabolism observed in the Biolog™ system.

  16. Gearbox gene expression and growth rate.

    Science.gov (United States)

    Aldea, M; Garrido, T; Tormo, A

    1993-07-01

    Regulation of gene expression in prokaryotic cells usually takes place at the level of transcription initiation. Different forms of RNA polymerase recognizing specific promoters are engaged in the control of many prokaryotic regulons. This also seems to be the case for some Escherichia coli genes that are induced at low growth rates and by nutrient starvation. Their gene products are synthesized at levels inversely proportional to growth rate, and this mode of regulation has been termed gearbox gene expression. This kind of growth-rate modulation is exerted by specific transcriptional initiation signals, the gearbox promoters, and some of them depend on a putative new σ factor (RpoS). Gearbox promoters drive expression of morphogenetic and cell division genes at constant levels per cell and cycle to meet the demands of cell division and septum formation. A mechanism is proposed that could sense the growth rate of the cell to alter gene expression by the action of specific σ factors.

  17. Pseudomonas aeruginosa uses type III secretion system to kill biofilm-associated amoebae

    DEFF Research Database (Denmark)

    Matz, Carsten; Moreno, Ana Maria; Alhede, Morten

    2008-01-01

    and killed biofilm-associated amoebae by a quorum-sensing independent mechanism. Analysis of the amoeba-induced transcriptome indicated the involvement of the P. aeruginosa type III secretion system (T3SS) in this interaction. A comparison of mutants with specific defects in the T3SS demonstrated the use...... of the secretion apparatus and the effectors ExoU, ExoS and ExoT in the killing process, of which ExoU had the greatest impact. T3SS-mediated virulence towards A. castellanii was found to be controlled by the global regulators RpoN and RpoS and through modulation of cAMP and alginate biosynthesis. Our findings...

  18. Importance of sigma factor mutations in increased triclosan resistance in Salmonella Typhimurium

    DEFF Research Database (Denmark)

    Gantzhorn, Mette Rørbæk; Olsen, John Elmerdahl; Thomsen, Line Elnif

    2015-01-01

    BACKGROUND: Salmonella enterica is the second most common foodborne pathogen. The use of biocides is crucial to prevent spread of foodborne pathogens, and it would be devastating for food safety if Salmonella would become resistant to the disinfectants used. Another concern is that exposure...... to disinfectants might lead to decreased susceptibility to antibiotics. The current study aimed to identify genetic changes causing high level triclosan resistance in S. enterica serovar Typhimurium and evaluate how these affected antibiotic resistance and efflux pump activity. RESULTS: Wild type strains S....... Typhimurium 4/74 and DTU3 were adapted to increasing concentrations of the biocide triclosan by serial passage. High level triclosan resistant isolates (MIC > 1000 μg/ml) were obtained. Strains were genome sequenced, and SNPs in fabI, rpoS and rpoD were found to be associated with high level resistance...

  19. dsdA Does Not Affect Colonization of the Murine Urinary Tract by Escherichia coli CFT073.

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    Andrew J Hryckowian

    Full Text Available The urinary tract environment provides many conditions that deter colonization by microorganisms. D-serine is thought to be one of these stressors and is present at high concentrations in urine. D-serine interferes with L-serine and pantothenate metabolism and is bacteriostatic to many species. Uropathogenic Escherichia coli commonly possess the dsdCXA genetic locus, which allows them to use D-serine as a sole carbon, nitrogen, and energy source. It was previously reported that in the model UPEC strain CFT073, a dsdA mutant outcompetes wild type in the murine model of urinary tract infection. This "hypercolonization" was used to propose a model whereby UPEC strains sense D-serine in the urinary tract and subsequently up-regulate genes necessary for pathogenesis. Here, we show that inactivation of dsdA does not lead to hypercolonization. We suggest that this previously observed effect is due to an unrecognized secondary mutation in rpoS and that some D-serine specific effects described in other studies may be affected by the rpoS status of the strains used. Inactivation of dsdA in the original clinical isolate of CFT073 gives CFT073 ΔdsdA a growth defect in human urine and renders it unable to grow on minimal medium containing D-serine as the sole carbon source. However, CFT073 ΔdsdA is able to colonize the urinary tracts of CBA/J mice indistinguishably from wild type. These findings indicate that D-serine catabolism, though it may play role(s during urinary tract infection, does not affect the ability of uropathogenic E. coli to colonize the murine urinary tract.

  20. Inactivation of Escherichia coli by citral.

    Science.gov (United States)

    Somolinos, M; García, D; Condón, S; Mackey, B; Pagán, R

    2010-06-01

    The aim was to evaluate (i) the resistance of Escherichia coli BJ4 to citral in a buffer system as a function of citral concentration, treatment medium pH, storage time and initial inoculum size, (ii) the role of the sigma factor RpoS on citral resistance of E. coli, (iii) the role of the cell envelope damage in the mechanism of microbial inactivation by citral and (iiii) possible synergistic effects of mild heat treatment and pulsed electric fields (PEF) treatment combined with citral. The initial inoculum size greatly affected the efficacy of citral against E. coli cells. Exposure to 200 microl l(-1) of citral at pH 4.0 for 24 h at 20 degrees C caused the inactivation of more than 5 log(10) cycles of cells starting at an inoculum size of 10(6) or 10(7) CFU ml(-1), whereas increasing the cell concentration to 10(9) CFU ml(-1) caused citral at pH 4.0 than pH 7.0. The rpoS null mutant strain E. coli BJ4L1 was less resistant to citral than the wild-type strain. Occurrence of sublethal injury to both the cytoplasmic and outer membranes was demonstrated by adding sodium chloride or bile salts to the recovery media. The majority of sublethally injured cells by citral required energy and lipid synthesis for repair. A strongly synergistic lethal effect was shown by mild heat treatment combined with citral but the presence of citral during the application of a PEF treatment did not show any advantage. This work confirms that cell envelope damage is an important event in citral inactivation of bacteria, and it describes the key factors on the inactivation of E. coli cells by citral. Knowledge about the mechanism of microbial inactivation by citral helps establish successful combined preservation treatments.

  1. Interdependence of environmental factors influencing reciprocal patterns of gene expression in virulent Borrelia burgdorferi.

    Science.gov (United States)

    Yang, X; Goldberg, M S; Popova, T G; Schoeler, G B; Wikel, S K; Hagman, K E; Norgard, M V

    2000-09-01

    The paradigm for differential antigen expression in Borrelia burgdorferi, the agent of Lyme disease, is the reciprocal expression of its outer surface (lipo)proteins (Osp) A and C; as B. burgdorferi transitions from its arthropod vector into mammalian tissue, ospC is upregulated, and ospA is downregulated. In the current study, using B. burgdorferi cultivated under varying conditions in BSK-H medium, we found that a decrease in pH, in conjunction with increases in temperature (e.g. 34 degrees C or 37 degrees C) and cell density, acted interdependently for the reciprocal expression of ospC and ospA. The lower pH (6.8), which induced the reciprocal expression of ospC and ospA in BSK-H medium, correlated with a drop in pH from 7.4 to 6.8 of tick midgut contents during tick feeding. In addition to ospC and ospA, other genes were found to be regulated in reciprocal fashion. Such genes were either ospC-like (e.g. ospF, mlp-8 and rpoS) (group I) or ospA-like (lp6.6 and p22) (group II); changes in expression occurred at the mRNA level. That the expression of rpoS, encoding a putative stress-related alternative sigma factor (sigma(s)), was ospC-like suggested that the expression of some of the group I genes may be controlled through sigma(s). The combined results prompt a model that allows for predicting the regulation of other B. burgdorferi genes that may be involved in spirochaete transmission, virulence or mammalian host immune responses.

  2. Role of the interplay between quorum sensing regulator VqsR and the Pseudomonas quinolone signal in mediating carbapenem tolerance in Pseudomonas aeruginosa.

    Science.gov (United States)

    Viducic, Darija; Murakami, Keiji; Amoh, Takashi; Ono, Tsuneko; Miyake, Yoichiro

    2017-06-01

    Pseudomonas aeruginosa coordinates its response to environmental conditions through activation of a quorum sensing (QS) system. In this study, we investigated the regulatory interaction between the QS transcriptional regulator VqsR and the Pseudomonas quinolone signal (PQS) through integration of sigma factor RpoS, and we addressed whether one of the pathways controlling carbapenem tolerance can be attributed to VqsR. We demonstrate that vqsR expression at the transcriptional level is regulated by pqsA, pqsR, and pqsE. Assessment of the transcriptional expression of vqsR, lasI, rhlI, and qscR in ΔpqsA and ΔpqsAΔrpoS mutants provided insight into pqsA- and rpoS-dependent regulation of vqsR and vqsR-controlled genes. Exogenously supplemented PQS reversed expression of vqsR and vqsR-controlled genes in the ΔpqsA mutant to wild-type levels, but failed to increase expression levels of lasI and qscR in the ΔpqsAΔrpoS mutant to levels observed in wild-type PAO1. The ΔvqsR mutant showed reduced survival when challenged with carbapenems compared to wild-type PAO1. Introduction of a pqsA mutation into the ΔvqsR mutant completely abolished its carbapenem-sensitive phenotype. We conclude that a regulatory link between PQS and vqsR exists, and that RpoS is important in their interaction. We also demonstrate that VqsR affects carbapenem tolerance. Copyright © 2017 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  3. The whole set of the constitutive promoters recognized by four minor sigma subunits of Escherichia coli RNA polymerase

    Science.gov (United States)

    Shimada, Tomohiro; Tanaka, Kan

    2017-01-01

    The promoter selectivity of Escherichia coli RNA polymerase (RNAP) is determined by the sigma subunit. The model prokaryote Escherichia coli K-12 contains seven species of the sigma subunit, each recognizing a specific set of promoters. For identification of the “constitutive promoters” that are recognized by each RNAP holoenzyme alone in the absence of other supporting factors, we have performed the genomic SELEX screening in vitro for their binding sites along the E. coli K-12 W3110 genome using each of the reconstituted RNAP holoenzymes and a collection of genome DNA segments of E. coli K-12. The whole set of constitutive promoters for each RNAP holoenzyme was then estimated based on the location of RNAP-binding sites. The first successful screening of the constitutive promoters was achieved for RpoD (σ70), the principal sigma for transcription of growth-related genes. As an extension, we performed in this study the screening of constitutive promoters for four minor sigma subunits, stationary-phase specific RpoS (σ38), heat-shock specific RpoH (σ32), flagellar-chemotaxis specific RpoF (σ28) and extra-cytoplasmic stress-response RpoE (σ24). The total number of constitutive promoters were: 129~179 for RpoS; 101~142 for RpoH; 34~41 for RpoF; and 77~106 for RpoE. The list of constitutive promoters were compared with that of known promoters identified in vivo under various conditions and using varieties of E. coli strains, altogether allowing the estimation of “inducible promoters” in the presence of additional supporting factors. PMID:28666008

  4. The strong efficiency of the Escherichia coli gapA P1 promoter depends on a complex combination of functional determinants.

    Science.gov (United States)

    Thouvenot, Benoit; Charpentier, Bruno; Branlant, Christiane

    2004-10-15

    The Escherichia coli multi-promoter region of the gapA gene ensures a high level of GAPDH (glyceraldehyde-3-phosphate dehydrogenase) production under various growth conditions. In the exponential phase of growth, gapA mRNAs are mainly initiated at the highly efficient gapA P1 promoter. In the present study, by using site-directed mutagenesis and chemical probing of the RPo (open complex) formed by Esigma70 (holoenzyme associated with sigma70) RNAP (RNA polymerase) at promoter gapA P1, we show that this promoter is an extended -10 promoter that needs a -35 sequence for activity. The -35 sequence compensates for the presence of a suboptimal -10 hexamer. A tract of thymine residues in the spacer region, which is responsible for a DNA distortion, is also required for efficient activity. We present the first chemical probing of an RPo formed at a promoter needing both a -10 extension and a -35 sequence. It reveals a complex array of RNAP-DNA interactions. In agreement with the fact that residue A-11 in the non-template strand is flipped out in a protein pocket in previously studied RPos, the corresponding A residue in gapA P1 promoter is protected in RPo and is essential for activity. However, in contrast with some of the previous findings on RPos formed at other promoters, the -12 A:T pair is opened. Strong contacts with RNAP occur both with the -35 sequence and the TG extension, so that the sigma4 and sigma2 domains may simultaneously contact the promoter DNA. RNAP-DNA interactions were also detected immediately downstream of the -35 hexamer and in a more distal upstream segment, reflecting a wrapping of RNAP by the core and upstream promoter DNA. Altogether, the data reveal that promoter gapA P1 is a very efficient promoter sharing common properties with both extended -10 and non-extended -10 promoters.

  5. Impact of a stress-inducible switch to mutagenic repair of DNA breaks on mutation in Escherichia coli.

    Science.gov (United States)

    Shee, Chandan; Gibson, Janet L; Darrow, Michele C; Gonzalez, Caleb; Rosenberg, Susan M

    2011-08-16

    Basic ideas about the constancy and randomness of mutagenesis that drives evolution were challenged by the discovery of mutation pathways activated by stress responses. These pathways could promote evolution specifically when cells are maladapted to their environment (i.e., are stressed). However, the clearest example--a general stress-response-controlled switch to error-prone DNA break (double-strand break, DSB) repair--was suggested to be peculiar to an Escherichia coli F' conjugative plasmid, not generally significant, and to occur by an alternative stress-independent mechanism. Moreover, mechanisms of spontaneous mutation in E. coli remain obscure. First, we demonstrate that this same mechanism occurs in chromosomes of starving F(-) E. coli. I-SceI endonuclease-induced chromosomal DSBs increase mutation 50-fold, dependent upon general/starvation- and DNA-damage-stress responses, DinB error-prone DNA polymerase, and DSB-repair proteins. Second, DSB repair is also mutagenic if the RpoS general-stress-response activator is expressed in unstressed cells, illustrating a stress-response-controlled switch to mutagenic repair. Third, DSB survival is not improved by RpoS or DinB, indicating that mutagenesis is not an inescapable byproduct of repair. Importantly, fourth, fully half of spontaneous frame-shift and base-substitution mutation during starvation also requires the same stress-response, DSB-repair, and DinB proteins. These data indicate that DSB-repair-dependent stress-induced mutation, driven by spontaneous DNA breaks, is a pathway that cells usually use and a major source of spontaneous mutation. These data also rule out major alternative models for the mechanism. Mechanisms that couple mutagenesis to stress responses can allow cells to evolve rapidly and responsively to their environment.

  6. Autocrine CSF-1R signaling drives mesothelioma chemoresistance via AKT activation.

    Science.gov (United States)

    Cioce, M; Canino, C; Goparaju, C; Yang, H; Carbone, M; Pass, H I

    2014-04-10

    Clinical management of malignant pleural mesothelioma (MPM) is very challenging because of the uncommon resistance of this tumor to chemotherapy. We report here increased expression of macrophage colony-stimulating-factor-1-receptor (M-CSF/CSF-1R) mRNA in mesothelioma versus normal tissue specimens and demonstrate that CSF-1R expression identifies chemoresistant cells of mesothelial nature in both primary cultures and mesothelioma cell lines. By using RNAi or ligand trapping, we demonstrate that the chemoresistance properties of those cells depend on autocrine CSF-1R signaling. At the single-cell level, the isolated CSF-1R(pos) cells exhibit a complex repertoire of pluripotency, epithelial-mesenchymal transition and detoxifying factors, which define a clonogenic, chemoresistant, precursor-like cell sub-population. The simple activation of CSF-1R in untransformed mesothelial cells is sufficient to confer clonogenicity and resistance to pemetrexed, hallmarks of mesothelioma. In addition, this induced a gene expression profile highly mimicking that observed in the MPM cells endogenously expressing the receptor and the ligands, suggesting that CSF-1R expression is mainly responsible for the phenotype of the identified cell sub-populations. The survival of CSF1R(pos) cells requires active AKT (v-akt murine thymoma viral oncogene homolog 1) signaling, which contributed to increased levels of nuclear, transcriptionally competent β-catenin. Inhibition of AKT reduced the transcriptional activity of β-catenin-dependent reporters and sensitized the cells to senescence-induced clonogenic death after pemetrexed treatment. This work expands what is known on the non-macrophage functions of CSF-1R and its role in solid tumors, and suggests that CSF-1R signaling may have a critical pathogenic role in a prototypical, inflammation-related cancer such as MPM and therefore may represent a promising target for therapeutic intervention.

  7. The whole set of the constitutive promoters recognized by four minor sigma subunits of Escherichia coli RNA polymerase.

    Directory of Open Access Journals (Sweden)

    Tomohiro Shimada

    Full Text Available The promoter selectivity of Escherichia coli RNA polymerase (RNAP is determined by the sigma subunit. The model prokaryote Escherichia coli K-12 contains seven species of the sigma subunit, each recognizing a specific set of promoters. For identification of the "constitutive promoters" that are recognized by each RNAP holoenzyme alone in the absence of other supporting factors, we have performed the genomic SELEX screening in vitro for their binding sites along the E. coli K-12 W3110 genome using each of the reconstituted RNAP holoenzymes and a collection of genome DNA segments of E. coli K-12. The whole set of constitutive promoters for each RNAP holoenzyme was then estimated based on the location of RNAP-binding sites. The first successful screening of the constitutive promoters was achieved for RpoD (σ70, the principal sigma for transcription of growth-related genes. As an extension, we performed in this study the screening of constitutive promoters for four minor sigma subunits, stationary-phase specific RpoS (σ38, heat-shock specific RpoH (σ32, flagellar-chemotaxis specific RpoF (σ28 and extra-cytoplasmic stress-response RpoE (σ24. The total number of constitutive promoters were: 129~179 for RpoS; 101~142 for RpoH; 34~41 for RpoF; and 77~106 for RpoE. The list of constitutive promoters were compared with that of known promoters identified in vivo under various conditions and using varieties of E. coli strains, altogether allowing the estimation of "inducible promoters" in the presence of additional supporting factors.

  8. Experimental measurements of motion cue effects on STOL approach tasks

    Science.gov (United States)

    Ringland, R. F.; Stapleford, R. L.

    1972-01-01

    An experimental program to investigate the effects of motion cues on STOL approach is presented. The simulator used was the Six-Degrees-of-Freedom Motion Simulator (S.01) at Ames Research Center of NASA which has ?2.7 m travel longitudinally and laterally and ?2.5 m travel vertically. Three major experiments, characterized as tracking tasks, were conducted under fixed and moving base conditions: (1) A simulated IFR approach of the Augmentor Wing Jet STOL Research Aircraft (AWJSRA), (2) a simulated VFR task with the same aircraft, and (3) a single-axis task having only linear acceleration as the motion cue. Tracking performance was measured in terms of the variances of several motion variables, pilot vehicle describing functions, and pilot commentary.

  9. Influences of the Tonga Subduction Zone on seafloor massive sulfide deposits along the Eastern Lau Spreading Center and Valu Fa Ridge

    Science.gov (United States)

    Evans, Guy N.; Tivey, Margaret K.; Seewald, Jeffrey S.; Wheat, C. Geoff

    2017-10-01

    This study investigates the morphology, mineralogy, and geochemistry of seafloor massive sulfide (SMS) deposits from six back-arc hydrothermal vent fields along the Eastern Lau Spreading Center (ELSC) and Valu Fa Ridge (VFR) in the context of endmember vent fluid chemistry and proximity to the Tonga Subduction Zone. To complement deposit geochemistry, vent fluid analyses of Cu, Zn, Ba, Pb and H2,(aq) were completed to supplement existing data and enable thermodynamic calculations of mineral saturation states at in situ conditions. Results document southward increases in the abundance of mantle-incompatible elements in hydrothermal fluids (Ba and Pb) and SMS deposits (Ba, Pb, As, and Sb), which is also expressed in the abundance of barite (BaSO4) and galena (PbS) in SMS deposits. These increases correspond to a decrease in distance between the ELSC/VFR and the Tonga Subduction Zone that correlates with a change in crustal lithology from back-arc basin basalt in the north to mixed andesite, rhyolite, and dacite in the south. Barite influences deposit morphology, contributing to the formation of horizontal flanges and squat terraces. Results are also consistent with a regional-scale lowering of hydrothermal reaction zone temperatures from north to south (except at the southernmost Mariner vent field) that leads to lower-temperature, higher-pH vent fluids relative to mid-ocean ridges of similar spreading rates (Mottl et al., 2011). These fluids are Cu- and Zn-poor and the deposits formed from these fluids are Cu-poor but Zn-rich. In contrast, at the Mariner vent field, higher-temperature and lower pH vent fluids are hypothesized to result from higher reaction zone temperatures and the localized addition of acidic magmatic volatiles (Mottl et al., 2011). The Mariner fluids are Cu- and Zn-rich and vent from SMS deposits that are rich in Cu but poor in Zn with moderate amounts of Pb. Thermodynamic calculations indicate that the contrasting metal contents of vent fluids

  10. Infecciones en viajeros internacionales

    Directory of Open Access Journals (Sweden)

    Perret P. Cecilia, Dra.

    2014-05-01

    Enfermedades gastrointestinales, síndrome febril y alteraciones dermatológicas son las principales causas de consulta en viajeros que retornan de un viaje. Giardia y Campylobacter son los agentes causales de diarrea más frecuentes. La causa de fiebre sistémica es la malaria, en aquellos que vienen de África Subsahariana; y dengue en aquellos que retornan de Sudamérica o Sudeste Asiático. Larva cutánea migrante es la principal consulta dermatológica. Especial atención se debe prestar a poblaciones de riesgo como los niños y VFR (visitas a familiares y parientes en la indicación de inmunizaciones y quimioprofilaxis.

  11. Flight management research utilizing an oculometer. [pilot scanning behavior during simulated approach and landing

    Science.gov (United States)

    Spady, A. A., Jr.; Kurbjun, M. C.

    1978-01-01

    This paper presents an overview of the flight management work being conducted using NASA Langley's oculometer system. Tests have been conducted in a Boeing 737 simulator to investigate pilot scan behavior during approach and landing for simulated IFR, VFR, motion versus no motion, standard versus advanced displays, and as a function of various runway patterns and symbology. Results of each of these studies are discussed. For example, results indicate that for the IFR approaches a difference in pilot scan strategy was noted for the manual versus coupled (autopilot) conditions. Also, during the final part of the approach when the pilot looks out-of-the-window he fixates on his aim or impact point on the runway and holds this point until flare initiation.

  12. How effective is aeration with vortex flow regulators? Pilot scale experiments

    Science.gov (United States)

    Wójtowicz, Patryk; Szlachta, Małgorzata

    2017-11-01

    Vortex flow regulators (VFR) are used in urban drainage systems as a replacement for traditional flow throttling devices. Vortex regulators are not only very efficient energy dissipators but also atomizers which are beneficial for sewer aeration. A deficit of dissolved oxygen can be a problem in both natural waters and sewerage. Hydrodynamic flow regulators can boost oxygen concentration preventing putrefaction and improving treatment of stormwater and wastewater. We were first to investigate the aeration efficiency of semi-commercial scale cylindrical vortex flow regulators to determine the potential of their application in environmental engineering and to propose modification to enhance the aeration capacity of basic designs. Different device geometries and arrangements of active outlets for both single and double discharge vortex regulators were tested in a recirculating system. In this study, we present a concise review of the current state of our extensive research on the aeration efficiency of vortex flow regulators and their application in sewerage systems.

  13. First optical identification of a supersoft X-ray source in M31

    Science.gov (United States)

    Nedialkov, P.; Orio, M.; Birkle, K.; Conselice, C.; Della Valle, M.; Greiner, J.; Magnier, E.; Tikhonov, N. A.

    2002-07-01

    We propose the first association of an optical counterpart with a luminous supersoft X-ray source in M31, RX J0044.0+4118, observed with ROSAT in July 1991. The PSPC position is at 1.6 arcsec angular distance from a candidate nova in outburst in September of 1990. This is interesting because the incidence of classical novae among supersoft X-ray sources is an open question. The proposed optical counterpart was measured at R =~ 17.7 in September of 1990, and it had faded to R> 19.2 when it was observed again after 70 days. The light curve was too sparsely monitored for definite conclusions on the speed class of the nova. No other variable objects with Vfr (130.79.128.5) or via http://cdsweb.u-strasbg.fr/cgi-bin/qcat?J/A+A/389/439

  14. Unusual temperature dependence of the dissociative electron attachment cross section of 2-thiouracil

    Energy Technology Data Exchange (ETDEWEB)

    Kopyra, Janina [Faculty of Science, Siedlce University, 3 Maja 54, 08-110 Siedlce (Poland); Abdoul-Carime, Hassan [Université de Lyon, F-69003 Lyon (France); Université Lyon 1, Villeurbanne (France); CNRS/IN2P3, UMR5822, Institut de Physique Nucléaire de Lyon, Lyon (France)

    2016-01-21

    At low energies (<3 eV), molecular dissociation is controlled by dissociative electron attachment for which the initial step, i.e., the formation of the transient negative ion, can be initiated by shape resonance or vibrational Feshbach resonance (VFR) mediated by the formation of a dipole bound anion. The temperature dependence for shape-resonances is well established; however, no experimental information is available yet on the second mechanism. Here, we show that the dissociation cross section for VFRs mediated by the formation of a dipole bound anion decreases as a function of a temperature. The change remains, however, relatively small in the temperature range of 370-440 K but it might be more pronounced at the extended temperature range.

  15. Pseudomonas aeruginosa quorum-sensing response in the absence of functional LasR and LasI proteins: the case of strain 148, a virulent dolphin isolate.

    Science.gov (United States)

    Morales, Estefanía; González-Valdez, Abigail; Servín-González, Luis; Soberón-Chávez, Gloria

    2017-07-03

    Pseudomonas aeruginosa is an opportunistic pathogen that presents a complex regulatory network called 'quorum-sensing', which is responsible for the transcription of genes coding for several traits implicated in its pathogenicity. Strain 148 is a dolphin isolate that has been shown to produce quorum-sensing-regulated virulence traits and to be virulent in a mouse model, despite the fact that it contains a 20-kbp deletion that eliminates from the chromosome the lasR gene and the lasI promoter. LasR is a key quorum-sensing transcriptional regulator that, when coupled with the autoinducer 3-oxo-dodecanoyl homoserine lactone (3O-C12-HSL) produced by LasI, activates transcription of genes coding for some virulence-associated traits such as elastase, lasI, rhlI and rhlR. RhlR is also a key quorum-sensing transcriptional regulator that, when interacting with the autoinducer butanoyl homoserine lactone (C4-HSL) that is produced by the synthase RhlI, activates the genes involved in the synthesis of some virulence-associated traits, as rhamnolipids and pyocyanin. We describe that in P. aeruginosa 148, the LasR/3O-C12-HSL-independent rhlR transcriptional activation is due to the release of the negative effect of Vfr (a CRP-ortholog) caused by the insertion of an IS element in vfr, and that rhlI transcription is driven from the rhlR promoter, forming the rhlR-I operon. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  16. CT-based compartmental quantification of adipose tissue versus body metrics in colorectal cancer patients

    Energy Technology Data Exchange (ETDEWEB)

    Nattenmueller, Johanna; Hoegenauer, Hanna; Grenacher, Lars; Kauczor, Hans-Ulrich [University Hospital, Department of Diagnostic and Interventional Radiology, Heidelberg (Germany); Boehm, Juergen; Ulrich, Cornelia [Huntsman Cancer Institute, Department of Population Health Sciences, Salt Lake City, UT (United States); Scherer, Dominique; Paskow, Michael; Gigic, Biljana; Schrotz-King, Petra [National Center for Tumor Diseases and German Cancer Research Center, Division of Preventive Oncology, Heidelberg (Germany)

    2016-11-15

    While obesity is considered a prognostic factor in colorectal cancer (CRC), there is increasing evidence that not simply body mass index (BMI) alone but specifically abdominal fat distribution is what matters. As part of the ColoCare study, this study measured the distribution of adipose tissue compartments in CRC patients and aimed to identify the body metric that best correlates with these measurements as a useful proxy for adipose tissue distribution. In 120 newly-diagnosed CRC patients who underwent multidetector computed tomography (CT), densitometric quantification of total (TFA), visceral (VFA), intraperitoneal (IFA), retroperitoneal (RFA), and subcutaneous fat area (SFA), as well as the M. erector spinae and psoas was performed to test the association with gender, age, tumor stage, metabolic equivalents, BMI, waist-to-height (WHtR) and waist-to-hip ratio (WHR). VFA was 28.8 % higher in men (p{sub VFA}<0.0001) and 30.5 % higher in patients older than 61 years (p{sub VFA}<0.0001). WHtR correlated best with all adipose tissue compartments (r{sub VFA}=0.69, r{sub TFA}=0.84, p<0.0001) and visceral-to-subcutaneous-fat-ratio (VFR, r{sub VFR}=0.22, p=<0.05). Patients with tumor stages III/IV showed significantly lower overall adipose tissue than I/II. Increased M. erector spinae mass was inversely correlated with all compartments. Densitometric quantification on CT is a highly reproducible and reliable method to show fat distribution across adipose tissue compartments. This distribution might be best reflected by WHtR, rather than by BMI or WHR. (orig.)

  17. Evaluation of Lipid Metabolism and Nutritional Status in Male Goalball Players.

    Science.gov (United States)

    Gawlik, Krystyna; Zwierzchowska, Anna; Rosołek, Barbara

    2015-11-22

    Lipid disorders, obesity and overweight are considered one of the most important modifiable cardiovascular risk factors. Population surveys carried out in Poland have demonstrated a tendency for lipid disorders to occur in 70% and overweight and obesity in more than half of Poles. No such studies have been conducted in groups of people with vision impairment so far. Yet, regular involvement of visually impaired people in sports is likely to reduce cardiovascular risk. Therefore, the authors attempted to evaluate the lipid profile and nutritional status of male goalball players. Thirty two blind or visually impaired male goalball players aged 20 to 45 years participated in the study during which somatic variables (BH, BM, WC, VFR, BMI) and the lipid profile (TC, LDL, HDL, TG) were evaluated. Overweight was found in 40.6% of athletes, with obesity being at the level of 9.3%. A high correlation was found between visceral fat and the BMI (r=0.7; p<0.001), as well as between visceral fat and WC (r=0.8; p<0.001). Abnormal total cholesterol levels were recorded for LDL (22% of study participants), HDL (17%) and triglycerides (13%). Lower levels of individual components of lipid profiles (and higher levels for HDL) were found in athletes with a normal BMI. A correlation was found between the BMI and TG (r=0.4, p<0.01), WC and TG (r=0.4, p<0.01), VFR and LDL ( r=0.4, p<0.05) and TG (r=0.5, p<0.001). The percentage of overweight and obese subjects with vision impairment was lower compared to the general population of men in Poland, with a more beneficial lipid profile. Regular physical activity of the study participants is likely to have a positive effect on their health.

  18. Evaluation of Lipid Metabolism and Nutritional Status in Male Goalball Players

    Directory of Open Access Journals (Sweden)

    Gawlik Krystyna

    2015-12-01

    Full Text Available Lipid disorders, obesity and overweight are considered one of the most important modifiable cardiovascular risk factors. Population surveys carried out in Poland have demonstrated a tendency for lipid disorders to occur in 70% and overweight and obesity in more than half of Poles. No such studies have been conducted in groups of people with vision impairment so far. Yet, regular involvement of visually impaired people in sports is likely to reduce cardiovascular risk. Therefore, the authors attempted to evaluate the lipid profile and nutritional status of male goalball players. Thirty two blind or visually impaired male goalball players aged 20 to 45 years participated in the study during which somatic variables (BH, BM, WC, VFR, BMI and the lipid profile (TC, LDL, HDL, TG were evaluated. Overweight was found in 40.6% of athletes, with obesity being at the level of 9.3%. A high correlation was found between visceral fat and the BMI (r=0.7; p<0.001, as well as between visceral fat and WC (r=0.8; p<0.001. Abnormal total cholesterol levels were recorded for LDL (22% of study participants, HDL (17% and triglycerides (13%. Lower levels of individual components of lipid profiles (and higher levels for HDL were found in athletes with a normal BMI. A correlation was found between the BMI and TG (r=0.4, p<0.01, WC and TG (r=0.4, p<0.01, VFR and LDL (r=0.4, p<0.05 and TG (r=0.5, p<0.001. The percentage of overweight and obese subjects with vision impairment was lower compared to the general population of men in Poland, with a more beneficial lipid profile. Regular physical activity of the study participants is likely to have a positive effect on their health.

  19. Mosquito Exposure and Chikungunya and Dengue Infection Among Travelers During the Chikungunya Outbreak in the Americas.

    Science.gov (United States)

    Lindholm, David A; Myers, Todd; Widjaja, Susana; Grant, Edward M; Telu, Kalyani; Lalani, Tahaniyat; Fraser, Jamie; Fairchok, Mary; Ganesan, Anuradha; Johnson, Mark D; Kunz, Anjali; Tribble, David R; Yun, Heather C

    2017-04-01

    AbstractTravelers are at risk for arbovirus infection. We prospectively enrolled 267 Department of Defense beneficiaries traveling to chikungunya-outbreak regions in the Americas between December 2013 and May 2015 and assessed travel characteristics and serologic exposure to chikungunya virus (CHIKV) and dengue virus (DENV). Ten ill-returning travelers were also assessed retrospectively. Self-reported mosquito exposure was common (64% of 198 evaluable travelers saw mosquitoes; 53% of 201 reported ≥ 1 bite). Increased exposure was associated with active-duty travelers (odds ratio [OR] = 2.6 [1.3-5.4] for seeing mosquitoes) or travelers visiting friends and relatives (VFR) (OR = 3.5 [1.0-10.0] for high-intensity bite exposure). Arbovirus infection was defined as seroconversion on plaque reduction neutralization testing (PRNT) of pre- and posttravel sera. For ill subjects enrolled posttravel, infection was defined by a positive convalescent PRNT and/or a positive reverse transcription polymerase chain reaction for CHIKV or DENV. We identified seven cases of arbovirus infection: four with CHIKV, five with DENV, and two with both. The composite attack rate for CHIKV and DENV infection was 3.7% of 108 evaluable, immunologically naïve, prospectively assessed travelers; there was serologic and/or polymerase chain reaction evidence of arbovirus infection in three of four evaluable (three of 10 total) ill-returning travelers. We identified both symptomatic and asymptomatic cases. Military purpose of travel and VFR travel accounted for five of seven cases. Pretravel counseling is important and should target higher risk groups. Given a shared vector between CHIKV, DENV, and Zika virus (ZIKV), this study can also help guide counseling for travelers to ZIKV-outbreak regions.

  20. The role of travel in measles outbreaks in Australia - An enhanced surveillance study.

    Science.gov (United States)

    MacIntyre, C R; Karki, S; Sheikh, M; Zwar, N; Heywood, A E

    2016-08-17

    Many developed countries, like Australia, maintain a high population level immunity against measles, however, there remains a risk of acquisition of measles in non-immune travellers and subsequent importation into Australia leading to localised outbreaks. In this study, we estimate the incidence of measles and describe characteristics including immunisation and pre-travel health seeking behaviour of notified cases of measles in New South Wales and Victoria, Australia between February 2013 and January 2014. Cases were followed up by telephone interview using a questionnaire to collect information of demographic and travel characteristics. In NSW, the incidence was highest in age group 0-9years (20/million population) whereas in Victoria the highest incidence was observed in 10-19 (23/million population) years group. Out of 44 cases interviewed, 25 (56.8%) had history of travel outside of Australia during or immediately prior to the onset of measles. Holiday (60%) was the main reason for travel with 44% (11/25) reporting visiting friends and relatives (VFR) during the trip. The major reason described for not seeking prior medical advice before travel were "no perceived risk of diseases" (41%) and "previous overseas travel without any problem" (41%). Of the 25 measles cases with recent overseas travel during the incubation period, one reported a measles vaccine prior to their recent trip. Four cases were children of parents who refused vaccination. Twenty out of 25 (80.0%) had attended mass gathering events. Young adults and VFR travellers should be a high priority for preventive strategies in order to maintain measles elimination status. Copyright © 2016. Published by Elsevier Ltd.

  1. A cost-minimization analysis of an RCT of three retention methods.

    Science.gov (United States)

    Tynelius, Gudrun Edman; Lilja-Karlander, Eva; Petrén, Sofia

    2014-08-01

    There are few cost evaluation studies of orthodontic retention treatment. The aim of this study was to compare the costs in a randomized controlled trial of three retention methods during 2 years of retention treatment. To determine which alternative has the lower cost, a cost-minimization analysis (CMA) was undertaken, based on that the outcome of the treatment alternatives was equivalent. The study comprised 75 patients in 3 groups consisting of 25 each. The first group had a vacuum-formed retainer (VFR) in the maxilla and a cuspid retainer in the mandible (group V-CTC), the second group had a VFR in the maxilla combined with stripping of the incisors and cuspids in the mandible (group V-S), and the third group had a prefabricated positioner (group P). Direct cost (premises, staff salaries, material and laboratory costs) and indirect costs (loss of time at school) were calculated. Societal costs were defined as the sum of direct and indirect costs. The societal costs/patient for scheduled appointments for 2 years of retention treatment in group V-CTC were €497, group V-S €451 and group P €420. Societal costs for unscheduled appointments in group V-CTC were €807 and in group V-S €303. In group P, there were no unscheduled appointments. After 2 years of retention in compliant patients, the cuspid retainer was the least cost-effective retention appliance. The CMA showed that for a clinically similar result, there were differences in societal costs, but treatment decisions should always be performed on an individual basis. © The Author 2013. Published by Oxford University Press on behalf of the European Orthodontic Society. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  2. Clinical research on the development of posterior vitreous detachments after coaxial microincision phacoemulsification

    Directory of Open Access Journals (Sweden)

    Rong Xu

    2013-09-01

    Full Text Available AIM: To explore the development process of when posterior vitreous detachments(PVDsafter coaxial micro-incision phacoemulsification and aspiration combined with foldable intraocular lens implantation.METHODS: Clinical data of 917 patients with cataract were collected after coaxial microincision phacoemulsification and aspiration combined with foldable intraocular lens implantation. The 917 operating eyes forms observation group and the 917 corresponding normal eyes serve as control group. All patients underwent a comprehensive ocular examination before surgery. The vitreous and retinal condition was tested by B-scan ultrasonography and biomicroscope with a 90-diopter(Dpreset lens after 1 week and 1, 3, 6, and 12 months postoperatively. RESULTS: The incident rate of eyes in the observation group that developed PVD after operation were 8(0.9%, 31(3.4%, 53(5.8%, 78(8.5%, 129(14.1%within 1 week and 1, 3, 6, 12 months, respectively. Ten(7.8%of the 129 eyes in which PVD developed during the one year follow-up period had new retinal breaks with or without a retinal detachment; One(0.1%of the 788 eyes in which a PVD did not developed during the follow-up period had new retinal tear with or without a retinal detachment; these percentages were statistically significant(P=0.009. Eight(14.8%of 54 eyes with lattice degeneration and 2(2.7%of 75 eyes without lattice degeneration had retinal tears associated with a PVD; the difference was statistically significant(P=0.004.CONCLUSION: Development of PVDs seems to accelerate after coaxial microincision phacoemulsification and linearly accumulated for 1 year postoperatively. Approximately 7.8% of eyes with a PVD had retinal tears. Compared with patients having no lattice degeneration, Patients' eyes with lattice degeneration had a 5.5-fold higher risk of developing retinal breaks associated with a PVD. This indicates long-term follow-up is necessary after phacoemulsification is performed.

  3. Biofilms promote survival and virulence of Salmonella enterica sv. Tennessee during prolonged dry storage and after passage through an in vitro digestion system.

    Science.gov (United States)

    Aviles, Bryan; Klotz, Courtney; Eifert, Joseph; Williams, Robert; Ponder, Monica

    2013-04-01

    Salmonella enterica serotypes have been linked to outbreaks associated with low water activity foods. While the biofilm-forming abilities of Salmonella improve its survival during thermal processing and sanitation it is unclear whether biofilms enhance survival to desiccation and gastric stresses. The purpose of this study was to quantify the effect of physiological state (planktonic versus biofilm) and prior exposure to desiccation and storage in dry milk powder on Salmonella survival and gene expression after passage through an in vitro digestion model. Planktonic cells of Salmonella enterica serotype Tennessee were deposited onto membranes while biofilms were formed on glass beads. The cells were subsequently dried at room temperature and stored in dried milk powder (a(w)=0.3) for up to 30 days. Salmonella survival was quantified by serial dilution onto Brilliant Green Agar before desiccation, after desiccation, after 1-day storage and after 30-day storage. At each sampling period both physiological states were tested for survival through a simulated gastrointestinal system. RNA was extracted at the identical time points and Quantitative Real-Time PCR was used to determine relative expression for genes associated with stress response (rpoS, otsB), virulence (hilA, invA, sipC) and a housekeeping gene 16S rRNA. The physiological state and length of storage affected the survival and gene expression of Salmonella within the desiccated milk powder environment and after passage through an in vitro digestion system (pSalmonella genomes detected by qPCR were not significantly different suggesting entry of the planktonic cells of S. Tennessee into a viable but non-culturable state. The increased expression of stress response genes rpoS and otsB correlated with survival, indicating cross-protection to low water activity and acid stress. Increased expression of virulence-associated genes was seen in cells exposed to dry storage for short periods, however the largest amount

  4. E Unibus Plurum: genomic analysis of an experimentally evolved polymorphism in Escherichia coli.

    Directory of Open Access Journals (Sweden)

    Margie A Kinnersley

    2009-11-01

    Full Text Available Microbial populations founded by a single clone and propagated under resource limitation can become polymorphic. We sought to elucidate genetic mechanisms whereby a polymorphism evolved in Escherichia coli under glucose limitation and persisted because of cross-feeding among multiple adaptive clones. Apart from a 29 kb deletion in the dominant clone, no large-scale genomic changes distinguished evolved clones from their common ancestor. Using transcriptional profiling on co-evolved clones cultured separately under glucose-limitation we identified 180 genes significantly altered in expression relative to the common ancestor grown under similar conditions. Ninety of these were similarly expressed in all clones, and many of the genes affected (e.g., mglBAC, mglD, and lamB are in operons coordinately regulated by CRP and/or rpoS. While the remaining significant expression differences were clone-specific, 93% were exhibited by the majority clone, many of which are controlled by global regulators, CRP and CpxR. When transcriptional profiling was performed on adaptive clones cultured together, many expression differences that distinguished the majority clone cultured in isolation were absent, suggesting that CpxR may be activated by overflow metabolites removed by cross-feeding strains in co-culture. Relative to their common ancestor, shared expression differences among adaptive clones were partly attributable to early-arising shared mutations in the trans-acting global regulator, rpoS, and the cis-acting regulator, mglO. Gene expression differences that distinguished clones may in part be explained by mutations in trans-acting regulators malT and glpK, and in cis-acting sequences of acs. In the founder, a cis-regulatory mutation in acs (acetyl CoA synthetase and a structural mutation in glpR (glycerol-3-phosphate repressor likely favored evolution of specialists that thrive on overflow metabolites. Later-arising mutations that led to specialization

  5. Uropathogenic Escherichia coli Metabolite-Dependent Quiescence and Persistence May Explain Antibiotic Tolerance during Urinary Tract Infection.

    Science.gov (United States)

    Leatham-Jensen, Mary P; Mokszycki, Matthew E; Rowley, David C; Deering, Robert; Camberg, Jodi L; Sokurenko, Evgeni V; Tchesnokova, Veronika L; Frimodt-Møller, Jakob; Krogfelt, Karen A; Leth Nielsen, Karen; Frimodt-Møller, Niels; Sun, Gongqin; Cohen, Paul S

    2016-01-01

    In the present study, it is shown that although Escherichia coli CFT073, a human uropathogenic (UPEC) strain, grows in liquid glucose M9 minimal medium, it fails to grow on glucose M9 minimal medium agar plates seeded with ≤10(6) CFU. The cells on glucose plates appear to be in a "quiescent" state that can be prevented by various combinations of lysine, methionine, and tyrosine. Moreover, the quiescent state is characteristic of ~80% of E. coli phylogenetic group B2 multilocus sequence type 73 strains, as well as 22.5% of randomly selected UPEC strains isolated from community-acquired urinary tract infections in Denmark. In addition, E. coli CFT073 quiescence is not limited to glucose but occurs on agar plates containing a number of other sugars and acetate as sole carbon sources. It is also shown that a number of E. coli CFT073 mini-Tn5 metabolic mutants (gnd, gdhA, pykF, sdhA, and zwf) are nonquiescent on glucose M9 minimal agar plates and that quiescence requires a complete oxidative tricarboxylic acid (TCA) cycle. In addition, evidence is presented that, although E. coli CFT073 quiescence and persistence in the presence of ampicillin are alike in that both require a complete oxidative TCA cycle and each can be prevented by amino acids, E. coli CFT073 quiescence occurs in the presence or absence of a functional rpoS gene, whereas maximal persistence requires a nonfunctional rpoS. Our results suggest that interventions targeting specific central metabolic pathways may mitigate UPEC infections by interfering with quiescence and persistence. IMPORTANCE Recurrent urinary tract infections (UTIs) affect 10 to 40% of women. In up to 77% of those cases, the recurrent infections are caused by the same uropathogenic E. coli (UPEC) strain that caused the initial infection. Upon infection of urothelial transitional cells in the bladder, UPEC appear to enter a nongrowing quiescent intracellular state that is thought to serve as a reservoir responsible for recurrent

  6. Toxicogenomic response of Pseudomonas aeruginosa to ortho-phenylphenol

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    Toghrol Freshteh

    2008-10-01

    Full Text Available Abstract Background Pseudomonas aeruginosa (P. aeruginosa is the most common opportunistic pathogen implicated in nosocomial infections and in chronic lung infections in cystic fibrosis patients. Ortho-phenylphenol (OPP is an antimicrobial agent used as an active ingredient in several EPA registered disinfectants. Despite its widespread use, there is a paucity of information on its target molecular pathways and the cellular responses that it elucidates in bacteria in general and in P. aeruginosa in particular. An understanding of the OPP-driven gene regulation and cellular response it elicits will facilitate more effective utilization of this antimicrobial and possibly lead to the development of more effective disinfectant treatments. Results Herein, we performed a genome-wide transcriptome analysis of the cellular responses of P. aeruginosa exposed to 0.82 mM OPP for 20 and 60 minutes. Our data indicated that OPP upregulated the transcription of genes encoding ribosomal, virulence and membrane transport proteins after both treatment times. After 20 minutes of exposure to 0.82 mM OPP, genes involved in the exhibition of swarming motility and anaerobic respiration were upregulated. After 60 minutes of OPP treatment, the transcription of genes involved in amino acid and lipopolysaccharide biosynthesis were upregulated. Further, the transcription of the ribosome modulation factor (rmf and an alternative sigma factor (rpoS of RNA polymerase were downregulated after both treatment times. Conclusion Results from this study indicate that after 20 minutes of exposure to OPP, genes that have been linked to the exhibition of anaerobic respiration and swarming motility were upregulated. This study also suggests that the downregulation of the rmf and rpoS genes may be indicative of the mechanism by which OPP causes decreases in cell viability in P. aeruginosa. Consequently, a protective response involving the upregulation of translation leading to the

  7. Uropathogenic Escherichia coli Metabolite-Dependent Quiescence and Persistence May Explain Antibiotic Tolerance during Urinary Tract Infection

    Science.gov (United States)

    Leatham-Jensen, Mary P.; Mokszycki, Matthew E.; Rowley, David C.; Deering, Robert; Camberg, Jodi L.; Sokurenko, Evgeni V.; Tchesnokova, Veronika L.; Frimodt-Møller, Jakob; Leth Nielsen, Karen; Sun, Gongqin

    2016-01-01

    ABSTRACT In the present study, it is shown that although Escherichia coli CFT073, a human uropathogenic (UPEC) strain, grows in liquid glucose M9 minimal medium, it fails to grow on glucose M9 minimal medium agar plates seeded with ≤106 CFU. The cells on glucose plates appear to be in a “quiescent” state that can be prevented by various combinations of lysine, methionine, and tyrosine. Moreover, the quiescent state is characteristic of ~80% of E. coli phylogenetic group B2 multilocus sequence type 73 strains, as well as 22.5% of randomly selected UPEC strains isolated from community-acquired urinary tract infections in Denmark. In addition, E. coli CFT073 quiescence is not limited to glucose but occurs on agar plates containing a number of other sugars and acetate as sole carbon sources. It is also shown that a number of E. coli CFT073 mini-Tn5 metabolic mutants (gnd, gdhA, pykF, sdhA, and zwf) are nonquiescent on glucose M9 minimal agar plates and that quiescence requires a complete oxidative tricarboxylic acid (TCA) cycle. In addition, evidence is presented that, although E. coli CFT073 quiescence and persistence in the presence of ampicillin are alike in that both require a complete oxidative TCA cycle and each can be prevented by amino acids, E. coli CFT073 quiescence occurs in the presence or absence of a functional rpoS gene, whereas maximal persistence requires a nonfunctional rpoS. Our results suggest that interventions targeting specific central metabolic pathways may mitigate UPEC infections by interfering with quiescence and persistence. IMPORTANCE Recurrent urinary tract infections (UTIs) affect 10 to 40% of women. In up to 77% of those cases, the recurrent infections are caused by the same uropathogenic E. coli (UPEC) strain that caused the initial infection. Upon infection of urothelial transitional cells in the bladder, UPEC appear to enter a nongrowing quiescent intracellular state that is thought to serve as a reservoir responsible

  8. Pyrrolnitrin and Hydrogen Cyanide Production by Pseudomonas chlororaphis Strain PA23 Exhibits Nematicidal and Repellent Activity against Caenorhabditis elegans.

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    Munmun Nandi

    Full Text Available Pseudomonas chlororaphis strain PA23 is a biocontrol agent able to suppress growth of the fungal pathogen Sclerotinia sclerotiorum. This bacterium produces an arsenal of exometabolites including pyrrolnitrin (PRN, phenazine (PHZ, hydrogen cyanide (HCN, and degradative enzymes. Production of these compounds is controlled at both the transcriptional and posttranscriptional levels by the Gac-Rsm system, RpoS, PsrA, and the Phz quorum-sensing system. Beyond pathogen-suppression, the success of a biocontrol agent is dependent upon its ability to establish itself in the environment where predation by bacterivorous organisms, including nematodes, may threaten persistence. The focus of this study was to investigate whether PA23 is able to resist grazing by Caenorhabditis elegans and to define the role played by exoproducts in the bacterial-nematode interaction. We discovered that both PRN and HCN contribute to fast- and slow-killing of C. elegans. HCN is well-established as having lethal effects on C. elegans; however, PRN has not been reported to be nematicidal. Exposure of L4 stage nematodes to purified PRN reduced nematode viability in a dose-dependent fashion and led to reduced hatching of eggs laid by gravid adults. Because bacterial metabolites can act as chemoattractants or repellents, we analyzed whether PA23 exhibited attractant or repulsive properties towards C. elegans. Both PRN and HCN were found to be potent repellents. Next we investigated whether the presence of C. elegans would elicit changes in PA23 gene activity. Co-culturing the two organisms increased expression of a number of genes associated with biocontrol, including phzA, hcnA, phzR, phzI, rpoS and gacS. Exoproduct analysis showed that PHZ and autoinducer signals were upregulated, consistent with the gene expression profiles. Collectively, these findings indicate that PA23 is able to sense the presence of C. elegans and it is able to both repel and kill the nematodes, which

  9. The novel sigma factor-like regulator RpoQ controls luminescence, chitinase activity, and motility in Vibrio fischeri.

    Science.gov (United States)

    Cao, Xiaodan; Studer, Sarah V; Wassarman, Karen; Zhang, Yuanxing; Ruby, Edward G; Miyashiro, Tim

    2012-01-01

    Vibrio fischeri, the bacterial symbiont of the Hawaiian bobtail squid, Euprymna scolopes, uses quorum sensing to control genes involved in bioluminescence, host colonization, and other biological processes. Previous work has shown that AinS/R-directed quorum sensing also regulates the expression of rpoQ (VF_A1015), a gene annotated as an RpoS-like sigma factor. In this study, we demonstrate using phylogenetics that RpoQ is related to, but distinct from, the stationary-phase sigma factor RpoS. Overexpression of rpoQ results in elevated chitinase activity but decreased motility and luminescence, three activities associated with symbiosis. The reduction in bacterial luminescence associated with the overexpression of rpoQ occurs both in culture and within the light-emitting organ of the squid host. This suppression of bioluminescence is due to the repression of the luxICDABEG promoter. Our results highlight RpoQ as a novel regulatory component, embedded in the quorum-signaling network that controls several biological processes in V. fischeri. Quorum signaling is a widely occurring phenomenon that functions in diverse bacterial taxa. It is most often found associated with species that interact with animal or plant hosts, either as mutualists or pathogens, and controls the expression of genes critical to tissue colonization. We present the discovery of rpoQ, which encodes a new regulatory component in the quorum-signaling pathway of Vibrio fischeri. RpoQ is a novel protein in the RpoS family of stationary-phase sigma factors. Unlike many other regulatory proteins involved in the quorum-signaling pathways of the Vibrionaceae, the distribution of RpoQ appears to be restricted to only two closely related species. The role of this regulator is to enhance some quorum-signaling outputs (motility) while suppressing others (luminescence). We propose that RpoQ may be a recently evolved or acquired component in V. fischeri that provides this organism with an additional level of

  10. Pyrrolnitrin and Hydrogen Cyanide Production by Pseudomonas chlororaphis Strain PA23 Exhibits Nematicidal and Repellent Activity against Caenorhabditis elegans

    Science.gov (United States)

    Nandi, Munmun; Selin, Carrie; Brassinga, Ann Karen C.; Belmonte, Mark F.; Fernando, W. G. Dilantha; Loewen, Peter C.; de Kievit, Teresa R.

    2015-01-01

    Pseudomonas chlororaphis strain PA23 is a biocontrol agent able to suppress growth of the fungal pathogen Sclerotinia sclerotiorum. This bacterium produces an arsenal of exometabolites including pyrrolnitrin (PRN), phenazine (PHZ), hydrogen cyanide (HCN), and degradative enzymes. Production of these compounds is controlled at both the transcriptional and posttranscriptional levels by the Gac-Rsm system, RpoS, PsrA, and the Phz quorum-sensing system. Beyond pathogen-suppression, the success of a biocontrol agent is dependent upon its ability to establish itself in the environment where predation by bacterivorous organisms, including nematodes, may threaten persistence. The focus of this study was to investigate whether PA23 is able to resist grazing by Caenorhabditis elegans and to define the role played by exoproducts in the bacterial-nematode interaction. We discovered that both PRN and HCN contribute to fast- and slow-killing of C. elegans. HCN is well-established as having lethal effects on C. elegans; however, PRN has not been reported to be nematicidal. Exposure of L4 stage nematodes to purified PRN reduced nematode viability in a dose-dependent fashion and led to reduced hatching of eggs laid by gravid adults. Because bacterial metabolites can act as chemoattractants or repellents, we analyzed whether PA23 exhibited attractant or repulsive properties towards C. elegans. Both PRN and HCN were found to be potent repellents. Next we investigated whether the presence of C. elegans would elicit changes in PA23 gene activity. Co-culturing the two organisms increased expression of a number of genes associated with biocontrol, including phzA, hcnA, phzR, phzI, rpoS and gacS. Exoproduct analysis showed that PHZ and autoinducer signals were upregulated, consistent with the gene expression profiles. Collectively, these findings indicate that PA23 is able to sense the presence of C. elegans and it is able to both repel and kill the nematodes, which should facilitate

  11. Transcriptomic and Metabolomic Analysis Revealed Multifaceted Effects of Phage Protein Gp70.1 on Pseudomonas aeruginosa

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    Xia Zhao

    2016-09-01

    Full Text Available The impact of phage infection on the host cell is severe. In order to take over the cellular machinery, some phage proteins were produced to shut off the host biosynthesis early in the phage infection. The discovery and identification of these phage-derived inhibitors have a significant prospect of application in antibacterial treatment. This work presented a phage protein, gp70.1, with nonspecific inhibitory effects on Pseudomonas aeruginosa (P. aeruginosa and Escherichia coli (E. coli. Gp70.1 was encoded by early gene – orf 70.1 from P. aeruginosa phage PaP3. The P. aeruginosa with a plasmid encoding gp70.1 showed with delayed growth and had the appearance of a small colony. The combination of multifaceted analysis including microarray-based transcriptomic analysis, RT-qPCR, nuclear magnetic resonance (NMR spectroscopy-based metabolomics and phenotype experiments were performed to investigate the effects of gp70.1 on P. aeruginosa. A total of 178 genes of P. aeruginosa mainly involved in extracellular function and metabolism were differentially expressed in the presence of gp70.1 at 3 examined time points. Furthermore, our results indicated that gp70.1 had an extensive impact on the extracellular phenotype of P. aeruginosa, such as motility, pyocyanin, extracellular protease, polysaccharide, and cellulase. For the metabolism of P. aeruginosa, the main effect of gp70.1 was the reduction of amino acid consumption. Finally, the RNA polymerase sigma factor RpoS was identified as a potential cellular target of gp70.1. Gp70.1 was the first bacterial inhibitor identified from Pseudomonas aeruginosa phage PaP3. It was also the first phage protein that interacted with the global regulator RpoS of bacteria. Our results indicated the potential value of gp70.1 in antibacterial applications. This study preliminarily revealed the biological function of gp70.1 and provided a reference for the study of other phage genes sharing similarities with orf70.1.

  12. Survival and expression of acid resistance genes in Shiga toxin-producing Escherichia coli acid adapted in pineapple juice and exposed to synthetic gastric fluid.

    Science.gov (United States)

    Kim, G-H; Fratamico, P; Breidt, F; Oh, D-H

    2016-11-01

    The aim of this research was to determine the ability of non-O157 Shiga toxin-producing Escherichia coli (STEC) serogroups to survive with exposure to synthetic gastric fluid (SGF) after adaptation to pineapple juice (PJ) at room and refrigerated temperatures compared to E. coli O157:H7 and to examine the relative transcriptional expression of acid resistance (AR) genes, rpoS, gadA and adiA. Resistant and sensitive strains belonging to five different STEC serogroups (O26, O103, O104, O111 and O157; n = 10) were used in this study. All strains were adapted in PJ (pH 3·8) stored at 4 and 20°C for 24 h, and then the relative transcription levels of genes in all strains were quantified using a real-time quantitative-PCR assay. After adaptation in PJ, the STEC strains were exposed to SGF (pH 1·5 and 2·0) at 37°C for 2 h. Generally, the STEC adapted in PJ at 4°C displayed enhanced survival compared to acid adaptation in PJ at 20°C and nonadapted controls with exposure to SGF (P < 0·05). Moreover, resistant strains exhibited higher survival rates compared to sensitive strains (P < 0·05). Overall, adaptation at 4°C resulted in significantly (P < 0·05) enhanced gene expression levels in PJ, and transcript levels of gadA were higher than those of the rpoS and adiA genes. The up-regulation of AR genes due to adaptation in PJ at low temperature may increase STEC survival in acidic environments such as the gastrointestinal tract. Some non-O157 STEC strains, including serotypes O103:H2 and O111:H8, showed relatively high AR levels similar to those of STEC O157:H7. Induction of AR genes in acidic fruit juice, and potentially in other acidic foods may increase the risk of foodborne illness by non-O157 STEC serogroups. © 2016 The Society for Applied Microbiology.

  13. Distribution of mega fauna on sulfide edifices on the Eastern Lau Spreading Center and Valu Fa Ridge

    Science.gov (United States)

    Sen, Arunima; Becker, Erin L.; Podowski, Elizabeth L.; Wickes, Leslie N.; Ma, Shufen; Mullaugh, Katherine M.; Hourdez, Stéphane; Luther, George W.; Fisher, Charles R.

    2013-02-01

    Hydrothermal vent sulfide edifices contain some of the most extreme thermal and chemical conditions in which animals are able to live. As a result, sulfide edifices in the East Pacific Rise, Juan de Fuca Ridge, and Mid Atlantic Ridge vent systems often contain distinct faunal assemblages. In this study, we used high-resolution imagery and in-situ physico-chemical measurements within the context of a Geographic Information System (GIS) to examine community structure and niche differentiation of dominant fauna on sulfide edifices in the Eastern Lau Spreading Center (ELSC) and Valu Fa Ridge (VFR) in the Western Pacific Ocean. Our results show that ELSC and VFR sulfide edifices host two distinct types of communities. One type, that covers the majority of sulfide edifice faces, is overall very similar to nearby lava communities and biomass is dominated by the same chemoautotrophic symbiont-containing molluscs that dominate lava communities, namely the provannid gastropods Alviniconcha spp. and Ifremeria nautilei and the mytilid bivalve Bathymodiolus brevior. The spatial distribution of the dominant molluscs is often a variation of the pattern of concentric rings observed on lavas, with Alviniconcha spp. at the tops of edifices where exposure to vent flow is the highest, and I. nautilei and B. brevior below. Our physico-chemical measurements indicate that because of rapid dispersion of vent fluid, habitable area for symbiont-containing fauna is quite limited on sulfide edifices, and the realized niches of the mollusc groups are narrower on sulfide edifices than on lavas. We suggest that competition plays an important role in determining the realized distributions of the mollusc groups on edifices. The other habitat, present in small patches of presumably hot, new anhydrite, is avoided by the dominant symbiont-containing molluscs and inhabited by crabs, shrimp and polynoids that are likely more heat tolerant. The ratio of sulfide concentration to temperature anomaly of

  14. Structural insights into the recognition of the internal A-rich linker from OxyS sRNA by Escherichia coli Hfq.

    Science.gov (United States)

    Wang, Lijun; Wang, Weiwei; Li, Fudong; Zhang, Jiahai; Wu, Jihui; Gong, Qingguo; Shi, Yunyu

    2015-02-27

    Small RNA OxyS is induced during oxidative stress in Escherichia coli and it is an Hfq-dependent negative regulator of mRNA translation. OxyS represses the translation of fhlA and rpoS mRNA, which encode the transcriptional activator and σ(s) subunit of RNA polymerase, respectively. However, little is known regarding how Hfq, an RNA chaperone, interacts with OxyS at the atomic level. Here, using fluorescence polarization and tryptophan fluorescence quenching assays, we verified that the A-rich linker region of OxyS sRNA binds Hfq at its distal side. We also report two crystal structures of Hfq in complex with A-rich RNA fragments from this linker region. Both of these RNA fragments bind to the distal side of Hfq and adopt a different conformation compared with those previously reported for the (A-R-N)n tripartite recognition motif. Furthermore, using fluorescence polarization, electrophoresis mobility shift assays and in vivo translation assays, we found that an Hfq mutant, N48A, increases the binding affinity of OxyS for Hfq in vitro but is defective in the negative regulation of fhlA translation in vivo, suggesting that the normal function of OxyS depends on the details of the interaction with Hfq that may be related to the rapid recycling of Hfq in the cell. © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

  15. Expression of signal transduction system encoding genes of Yersinia pseudotuberculosis IP32953 at 28°C and 3°C.

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    Eveliina Palonen

    Full Text Available Yersinia pseudotuberculosis is a significant psychrotrophic food pathogen whose cold tolerance mechanisms are poorly understood. Signal transduction systems serve to monitor the environment, but no systematic investigation of their role at cold temperatures in Y. pseudotuberculosis has yet been undertaken. The relative expression levels of 54 genes predicted to encode proteins belonging to signal transduction systems in Y. pseudotuberculosis IP32953 were determined at 28°C and 3°C by quantitative real-time reverse transcription-PCR. The relative expression levels of 44 genes were significantly (p<0.05 higher at 3°C than at 28°C. Genes encoding the two-component system CheA/CheY had the highest relative expression levels at 3°C. Mutational analysis revealed that cheA is important for growth and motility at 3°C. The relative expression level of one gene, rssB, encoding an RpoS regulator, was significantly (p<0.05 lower at 3°C than at 28°C. The results suggest that several signal transduction systems might be used during growth at low temperature, and at least, CheA/CheY two-component system is important for low-temperature growth.

  16. Vibrio cholerae classical biotype is converted to the viable non-culturable state when cultured with the El Tor biotype.

    Science.gov (United States)

    Pradhan, Subhra; Mallick, Sanjaya K; Chowdhury, Rukhsana

    2013-01-01

    A unique event in bacterial epidemiology was the emergence of the El Tor biotype of Vibrio cholerae O1 and the subsequent rapid displacement of the existing classical biotype as the predominant cause of epidemic cholera. We demonstrate that when the El Tor and classical biotypes were cocultured in standard laboratory medium a precipitous decline in colony forming units (CFU) of the classical biotype occurred in a contact dependent manner. Several lines of evidence including DNA release, microscopy and flow cytometric analysis indicated that the drastic reduction in CFU of the classical biotype in cocultures was not accompanied by lysis, although when the classical biotype was grown individually in monocultures, lysis of the cells occurred concomitant with decrease in CFU starting from late stationary phase. Furthermore, uptake of a membrane potential sensitive dye and protection of genomic DNA from extracellular DNase strongly suggested that the classical biotype cells in cocultures retained viability in spite of loss of culturability. These results suggest that coculturing the classical biotype with the El Tor biotype protects the former from lysis allowing the cells to remain viable in spite of the loss of culturability. The stationary phase sigma factor RpoS may have a role in the loss of culturability of the classical biotype in cocultures. Although competitive exclusion of closely related strains has been reported for several bacterial species, conversion of the target bacterial population to the viable non-culturable state has not been demonstrated previously and may have important implications in the evolution of bacterial strains.

  17. Type VI secretion system contributes to Enterohemorrhagic Escherichia coli virulence by secreting catalase against host reactive oxygen species (ROS).

    Science.gov (United States)

    Wan, Baoshan; Zhang, Qiufen; Ni, Jinjing; Li, Shuxian; Wen, Donghua; Li, Jun; Xiao, Haihan; He, Ping; Ou, Hong-Yu; Tao, Jing; Teng, Qihui; Lu, Jie; Wu, Wenjuan; Yao, Yu-Feng

    2017-03-01

    Enterohemorrhagic Escherichia coli (EHEC) is one major type of contagious and foodborne pathogens. The type VI secretion system (T6SS) has been shown to be involved in the bacterial pathogenicity and bacteria-bacteria competition. Here, we show that EHEC could secrete a novel effector KatN, a Mn-containing catalase, in a T6SS-dependent manner. Expression of katN is promoted by RpoS and OxyR and repressed by H-NS, and katN contributes to bacterial growth under oxidative stress in vitro. KatN could be secreted into host cell cytosol after EHEC is phagocytized by macrophage, which leads to decreased level of intracellular reactive oxygen species (ROS) and facilitates the intramacrophage survival of EHEC. Finally, animal model results show that the deletion mutant of T6SS was attenuated in virulence compared with the wild type strain, while the deletion mutant of katN had comparable virulence to the wild type strain. Taken together, our findings suggest that EHEC could sense oxidative stress in phagosome and decrease the host cell ROS by secreting catalase KatN to facilitate its survival in the host cells.

  18. The P1 promoter of the Escherichia coli rpoH gene is utilized by sigma 70 -RNAP or sigma s -RNAP depending on growth phase.

    Science.gov (United States)

    Janaszak, Anna; Nadratowska-Wesołowska, Beata; Konopa, Grazyna; Taylor, Alina

    2009-02-01

    The P1 promoter of the Escherichia coli rpoH gene has been known as a sigma(70)-dependent promoter (RNAP). We show here that it is also recognized by sigma(S). The sigma(70) and sigma(S) RNA polymerase subunits direct transcription from the P1 promoter in the exponential and stationary growth phases, respectively, and transcriptional start sites for the two holoenzymes differ by 1 nt. The transcription after heat shock is sigma(70)-dependent. The results are based on (1) sequence analysis that revealed features of promoters responsive to both, sigma(70)- and sigma(S)-RNAP, (2) in vitro transcription experiments verifying that both holoenzymes are able to transcribe the promoter, (3) electron microscopy results indicating that both holoenzymes bind the same site, (4) primer extension test performed with RNA isolated from the wild-type and rpoS mutant strains, demonstrating that transcription from the P1 promoter in the stationary phase is sigma(S)-dependent. These and previous results point to cooperation of sigma(70), sigma(24), sigma(S) and sigma(54) regulons in the expression of the rpoH gene, coding for the main regulator of the stress response, thus rendering it active in a variety of conditions.

  19. An unusual primary sigma factor in the Bacteroidetes phylum.

    Science.gov (United States)

    Vingadassalom, Didier; Kolb, Annie; Mayer, Claudine; Rybkine, Tania; Collatz, Ekkehard; Podglajen, Isabelle

    2005-05-01

    The presence of housekeeping gene promoters with a unique consensus sequence in Bacteroides fragilis, previously described by Bayley et al. (2000, FEMS Microbiol Lett 193: 149-154), suggested the existence of a particular primary sigma factor. The single rpoD-like gene observed in the B. fragilis genome, and similarly in those of other members of the Bacteroidetes phylum, was found to be essential. It encodes a protein, sigma(ABfr), of only 32.7 kDa that is produced with equal abundance during all phases of growth and was concluded to be the primary sigma factor. sigma(ABfr) and its orthologues in the Bacteroidetes are unusual primary sigma factors in that they lack region 1.1, have a unique signature made up of 29 strictly identical amino acids and are the only RpoD factors that cluster with the RpoS factors. Although binding to the Escherichia coli core RNA polymerase, sigma(ABfr) does not support transcription initiation from any promoter when it is part of the heterologous holoenzyme, while in the reconstituted homologous holoenzyme it does so only from typical B. fragilis, including rrs, promoters but not from the lacUV5 or RNA I promoters.

  20. Effect of salt addition on acid resistance response of Escherichia coli O157:H7 against acetic acid.

    Science.gov (United States)

    Bae, Young-Min; Lee, Sun-Young

    2017-08-01

    A combination of salt and acid is commonly used in the production of many foods, such as pickles and fermented foods. However, in our previous studies, addition of salt significantly reduced the inhibitory effect of acetic acid against E. coli O157:H7 in laboratory media and pickled cucumbers. Therefore, this study was conducted to determine the effect of salt addition on the acid resistance (AR) response of E. coli O157:H7 after treatment with acetic acid. The combined effect of acetic acid and salt showed different results depending on media tested. Organic compounds such as yeast extract and tryptone were required to observe the antagonistic effect of salt and acetic acid in combination. However, use of an rpoS mutant or addition of chloramphenicol resulted in no changes in the antagonistic effect of acetic acid and salt. The addition of glutamate to phosphate buffer significantly increased the survival levels of E. coli O157:H7 after the acetic acid treatment; however, the survival levels were lower than those after the treatment with acetic acid alone. Thus, the addition of salt may increase the AR response of E. coli O157:H7; however, these survival mechanisms were not proven clearly. Therefore, further studies need to be performed to better understand the antagonism of acetic acid salt against E. coli O157:H7. Copyright © 2017 Elsevier Ltd. All rights reserved.

  1. Constructed molecular sensor to enhance metal detection by bacterial ribosomal switch-ion channel protein interaction.

    Science.gov (United States)

    Cuero, Raul; Lilly, J; McKay, David S

    2012-03-31

    Molecular biosensors are useful tools that detect metal ions or other potentially toxic chemicals. However, the efficiency of conventional sensors is limited in mixed metals substrates, which is the common way they are found in nature. The use of biosensors constructed from genetically modified living microbial systems has the potential of providing sensitive detection systems for specific toxic targets. Consequently, our investigation was aimed at assembling different genetic building blocks to produce a focused microbial biosensor with the ability to detect specific metals. This objective was achieved by using a synthetic biology approach. Our genetic building blocks, including a synchronized ribosomal switch-iron ion channel, along with sequences of promoters, metal-binding proteins (Fe, Pb), ribosomal binding sites, yellow fluorescence reporter protein (YFRP), and terminators, were constructed within the same biobrick in Escherichia coli. We used an rpoS ribosomal switch containing an aptamer, which responds to the specific metal ligands, in synchronization with an iron ion channel, TonB. This switch significantly stimulates translation, as expressed by higher fluorescence, number of colonies, and concentration of RNA in E. coli. The positive results show the effectiveness of using genetically tailored synchronized ribosomal switch-ion channels to construct microbial biosensors to detect specific metals, as tested in iron solutions. Copyright © 2012 Elsevier B.V. All rights reserved.

  2. The adaptive response of bacterial food-borne pathogens in the environment, host and food: Implications for food safety.

    Science.gov (United States)

    Alvarez-Ordóñez, Avelino; Broussolle, Véronique; Colin, Pierre; Nguyen-The, Christophe; Prieto, Miguel

    2015-11-20

    Bacteria are constantly faced to stress situations in their ecological niches, the food and the host gastrointestinal tract. The capacity to detect and respond to surrounding changes is crucial for bacterial pathogens to survive or grow in changing environments. To this purpose, cells have evolved various sophisticated networks designed to protect against stressors or repair damage caused by them. Challenges can occur during production of foods when subjected to processing, and after food ingestion when confronted with host defensive barriers. Some pathogenic bacteria have shown the capacity to develop stable resistance against extreme conditions within a defined genomic context and a limited number of generations. On the other hand, bacteria can also respond to adverse conditions in a transient manner, through the so-called stress tolerance responses. Bacterial stress tolerance responses include both structural and physiological modifications in the cell and are mediated by complex genetic regulatory machinery. Major aspects in the adaptive response are the sensing mechanisms, the characterization of cell defensive systems, such as the operation of regulatory proteins (e.g. RpoS), the induction of homeostatic and repair systems, the synthesis of shock response proteins, and the modifications of cell membranes, particularly in their fatty acid composition and physical properties. This article reviews certain strategies used by food-borne bacteria to respond to particular stresses (acid, cold stress, extreme pressure) in a permanent or transient manner and discusses the implications that such adaptive responses pose for food safety. Copyright © 2015 Elsevier B.V. All rights reserved.

  3. Regulatory design governing progression of population growth phases in bacteria.

    Directory of Open Access Journals (Sweden)

    Agustino Martínez-Antonio

    Full Text Available It has long been noted that batch cultures inoculated with resting bacteria exhibit a progression of growth phases traditionally labeled lag, exponential, pre-stationary and stationary. However, a detailed molecular description of the mechanisms controlling the transitions between these phases is lacking. A core circuit, formed by a subset of regulatory interactions involving five global transcription factors (FIS, HNS, IHF, RpoS and GadX, has been identified by correlating information from the well- established transcriptional regulatory network of Escherichia coli and genome-wide expression data from cultures in these different growth phases. We propose a functional role for this circuit in controlling progression through these phases. Two alternative hypotheses for controlling the transition between the growth phases are first, a continuous graded adjustment to changing environmental conditions, and second, a discontinuous hysteretic switch at critical thresholds between growth phases. We formulate a simple mathematical model of the core circuit, consisting of differential equations based on the power-law formalism, and show by mathematical and computer-assisted analysis that there are critical conditions among the parameters of the model that can lead to hysteretic switch behavior, which--if validated experimentally--would suggest that the transitions between different growth phases might be analogous to cellular differentiation. Based on these provocative results, we propose experiments to test the alternative hypotheses.

  4. Effects of Chlorine Stress on Pseudomonas aeruginosa Biofilm and Analysis of Related Gene Expressions.

    Science.gov (United States)

    Kekeç, Özge; Gökalsın, Barış; Karaltı, İskender; Kayhan, Figen Esin; Sesal, Nüzhet Cenk

    2016-08-01

    Chlorine is deployed worldwide to clean waters and prevent water-originated illnesses. However, chlorine has a limited disinfection capacity against biofilms. Microorganisms form biofilms to protect themselves from biological threats such as disinfectant chemicals. Pseudomonas aeruginosa is an opportunistic pathogen and its biofilm form attaches to surfaces, living buried into exopolysaccharides, can be present in all watery environments including tap water and drinking water. This research aimed to study the biofilm trigger mechanism of the opportunistic pathogen P. aeruginosa PAO1 strain, which is known to form biofilm in water supply systems and human body, under chlorine stress levels. In addition to biofilm staining, certain genes that are relevant to the stress condition were selected for gene expression analysis. The bacteria cultures were grown under chlorine stress with concentrations of 0.5, 0.7 and 1 mg/l. Six gene regions were determined related to biofilm and stress response: rpoS, bifA, migA, katB, soxR, and algC. Biofilm formation was analyzed by basic fuchsin staining, and gene expressions were quantified by quantitative real-time PCR. According to the results, highest biofilm production was observed in P. aeruginosa PAO1 wild strain under no stress conditions. Higher biofilm amounts were observed for bacteria under 0.5 and 0.7 mg/l chlorine stress compared to 1 mg/l chlorine stress.

  5. Weak Organic Acids Decrease Borrelia burgdorferi Cytoplasmic pH, Eliciting an Acid Stress Response and Impacting RpoN- and RpoS-Dependent Gene Expression

    Directory of Open Access Journals (Sweden)

    Daniel P. Dulebohn

    2017-09-01

    Full Text Available The spirochete Borrelia burgdorferi survives in its tick vector, Ixodes scapularis, or within various hosts. To transition between and survive in these distinct niches, B. burgdorferi changes its gene expression in response to environmental cues, both biochemical and physiological. Exposure of B. burgdorferi to weak monocarboxylic organic acids, including those detected in the blood meal of fed ticks, decreased the cytoplasmic pH of B. burgdorferi in vitro. A decrease in the cytoplasmic pH induced the expression of genes encoding enzymes that have been shown to restore pH homeostasis in other bacteria. These include putative coupled proton/cation exchangers, a putative Na+/H+ antiporter, a neutralizing buffer transporter, an amino acid deaminase and a proton exporting vacuolar-type VoV1 ATPase. Data presented in this report suggested that the acid stress response triggered the expression of RpoN- and RpoS-dependent genes including important virulence factors such as outer surface protein C (OspC, BBA66, and some BosR (Borreliaoxidative stress regulator-dependent genes. Because the expression of virulence factors, like OspC, are so tightly connected by RpoS to general cellular stress responses and cell physiology, it is difficult to separate transmission-promoting conditions in what is clearly a multifactorial and complex regulatory web.

  6. The effect of 20-hydroxyecdysone on the susceptibility of Escherichia coli to different antibiotics.

    Science.gov (United States)

    Smirnova, G; Bezmaternykh, K; Oktyabrsky, O N

    2016-12-01

    The aim of this study was to investigate the influence of 20-hydroxyecdysone (20E) on the susceptibility of growing Escherichia coli to antibiotics. Susceptibility of E. coli to antibiotics in the presence of 20E was estimated by determination of the colony-forming ability and the specific growth rate. Pretreatment with 20E decreased the bactericidal effect of ciprofloxacin (0·3 and 3·0 μg ml-1 ), streptomycin (10 and 40 μg ml-1 ) and kanamycin (10 μg ml-1 ) and increased the bactericidal action of 0·03 μg ml-1 ciprofloxacin. To study the influence of 20E on gene expression, we used strains of E. coli carrying fusions of promoters of relevant genes with the structural gene of β-galactosidase. 20E had no marked effect on the expression of antioxidant genes katG, katE, sodA and the rpoS gene (general stress response), while it (alone or combined with ciprofloxacin) markedly stimulated expression of the sulA gene belonging to the SOS regulon (DNA damage response). In growing E. coli, 20E modulated the bactericidal action of antibiotics and stimulated the SOS response. The results of this study may be used to enhance the efficiency of antibacterial therapies. © 2016 The Society for Applied Microbiology.

  7. Diversity and functions of bacterial community in drinking water biofilms revealed by high-throughput sequencing

    Science.gov (United States)

    Chao, Yuanqing; Mao, Yanping; Wang, Zhiping; Zhang, Tong

    2015-06-01

    The development of biofilms in drinking water (DW) systems may cause various problems to water quality. To investigate the community structure of biofilms on different pipe materials and the global/specific metabolic functions of DW biofilms, PCR-based 454 pyrosequencing data for 16S rRNA genes and Illumina metagenomic data were generated and analysed. Considerable differences in bacterial diversity and taxonomic structure were identified between biofilms formed on stainless steel and biofilms formed on plastics, indicating that the metallic materials facilitate the formation of higher diversity biofilms. Moreover, variations in several dominant genera were observed during biofilm formation. Based on PCA analysis, the global functions in the DW biofilms were similar to other DW metagenomes. Beyond the global functions, the occurrences and abundances of specific protective genes involved in the glutathione metabolism, the SoxRS system, the OxyR system, RpoS regulated genes, and the production/degradation of extracellular polymeric substances were also evaluated. A near-complete and low-contamination draft genome was constructed from the metagenome of the DW biofilm, based on the coverage and tetranucleotide frequencies, and identified as a Bradyrhizobiaceae-like bacterium according to a phylogenetic analysis. Our findings provide new insight into DW biofilms, especially in terms of their metabolic functions.

  8. The Legionella pneumophila GIG operon responds to gold and copper in planktonic and biofilm cultures.

    Directory of Open Access Journals (Sweden)

    Kathleen Jwanoswki

    Full Text Available Legionella pneumophila contaminates man-made water systems and creates numerous exposure risks for Legionnaires' Disease. Because copper/silver ionization is commonly used to control L. pneumophila, its mechanisms of metal response and detoxification are of significant interest. Here we describe an L. pneumophila operon with significant similarity to the GIG operon of Cupriavidus metallidurans. The Legionella GIG operon is present in a subset of strains and has been acquired as part of the ICE-βox 65-kB integrative conjugative element. We assessed GIG promoter activity following exposure of L. pneumophila to multiple concentrations of HAuCl4, CuSO4 and AgNO3. At 37°C, control stationary phase cultures exhibited GIG promoter activity. This activity increased significantly in response to 20 and 50uM HAuCl4 and CuSO4 but not in response to AgNO3. Conversely, at 26°C, cultures exhibited decreased promoter response to copper. GIG promoter activity was also induced by HAuCl4 or CuSO4 during early biofilm establishment at both temperatures. When an L. pneumophila GIG promoter construct was transformed into E. coli DH5α, cultures showed baseline expression levels that did not increase following metal addition. Analysis of L. pneumophila transcriptional regulatory mutants suggested that GIG up-regulation in the presence of metal ions may be influenced by the stationary phase sigma factor, RpoS.

  9. The Legionella pneumophila GIG operon responds to gold and copper in planktonic and biofilm cultures.

    Science.gov (United States)

    Jwanoswki, Kathleen; Wells, Christina; Bruce, Terri; Rutt, Jennifer; Banks, Tabitha; McNealy, Tamara L

    2017-01-01

    Legionella pneumophila contaminates man-made water systems and creates numerous exposure risks for Legionnaires' Disease. Because copper/silver ionization is commonly used to control L. pneumophila, its mechanisms of metal response and detoxification are of significant interest. Here we describe an L. pneumophila operon with significant similarity to the GIG operon of Cupriavidus metallidurans. The Legionella GIG operon is present in a subset of strains and has been acquired as part of the ICE-βox 65-kB integrative conjugative element. We assessed GIG promoter activity following exposure of L. pneumophila to multiple concentrations of HAuCl4, CuSO4 and AgNO3. At 37°C, control stationary phase cultures exhibited GIG promoter activity. This activity increased significantly in response to 20 and 50uM HAuCl4 and CuSO4 but not in response to AgNO3. Conversely, at 26°C, cultures exhibited decreased promoter response to copper. GIG promoter activity was also induced by HAuCl4 or CuSO4 during early biofilm establishment at both temperatures. When an L. pneumophila GIG promoter construct was transformed into E. coli DH5α, cultures showed baseline expression levels that did not increase following metal addition. Analysis of L. pneumophila transcriptional regulatory mutants suggested that GIG up-regulation in the presence of metal ions may be influenced by the stationary phase sigma factor, RpoS.

  10. Tailoring of global transcription sigma D factor by random mutagenesis to improve Escherichia coli tolerance towards low-pHs.

    Science.gov (United States)

    Gao, Xi; Jiang, Ling; Zhu, Liying; Xu, Qing; Xu, Xian; Huang, He

    2016-04-20

    Bioconversion processes of organic acid or acid hydrolysis of raw material for microbial metabolism often suffer limitations as a result of microbial sensitivity in low-pH conditions. We adopted a three-step method called RAndom Insertional-deletional Strand Exchange mutagenesis (RAISE) to engineer the components of global regulator Sigma D factor (RpoD) of Escherichia coli to improve its acid tolerance. The best strain Mutant VII was identified from random mutagenesis libraries based on the growth performance, which exhibited much higher growth rate than the control (0.22h(-1) vs. 0.15h(-1)) at pH as low as 3.17. Combined transcriptome and phenome analysis of E. coli was carried out to better understand the global effects of RpoD on the regulatory networks. Our analysis showed that 95 (2.1%) of all E. coli genes were induced and 178 (4.0%) genes were repressed, including those for trehalose biosynthesis, nucleotides biosynthesis, carbon metabolism, amino acid utilization, except for acid resistance. Also regulated were the master regulators (ArcA, EvgA, H-NS and RpoS) and gene/operon-specific transcription factors (GadX, GadW, AppY, YdeO, KdgR). These results demonstrated that RpoD acts as global regulator in the growth phase of E. coli and consequently improves acid tolerances. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. Role of Burkholderia pseudomallei Sigma N2 in Amino Acids Utilization and in Regulation of Catalase E Expression at the Transcriptional Level

    Directory of Open Access Journals (Sweden)

    Duong Thi Hong Diep

    2015-01-01

    Full Text Available Burkholderia pseudomallei is the causative agent of melioidosis. The complete genome sequences of this pathogen have been revealed, which explain some pathogenic mechanisms. In various hostile conditions, for example, during nitrogen and amino acid starvation, bacteria can utilize alternative sigma factors such as RpoS and RpoN to modulate genes expression for their adaptation and survival. In this study, we demonstrate that mutagenesis of rpoN2, which lies on chromosome 2 of B. pseudomallei and encodes a homologue of the sigma factor RpoN, did not alter nitrogen and amino acid utilization of the bacterium. However, introduction of B. pseudomallei rpoN2 into E. coli strain deficient for rpoN restored the ability to utilize amino acids. Moreover, comparative partial proteomic analysis of the B. pseudomallei wild type and its rpoN2 isogenic mutant was performed to elucidate its amino acids utilization property which was comparable to its function found in the complementation assay. By contrast, the rpoN2 mutant exhibited decreased katE expression at the transcriptional and translational levels. Our finding indicates that B. pseudomallei RpoN2 is involved in a specific function in the regulation of catalase E expression.

  12. Biochemical Mechanisms and Energy Strategies of Geobacter sulfurreducens for Long- Term Survival

    Science.gov (United States)

    Helmus, R. A.; Liermann, L. J.; Brantley, S. L.; Tien, M.

    2008-12-01

    Numerous species of bacteria have been observed to exhibit a growth advantage in stationary phase (GASP) phenotype, indicating that microorganisms starved of an energy source may adapt to allow for long-term survival. Understanding how Geobacter sulfurreducens persists using various metal forms as energy sources and whether a GASP phenotype develops during long-term growth are important for efficient application of this bacterium to sites requiring engineered bioremediation of soluble metals. Thus, we investigated the growth kinetics and survival of G. sulfurreducens. The growth rate of G. sulfurreducens was highest when cultured with soluble iron and generally higher on iron oxide than manganese oxide, suggesting that soluble metal forms are more readily utilized as energy sources by G. sulfurreducens. By monitoring the abundance of G. sulfurreducens in batch cultures for >6 months, distinct growth, stationary, and prolonged starvation phases were observed and a cell density of 105- 106 cells/mL persisted under long-term starvation conditions. The outgrowth of an aged G. sulfurreducens strain co-cultured with a young strain was monitored as a measure of the existence of the GASP phenotype. As the strains aged, the rpoS gene was cloned and sequenced at different stages of growth to identify mutations corresponding to a growth advantage. The results of these studies provide insight into the use of various metal forms for growth by G. sulfurreducens and its ability to persist when starved of energy sources.

  13. Several Hfq-dependent alterations in physiology of Yersinia enterocolitica O:3 are mediated by derepression of the transcriptional regulator RovM.

    Science.gov (United States)

    Leskinen, Katarzyna; Pajunen, Maria I; Varjosalo, Markku; Fernández-Carrasco, Helena; Bengoechea, José A; Skurnik, Mikael

    2017-03-01

    In bacteria, the RNA chaperone Hfq enables pairing of small regulatory RNAs with their target mRNAs and therefore is a key player of post-transcriptional regulation network. As a global regulator, Hfq is engaged in the adaptation to external environment, regulation of metabolism and bacterial virulence. In this study we used RNA-sequencing and quantitative proteomics (LC-MS/MS) to elucidate the role of this chaperone in the physiology and virulence of Yersinia enterocolitica serotype O:3. This global approach revealed the profound impact of Hfq on gene and protein expression. Furthermore, the role of Hfq in the cell morphology, metabolism, cell wall integrity, resistance to external stresses and pathogenicity was evaluated. Importantly, our results revealed that several alterations typical for the hfq-negative phenotype were due to derepression of the transcriptional factor RovM. The overexpression of RovM caused by the loss of Hfq chaperone resulted in extended growth defect, alterations in the lipid A structure, motility and biofilm formation defects, as well as changes in mannitol utilization. Furthermore, in Y. enterocolitica RovM only in the presence of Hfq affected the abundance of RpoS. Finally, the impact of hfq and rovM mutations on the virulence was assessed in the mouse infection model. © 2016 John Wiley & Sons Ltd.

  14. Stressing Escherichia coli to educate students about research: A CURE to investigate multiple levels of gene regulation.

    Science.gov (United States)

    McDonough, Janet; Goudsouzian, Lara K; Papaj, Agllai; Maceli, Ashley R; Klepac-Ceraj, Vanja; Peterson, Celeste N

    2017-09-01

    Course-based undergraduate research experiences (CUREs) have been shown to increase student retention and learning in the biological sciences. Most CURES cover only one aspect of gene regulation, such as transcriptional control. Here we present a new inquiry-based lab that engages understanding of gene expression from multiple perspectives. Students carry out a forward genetic screen to identify regulators of the stationary phase master regulator RpoS in the model organism Escherichia coli and then use a series of reporter fusions to determine if the regulation is at the level of transcription or the post-transcription level. This easy-to-implement course has been run both as a 9-week long project and a condensed 5-6 week version in three different schools and types of courses. A majority of the genes found in the screen are novel, thus giving students the opportunity to contribute to original findings to the field. Assessments of this CURE show student gains in learning in many knowledge areas. In addition, attitudinal surveys suggest the students are enthusiastic about the screen and their learning about gene regulation. In summary, this lab would be an appropriate addition to an intermediate or advanced level Molecular Biology, Genetics, or Microbiology curriculum. © 2017 by The International Union of Biochemistry and Molecular Biology, 45(5):449-458, 2017. © 2017 The International Union of Biochemistry and Molecular Biology.

  15. The role of the 'gearbox' in the transcription of essential genes.

    Science.gov (United States)

    Vicente, M; Kushner, S R; Garrido, T; Aldea, M

    1991-09-01

    Regulation of transcription occurs at different levels, one being in the presence of sequences specifically recognized by different forms of RNA polymerase, i.e. the promoters. Three different kinds of promoter are defined according, among other things, to their dependence on the growth rate of the cell: the 'house-keeper' promoter of many metabolic genes, the stringent promoter found at several rRNA and ribosomal protein genes, and the 'gearbox' at genes whose products are required at higher relative amounts at lower growth rates. The identified gearbox promoters of Escherichia coli share specific homologies in the -10, -35 and upstream regions. Although there may be different types of gearbox promoters, the -10 sequence of one of these promoters has been found to be essential for functioning as a gearbox. This suggests the existence of specific sigma factors for its transcription. RpoS (KatF) is a likely candidate for being one of these sigma factors. Computer simulation allows us to predict that such sigma factors should, in turn, be expressed following a gearbox mode, which would then imply the existence of self-regulated loops contributing to the expression of some genes of bacterial division.

  16. The RNA chaperone Hfq enables the environmental stress tolerance super-phenotype of Pseudomonas putida.

    Science.gov (United States)

    Arce-Rodríguez, Alejandro; Calles, Belén; Nikel, Pablo I; de Lorenzo, Víctor

    2016-10-01

    The natural physiological regime of the soil bacterium Pseudomonas putida involves incessant exposure to endogenous metabolic conflicts and environmental physicochemical insults. Yet, the role of assisted small RNA-mRNA pairing in the stress tolerance super-phenotype that is the trademark of this bacterium has not been accredited. We have thoroughly explored the physiological consequences -in particular those related to exogenous stress - of deleting the hfq gene of P. putida, which encodes the major RNA chaperone that promotes sRNA-target mRNA interactions. While the overall trend was a general weakening of every robustness descriptor of the Δhfq strain, growth parameters and production of central metabolic enzymes were comparatively less affected than other qualities that depend directly on energy status (e.g. motility, DNA repair). The overall catalytic vigour of the mutant decreased to < 20% than the wild-type strain, as estimated from the specific growth rate of cells carrying the catabolic TOL plasmid pWW0 for m-xylene biodegradation. Several loss-of-function phenotypes could be traced to the effect of the Δhfq deletion on the intracellular contents of the stationary sigma factor RpoS. It thus seems that Hfq, while not indispensable for any essential function, contributes to shape the environmental lifestyle of P. putida. © 2015 Society for Applied Microbiology and John Wiley & Sons Ltd.

  17. Medium-chain-length polyhydroxyalkanoates synthesis by Pseudomonas putida KT2440 relA/spoT mutant: bioprocess characterization and transcriptome analysis.

    Science.gov (United States)

    Mozejko-Ciesielska, Justyna; Dabrowska, Dorota; Szalewska-Palasz, Agnieszka; Ciesielski, Slawomir

    2017-12-01

    Pseudomonas putida KT2440 is a model bacteria used commonly for medium-chain-length polyhydroxyalkanoates (mcl-PHAs) production using various substrates. However, despite many studies conducted on P. putida KT2440 strain, the molecular mechanisms of leading to mcl-PHAs synthesis in reaction to environmental stimuli are still not clear. The rearrangement of the metabolism in response to environmental stress could be controlled by stringent response that modulates the transcription of many genes in order to promote survival under nutritional deprivation conditions. Therefore, in this work we investigated the relation between mcl-PHAs synthesis and stringent response. For this study, a relA/spoT mutant of P. putida KT2440, unable to induce the stringent response, was used. Additionally, the transcriptome of this mutant was analyzed using RNA-seq in order to examine rearrangements of the metabolism during cultivation. The results show that the relA/spoT mutant of P. putida KT2440 is able to accumulate mcl-PHAs in both optimal and nitrogen limiting conditions. Nitrogen starvation did not change the efficiency of mcl-PHAs synthesis in this mutant. The transition from exponential growth to stationary phase caused significant upregulation of genes involved in transport system and nitrogen metabolism. Transcriptional regulators, including rpoS, rpoN and rpoD, did not show changes in transcript abundance when entering the stationary phase, suggesting their limited role in mcl-PHAs accumulation during stationary phase.

  18. Natural plant products inhibits growth and alters the swarming motility, biofilm formation, and expression of virulence genes in enteroaggregative and enterohemorrhagic Escherichia coli.

    Science.gov (United States)

    García-Heredia, Alam; García, Santos; Merino-Mascorro, José Ángel; Feng, Peter; Heredia, Norma

    2016-10-01

    The purpose of this study was to determine the effects of plant products on the growth, swarming motility, biofilm formation and virulence gene expression in enterohemorrhagic Escherichia coli O157:H7 and enteroaggregative E. coli strain 042 and a strain of O104:H4 serotype. Extracts of Lippia graveolens and Haematoxylon brassiletto, and carvacrol, brazilin were tested by an antimicrobial microdilution method using citral and rifaximin as controls. All products showed bactericidal activity with minimal bactericidal concentrations ranging from 0.08 to 8.1 mg/ml. Swarming motility was determined in soft LB agar. Most compounds reduced swarming motility by 7%-100%; except carvacrol which promoted motility in two strains. Biofilm formation studies were done in microtiter plates. Rifaximin inhibited growth and reduced biofilm formation, but various concentrations of other compounds actually induced biofilm formation. Real time PCR showed that most compounds decreased stx2 expression. The expression of pic and rpoS in E. coli 042 were suppressed but in E. coli O104:H4 they varied depending on compounds. In conclusion, these extracts affect E. coli growth, swarming motility and virulence gene expression. Although these compounds were bactericidal for pathogenic E. coli, sublethal concentrations had varied effects on phenotypic and genotypic traits, and some increased virulence gene expression. Copyright © 2016 Elsevier Ltd. All rights reserved.

  19. Type VI secretion system contributes to Enterohemorrhagic Escherichia coli virulence by secreting catalase against host reactive oxygen species (ROS.

    Directory of Open Access Journals (Sweden)

    Baoshan Wan

    2017-03-01

    Full Text Available Enterohemorrhagic Escherichia coli (EHEC is one major type of contagious and foodborne pathogens. The type VI secretion system (T6SS has been shown to be involved in the bacterial pathogenicity and bacteria-bacteria competition. Here, we show that EHEC could secrete a novel effector KatN, a Mn-containing catalase, in a T6SS-dependent manner. Expression of katN is promoted by RpoS and OxyR and repressed by H-NS, and katN contributes to bacterial growth under oxidative stress in vitro. KatN could be secreted into host cell cytosol after EHEC is phagocytized by macrophage, which leads to decreased level of intracellular reactive oxygen species (ROS and facilitates the intramacrophage survival of EHEC. Finally, animal model results show that the deletion mutant of T6SS was attenuated in virulence compared with the wild type strain, while the deletion mutant of katN had comparable virulence to the wild type strain. Taken together, our findings suggest that EHEC could sense oxidative stress in phagosome and decrease the host cell ROS by secreting catalase KatN to facilitate its survival in the host cells.

  20. Hemodynamic alterations measured with phase-contrast MRI in a giant cerebral aneurysm treated with a flow-diverting stent

    Directory of Open Access Journals (Sweden)

    Matthew Ethan MacDonald, PhD

    2015-01-01

    Full Text Available Many risk factors have been proposed in the development of the cerebral aneurysms. Hemodynamics including blood velocity, volume flow rate (VFR, and intravascular pressure are thought to be prognostic indicators of aneurysm development. We hypothesize that treatment of cerebral aneurysm using a flow-diverting stent will bring these hemodynamic parameters closer to those observed on the contralateral side. In the current study, a patient with a giant cerebral aneurysm was studied pre- and postoperatively using phase contrast MRI (PC-MRI to measure the hemodynamic changes resulting from the deployment of a flow-diverting stent. PC-MRI was used to calculate intravascular pressure, which was compared to more invasive endovascular catheter-derived measurements. After stent placement, the measured VFRs in vessels of the treated hemisphere approached those measured on the contralateral side, and flow symmetry changed from a laterality index of -0.153 to 0.116 in the middle cerebral artery. Pressure estimates derived from the PC-MRI velocity data had an average difference of 6.1% as compared to invasive catheter transducer measurements. PC-MRI can measure the hemodynamic parameters with the same accuracy as invasive methods pre- and postoperatively.

  1. Fast scanning probe for tokamak plasmas

    Science.gov (United States)

    Boedo, J.; Gray, D.; Chousal, L.; Conn, R.; Hiller, B.; Finken, K. H.

    1998-07-01

    We describe a fast reciprocating probe drive, which has three main new features: (1) a detachable and modular probe head for easy maintenance, (2) a combination of high heat flux capability, high bandwidth, and low-Z materials construction, and (3) low weight, compact, inexpensive construction. The probe is mounted in a fast pneumatic drive in order to reach plasma regions of interest and remain inserted long enough to obtain good statistics while minimizing the heat flux to the tips and head. The drive is pneumatic and has been designed to be compact and reliable to comply with space and maintenance requirements of tokamaks. The probe described here has five tips which obtain a full spectrum of plasma parameters: electron temperature profile Te(r), electron density profile ne(r), floating potential profile Vf(r), poloidal electric field profile Eθ(r), saturation current profile Isat(r), and their fluctuations up to 3 MHz. We describe the probe show radial profiles of various parameters. We compare the density and temperature data to that obtained with a helium beam. We also discuss the techniques to process the data optimally, particularly double probe data and profile fits.

  2. Flight Deck-Based Delegated Separation: Evaluation of an On-Board Interval Management System with Synthetic and Enhanced Vision Technology

    Science.gov (United States)

    Prinzel, Lawrence J., III; Shelton, Kevin J.; Kramer, Lynda J.; Arthur, Jarvis J.; Bailey, Randall E.; Norman, Rober M.; Ellis, Kyle K. E.; Barmore, Bryan E.

    2011-01-01

    An emerging Next Generation Air Transportation System concept - Equivalent Visual Operations (EVO) - can be achieved using an electronic means to provide sufficient visibility of the external world and other required flight references on flight deck displays that enable the safety, operational tempos, and visual flight rules (VFR)-like procedures for all weather conditions. Synthetic and enhanced flight vision system technologies are critical enabling technologies to EVO. Current research evaluated concepts for flight deck-based interval management (FIM) operations, integrated with Synthetic Vision and Enhanced Vision flight-deck displays and technologies. One concept involves delegated flight deck-based separation, in which the flight crews were paired with another aircraft and responsible for spacing and maintaining separation from the paired aircraft, termed, "equivalent visual separation." The operation required the flight crews to acquire and maintain an "equivalent visual contact" as well as to conduct manual landings in low-visibility conditions. The paper describes results that evaluated the concept of EVO delegated separation, including an off-nominal scenario in which the lead aircraft was not able to conform to the assigned spacing resulting in a loss of separation.

  3. Effect of myocyte-fibroblast coupling on the onset of pathological dynamics in a model of ventricular tissue

    Science.gov (United States)

    Sridhar, S.; Vandersickel, Nele; Panfilov, Alexander V.

    2017-01-01

    Managing lethal cardiac arrhythmias is one of the biggest challenges in modern cardiology, and hence it is very important to understand the factors underlying such arrhythmias. While early afterdepolarizations (EAD) of cardiac cells is known to be one such arrhythmogenic factor, the mechanisms underlying the emergence of tissue level arrhythmias from cellular level EADs is not fully understood. Another known arrhythmogenic condition is fibrosis of cardiac tissue that occurs both due to aging and in many types of heart diseases. In this paper we describe the results of a systematic in-silico study, using the TNNP model of human cardiac cells and MacCannell model for (myo)fibroblasts, on the possible effects of diffuse fibrosis on arrhythmias occurring via EADs. We find that depending on the resting potential of fibroblasts (VFR), M-F coupling can either increase or decrease the region of parameters showing EADs. Fibrosis increases the probability of occurrence of arrhythmias after a single focal stimulation and this effect increases with the strength of the M-F coupling. While in our simulations, arrhythmias occur due to fibrosis induced ectopic activity, we do not observe any specific fibrotic pattern that promotes the occurrence of these ectopic sources.

  4. The cyclic nucleotide monophosphate domain of Xanthomonas campestris global regulator Clp defines a new class of cyclic di-GMP effectors.

    Science.gov (United States)

    Tao, Fei; He, Ya-Wen; Wu, Dong-Hui; Swarup, Sanjay; Zhang, Lian-Hui

    2010-02-01

    The widely conserved second messenger cyclic diguanosine monophosphate (c-di-GMP) plays a key role in quorum-sensing (QS)-dependent production of virulence factors in Xanthomonas campestris pv. campestris. The detection of QS diffusible signal factor (DSF) by the sensor RpfC leads to the activation of response regulator RpfG, which activates virulence gene expression by degrading c-di-GMP. Here, we show that a global regulator in the X. campestris pv. campestris QS regulatory pathway, Clp, is a c-di-GMP effector. c-di-GMP specifically binds to Clp with high affinity and induces allosteric conformational changes that abolish the interaction between Clp and its target gene promoter. Clp is similar to the cyclic AMP (cAMP) binding proteins Crp and Vfr and contains a conserved cyclic nucleotide monophosphate (cNMP) binding domain. Using site-directed mutagenesis, we found that the cNMP binding domain of Clp contains a glutamic acid residue (E99) that is essential for c-di-GMP binding. Substituting the residue with serine (E99S) resulted in decreased sensitivity to changes in the intracellular c-di-GMP level and attenuated bacterial virulence. These data establish the direct role of Clp in the response to fluctuating c-di-GMP levels and depict a novel mechanism by which QS links the second messenger with the X. campestris pv. campestris virulence regulon.

  5. A 3D numerical study of the collateral capacity of the Circle of Willis with anatomical variation in the posterior circulation.

    Science.gov (United States)

    Ren, Yuan; Chen, Qiang; Li, Zhi-Yong

    2015-01-01

    The Circle of Willis (CoW) is the most important collateral pathway of the cerebral artery. The present study aims to investigate the collateral capacity of CoW with anatomical variation when unilateral internalcarotid artery (ICA) is occluded. Basing on MRI data, we have reconstructed eight 3D models with variations in the posterior circulation of the CoW and set four different degrees of stenosis in the right ICA, namely 24%, 43%, 64% and 79%, respectively. Finally, a total of 40 models are performed with computational fluid dynamics simulations. All of the simulations share the same boundary condition with static pressure and the volume flow rate (VFR) are obtained to evaluate their collateral capacity. As for the middle cerebral artery (MCA) and the anterior cerebral artery (ACA), the transitional-type model possesses the best collateral capacity. But for the posterior cerebral artery (PCA), unilateral stenosis of ICA has the weakest influence on the unilateral posterior communicating artery (PCoA) absent model. We also find that the full fetal-type posterior circle of Willis is an utmost dangerous variation which must be paid more attention. The results demonstrate that different models have different collateral capacities in coping stenosis of unilateral ICA and these differences can be reflected by different outlets. The study could be used as a reference for neurosurgeon in choosing the best treatment strategy.

  6. Airline pilot scanning behavior during approaches and landing in a Boeing 737 simulator

    Science.gov (United States)

    Spady, A. A., Jr.

    1978-01-01

    A series of approaches using airline-rated Boeing 737 pilots in an FAA qualified simulator was conducted. The test matrices include both manual and coupled approaches for VFR, Category 1 and Category 2 conditions. A nonintrusive oculometer system was used to track the pilot's eye-point-of-regard throughout the approach. The results indicate that, in general, the pilots use a different scan technique for the manual and coupled (auto-pilot with manual throttle) conditions. For the manual approach 73 percent of the time was spent on the Flight Director and 13 percent on airspeed as opposed to 50 percent on Flight Director and 23 percent on airspeed for the coupled approaches. For the visual portion of approach from less than 100m to touchdown or when the touchdown point came into view, the pilots tend to fixate on their aim or touchdown area until the flare initiation, at which time they let their eye-point-of-regard move up the runway to use the centerline lights for rollout guidance.

  7. Estimation of mass transfer parameters in a Taylor-Couette-Poiseuille heterogeneous reactor

    Directory of Open Access Journals (Sweden)

    Resende M. M.

    2004-01-01

    Full Text Available A bench-scale, continuous vortex flow reactor (VFR, with a radius ratio, h, equal to 0.48 and an aspect ratio, G, equal to 11.19 was studied. This reactor may be used in the enzymatic hydrolysis of polypeptides obtained from sweet cheese whey with enzymes immobilized on agarose gel. Operational conditions were 2410 < Re q < 11793 and 30-min residence time for glycerol-water, 14% w/w, 27ºC (Re ax = 1.1 and for water, 38ºC (Re ax = 1.5. Residence time distributions (RTDs were obtained after pulse injections of different tracers (including dyed solid particles. Mass transfer coefficients of a lumped-parameter model of the reactor were estimated from these data. Model fitting to experimental data was accurate. Working conditions were selected so that transport properties of the fluids would be similar to the ones in the actual process at the final stages of whey hydrolysis.

  8. The Presence of Behavioral Traps in U.S. Airline Accidents: A Qualitative Analysis

    Directory of Open Access Journals (Sweden)

    Jonathan Velazquez

    2018-01-01

    Full Text Available Behavioral traps are accident-inducing operational pitfalls aviators may encounter as a result of poor decision making. The Federal Aviation Administration (FAA identifies the existence of twelve of these negative pilot behaviors. These are: Peer Pressure; Get-There-Itis; Loss of Situational Awareness; Descent Below the Minimum En Route Altitude (MEA; Mind Set; Duck-Under Syndrome; Getting Behind the Aircraft; Continuing Visual Flight Rules (VFR into Instrument Conditions; Scud Running; Operating Without Adequate Fuel Reserves; Flying Outside the Envelope; and Neglect of Flight Planning, Preflight Inspections, and Checklists. The purpose of this paper was to study the nature of their occurrence in the airline domain. Four Subject Matter Experts (SMEs analyzed 34 National Transportation Safety Board (NTSB accident reports. The SMEs were able to identify many pilot actions that were representative of the behavioral traps. Behavioral traps were found in all accidents with Loss of Situational Awareness and Neglect of Flight Planning, Preflight Inspections, and Checklists dominant. Various themes began to emerge, which played important roles in many accidents. These themes included Crew Resource Management (CRM issues, airline management and fatigue. The findings of this study indicated that behavioral traps were prevalent in airline accidents including habitual noncompliance by pilots. Attitude management training is recommended.

  9. Risk factors and pre-travel healthcare of international travellers attending a Dutch travel clinic: a cross-sectional analysis.

    Science.gov (United States)

    Wieten, Rosanne W; van der Schalie, Maurice; Visser, Benjamin J; Grobusch, Martin P; van Vugt, Michèle

    2014-01-01

    The number of international travellers is currently estimated to exceed one billion annually. To address travel related health risks and facilitate risk reduction strategies, detailed knowledge of travellers' characteristics is important. In this cross-sectional study, data of a 20% sample of travellers visiting the Academic Medical Center (AMC) travel clinic Amsterdam from July 2011 to July 2012 was collected. Itineraries and protection versus exposure rates of preventable infectious diseases were mapped and reported according to STROBE guidelines. 1749 travellers were included. South-Eastern Asia, South-America and West-Africa were most frequently visited. 26.2% of the population had pre-existing medical conditions (often cardiovascular). Young and VFR travellers had a longer median travel time (28 and 30 days) compared to the overall population (21 days). Young adult travellers were relatively often vaccinated against hepatitis B (43.9% vs. 20.5%, p travellers. Pre-travel guidelines were well adhered to. Young adult travellers had high-risk itineraries but were adequately protected. Improvement of hepatitis B and rabies protection would be desirable, specifically for VFRs. Copyright © 2014 Elsevier Ltd. All rights reserved.

  10. Mefloquine at the crossroads? Implications for malaria chemoprophylaxis in Europe.

    Science.gov (United States)

    Schlagenhauf, Patricia; Hatz, Christoph; Behrens, Ron; Visser, Leo; Funk, Maia; Holzer, Benedikt; Beck, Bernhard; Bourquin, Cathérine; Etter, Hermann; Furrer, Hansjakob; Genton, Blaise; Landry, Pierre; Chappuis, Francois; Loutan, Louis; Stössel, Ulrich; Jeschko, Eva; Rossanese, Andrea; Nothdurft, Hans Dieter

    2015-01-01

    Since its introduction to the market in 1985, mefloquine has been used for malaria chemoprophylaxis by more than 35 million travellers. In Europe, in 2014, the European Medicines Agency (EMA) issued recommendations on strengthened warnings, prescribing checklists and updates to the product information of mefloquine. Some malaria prevention advisors question the scientific basis for the restrictions and suggest that this cost-effective, anti-malarial drug will be displaced as a first-line anti-malaria medication with the result that vulnerable groups such as VFR and long-term travellers, pregnant travellers and young children are left without a suitable alternative chemoprophylaxis. This commentary looks at the current position of mefloquine prescribing and the rationale of the new EMA recommendations and restrictions. It also describes the new recommendations for malaria prophylaxis that have been adapted by Switzerland, Germany, Austria and Italy where chemoprophylaxis use is restricted to high-risk malaria-endemic areas. Copyright © 2015 Elsevier Ltd. All rights reserved.

  11. CO2 utilization via a novel anaerobic bioprocess configuration with simulated gas mixture and real stack gas samples.

    Science.gov (United States)

    Daglioglu, S Tugce; Karabey, Burcin; Ozdemir, Guven; Azbar, Nuri

    2017-11-28

    CO2, which is considered to be one of the major causes of climate change, has reached to critical levels in the atmosphere due to tremendous consumption of fossil fuels all over the world. In this study, anaerobic bioconversion of CO2 into bio-methane using a novel bioprocess configuration (HYBRID bioreactor) was studied under mesophilic conditions. Varying ratios of H2/CO2 gas mixture and volumetric feeding rates were investigated and no additional organic matter and trace element were needed throughout the study. The maximum methane production of 19 m3 CH4/m3reactor/d was achieved at a H2/CO2 ratio of 4:1 and feeding rate of 24 m3 gas/m3reactor/d. It was determined that H2 conversion rate is about 96%. For demonstration purpose, real stack gas sample from a petrochemical industry was also tested under optimized operational conditions. No inhibitory effect from stack gas mixture was observed. This study provided an environmentally friendly and sustainable solution for industries such as petrochemical industry in order to produce extra energy while capturing their waste CO2. Thereby, a sustainable and environmentally friendly model solution was presented for industries with high CO2 emissions. COV: coefficient of variation; Gt: gigatone; IEA: International Energy Agency; IPCC: International Panel on Climate Change; MBBR: moving bed biofilm reactor; MJ: Megajoule; UASB: upflow anaerobic sludge blanket; VFR: volumetric feeding rate.

  12. The Odyssey of the Ancestral Escherich Strain through Culture Collections: an Example of Allopatric Diversification.

    Science.gov (United States)

    Desroches, M; Royer, G; Roche, D; Mercier-Darty, M; Vallenet, D; Médigue, C; Bastard, K; Rodriguez, C; Clermont, O; Denamur, E; Decousser, J-W

    2018-01-01

    More than a century ago, Theodor Escherich isolated the bacterium that was to become Escherichia coli , one of the most studied organisms. Not long after, the strain began an odyssey and landed in many laboratories across the world. As laboratory culture conditions could be responsible for major changes in bacterial strains, we conducted a genome analysis of isolates of this emblematic strain from different culture collections (England, France, the United States, Germany). Strikingly, many discrepancies between the isolates were observed, as revealed by multilocus sequence typing (MLST), the presence of virulence-associated genes, core genome MLST, and single nucleotide polymorphism/indel analyses. These differences are correlated with the phylogeographic history of the strain and were due to an unprecedented number of mutations in coding DNA repair functions such as mismatch repair (MutL) and oxidized guanine nucleotide pool cleaning (MutT), conferring a specific mutational spectrum and leading to a mutator phenotype. The mutator phenotype was probably acquired during subculturing and corresponded to second-order selection. Furthermore, all of the isolates exhibited hypersusceptibility to antibiotics due to mutations in efflux pump- and porin-encoding genes, as well as a specific mutation in the sigma factor-encoding gene rpoS . These defects reflect a self-preservation and nutritional competence tradeoff allowing survival under the starvation conditions imposed by storage. From a clinical point of view, dealing with such mutator strains can lead microbiologists to draw false conclusions about isolate relatedness and may impact therapeutic effectiveness. IMPORTANCE Mutator phenotypes have been described in laboratory-evolved bacteria, as well as in natural isolates. Several genes can be impacted, each of them being associated with a typical mutational spectrum. By studying one of the oldest strains available, the ancestral Escherich strain, we were able to

  13. The E. coli anti-sigma factor Rsd: studies on the specificity and regulation of its expression.

    Science.gov (United States)

    Hofmann, Nina; Wurm, Reinhild; Wagner, Rolf

    2011-05-06

    Among the seven different sigma factors in E. coli σ(70) has the highest concentration and affinity for the core RNA polymerase. The E. coli protein Rsd is regarded as an anti-sigma factor, inhibiting σ(70)-dependent transcription at the onset of stationary growth. Although binding of Rsd to σ(70) has been shown and numerous structural studies on Rsd have been performed the detailed mechanism of action is still unknown. We have performed studies to unravel the function and regulation of Rsd expression in vitro and in vivo. Cross-linking and affinity binding revealed that Rsd is able to interact with σ(70), with the core enzyme of RNA polymerase and is able to form dimers in solution. Unexpectedly, we find that Rsd does also interact with σ(38), the stationary phase-specific sigma factor. This interaction was further corroborated by gel retardation and footprinting studies with different promoter fragments and σ(38)- or σ(70)-containing RNA polymerase in presence of Rsd. Under competitive in vitro transcription conditions, in presence of both sigma factors, a selective inhibition of σ(70)-dependent transcription was prevailing, however. Analysis of rsd expression revealed that the nucleoid-associated proteins H-NS and FIS, StpA and LRP bind to the regulatory region of the rsd promoters. Furthermore, the major promoter P2 was shown to be down-regulated in vivo by RpoS, the stationary phase-specific sigma factor and the transcription factor DksA, while induction of the stringent control enhanced rsd promoter activity. Most notably, the dam-dependent methylation of a cluster of GATC sites turned out to be important for efficient rsd transcription. The results contribute to a better understanding of the intricate mechanism of Rsd-mediated sigma factor specificity changes during stationary phase.

  14. Autoinducer 2 activity in Escherichia coli culture supernatants can be actively reduced despite maintenance of an active synthase, LuxS.

    Science.gov (United States)

    Hardie, Kim R; Cooksley, Clare; Green, Andrew D; Winzer, Klaus

    2003-03-01

    Production of the signalling molecule (autoinducer-2) synthesized by LuxS has been proposed to be pivotal to a universal mechanism of inter-species bacterial cell-cell communication (quorum sensing); however recently the function of LuxS has been noted to be integral to central metabolism since it contributes to the activated methyl cycle. This paper shows that when Helicobacter pylori LuxS is overproduced in Escherichia coli, it forms cross-linkable multimers. These multimers persist at comparable levels after 24 h of growth if glucose is omitted from the growth medium; however, the levels of extracellular autoinducer-2 decline (Glucose Retention of AI-2 Levels: GRAIL). Glycerol, maltose, galactose, ribose and L-arabinose could substitute for glucose, but lactose, D-arabinose, acetate, citrate and pyruvate could not. Mutations in (i). metabolic pathways (glycolytic enzymes eno, pgk, pgm; galactose epimerase; the Pta-AckA pathway), (ii). sugar transport (pts components, rbs operon, mgl, trg), and (iii). regulators involved in conventional catabolic repression (crp, cya), cAMP-independent catabolite repression (creC, fruR, rpoS,) the stringent response (relA, spoT) and the global carbon storage regulator (csrA) did not prevent GRAIL. Although the basis of GRAIL remains uncertain, it is clear that the mechanism is distinct from conventional catabolite repression. Moreover, GRAIL is not due to inactivation of the enzymic activity of LuxS, since in E. coli, LuxS contained within stationary-phase cells grown in the absence of glucose maintains its activity in vitro.

  15. Dickeya solani sp. nov., a pectinolytic plant-pathogenic bacterium isolated from potato (Solanum tuberosum).

    Science.gov (United States)

    van der Wolf, Jan M; Nijhuis, Els H; Kowalewska, Malgorzata J; Saddler, Gerry S; Parkinson, Neil; Elphinstone, John G; Pritchard, Leighton; Toth, Ian K; Lojkowska, Ewa; Potrykus, Marta; Waleron, Malgorzata; de Vos, Paul; Cleenwerck, Ilse; Pirhonen, Minna; Garlant, Linda; Hélias, Valérie; Pothier, Joël F; Pflüger, Valentin; Duffy, Brion; Tsror, Leah; Manulis, Shula

    2014-03-01

    Pectinolytic bacteria have been recently isolated from diseased potato plants exhibiting blackleg and slow wilt symptoms found in a number of European countries and Israel. These Gram-reaction-negative, motile, rods were identified as belonging to the genus Dickeya, previously the Pectobacterium chrysanthemi complex (Erwinia chrysanthemi), on the basis of production of a PCR product with the pelADE primers, 16S rRNA gene sequence analysis, fatty acid methyl esterase analysis, the production of phosphatases and the ability to produce indole and acids from α-methylglucoside. Differential physiological assays used previously to differentiate between strains of E. chrysanthemi, showed that these isolates belonged to biovar 3. Eight of the isolates, seven from potato and one from hyacinth, were analysed together with 21 reference strains representing all currently recognized taxa within the genus Dickeya. The novel isolates formed a distinct genetic clade in multilocus sequence analysis (MLSA) using concatenated sequences of the intergenic spacer (IGS), as well as dnaX, recA, dnaN, fusA, gapA, purA, rplB, rpoS and gyrA. Characterization by whole-cell MALDI-TOF mass spectrometry, pulsed field gel electrophoresis after digestion of whole-genome DNA with rare-cutting restriction enzymes, average nucleotide identity analysis and DNA-DNA hybridization studies, showed that although related to Dickeya dadantii, these isolates represent a novel species within the genus Dickeya, for which the name Dickeya solani sp. nov. (type strain IPO 2222(T) = LMG25993(T) = NCPPB4479(T)) is proposed.

  16. Payload hardware and experimental protocol development to enable future testing of the effect of space microgravity on the resistance to gentamicin of uropathogenic Escherichia coli and its σs-deficient mutant

    Science.gov (United States)

    Matin, A. C.; Wang, J.-H.; Keyhan, Mimi; Singh, Rachna; Benoit, Michael; Parra, Macarena P.; Padgen, Michael R.; Ricco, Antonio J.; Chin, Matthew; Friedericks, Charlie R.; Chinn, Tori N.; Cohen, Aaron; Henschke, Michael B.; Snyder, Timothy V.; Lera, Matthew P.; Ross, Shannon S.; Mayberry, Christina M.; Choi, Sungshin; Wu, Diana T.; Tan, Ming X.; Boone, Travis D.; Beasley, Christopher C.; Piccini, Matthew E.; Spremo, Stevan M.

    2017-11-01

    Human immune response is compromised and bacteria can become more antibiotic resistant in space microgravity (MG). We report that under low-shear modeled microgravity (LSMMG), stationary-phase uropathogenic Escherichia coli (UPEC) become more resistant to gentamicin (Gm), and that this increase is dependent on the presence of σs (a transcription regulator encoded by the rpoS gene). UPEC causes urinary tract infections (UTIs), reported to afflict astronauts; Gm is a standard treatment, so these findings could impact astronaut health. Because LSMMG findings can differ from MG, we report preparations to examine UPEC's Gm sensitivity during spaceflight using the E. coli Anti-Microbial Satellite (EcAMSat) as a free-flying "nanosatellite" in low Earth orbit. Within EcAMSat's payload, a 48-microwell fluidic card contains and supports study of bacterial cultures at constant temperature; optical absorbance changes in cell suspensions are made at three wavelengths for each microwell and a fluid-delivery system provides growth medium and predefined Gm concentrations. Performance characterization is reported here for spaceflight prototypes of this payload system. Using conventional microtiter plates, we show that Alamar Blue (AB) absorbance changes can assess the Gm effect on E. coli viability, permitting telemetric transfer of the spaceflight data to Earth. Laboratory results using payload prototypes are consistent with wellplate and flask findings of differential sensitivity of UPEC and its ΔrpoS strain to Gm. if σs plays the same role in space MG as in LSMMG and Earth gravity, countermeasures discovered in recent Earth studies (aimed at weakening the UPEC antioxidant defense) to control UPEC infections would prove useful also in space flights. Further, EcAMSat results should clarify inconsistencies from previous space experiments on bacterial antibiotic sensitivity and other issues.

  17. DNA methylation-dependent regulation of pef expression in Salmonella typhimurium.

    Science.gov (United States)

    Nicholson, B; Low, D

    2000-02-01

    Plasmid-encoded fimbriae (Pef) expressed by Salmonella typhimurium mediate adhesion to mouse intestinal epithelium. The pef operon shares features with the Escherichia coli pyelonephritis-associated pilus (pap) operon, which is under methylation-dependent transcriptional regulation. These features include conserved DNA GATC box sites in the upstream regulatory region as well as homologues of the PapI and PapB regulatory proteins. Unlike Pap fimbriae, which are expressed in a variety of laboratory media, Pef fimbriae were expressed only in acidic, rich broth under standing culture conditions. Analysis of S. typhimurium grown under these conditions indicated that Pef production was regulated by a phase variation mechanism, in which the bacterial population was skewed between fimbrial expression (phase ON) and non-expression (phase OFF) states. Leucine-responsive regulatory protein (Lrp) and DNA adenine methylase (Dam) were required for pef transcription. In contrast, the histone-like protein (H-NS) and the stationary-phase sigma factor (RpoS) repressed pef transcription. Methylation of the pef GATC II site appeared to be required for pef fimbrial expression based on analysis of a GCTC II mutant that did not express Pef fimbriae. Analysis of the DNA methylation states of pef GATC sites indicated that, under acidic growth conditions, which induced Pef production, most GATC I sites were non-methylated, whereas GATC II and GATC X were predominantly methylated. The methylation protection at GATC I and GATC II was dependent upon Lrp and was modulated by PefI. Together, these results indicate that Pef production is regulated by DNA methylation, which is the first example of methylation-dependent gene regulation outside of E. coli.

  18. Effect of levels of acetate on the mevalonate pathway of Borrelia burgdorferi.

    Directory of Open Access Journals (Sweden)

    Tricia A Van Laar

    Full Text Available Borrelia burgdorferi, the agent of Lyme disease, is a spirochetal pathogen with limited metabolic capabilities that survives under highly disparate host-specific conditions. However, the borrelial genome encodes several proteins of the mevalonate pathway (MP that utilizes acetyl-CoA as a substrate leading to intermediate metabolites critical for biogenesis of peptidoglycan and post-translational modifications of proteins. In this study, we analyzed the MP and contributions of acetate in modulation of adaptive responses in B. burgdorferi. Reverse-transcription PCR revealed that components of the MP are transcribed as individual open reading frames. Immunoblot analysis using monospecific sera confirmed synthesis of members of the MP in B. burgdorferi. The rate-limiting step of the MP is mediated by HMG-CoA reductase (HMGR via conversion of HMG-CoA to mevalonate. Recombinant borrelial HMGR exhibited a K(m value of 132 µM with a V(max of 1.94 µmol NADPH oxidized minute(-1 (mg protein(-1 and was inhibited by statins. Total protein lysates from two different infectious, clonal isolates of B. burgdorferi grown under conditions that mimicked fed-ticks (pH 6.8/37°C exhibited increased levels of HMGR while other members of the MP were elevated under unfed-tick (pH 7.6/23°C conditions. Increased extra-cellular acetate gave rise to elevated levels of MP proteins along with RpoS, CsrA(Bb and their respective regulons responsible for mediating vertebrate host-specific adaptation. Both lactone and acid forms of two different statins inhibited growth of B. burgdorferi strain B31, while overexpression of HMGR was able to partially overcome that inhibition. In summary, these studies on MP and contributions of acetate to host-specific adaptation have helped identify potential metabolic targets that can be manipulated to reduce the incidence of Lyme disease.

  19. Inhibition of the colony-stimulating-factor-1 receptor affects the resistance of lung cancer cells to cisplatin.

    Science.gov (United States)

    Pass, Harvey I; Lavilla, Carmencita; Canino, Claudia; Goparaju, Chandra; Preiss, Jordan; Noreen, Samrah; Blandino, Giovanni; Cioce, Mario

    2016-08-30

    In the present work we show that multiple lung cancer cell lines contain cisplatin resistant cell subpopulations expressing the Colony-Stimulating-Factor-Receptor-1 (CSF-1R) and surviving chemotherapy-induced stress. By exploiting siRNA-mediated knock down in vitro and the use of an investigational CSF-1R TKI (JNJ-40346527) in vitro and in vivo, we show that expression and function of the receptor are required for the clonogenicity and chemoresistance of the cell lines. Thus, inhibition of the kinase activity of the receptor reduced the levels of EMT-associated genes, stem cell markers and chemoresistance genes. Additionally, the number of high aldehyde dehydrogenase (ALDH) expressing cells was reduced, consequent to the lack of cisplatin-induced increase of ALDH isoforms. This affected the collective chemoresistance of the treated cultures. Treatment of tumor bearing mice with JNJ-40346527, at pharmacologically relevant doses, produced strong chemo-sensitizing effects in vivo. These anticancer effects correlated with a reduced number of CSF-1Rpos cells, in tumors excised from the treated mice. Depletion of the CD45pos cells within the treated tumors did not, apparently, play a major role in mediating the therapeutic response to the TKI. Thus, lung cancer cells express a functional CSF-1 and CSF-1R duo which mediates pro-tumorigenic effects in vivo and in vitro and can be targeted in a therapeutically relevant way. These observations complement the already known role for the CSF-1R at mediating the pro-tumorigenic properties of tumor-infiltrating immune components.

  20. Contribution of Stress Responses to Antibiotic Tolerance in Pseudomonas aeruginosa Biofilms

    Science.gov (United States)

    Franklin, Michael J.; Williamson, Kerry S.; Folsom, James P.; Boegli, Laura; James, Garth A.

    2015-01-01

    Enhanced tolerance of biofilm-associated bacteria to antibiotic treatments is likely due to a combination of factors, including changes in cell physiology as bacteria adapt to biofilm growth and the inherent physiological heterogeneity of biofilm bacteria. In this study, a transcriptomics approach was used to identify genes differentially expressed during biofilm growth of Pseudomonas aeruginosa. These genes were tested for statistically significant overlap, with independently compiled gene lists corresponding to stress responses and other putative antibiotic-protective mechanisms. Among the gene groups tested were those associated with biofilm response to tobramycin or ciprofloxacin, drug efflux pumps, acyl homoserine lactone quorum sensing, osmotic shock, heat shock, hypoxia stress, and stationary-phase growth. Regulons associated with Anr-mediated hypoxia stress, RpoS-regulated stationary-phase growth, and osmotic stress were significantly enriched in the set of genes induced in the biofilm. Mutant strains deficient in rpoS, relA and spoT, or anr were cultured in biofilms and challenged with ciprofloxacin and tobramycin. When challenged with ciprofloxacin, the mutant strain biofilms had 2.4- to 2.9-log reductions in viable cells compared to a 0.9-log reduction of the wild-type strain. Interestingly, none of the mutants exhibited a statistically significant alteration in tobramycin susceptibility compared to that with the wild-type biofilm. These results are consistent with a model in which multiple genes controlled by overlapping starvation or stress responses contribute to the protection of a P. aeruginosa biofilm from ciprofloxacin. A distinct and as yet undiscovered mechanism protects the biofilm bacteria from tobramycin. PMID:25870065

  1. PtrA Is Functionally Intertwined with GacS in Regulating the Biocontrol Activity of Pseudomonas chlororaphis PA23

    Science.gov (United States)

    Shah, Nidhi; Klaponski, Natasha; Selin, Carrie; Rudney, Rachel; Fernando, W. G. Dilantha; Belmonte, Mark F.; de Kievit, Teresa R.

    2016-01-01

    In vitro inhibition of the fungal pathogen Sclerotinia sclerotiorum by Pseudomonas chlororaphis PA23 is reliant upon a LysR-type transcriptional regulator (LTTR) called PtrA. In the current study, we show that Sclerotinia stem rot and leaf infection are significantly increased in canola plants inoculated with the ptrA-mutant compared to the wild type, establishing PtrA as an essential regulator of PA23 biocontrol. LTTRs typically regulate targets that are upstream of and divergently transcribed from the LTTR locus. We identified a short chain dehydrogenase (scd) gene immediately upstream of ptrA. Characterization of a scd mutant revealed that it is phenotypically identical to the wild type. Moreover, scd transcript abundance was unchanged in the ptrA mutant. These findings indicate that PtrA regulation does not involve scd, rather this LTTR controls genes located elsewhere on the chromosome. Employing a combination of complementation and transcriptional analysis we investigated whether connections exist between PtrA and other regulators of biocontrol. Besides ptrA, gacS was the only gene able to partially rescue the wild-type phenotype, establishing a connection between PtrA and the sensor kinase GacS. Transcriptomic analysis revealed decreased expression of biosynthetic (phzA, prnA) and regulatory genes (phzI, phzR, rpoS, gacA, rsmX, rsmZ, retS) in the ptrA mutant; conversely, rsmE, and rsmY were markedly upregulated. The transcript abundance of ptrA was nine-fold higher in the mutant background indicating that this LTTR negatively autoregulates itself. In summary, PtrA is an essential regulator of genes required for PA23 biocontrol that is functionally intertwined with GacS. PMID:27713742

  2. Effect of heparin and heparan sulphate on open promoter complex formation for a simple tandem gene model using ex situ atomic force microscopy.

    Science.gov (United States)

    Chammas, Oliver; Bonass, William A; Thomson, Neil H

    2017-05-01

    The influence of heparin and heparan sulphate (HepS) on the appearance and analysis of open promoter complex (RPo) formation by E. coli RNA polymerase (RNAP) holoenzyme (σ70RNAP) on linear DNA using ex situ imaging by atomic force microscopy (AFM) has been investigated. Introducing heparin or HepS into the reaction mix significantly reduces non-specific interactions of the σ70RNAP and RNAP after RPo formation allowing for better interpretation of complexes shown within AFM images, particularly on DNA templates containing more than one promoter. Previous expectation was that negatively charged polysaccharides, often used as competitive inhibitors of σRNAP binding and RPo formation, would also inhibit binding of the DNA template to the mica support surface and thereby lower the imaging yield of active RNAP-DNA complexes. We found that the reverse of this was true, and that the yield of RPo formation detected by AFM, for a simple tandem gene model containing two λPR promoters, increased. Moreover and unexpectedly, HepS was more efficient than heparin, with both of them having a dispersive effect on the sample, minimising unwanted RNAP-RNAP interactions as well as non-specific interactions between the RNAP and DNA template. The success of this method relied on the observation that E. coli RNAP has the highest affinity for the mica surface of all the molecular components. For our system, the affinity of the three constituent biopolymers to muscovite mica was RNAP>Heparin or HepS>DNA. While we observed that heparin and HepS can inhibit DNA binding to the mica, the presence of E. coli RNAP overcomes this effect allowing a greater yield of RPos for AFM analysis. This method can be extended to other DNA binding proteins and enzymes, which have an affinity to mica higher than DNA, to improve sample preparation for AFM studies. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

  3. Activation of the Glutamic Acid-Dependent Acid Resistance System in Escherichia coli BL21(DE3 Leads to Increase of the Fatty Acid Biotransformation Activity.

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    Ji-Min Woo

    Full Text Available The biosynthesis of carboxylic acids including fatty acids from biomass is central in envisaged biorefinery concepts. The productivities are often, however, low due to product toxicity that hamper whole-cell biocatalyst performance. Here, we have investigated factors that influence the tolerance of Escherichia coli to medium chain carboxylic acid (i.e., n-heptanoic acid-induced stress. The metabolic and genomic responses of E. coli BL21(DE3 and MG1655 grown in the presence of n-heptanoic acid indicated that the GadA/B-based glutamic acid-dependent acid resistance (GDAR system might be critical for cellular tolerance. The GDAR system, which is responsible for scavenging intracellular protons by catalyzing decarboxylation of glutamic acid, was inactive in E. coli BL21(DE3. Activation of the GDAR system in this strain by overexpressing the rcsB and dsrA genes, of which the gene products are involved in the activation of GadE and RpoS, respectively, resulted in acid tolerance not only to HCl but also to n-heptanoic acid. Furthermore, activation of the GDAR system allowed the recombinant E. coli BL21(DE3 expressing the alcohol dehydrogenase of Micrococcus luteus and the Baeyer-Villiger monooxygenase of Pseudomonas putida to reach 60% greater product concentration in the biotransformation of ricinoleic acid (i.e., 12-hydroxyoctadec-9-enoic acid (1 into n-heptanoic acid (5 and 11-hydroxyundec-9-enoic acid (4. This study may contribute to engineering E. coli-based biocatalysts for the production of carboxylic acids from renewable biomass.

  4. Enhancing E. coli tolerance towards oxidative stress via engineering its global regulator cAMP receptor protein (CRP.

    Directory of Open Access Journals (Sweden)

    Souvik Basak

    Full Text Available Oxidative damage to microbial hosts often occurs under stressful conditions during bioprocessing. Classical strain engineering approaches are usually both time-consuming and labor intensive. Here, we aim to improve E. coli performance under oxidative stress via engineering its global regulator cAMP receptor protein (CRP, which can directly or indirectly regulate redox-sensing regulators SoxR and OxyR, and other ~400 genes in E. coli. Error-prone PCR technique was employed to introduce modifications to CRP, and three mutants (OM1~OM3 were identified with improved tolerance via H(2O(2 enrichment selection. The best mutant OM3 could grow in 12 mM H(2O(2 with the growth rate of 0.6 h(-1, whereas the growth of wild type was completely inhibited at this H(2O(2 concentration. OM3 also elicited enhanced thermotolerance at 48°C as well as resistance against cumene hydroperoxide. The investigation about intracellular reactive oxygen species (ROS, which determines cell viability, indicated that the accumulation of ROS in OM3 was always lower than in WT with or without H(2O(2 treatment. Genome-wide DNA microarray analysis has shown not only CRP-regulated genes have demonstrated great transcriptional level changes (up to 8.9-fold, but also RpoS- and OxyR-regulated genes (up to 7.7-fold. qRT-PCR data and enzyme activity assay suggested that catalase (katE could be a major antioxidant enzyme in OM3 instead of alkyl hydroperoxide reductase or superoxide dismutase. To our knowledge, this is the first work on improving E. coli oxidative stress resistance by reframing its transcription machinery through its native global regulator. The positive outcome of this approach may suggest that engineering CRP can be successfully implemented as an efficient strain engineering alternative for E. coli.

  5. Activation of the Glutamic Acid-Dependent Acid Resistance System in Escherichia coli BL21(DE3) Leads to Increase of the Fatty Acid Biotransformation Activity.

    Science.gov (United States)

    Woo, Ji-Min; Kim, Ji-Won; Song, Ji-Won; Blank, Lars M; Park, Jin-Byung

    The biosynthesis of carboxylic acids including fatty acids from biomass is central in envisaged biorefinery concepts. The productivities are often, however, low due to product toxicity that hamper whole-cell biocatalyst performance. Here, we have investigated factors that influence the tolerance of Escherichia coli to medium chain carboxylic acid (i.e., n-heptanoic acid)-induced stress. The metabolic and genomic responses of E. coli BL21(DE3) and MG1655 grown in the presence of n-heptanoic acid indicated that the GadA/B-based glutamic acid-dependent acid resistance (GDAR) system might be critical for cellular tolerance. The GDAR system, which is responsible for scavenging intracellular protons by catalyzing decarboxylation of glutamic acid, was inactive in E. coli BL21(DE3). Activation of the GDAR system in this strain by overexpressing the rcsB and dsrA genes, of which the gene products are involved in the activation of GadE and RpoS, respectively, resulted in acid tolerance not only to HCl but also to n-heptanoic acid. Furthermore, activation of the GDAR system allowed the recombinant E. coli BL21(DE3) expressing the alcohol dehydrogenase of Micrococcus luteus and the Baeyer-Villiger monooxygenase of Pseudomonas putida to reach 60% greater product concentration in the biotransformation of ricinoleic acid (i.e., 12-hydroxyoctadec-9-enoic acid (1)) into n-heptanoic acid (5) and 11-hydroxyundec-9-enoic acid (4). This study may contribute to engineering E. coli-based biocatalysts for the production of carboxylic acids from renewable biomass.

  6. Representation of planar motion of complex joints by means of rolling pairs. Application to neck motion.

    Science.gov (United States)

    Page, Alvaro; de Rosario, Helios; Gálvez, José A; Mata, Vicente

    2011-02-24

    We propose to model planar movements between two human segments by means of rolling-without-slipping kinematic pairs. We compute the path traced by the instantaneous center of rotation (ICR) as seen from the proximal and distal segments, thus obtaining the fixed and moving centrodes, respectively. The joint motion is then represented by the rolling-without-slipping of one centrode on the other. The resulting joint kinematic model is based on the real movement and accounts for nonfixed axes of rotation; therefore it could improve current models based on revolute pairs in those cases where joint movement implies displacement of the ICR. Previous authors have used the ICR to characterize human joint motion, but they only considered the fixed centrode. Such an approach is not adequate for reproducing motion because the fixed centrode by itself does not convey information about body position. The combination of the fixed and moving centrodes gathers the kinematic information needed to reproduce the position and velocities of moving bodies. To illustrate our method, we applied it to the flexion-extension movement of the head relative to the thorax. The model provides a good estimation of motion both for position variables (mean R(pos)=0.995) and for velocities (mean R(vel)=0.958). This approach is more realistic than other models of neck motion based on revolute pairs, such as the dual-pivot model. The geometry of the centrodes can provide some information about the nature of the movement. For instance, the ascending and descending curves of the fixed centrode suggest a sequential movement of the cervical vertebrae. Copyright © 2010 Elsevier Ltd. All rights reserved.

  7. Global genome response of Escherichia coli O157∶H7 Sakai during dynamic changes in growth kinetics induced by an abrupt downshift in water activity.

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    Chawalit Kocharunchitt

    Full Text Available The present study was undertaken to investigate growth kinetics and time-dependent change in global expression of Escherichia coli O157∶H7 Sakai upon an abrupt downshift in water activity (aw. Based on viable count data, shifting E. coli from aw 0.993 to aw 0.985 or less caused an apparent loss, then recovery, of culturability. Exponential growth then resumed at a rate characteristic for the aw imposed. To understand the responses of this pathogen to abrupt osmotic stress, we employed an integrated genomic and proteomic approach to characterize its cellular response during exposure to a rapid downshift but still within the growth range from aw 0.993 to aw 0.967. Of particular interest, genes and proteins with cell envelope-related functions were induced during the initial loss and subsequent recovery of culturability. This implies that cells undergo remodeling of their envelope composition, enabling them to adapt to osmotic stress. Growth at low aw, however, involved up-regulating additional genes and proteins, which are involved in the biosynthesis of specific amino acids, and carbohydrate catabolism and energy generation. This suggests their important role in facilitating growth under such stress. Finally, we highlighted the ability of E. coli to activate multiple stress responses by transiently inducing the RpoE and RpoH regulons to control protein misfolding, while simultaneously activating the master stress regulator RpoS to mediate long-term adaptation to hyperosmolality. This investigation extends our understanding of the potential mechanisms used by pathogenic E. coli to adapt, survive and grow under osmotic stress, which could potentially be exploited to aid the selection and/or development of novel strategies to inactivate this pathogen.

  8. The membrane protein LasM Promotes the Culturability of Legionella pneumophila in Water

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    Laam Li

    2016-09-01

    Full Text Available The water-borne pathogen Legionella pneumophila (Lp strongly expresses the lpg1659 gene in water. This gene encodes a hypothetical protein predicted to be a membrane protein using in silico analysis. While no conserved domains were identified in Lpg1659, similar proteins are found in many Legionella species and other aquatic bacteria. RT-qPCR showed that lpg1659 is positively regulated by the alternative sigma factor RpoS, which is essential for Lp to survive in water. These observations suggest an important role of this novel protein in the survival of Lp in water. Deletion of lpg1659 did not affect cell morphology, membrane integrity or tolerance to high temperature. Moreover, lpg1659 was dispensable for growth of Lp in rich medium, and during infection of the amoeba Acanthamoeba castellanii and of THP-1 human macrophages. However, deletion of lpg1659 resulted in an early loss of culturability in water, while over-expression of this gene promoted the culturability of Lp. Therefore, these results suggest that lpg1659 is required for Lp to maintain culturability, and possibly long-term survival, in water. Since the loss of culturability observed in the absence of Lpg1659 was complemented by the addition of trace metals into water, this membrane protein is likely a transporter for acquiring essential trace metal for maintaining culturability in water and potentially in other metal-deprived conditions. Given its role in the survival of Lp in water, Lpg1659 was named LasM for Legionella aquatic survival membrane protein.

  9. Toxin YafQ Reduces Escherichia coli Growth at Low Temperatures.

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    Yueju Zhao

    Full Text Available Toxin/antitoxin (TA systems reduce metabolism under stress; for example, toxin YafQ of the YafQ/DinJ Escherichia coli TA system reduces growth by cleaving transcripts with in-frame 5'-AAA-G/A-3' sites, and antitoxin DinJ is a global regulator that represses its locus as well as controls levels of the stationary sigma factor RpoS. Here we investigated the influence on cell growth at various temperatures and found that deletion of the antitoxin gene, dinJ, resulted in both reduced metabolism and slower growth at 18°C but not at 37°C. The reduction in growth could be complemented by producing DinJ from a plasmid. Using a transposon screen to reverse the effect of the absence of DinJ, two mutations were found that inactivated the toxin YafQ; hence, the toxin caused the slower growth only at low temperatures rather than DinJ acting as a global regulator. Corroborating this result, a clean deletion of yafQ in the ΔdinJ ΔKmR strain restored both metabolism and growth at 18°C. In addition, production of YafQ was more toxic at 18°C compared to 37°C. Furthermore, by overproducing all the E. coli proteins, the global transcription repressor Mlc was found that counteracts YafQ toxicity only at 18°C. Therefore, YafQ is more effective at reducing metabolism at low temperatures, and Mlc is its putative target.

  10. A role for the RNA chaperone Hfq in controlling adherent-invasive Escherichia coli colonization and virulence.

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    Karina T Simonsen

    Full Text Available Adherent-invasive Escherichia coli (AIEC has been linked with the onset and perpetuation of inflammatory bowel diseases. The AIEC strain LF82 was originally isolated from an ileal biopsy from a patient with Crohn's disease. The pathogenesis of LF82 results from its abnormal adherence to and subsequent invasion of the intestinal epithelium coupled with its ability to survive phagocytosis by macrophages once it has crossed the intestinal barrier. To gain further insight into AIEC pathogenesis we employed the nematode Caenorhabditis elegans as an in vivo infection model. We demonstrate that AIEC strain LF82 forms a persistent infection in C. elegans, thereby reducing the host lifespan significantly. This host killing phenotype was associated with massive bacterial colonization of the nematode intestine and damage to the intestinal epithelial surface. C. elegans killing was independent of known LF82 virulence determinants but was abolished by deletion of the LF82 hfq gene, which encodes an RNA chaperone involved in mediating posttranscriptional gene regulation by small non-coding RNAs. This finding reveals that important aspects of LF82 pathogenesis are controlled at the posttranscriptional level by riboregulation. The role of Hfq in LF82 virulence was independent of its function in regulating RpoS and RpoE activity. Further, LF82Δhfq mutants were non-motile, impaired in cell invasion and highly sensitive to various chemical stress conditions, reinforcing the multifaceted function of Hfq in mediating bacterial adaptation. This study highlights the usefulness of simple non-mammalian infection systems for the identification and analysis of bacterial virulence factors.

  11. Improving ethanol tolerance of Escherichia coli by rewiring its global regulator cAMP receptor protein (CRP.

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    Huiqing Chong

    Full Text Available A major challenge in bioethanol fermentation is the low tolerance of the microbial host towards the end product bioethanol. Here we report to improve the ethanol tolerance of E. coli from the transcriptional level by engineering its global transcription factor cAMP receptor protein (CRP, which is known to regulate over 400 genes in E. coli. Three ethanol tolerant CRP mutants (E1- E3 were identified from error-prone PCR libraries. The best ethanol-tolerant strain E2 (M59T had the growth rate of 0.08 h(-1 in 62 g/L ethanol, higher than that of the control at 0.06 h(-1. The M59T mutation was then integrated into the genome to create variant iE2. When exposed to 150 g/l ethanol, the survival of iE2 after 15 min was about 12%, while that of BW25113 was <0.01%. Quantitative real-time reverse transcription PCR analysis (RT-PCR on 444 CRP-regulated genes using OpenArray® technology revealed that 203 genes were differentially expressed in iE2 in the absence of ethanol, whereas 92 displayed differential expression when facing ethanol stress. These genes belong to various functional groups, including central intermediary metabolism (aceE, acnA, sdhD, sucA, iron ion transport (entH, entD, fecA, fecB, and general stress response (osmY, rpoS. Six up-regulated and twelve down-regulated common genes were found in both iE2 and E2 under ethanol stress, whereas over one hundred common genes showed differential expression in the absence of ethanol. Based on the RT-PCR results, entA, marA or bhsA was knocked out in iE2 and the resulting strains became more sensitive towards ethanol.

  12. The NlpD lipoprotein is a novel Yersinia pestis virulence factor essential for the development of plague.

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    Avital Tidhar

    Full Text Available Yersinia pestis is the causative agent of plague. Previously we have isolated an attenuated Y. pestis transposon insertion mutant in which the pcm gene was disrupted. In the present study, we investigated the expression and the role of pcm locus genes in Y. pestis pathogenesis using a set of isogenic surE, pcm, nlpD and rpoS mutants of the fully virulent Kimberley53 strain. We show that in Y. pestis, nlpD expression is controlled from elements residing within the upstream genes surE and pcm. The NlpD lipoprotein is the only factor encoded from the pcm locus that is essential for Y. pestis virulence. A chromosomal deletion of the nlpD gene sequence resulted in a drastic reduction in virulence to an LD(50 of at least 10(7 cfu for subcutaneous and airway routes of infection. The mutant was unable to colonize mouse organs following infection. The filamented morphology of the nlpD mutant indicates that NlpD is involved in cell separation; however, deletion of nlpD did not affect in vitro growth rate. Trans-complementation experiments with the Y. pestis nlpD gene restored virulence and all other phenotypic defects. Finally, we demonstrated that subcutaneous administration of the nlpD mutant could protect animals against bubonic and primary pneumonic plague. Taken together, these results demonstrate that Y. pestis NlpD is a novel virulence factor essential for the development of bubonic and pneumonic plague. Further, the nlpD mutant is superior to the EV76 prototype live vaccine strain in immunogenicity and in conferring effective protective immunity. Thus it could serve as a basis for a very potent live vaccine against bubonic and pneumonic plague.

  13. Escherichia coli O157:H7 Acid Sensitivity Correlates with Flocculation Phenotype during Nutrient Limitation

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    Kathryn L. Kay

    2017-07-01

    Full Text Available Shiga toxin producing Escherichia coli (STEC strains vary in acid resistance; however, little is known about the underlying mechanisms that result in strain specific differences. Among 25 STEC O157:H7 strains tested, 7 strains flocculated when grown statically for 18 h in minimal salts medium at 37°C, while 18 strains did not. Interestingly, the flocculation phenotype (cells came out of suspension was found to correlate with degree of acid sensitivity in an assay with 400 mM acetic acid solution at pH 3.3 targeting acidified foods. Strains exhibiting flocculation were more acid sensitive and were designated FAS, for flocculation acid sensitive, while the acid resistant strain designated PAR for planktonic acid resistant. Flocculation was not observed for any strains during growth in complex medium (Luria Bertani broth. STEC strains B201 and B241 were chosen as representative FAS (2.4 log reduction and PAR (0.15 log reduction strains, respectively, due to differences in acid resistance and flocculation phenotype. Results from electron microscopy showed evidence of fimbriae production in B201, whereas fimbriae were not observed in B241.Curli fimbriae production was identified through plating on Congo red differential medium, and all FAS strains showed curli fimbriae production. Surprisingly, 5 PAR strains also had evidence of curli production. Transcriptomic and targeted gene expression data for B201 and B241indicated that csg and hde (curli and acid induced chaperone genes, respectively expression positively correlated with the phenotypic differences observed for these strains. These data suggest that FAS strains grown in minimal medium express curli, resulting in a flocculation phenotype. This may be regulated by GcvB, which positively regulates curli fimbriae production and represses acid chaperone proteins. RpoS and other regulatory mechanisms may impact curli fimbriae production, as well. These findings may help elucidate mechanisms

  14. Mechanism of bacterial inactivation by (+-limonene and its potential use in food preservation combined processes.

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    Laura Espina

    Full Text Available This work explores the bactericidal effect of (+-limonene, the major constituent of citrus fruits' essential oils, against E. coli. The degree of E. coli BJ4 inactivation achieved by (+-limonene was influenced by the pH of the treatment medium, being more bactericidal at pH 4.0 than at pH 7.0. Deletion of rpoS and exposure to a sub-lethal heat or an acid shock did not modify E. coli BJ4 resistance to (+-limonene. However, exposure to a sub-lethal cold shock decreased its resistance to (+-limonene. Although no sub-lethal injury was detected in the cell envelopes after exposure to (+-limonene by the selective-plating technique, the uptake of propidium iodide by inactivated E. coli BJ4 cells pointed out these structures as important targets in the mechanism of action. Attenuated Total Reflectance Infrared Microspectroscopy (ATR-IRMS allowed identification of altered E. coli BJ4 structures after (+-limonene treatments as a function of the treatment pH: β-sheet proteins at pH 4.0 and phosphodiester bonds at pH 7.0. The increased sensitivity to (+-limonene observed at pH 4.0 in an E. coli MC4100 lptD4213 mutant with an increased outer membrane permeability along with the identification of altered β-sheet proteins by ATR-IRMS indicated the importance of this structure in the mechanism of action of (+-limonene. The study of mechanism of inactivation by (+-limonene led to the design of a synergistic combined process with heat for the inactivation of the pathogen E. coli O157:H7 in fruit juices. These results show the potential of (+-limonene in food preservation, either acting alone or in combination with lethal heat treatments.

  15. Mechanism of Bacterial Inactivation by (+)-Limonene and Its Potential Use in Food Preservation Combined Processes

    Science.gov (United States)

    Espina, Laura; Gelaw, Tilahun K.; de Lamo-Castellví, Sílvia; Pagán, Rafael; García-Gonzalo, Diego

    2013-01-01

    This work explores the bactericidal effect of (+)-limonene, the major constituent of citrus fruits' essential oils, against E. coli. The degree of E. coli BJ4 inactivation achieved by (+)-limonene was influenced by the pH of the treatment medium, being more bactericidal at pH 4.0 than at pH 7.0. Deletion of rpoS and exposure to a sub-lethal heat or an acid shock did not modify E. coli BJ4 resistance to (+)-limonene. However, exposure to a sub-lethal cold shock decreased its resistance to (+)-limonene. Although no sub-lethal injury was detected in the cell envelopes after exposure to (+)-limonene by the selective-plating technique, the uptake of propidium iodide by inactivated E. coli BJ4 cells pointed out these structures as important targets in the mechanism of action. Attenuated Total Reflectance Infrared Microspectroscopy (ATR-IRMS) allowed identification of altered E. coli BJ4 structures after (+)-limonene treatments as a function of the treatment pH: β-sheet proteins at pH 4.0 and phosphodiester bonds at pH 7.0. The increased sensitivity to (+)-limonene observed at pH 4.0 in an E. coli MC4100 lptD4213 mutant with an increased outer membrane permeability along with the identification of altered β-sheet proteins by ATR-IRMS indicated the importance of this structure in the mechanism of action of (+)-limonene. The study of mechanism of inactivation by (+)-limonene led to the design of a synergistic combined process with heat for the inactivation of the pathogen E. coli O157:H7 in fruit juices. These results show the potential of (+)-limonene in food preservation, either acting alone or in combination with lethal heat treatments. PMID:23424676

  16. New Stereo Vision Digital Camera System for Simultaneous Measurement of Cloud Base Height and Atmospheric Visibility

    Science.gov (United States)

    Janeiro, F. M.; Carretas, F.; Palma, N.; Ramos, P. M.; Wagner, F.

    2013-12-01

    Clouds play an important role in many aspects of everyday life. They affect both the local weather as well as the global climate and are an important parameter on climate change studies. Cloud parameters are also important for weather prediction models which make use of actual measurements. It is thus important to have low-cost instrumentation that can be deployed in the field to measure those parameters. This kind of instruments should also be automated and robust since they may be deployed in remote places and be subject to adverse weather conditions. Although clouds are very important in environmental systems, they are also an essential component of airplane safety when visual flight rules (VFR) are enforced, such as in most small aerodromes where it is not economically viable to install instruments for assisted flying. Under VFR there are strict limits on the height of the cloud base, cloud cover and atmospheric visibility that ensure the safety of the pilots and planes. Although there are instruments, available in the market, to measure those parameters, their relatively high cost makes them unavailable in many local aerodromes. In this work we present a new prototype which has been recently developed and deployed in a local aerodrome as proof of concept. It is composed by two digital cameras that capture photographs of the sky and allow the measurement of the cloud height from the parallax effect. The new developments consist on having a new geometry which allows the simultaneous measurement of cloud base height, wind speed at cloud base height and atmospheric visibility, which was not previously possible with only two cameras. The new orientation of the cameras comes at the cost of a more complex geometry to measure the cloud base height. The atmospheric visibility is calculated from the Lambert-Beer law after the measurement of the contrast between a set of dark objects and the background sky. The prototype includes the latest hardware developments that

  17. Terpenoids from Platostoma rotundifolium (Briq.) A. J. Paton Alter the Expression of Quorum Sensing-Related Virulence Factors and the Formation of Biofilm in Pseudomonas aeruginosa PAO1

    Science.gov (United States)

    Rasamiravaka, Tsiry; Ngezahayo, Jérémie; Pottier, Laurent; Oliveira Ribeiro, Sofia; Souard, Florence; Hari, Léonard; Stévigny, Caroline; El Jaziri, Mondher; Duez, Pierre

    2017-01-01

    Platostoma rotundifolium (Briq.) A. J. Paton aerial parts are widely used in Burundi traditional medicine to treat infectious diseases. In order to investigate their probable antibacterial activities, crude extracts from P. rotundifolium were assessed for their bactericidal and anti-virulence properties against an opportunistic bacterial model, Pseudomonas aeruginosa PAO1. Whereas none of the tested extracts exert bacteriostatic and/or bactericidal proprieties, the ethyl acetate and dichloromethane extracts exhibit anti-virulence properties against Pseudomonas aeruginosa PAO1 characterized by an alteration in quorum sensing gene expression and biofilm formation without affecting bacterial viability. Bioguided fractionation of the ethyl acetate extract led to the isolation of major anti-virulence compounds that were identified from nuclear magnetic resonance and high-resolution molecular spectroscopy spectra as cassipourol, β-sitosterol and α-amyrin. Globally, cassipourol and β-sitosterol inhibit quorum sensing-regulated and -regulatory genes expression in las and rhl systems without affecting the global regulators gacA and vfr, whereas α-amyrin had no effect on the expression of these genes. These terpenoids disrupt the formation of biofilms at concentrations down to 12.5, 50 and 50 µM for cassipourol, β-sitosterol and α-amyrin, respectively. Moreover, these terpenoids reduce the production of total exopolysaccharides and promote flagella-dependent motilities (swimming and swarming). The isolated terpenoids exert a wide range of inhibition processes, suggesting a complex mechanism of action targeting P. aeruginosa virulence mechanisms which support the wide anti-infectious use of this plant species in traditional Burundian medicine. PMID:28613253

  18. Hepatitis B risks and immunisation coverage amongst Australians travelling to southeast Asia and east Asia.

    Science.gov (United States)

    Leggat, Peter A; Zwar, Nicholas A; Hudson, Bernard J

    2009-11-01

    Southeast Asia and East Asia are regarded as highly endemic regions for hepatitis B virus (HBV) and include many popular destinations for Australian travellers. The objectives of this survey were to evaluate the extent of pre-travel health advice, the prevalence of behaviours with HBV infection risks and the prevalence of HBV vaccination amongst Australian travellers to Southeast Asia and East Asia. In 2004, a telephone survey was conducted amongst Australians, who had travelled overseas to Southeast Asia and East Asia in the past three years for three nights or more. Three hundred and nine travellers aged 14 years and over completed the interview, including 138 males (45%) and 171 females (55%). Respondents travelled for leisure (64%), business (20%), and visiting friends and relatives (VFR) (16%). The most common destinations were Indonesia (34%), Thailand (32%), and China (27%). About half of the travellers (54%) sought vaccination specific health advice before travel of which about half (56%) had sought this advice more than six weeks before travel. Just over one quarter of travellers reported receiving HBV vaccination (28%) of whom most (70%) were vaccinated at least three weeks before travel. About half of the travellers (49%) had participated in at least one activity with HBV risk during their last overseas trip. Of those travellers aged 18 years and over who either had no HBV vaccination or who were unsure, about half (49%) had participated in at least one activity with HBV risk during their last overseas trip. Australian travellers to Southeast Asia and East Asia commonly undertake activities with a risk of exposure to HBV. Hepatitis B vaccination coverage amongst this group remains low. It is important that travellers to this region seek travel health advice from a qualified source, which will include a risk assessment for hepatitis B vaccination.

  19. German travelers' preferences for travel vaccines assessed by a discrete choice experiment.

    Science.gov (United States)

    Poulos, Christine; Curran, Desmond; Anastassopoulou, Anastassia; De Moerlooze, Laurence

    2018-02-08

    Many travelers to regions with endemic infectious diseases do not follow health authorities' recommendations regarding vaccination against vaccine-preventable infectious diseases, before traveling. The determinants of individual travelers' decisions to vaccinate before traveling are largely unknown. This study aimed to provide this information using a discrete choice experiment (DCE) administered to four types of German travelers: (1) business travelers; (2) travelers visiting friends and relatives (VFR); (3) leisure travelers; and (4) backpackers. A DCE survey was developed, pretested and administered online. It included a series of choice questions in which respondents chose between two hypothetical vaccines, each characterized by four disease attributes with varying levels describing the of risk, health impact, curability and transmissibility of the disease they would prevent (described with four disease attributes with varying levels of risk, health impact, curability and transmissibility), and varying levels of four vaccine attributes (duration of protection, number of doses required, time required for vaccination, and vaccine cost). A random-parameters logit model was used to estimate the importance weights each traveler type placed on the various attribute levels. These weights were used to calculate mean monetary equivalents (MMEs) of changes in each attribute (holding all others constant) and of hypothetical disease-vaccine combinations. All traveler types' choices indicated that they attached the greatest importance to the risk and health impact of disease and to the vaccine cost whereas the other disease and vaccine attributes were less important for their decisions about travel vaccines. An option of not choosing any of the vaccine-pairs presented was rarely selected indicating that travelers' generally prefer to be vaccinated rather than not. The MMEs of changes in vaccine attributes indicated a very high variability between the individual travelers

  20. Imported pediatric malaria at the Hospital for Sick Children, Toronto, Canada: a 16 year review.

    Science.gov (United States)

    Evans, Andrea B; Kulik, Dina; Banerji, Anna; Boggild, Andrea; Kain, Kevin C; Abdelhaleem, Mohamed; Morris, Shaun K

    2014-10-04

    Children under 5 represent 86% of annual malaria deaths in the world. Following increasing trends in international travel, cases of imported malaria are rising in North America. We describe the epidemiology of malaria diagnosed at a tertiary care pediatric center in the multicultural city of Toronto. Retrospective chart review of all laboratory confirmed malaria from birth to travel history, chemoprophylaxis history, as well as clinical presentation, diagnosis and treatment were extracted. In total 107 children were diagnosed with malaria in the 16 year time period. Plasmodium falciparum malaria was identified in 76 (71%), Plasmodium vivax in 28 (26%). Median age of infected children was 6.7 years where 35% of children were born in Canada, 63% were recent or previous immigrants. Of those who resided in Canada, reason for travel included visiting friends or relatives (VFR) 95% and tourism or education (5%). Most common countries of infection were Ghana (22%), Nigeria (20%) and India (14%). Median parasitemia at presentation to our institution was 0.4% (IQR 0.1-2.3) with a maximum parasitemia of 31%. Nineteen (18%) met the WHO criteria for severe malaria due to hyperparasitemia, with 3 of these cases also meeting clinical criteria for severe malaria. One third of patients had a delay in treatment of 2 or more days. Ten percent of children had seen two or more primary health care professionals prior to admission. Prophylaxis was documented in 22 (21%), and out of those, 6 (27%) were appropriate for the region of travel and only 1 case was documented as adherent to their prescription. There were no cases of fatality. Malaria continues to be a significant disease in returning travelers and immigrant or refugee populations. An increase in physician awareness is required. Appropriate pre-travel advice, insect protection measures, effective chemoprophylaxis is needed to reduce the incidence and improve the management of imported pediatric malaria.

  1. Spatial and temporal pollutant budget analyses toward the total maximum daily loads management for the Yeongsan watershed in Korea.

    Science.gov (United States)

    Ki, S J; Lee, Y G; Kim, S W; Lee, Y J; Kim, J H

    2007-01-01

    This paper delivers two issues: water quality in the Yeongsan (YS) watershed which is one of the major watersheds in Korea and new watershed management plans with respect to the total maximum daily loads (TMDL) management. Field studies were conducted to estimate the pollutant loads according to the spatial and temporal distribution based on the biochemical oxygen demand (BOD) concentration and the volumetric flow rate (VFR) data from YS watershed. The results of both spatial and temporal analyses show the main pollutant source was originated from the city of Gwangju and the pollutant load from the city to YS watershed was the most out of five cities during this study period. Concerning YS reservoir located downstream of YS watershed, it also shows the worst water quality in the entire watershed during the study period. These results collectively demonstrate that the city of Gwangju is a main region which generates numerous point and non-point pollutant sources and eventually the pollutants are accumulated in YS reservoir. Based on the results, we suggest two different management plans for YS watershed. One is the flow-control approach that is to increase the amount of dam discharge in order to guarantee the river management flow for the midstream region. The other is the mass-control approach that is to dredge the contaminated sediments in YS reservoir for removing pollutants chronically accumulated in the sediment. Simulations for the former and the latter provide the pollution mitigation rate in the watershed up to 6 and 8% for BOD5, respectively. The methodology proposed here for TMDL management can be applied to a wide range of watersheds in Korea.

  2. Pseudomonas fluorescens F113 can produce a second flagellar apparatus, which is important for root colonization.

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    Emma Barahona

    2016-09-01

    Full Text Available The genomic sequence of Pseudomonas fluorescens F113 has shown the presence of a 41 kb cluster of genes that encode the production of a second flagellar apparatus. Among 2535 pseudomonads strains with sequenced genomes, these genes are only present in the genomes of F113 and other six strains, all but one belonging to the P. fluorescens cluster of species, in the form of a genetic island. The genes are homologous to the flagellar genes of the soil bacterium Azotobacter vinelandii. Regulation of these genes is mediated by the flhDC master operon, instead of the typical regulation in pseudomonads, which is through fleQ. Under laboratory conditions, F113 does not produce this flagellum and the flhDC operon is not expressed. However, ectopic expression of the flhDC operon is enough for its production, resulting in a hypermotile strain. This flagellum is also produced under laboratory conditions by the kinB and algU mutants. Genetic analysis has shown that kinB strongly represses the expression of the flhDC operon. This operon is activated by the Vfr protein probably in a c-AMP dependent way. The strains producing this second flagellum are all hypermotile and present a tuft of polar flagella instead of the single polar flagellum produced by the wild-type strain. Phenotypic variants isolated from the rhizosphere produce this flagellum and mutation of the genes encoding it, results in a defect in competitive colonization, showing its importance for root colonization.

  3. Imported malaria among people who travel to visit friends and relatives: is current UK policy effective or does it need a strategic change?

    Science.gov (United States)

    Behrens, Ron H; Neave, Penny E; Jones, Caroline O H

    2015-04-09

    The proportion of all imported malaria reported in travellers visiting friends and relatives (VFRs) in the UK has increased over the past decade and the proportion of Plasmodium falciparum malaria affecting this group has remained above 80% during that period. The epidemiological data suggest that the strategies employed in the UK to prevent imported malaria have been ineffective for VFRs. This paper attempts to identify possible reasons for the failure of the malaria prevention strategy among VFRs and suggest potential alternatives. A review of the current UK malaria prevention guidelines was undertaken and their approach was compared to the few data that are available on malaria perceptions and practices among VFRs. The current UK malaria prevention guidelines focus on educating travellers and health professionals using messages based on the personal threat of malaria and promoting the benefits of avoiding disease through the use of chemoprophylaxis. While malaria morbidity disproportionately affects VFRs, the mortality rates from malaria in VFRs is eight times, and severe disease eight times lower than in tourist and business travellers. Recent research into VFR malaria perceptions and practices has highlighted the complex socio-ecological context within which VFRs make their decisions about malaria. These data suggest that alternative strategies that move beyond a knowledge-deficit approach are required to address the burden of malaria in VFRs. Potential alternative strategies include the use of standby emergency-treatment (SBET) for the management of fevers with an anti-malarial provided pre-travel, the provision of rapid diagnostic testing and treatment regimen based in general-practitioner surgeries, and urgent and walk-in care centres and local accident and emergency (A&E) departments to provide immediate diagnosis and accessible ambulatory treatment for malaria patients. This latter approach would potentially address some of the practical barriers to

  4. The mucoid switch in Pseudomonas aeruginosa represses quorum sensing systems and leads to complex changes to stationary phase virulence factor regulation.

    Directory of Open Access Journals (Sweden)

    Ben Ryall

    Full Text Available The opportunistic pathogen Pseudomonas aeruginosa chronically infects the airways of Cystic Fibrosis (CF patients during which it adapts and undergoes clonal expansion within the lung. It commonly acquires inactivating mutations of the anti-sigma factor MucA leading to a mucoid phenotype, caused by excessive production of the extracellular polysaccharide alginate that is associated with a decline in lung function. Alginate production is believed to be the key benefit of mucA mutations to the bacterium in the CF lung. A phenotypic and gene expression characterisation of the stationary phase physiology of mucA22 mutants demonstrated complex and subtle changes in virulence factor production, including cyanide and pyocyanin, that results in their down-regulation upon entry into stationary phase but, (and in contrast to wildtype strains continued production in prolonged stationary phase. These findings may have consequences for chronic infection if mucoid P. aeruginosa were to continue to make virulence factors under non-growing conditions during infection. These changes resulted in part from a severe down-regulation of both AHL-and AQ (PQS-dependent quorum sensing systems. In trans expression of the cAMP-dependent transcription factor Vfr restored both quorum sensing defects and virulence factor production in early stationary phase. Our findings have implications for understanding the evolution of P. aeruginosa during CF lung infection and it demonstrates that mucA22 mutation provides a second mechanism, in addition to the commonly occurring lasR mutations, of down-regulating quorum sensing during chronic infection this may provide a selection pressure for the mucoid switch in the CF lung.

  5. Enhanced/Synthetic Vision and Head-Worn Display Technologies for Terminal Maneuvering Area NextGen Operations

    Science.gov (United States)

    Arthur, Jarvis J., III; Prinzell, Lawrence J.; Williams, Steven P.; Bailey, Randall E.; Shelton, Kevin J.; Norman, R. Mike

    2011-01-01

    NASA is researching innovative technologies for the Next Generation Air Transportation System (NextGen) to provide a "Better-Than-Visual" (BTV) capability as adjunct to "Equivalent Visual Operations" (EVO); that is, airport throughputs equivalent to that normally achieved during Visual Flight Rules (VFR) operations rates with equivalent and better safety in all weather and visibility conditions including Instrument Meteorological Conditions (IMC). These new technologies build on proven flight deck systems and leverage synthetic and enhanced vision systems. Two piloted simulation studies were conducted to access the use of a Head-Worn Display (HWD) with head tracking for synthetic and enhanced vision systems concepts. The first experiment evaluated the use a HWD for equivalent visual operations to San Francisco International Airport (airport identifier: KSFO) compared to a visual concept and a head-down display concept. A second experiment evaluated symbology variations under different visibility conditions using a HWD during taxi operations at Chicago O'Hare airport (airport identifier: KORD). Two experiments were conducted, one in a simulated San Francisco airport (KSFO) approach operation and the other, in simulated Chicago O'Hare surface operations, evaluating enhanced/synthetic vision and head-worn display technologies for NextGen operations. While flying a closely-spaced parallel approach to KSFO, pilots rated the HWD, under low-visibility conditions, equivalent to the out-the-window condition, under unlimited visibility, in terms of situational awareness (SA) and mental workload compared to a head-down enhanced vision system. There were no differences between the 3 display concepts in terms of traffic spacing and distance and the pilot decision-making to land or go-around. For the KORD experiment, the visibility condition was not a factor in pilot's rating of clutter effects from symbology. Several concepts for enhanced implementations of an unlimited field

  6. Travel-associated disease among US residents visiting US GeoSentinel clinics after return from international travel.

    Science.gov (United States)

    Hagmann, Stefan H F; Han, Pauline V; Stauffer, William M; Miller, Andy O; Connor, Bradley A; Hale, DeVon C; Coyle, Christina M; Cahill, John D; Marano, Cinzia; Esposito, Douglas H; Kozarsky, Phyllis E

    2014-12-01

    US residents make 60 million international trips annually. Family practice providers need to be aware of travel-associated diseases affecting this growing mobile population. To describe demographics, travel characteristics and clinical diagnoses of US residents who present ill after international travel. Descriptive analysis of travel-associated morbidity and mortality among US travellers seeking care at 1 of the 22 US practices and clinics participating in the GeoSentinel Global Surveillance Network from January 2000 to December 2012. Of the 9624 ill US travellers included in the analysis, 3656 (38%) were tourist travellers, 2379 (25%) missionary/volunteer/research/aid workers (MVRA), 1580 (16%) travellers visiting friends and relatives (VFRs), 1394 (15%) business travellers and 593 (6%) student travellers. Median (interquartile range) travel duration was 20 days (10-60 days). Pre-travel advice was sought by 45%. Hospitalization was required by 7%. Compared with other groups of travellers, ill MVRA travellers returned from longer trips (median duration 61 days), while VFR travellers disproportionately required higher rates of inpatient care (24%) and less frequently had received pre-travel medical advice (20%). Illnesses of the gastrointestinal tract were the most common (58%), followed by systemic febrile illnesses (18%) and dermatologic disorders (17%). Three deaths were reported. Diagnoses varied according to the purpose of travel and region of exposure. Returning ill US international travellers present with a broad spectrum of travel-associated diseases. Destination and reason for travel may help primary health care providers to generate an accurate differential diagnosis for the most common disorders and for those that may be life-threatening. © The Author 2014. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  7. Pilot behaviors in the face of adverse weather: A new look at an old problem.

    Science.gov (United States)

    Batt, Richard; O'Hare, David

    2005-06-01

    Weather-related general aviation accidents remain one of the most significant causes for concern in aviation safety. Previous studies have typically compared accident and non-accident cases. In contrast, the current study does not concentrate on occurrence outcome. Instead, the emphasis is on the different behaviors that pilots exhibit in the face of adverse weather and, by inference, on the decision-making processes that underlie those behaviors. This study compares three weather-related behaviors that reflect different levels of risk: visual flight rules flight into instrument meteorological conditions ('VFR into IMC'); precautionary landing; and other significant weather avoidance actions. Occurrence data (n=491) were drawn from the Australian Transport Safety Bureau database of aviation occurrences, and included weather-related accidents, incidents, and 'normal operationsd.' There were few significant differences between the three weather-related behavior groups in terms of pilot demographics, aircraft characteristics, geographic or environmental factors, or absolute flight distances. The pattern of relative flight distances (a psychological construct) was markedly different for the three groups, with pilots in the weather avoidance group being distinguished by taking timely action. The relative distance results suggest that the mid-point of the flight can be a 'psychological turning point' for pilots, irrespective of the absolute flight distance involved. Hence, pilots' behavior was sometimes influenced by psychological factors not related to any particular operational aspect of the flight. The results of the weather avoidance group indicate that a safe pilot is a proactive pilot. Dealing with adverse weather is not a one-off decision but a continually evolving process. This aspect is discussed in terms of the concept of 'mindfulness'.

  8. Persistent damaged bases in DNA allow mutagenic break repair in Escherichia coli.

    Directory of Open Access Journals (Sweden)

    Jessica M Moore

    2017-07-01

    Full Text Available Bacteria, yeast and human cancer cells possess mechanisms of mutagenesis upregulated by stress responses. Stress-inducible mutagenesis potentially accelerates adaptation, and may provide important models for mutagenesis that drives cancers, host pathogen interactions, antibiotic resistance and possibly much of evolution generally. In Escherichia coli repair of double-strand breaks (DSBs becomes mutagenic, using low-fidelity DNA polymerases under the control of the SOS DNA-damage response and RpoS general stress response, which upregulate and allow the action of error-prone DNA polymerases IV (DinB, II and V to make mutations during repair. Pol IV is implied to compete with and replace high-fidelity DNA polymerases at the DSB-repair replisome, causing mutagenesis. We report that up-regulated Pol IV is not sufficient for mutagenic break repair (MBR; damaged bases in the DNA are also required, and that in starvation-stressed cells, these are caused by reactive-oxygen species (ROS. First, MBR is reduced by either ROS-scavenging agents or constitutive activation of oxidative-damage responses, both of which reduce cellular ROS levels. The ROS promote MBR other than by causing DSBs, saturating mismatch repair, oxidizing proteins, or inducing the SOS response or the general stress response. We find that ROS drive MBR through oxidized guanines (8-oxo-dG in DNA, in that overproduction of a glycosylase that removes 8-oxo-dG from DNA prevents MBR. Further, other damaged DNA bases can substitute for 8-oxo-dG because ROS-scavenged cells resume MBR if either DNA pyrimidine dimers or alkylated bases are induced. We hypothesize that damaged bases in DNA pause the replisome and allow the critical switch from high fidelity to error-prone DNA polymerases in the DSB-repair replisome, thus allowing MBR. The data imply that in addition to the indirect stress-response controlled switch to MBR, a direct cis-acting switch to MBR occurs independently of DNA breakage

  9. Persistent damaged bases in DNA allow mutagenic break repair in Escherichia coli.

    Science.gov (United States)

    Moore, Jessica M; Correa, Raul; Rosenberg, Susan M; Hastings, P J

    2017-07-01

    Bacteria, yeast and human cancer cells possess mechanisms of mutagenesis upregulated by stress responses. Stress-inducible mutagenesis potentially accelerates adaptation, and may provide important models for mutagenesis that drives cancers, host pathogen interactions, antibiotic resistance and possibly much of evolution generally. In Escherichia coli repair of double-strand breaks (DSBs) becomes mutagenic, using low-fidelity DNA polymerases under the control of the SOS DNA-damage response and RpoS general stress response, which upregulate and allow the action of error-prone DNA polymerases IV (DinB), II and V to make mutations during repair. Pol IV is implied to compete with and replace high-fidelity DNA polymerases at the DSB-repair replisome, causing mutagenesis. We report that up-regulated Pol IV is not sufficient for mutagenic break repair (MBR); damaged bases in the DNA are also required, and that in starvation-stressed cells, these are caused by reactive-oxygen species (ROS). First, MBR is reduced by either ROS-scavenging agents or constitutive activation of oxidative-damage responses, both of which reduce cellular ROS levels. The ROS promote MBR other than by causing DSBs, saturating mismatch repair, oxidizing proteins, or inducing the SOS response or the general stress response. We find that ROS drive MBR through oxidized guanines (8-oxo-dG) in DNA, in that overproduction of a glycosylase that removes 8-oxo-dG from DNA prevents MBR. Further, other damaged DNA bases can substitute for 8-oxo-dG because ROS-scavenged cells resume MBR if either DNA pyrimidine dimers or alkylated bases are induced. We hypothesize that damaged bases in DNA pause the replisome and allow the critical switch from high fidelity to error-prone DNA polymerases in the DSB-repair replisome, thus allowing MBR. The data imply that in addition to the indirect stress-response controlled switch to MBR, a direct cis-acting switch to MBR occurs independently of DNA breakage, caused by ROS

  10. Genetic selection of peptide aptamers that interact and inhibit both Small protein B and alternative ribosome-rescue factor A of Aeromonas veronii C4

    Directory of Open Access Journals (Sweden)

    Peng Liu

    2016-08-01

    Full Text Available Aeromonas veronii is a pathogenic gram-negative bacterium, which infects a variety of animals and results in mass mortality. The stalled-ribosome rescues are reported to ensure viability and virulence under stress conditions, of which primarily include trans-translation and alternative ribosome-rescue factor A (ArfA in A. veronii. For identification of specific peptides that interact and inhibit the stalled-ribosome rescues, peptide aptamer library (pTRG-SN-peptides was constructed using pTRG as vector and Staphylococcus aureus nuclease (SN as scaffold protein, in which 16 random amino acids were introduced to form an exposed surface loop. In the meantime both Small Protein B (SmpB which acts as one of the key components in trans-translation, and alternative ribosome-rescue factor A (ArfA were inserted to pBT to constitute pBT-SmpB and pBT-ArfA, respectively. The peptide aptamer PA-2 was selected from pTRG-SN-peptides by bacterial two-hybrid system (B2H employing pBT-SmpB or pBT-ArfA as baits. The conserved sites G133K134 and D138K139R140 of C-terminal SmpB were identified by interacting with N-terminal SN, and concurrently the residue K62 of ArfA was recognized by interacting with the surface loop of the specific peptide aptamer PA-2. The expression plasmids pN-SN or pN-PA-2, which combined the duplication origin of pRE112 with the neokanamycin promoter expressing SN or PA-2, were created and transformed into A. veronii C4, separately. The engineered A. veronii C4 which endowing SN or PA-2 expression impaired growth capabilities under stress conditions including temperatures, sucrose, glucose, potassium chloride (KCl and antibiotics, and the stress-related genes rpoS and nhaP were down-regulated significantly by Quantitative Real-time PCR (qRT-PCR when treating in 2.0% KCl. Thus,the engineered A. veronii C4 conferring PA-2 expression might be potentially attenuated vaccine, and also the peptide aptamer PA-2 could develop as anti

  11. Transcriptional Control of Dual Transporters Involved in α-Ketoglutarate Utilization Reveals Their Distinct Roles in Uropathogenic Escherichia coli.

    Science.gov (United States)

    Cai, Wentong; Cai, Xuwang; Yang, Yongwu; Yan, Shigan; Zhang, Haibin

    2017-01-01

    Uropathogenic Escherichia coli (UPEC) are the primary causative agents of urinary tract infections. Some UPEC isolates are able to infect renal proximal tubule cells, and can potentially cause pyelonephritis. We have previously shown that to fulfill their physiological roles renal proximal tubule cells accumulate high concentrations of α-ketoglutarate (KG) and that gene cluster c5032-c5039 contribute to anaerobic utilization of KG by UPEC str. CFT073, thereby promoting its in vivo fitness. Given the importance of utilizing KG for UPEC, this study is designed to investigate the roles of two transporters KgtP and C5038 in KG utilization, their transcriptional regulation, and their contributions to UPEC fitness in vivo. Our phylogenetic analyses support that kgtP is a widely conserved locus in commensal and pathogenic E. coli, while UPEC-associated c5038 was acquired through horizontal gene transfer. Global anaerobic transcriptional regulators Fumarate and nitrate reduction (FNR) and ArcA induced c5038 expression in anaerobiosis, and C5038 played a major role in anaerobic growth on KG. KgtP was required for aerobic growth on KG, and its expression was repressed by FNR and ArcA under anaerobic conditions. Analyses of FNR and ArcA binding sites and results of EMS assays suggest that FNR and ArcA likely inhibit kgtP expression through binding to the -35 region of kgtP promoter and occluding the occupancy of RNA polymerases. Gene c5038 can be specifically induced by KG, whereas the expression of kgtP does not respond to KG, yet can be stimulated during growth on glycerol. In addition, c5038 and kgtP expression were further shown to be controlled by different alternative sigma factors RpoN and RpoS, respectively. Furthermore, dual-strain competition assays in a murine model showed that c5038 mutant but not kgtP mutant was outcompeted by the wild-type strain during the colonization of murine bladders and kidneys, highlighting the importance of C5038 under in vivo

  12. Profiling of Campylobacter jejuni Proteome in Exponential and Stationary Phase of Growth

    Directory of Open Access Journals (Sweden)

    Hana Turonova

    2017-05-01

    Full Text Available Campylobacter jejuni has been reported as a major cause of bacterial food-borne enteritides in developed countries during the last decade. Despite its fastidious growth requirements, including low level of oxygen and high level of CO2, this pathogen is able to persist in the environment without permanent loss of its viability and virulence. As C. jejuni is not able to multiply outside a host, the cells spend significant amount of time in stationary phase of growth. The entry into the stationary phase is often correlated to resistance to various stresses in bacteria. The switching between exponential and stationary phases is frequently mediated by the regulator sigma S (RpoS. However, this factor is absent in C. jejuni and molecular mechanisms responsible for transition of cells to the stationary phase remain elusive. In this work, proteomic profiles of cells from exponential and stationary phases were compared using 2-D electrophoresis (2DE fingerprinting combined with mass spectrometry analysis and qRT-PCR. The identified proteins, whose expression differed between the two phases, are mostly involved in protein biosynthesis, carbon metabolism, stress response and motility. Altered expression was observed also in the pleiotropic regulator CosR that was over-expressed during stationary phase. A shift between transcript and protein level evolution of CosR throughout the growth of C. jejuni was observed using qRT-PCR and (2DE. From these data, we hypothesized that CosR could undergo a negative autoregulation in stationary phase. A consensus sequence resulting from promoter sequence alignment of genes potentially regulated by CosR, including its own upstream region, among C. jejuni strains is proposed. To verify experimentally the potential autoregulation of CosR at the DNA level, electrophoretic mobility shift assay was performed with DNA fragments of CosR promoter region and rCosR. Different migration pattern of the promoter fragments indicates

  13. The yhiM gene codes for an inner membrane protein involved in GABA export in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Angela Tramonti

    2017-02-01

    Full Text Available In order to survive the exposure to acid pH, Escherichia coli activates molecular circuits leading from acid tolerance to extreme acid resistance (AR. The activation of the different circuits involves several global and specific regulators affecting the expression of membrane, periplasmic and cytosolic proteins acting at different levels to dampen the harmful consequences of the uncontrolled entry of protons intracellularly. Many genes coding for the structural components of the AR circuits (protecting from pH ≤ 2.5 and their specific transcriptional regulators cluster in a genomic region named AFI (acid fitness island and respond in the same way to global regulators (such as RpoS and H-NS as well as to anaerobiosis, alkaline, cold and respiratory stresses, in addition to the acid stress. Notably some genes coding for structural components of AR, though similarly regulated, are non-AFI localised. Amongst these the gadBC operon, coding for the major structural components of the glutamate-based AR system, and the ybaS gene, coding for a glutaminase required for the glutamine-based AR system. The yhiM gene, a non-AFI gene, appears to belong to this group. We mapped the transcription start of the 1.1 kb monocistronic yhiM transcript: it is an adenine residue located 22 nt upstream a GTG start codon. By real-time PCR we show that GadE and GadX equally affect the expression of yhiM under oxidative growth conditions. While YhiM is partially involved in the RpoS-dependent AR, we failed to detect a significant involvement in the glutamate- or glutamine-dependent AR at pH ≤ 2.5. However, when grown in EG at pH 5.0, the yhiM mutant displays impaired GABA export, whereas when YhiM is overexpressed, an increases of GABA export in EG medium in the pH range 2.5–5.5 is observed. Our data suggest that YhiM is a GABA transporter with a physiological role more relevant at mildly acidic pH, but not a key component of AR at pH < 2.5.

  14. Piecewise linear approximations to model the dynamics of adaptation to osmotic stress by food-borne pathogens.

    Science.gov (United States)

    Métris, Aline; George, Susie M; Ropers, Delphine

    2017-01-02

    Addition of salt to food is one of the most ancient and most common methods of food preservation. However, little is known of how bacterial cells adapt to such conditions. We propose to use piecewise linear approximations to model the regulatory adaptation of Escherichiacoli to osmotic stress. We apply the method to eight selected genes representing the functions known to be at play during osmotic adaptation. The network is centred on the general stress response factor, sigma S, and also includes a module representing the catabolic repressor CRP-cAMP. Glutamate, potassium and supercoiling are combined to represent the intracellular regulatory signal during osmotic stress induced by salt. The output is a module where growth is represented by the concentration of stable RNAs and the transcription of the osmotic gene osmY. The time course of gene expression of transport of osmoprotectant represented by the symporter proP and of the osmY is successfully reproduced by the network. The behaviour of the rpoS mutant predicted by the model is in agreement with experimental data. We discuss the application of the model to food-borne pathogens such as Salmonella; although the genes considered have orthologs, it seems that supercoiling is not regulated in the same way. The model is limited to a few selected genes, but the regulatory interactions are numerous and span different time scales. In addition, they seem to be condition specific: the links that are important during the transition from exponential to stationary phase are not all needed during osmotic stress. This model is one of the first steps towards modelling adaptation to stress in food safety and has scope to be extended to other genes and pathways, other stresses relevant to the food industry, and food-borne pathogens. The method offers a good compromise between systems of ordinary differential equations, which would be unmanageable because of the size of the system and for which insufficient data are available

  15. pqiABC and yebST, Putative mce Operons of Escherichia coli, Encode Transport Pathways and Contribute to Membrane Integrity.

    Science.gov (United States)

    Nakayama, Takayuki; Zhang-Akiyama, Qiu-Mei

    2017-01-01

    The membranes of single-cell organisms are crucial as the first line of defense. The outer membrane of Gram-negative bacteria is an asymmetric bilayer in which lipopolysaccharides (LPSs) and phospholipids are localized in the outer and inner leaflet, respectively. This asymmetry is important for membrane integrity. In Escherichia coli, the Mla transport pathway maintains this asymmetry by removing phospholipids from the outer leaflet. The MlaD component of this system is a mammalian cell entry (MCE) domain protein, and E. coli has two other MCE domain proteins of unknown function (PqiB and YebT). Here, we show that these two proteins are components of novel transport pathways that contribute to membrane integrity. The pqiAB operon is regulated by SoxS and RpoS. The yebST operon contains pqiAB homologues. Here, we found a third member of the pqi operon, ymbA (pqiC). A PqiB-PqiC complex bridges the inner and the outer membrane, and in other bacteria, pqiBC genes are located in operons together with transporter proteins. We show here that simultaneous deletion of pqiABC and yebST operons in an Δmla background rendered cells more sensitive to SDS-EDTA, and the SDS-EDTA sensitivity of mla mutants was rescued by additional copies of pqiABC We also found that the yebST operon was induced by a defect in LPS molecules. In conclusion, PqiABC and YebST are novel transport pathways related to the Mla transport pathway and important for membrane integrity. Membranes of bacteria are crucial for stress resistance. The composition of the E. coli outer membrane is asymmetric, with asymmetry maintained by the Mla ABC transport pathway. We propose that the stress-inducible pqiABC operon and homologous yebST operon, both of previously unknown function, encode transport pathway proteins related to the Mla transport pathway. Deletion of these operons rendered cells more sensitive to membrane stress, and additional copies of pqiABC suppressed the SDS-EDTA sensitivity of mla mutant

  16. Identification of Fitness Determinants during Energy-Limited Growth Arrest in Pseudomonas aeruginosa.

    Science.gov (United States)

    Basta, David W; Bergkessel, Megan; Newman, Dianne K

    2017-11-28

    Microbial growth arrest can be triggered by diverse factors, one of which is energy limitation due to scarcity of electron donors or acceptors. Genes that govern fitness during energy-limited growth arrest and the extent to which they overlap between different types of energy limitation are poorly defined. In this study, we exploited the fact that Pseudomonas aeruginosa can remain viable over several weeks when limited for organic carbon (pyruvate) as an electron donor or oxygen as an electron acceptor. ATP values were reduced under both types of limitation, yet more severely in the absence of oxygen. Using transposon-insertion sequencing (Tn-seq), we identified fitness determinants in these two energy-limited states. Multiple genes encoding general functions like transcriptional regulation and energy generation were required for fitness during carbon or oxygen limitation, yet many specific genes, and thus specific activities, differed in their relevance between these states. For instance, the global regulator RpoS was required during both types of energy limitation, while other global regulators such as DksA and LasR were required only during carbon or oxygen limitation, respectively. Similarly, certain ribosomal and tRNA modifications were specifically required during oxygen limitation. We validated fitness defects during energy limitation using independently generated mutants of genes detected in our screen. Mutants in distinct functional categories exhibited different fitness dynamics: regulatory genes generally manifested a phenotype early, whereas genes involved in cell wall metabolism were required later. Together, these results provide a new window into how P. aeruginosa survives growth arrest.IMPORTANCE Growth-arrested bacteria are ubiquitous in nature and disease yet understudied at the molecular level. For example, growth-arrested cells constitute a major subpopulation of mature biofilms, serving as an antibiotic-tolerant reservoir in chronic infections

  17. Phenotypic and Genotypic Features of a Salmonella Heidelberg Strain Isolated in Broilers in Brazil and Their Possible Association to Antibiotics and Short-Chain Organic Acids Resistance and Susceptibility.

    Science.gov (United States)

    Santin, Elizabeth; Hayashi, Ricardo Mitsuo; Wammes, Jessica Caroline; Gonzalez-Esquerra, Ricardo; Carazzolle, Marcelo Falsarella; Freire, Caio César de Melo; Monzani, Paulo Sérgio; da Cunha, Anderson Ferreira

    2017-01-01

    Salmonella enterica serovar Heidelberg is a human pathogen also found in broilers. A strain (UFPR1) has been associated with field reports of resistance to short-chain organic acids (SCOA) in broilers in the South of Brazil, but was susceptible to a Bacillus subtilis-based probiotic added in feed in a related study. This work aimed to (i) report clinical symptoms caused by SH UFPR1 in broilers, (ii) study its susceptibility to some antibiotics in vitro, and (iii) SCOA in vivo; and (iv) relate these phenotypic observations with its genome characteristics. Two in vivo trials used 1-day-old chicks housed for 21 days in 8 sterilized isolated negative pressure rooms with 4 battery cages of 12 birds each. Birds were challenged or not with 107 CFU/bird of SH UFPR1 orally and exposed or not to SCOA in a 2 × 2 factorial design. Zootechnical parameters were unaffected (P > 0.05), no clinical signs were observed, and few cecal and hepatic histologic and immune-related alterations were seen, in birds challenged with SH. Formic and propionic acids added together in drinking water, fumaric and benzoic acid in feed (Trial 1), and coated calcium butyrate in feed (Trial 2) did not reduce the SH isolation frequencies seen in cecum and liver in broilers after SH challenge (P > 0.05). SH UFPR1 was susceptible to amikacin, amoxicillin + clavulanate, ceftiofur, cephalexin, doxycycline and oxytetracycline; and mildly susceptible to ampicillin + sulbactam, cephalothin, ciprofloxacin, enrofloxacin, and gentamycin in an in vitro minimum inhibitory concentration model using Mueller-Hinton agar. The whole genome of SH UFPR1 was sequenced and consisted of a circular chromosome, spanning 4,760,321 bp with 52.18% of GC-content encoding 84 tRNA, 22 rRNA, and 4,427 protein-coding genes. The comparison between SH UFPR1 genome and a multidrug-resistant SL476 strain revealed 11 missing genomic fragments and 5 insertions related to bgt, bgr, and rpoS genes. The deleted genes

  18. Changes in Gene Transcription Induced by Hydrogen Peroxide Treatment of Verotoxin-Producing Escherichia coli O157:H7 and Non-O157 Serotypes on Romaine Lettuce.

    Science.gov (United States)

    Mei, Gui-Ying; Tang, Joshua; Bach, Susan; Kostrzynska, Magdalena

    2017-01-01

    Disease outbreaks of verotoxin-producing Escherichia coli (VTEC) O157:H7 and non-O157 serotypes associated with leafy green vegetables are becoming a growing concern. A better understanding of the behavior of VTEC, particularly non-O157 serotypes, on lettuce under stress conditions is necessary for designing more effective control strategies. Hydrogen peroxide (H2O2) can be used as a sanitizer to reduce the microbial load in leafy green vegetables, particularly in fresh produce destined for the organic market. In this study, we tested the hypothesis that H2O2 treatment of contaminated lettuce affects in the same manner transcription of stress-associated and virulence genes in VTEC strains representing O157 and non-O157 serotypes. Six VTEC isolates representing serotypes O26:H11, O103:H2, O104:H4, O111:NM, O145:NM, and O157:H7 were included in this study. The results indicate that 50 mM H2O2 caused a population reduction of 2.4-2.8 log10 (compared to non-treated control samples) in all six VTEC strains present on romaine lettuce. Following the treatment, the transcription of genes related to oxidative stress (oxyR and sodA), general stress (uspA and rpoS), starvation (phoA), acid stress (gadA, gadB, and gadW), and virulence (stx1A, stx2A, and fliC) were dramatically downregulated in all six VTEC serotypes (P ≤ 0.05) compared to not treated control samples. Therefore, VTEC O157:H7 and non-O157 serotypes on lettuce showed similar survival rates and gene transcription profiles in response to 50 mM H2O2 treatment. Thus, the results derived from this study provide a basic understanding of the influence of H2O2 treatment on the survival and virulence of VTEC O157:H7 and non-O157 serotypes on lettuce.

  19. Stress-Induced Evolution of Heat Resistance and Resuscitation Speed in Escherichia coli O157:H7 ATCC 43888.

    Science.gov (United States)

    Gayán, Elisa; Cambré, Alexander; Michiels, Chris W; Aertsen, Abram

    2016-11-15

    The development of resistance in foodborne pathogens to food preservation techniques is an issue of increasing concern, especially in minimally processed foods where safety relies on hurdle technology. In this context, mild heat can be used in combination with so-called nonthermal processes, such as high hydrostatic pressure (HHP), at lower individual intensities to better retain the quality of the food. However, mild stresses may increase the risk of (cross-)resistance development in the surviving population, which in turn might compromise food safety. In this investigation, we examined the evolution of Escherichia coli O157:H7 strain ATCC 43888 after recurrent exposure to progressively intensifying mild heat shocks (from 54.0°C to 60.0°C in 0.5°C increments) with intermittent resuscitation and growth of survivors. As such, mutant strains were obtained after 10 cycles of selection with ca. 106-fold higher heat resistance than that for the parental strain at 58.0°C, although this resistance did not extend to temperatures exceeding 60.0°C. Moreover, these mutant strains typically displayed cross-resistance against HHP shock and displayed signs of enhanced RpoS and RpoH activity. Interestingly, additional cycles of selection maintaining the intensity of the heat shock constant (58.5°C) selected for mutant strains in which resuscitation speed, rather than resistance, appeared to be increased. Therefore, it seems that resistance and resuscitation speed are rapidly evolvable traits in E. coli ATCC 43888 that can compromise food safety. In this investigation, we demonstrated that Escherichia coli O157:H7 ATCC 43888 rapidly acquires resistance to mild heat exposure, with this resistance yielding cross-protection to high hydrostatic pressure treatment. In addition, mutants of E. coli ATCC 43888 in which resuscitation speed, rather than resistance, appeared to be improved were selected. As such, both resistance and resuscitation speed seem to be rapidly evolvable

  20. Temporal transcriptomic analysis of Desulfovibrio vulgaris Hildenborough transition into stationary phase growth during electrondonor depletion

    Energy Technology Data Exchange (ETDEWEB)

    Clark, M.E.; He, Q.; He, Z.; Huang, K.H.; Alm, E.J.; Wan, X.-F.; Hazen, T.C.; Arkin, A.P.; Wall, J.D.; Zhou, J.-Z.; Fields, M.W.

    2006-08-01

    Desulfovibrio vulgaris was cultivated in a defined medium, and biomass was sampled for approximately 70 h to characterize the shifts in gene expression as cells transitioned from the exponential to the stationary phase during electron donor depletion. In addition to temporal transcriptomics, total protein, carbohydrate, lactate, acetate, and sulfate levels were measured. The microarray data were examined for statistically significant expression changes, hierarchical cluster analysis, and promoter element prediction and were validated by quantitative PCR. As the cells transitioned from the exponential phase to the stationary phase, a majority of the down-expressed genes were involved in translation and transcription, and this trend continued at the remaining times. There were general increases in relative expression for intracellular trafficking and secretion, ion transport, and coenzyme metabolism as the cells entered the stationary phase. As expected, the DNA replication machinery was down-expressed, and the expression of genes involved in DNA repair increased during the stationary phase. Genes involved in amino acid acquisition, carbohydrate metabolism, energy production, and cell envelope biogenesis did not exhibit uniform transcriptional responses. Interestingly, most phage-related genes were up-expressed at the onset of the stationary phase. This result suggested that nutrient depletion may affect community dynamics and DNA transfer mechanisms of sulfate-reducing bacteria via the phage cycle. The putative feoAB system (in addition to other presumptive iron metabolism genes) was significantly up-expressed, and this suggested the possible importance of Fe{sup 2+} acquisition under metal-reducing conditions. The expression of a large subset of carbohydrate-related genes was altered, and the total cellular carbohydrate levels declined during the growth phase transition. Interestingly, the D. vulgaris genome does not contain a putative rpoS gene, a common attribute

  1. Enzootic origins for clinical manifestations of Lyme borreliosis.

    Science.gov (United States)

    Jahfari, Setareh; Krawczyk, Aleksandra; Coipan, E Claudia; Fonville, Manoj; Hovius, Joppe W; Sprong, Hein; Takumi, Katsuhisa

    2017-04-01

    Both early localized and late disseminated forms of Lyme borreliosis are caused by Borrelia burgdorferi senso lato. Differentiating between the spirochetes that only cause localized skin infection from those that cause disseminated infection, and tracing the group of medically-important spirochetes to a specific vertebrate host species, are two critical issues in disease risk assessment and management. Borrelia burgdorferi senso lato isolates from Lyme borreliosis cases with distinct clinical manifestations (erythema migrans, neuroborreliosis, acrodermatitis chronica atrophicans, and Lyme arthritis) and isolates from Ixodes ricinus ticks feeding on rodents, birds and hedgehogs were typed to the genospecies level by sequencing part of the intergenic spacer region. In-depth molecular typing was performed by sequencing eight additional loci with different characteristics (plasmid-bound, regulatory, and housekeeping genes). The most abundant genospecies and genotypes in the clinical isolates were identified by using odds ratio as a measure of dominance. Borrelia afzelii was the most common genospecies in acrodermatitis patients and engorged ticks from rodents. Borrelia burgdorferi senso stricto was widespread in erythema migrans patients. Borrelia bavariensis was widespread in neuroborreliosis patients and in ticks from hedgehogs, but rare in erythema migrans patients. Borrelia garinii was the dominant genospecies in ticks feeding on birds. Spirochetes in ticks feeding on hedgehogs were overrepresented in genotypes of the plasmid gene ospC from spirochetes in erythema migrans patients. Spirochetes in ticks feeding on hedgehogs were overrepresented in genotypes of ospA from spirochetes in acrodermatitis patients. Spirochetes from ticks feeding on birds were overrepresented in genotypes of the plasmid and regulatory genes dbpA, rpoN and rpoS from spirochetes in neuroborreliosis patients. Overall, the analyses of our datasets support the existence of at least three

  2. Characteristics of HIV infected individuals traveling abroad. Results from the +REDIVI Collaborative Network.

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    Pérez-Molina, Jose A; Martinez-Perez, Angela; Serre, Nuria; Treviño, Begoña; Ruiz-Giardín, José Manuel; Torrús, Diego; Goikoetxea, Josune; Echevarría, Esteban Martín; Malmierca, Eduardo; Rojo, Gerardo; Calabuig, Eva; Gutierrez, Belén; Norman, Francesca; Lopez-Velez, Rogelio

    2016-02-01

    The improvement in the prognosis of HIV infection, coupled with the increase in international travel and migration, has led to a rising number of HIV infected travelers. The objective of this study was to describe the epidemiological and clinical features of returning travelers, according to their HIV status. An observational prospective study was conducted including travelers and immigrants who traveled to visit friends and relatives (VFRs) registered in the +REDIVI collaborative network (January-2009; October-2014). +REDIVI is a national network that registers information regarding infections imported by travelers and immigrants at 21 different centers using a standardized protocol. A total of 3464 travellers were identified: 72 were HIV+ (2.1%) and 3.392 HIV- (98%). HIV+ vs. HIV- travelers were often older (40.5y vs. 34.2y P=.001), VFRs (79.1% vs. 44.4%; P<.001), and consulted less for pre-travel advice (27% vs. 37%; P=.078). The main destinations for both groups were sub-Saharan Africa and Latin America. The most frequent reasons for consultation after travel were fever, request for a health examination, gastrointestinal complaints, and abnormal laboratory tests (mainly eosinophilia and anemia), which differed between groups. The most frequent diagnoses in HIV+ travelers were malaria (38.8%), newly diagnosed HIV infection (25%), and intestinal parasites (19.4%), while for HIV- travelers the main diagnoses were "healthy" (17.9%), malaria (14%), and intestinal parasites (17.3%). The typical profile of an HIV+ traveler in +REDIVI was that of a VFR traveler who did not seek pre-travel advice and made high-risk trips. This may increase the chance of acquiring travel-related infections which may pose a special risk for HIV-infected travelers. The post-travel visit was a good opportunity for HIV infection screening. Copyright © 2015 Elsevier España, S.L.U. y Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.

  3. NextGen Operations in a Simulated NY Area Airspace

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    Smith, Nancy M.; Parke, Bonny; Lee, Paul; Homola, Jeff; Brasil, Connie; Buckley, Nathan; Cabrall, Chris; Chevalley, Eric; Lin, Cindy; Morey, Susan; hide

    2013-01-01

    A human-in-the-loop simulation conducted in the Airspace Operations Laboratory (AOL) at NASA Ames Research Center explored the feasibility of a Next Generation Air Transportation System (NextGen) solution to address airspace and airport capacity limitations in and around the New York metropolitan area. A week-long study explored the feasibility of a new Optimal Profile Descent (OPD) arrival into the airspace as well as a novel application of a Terminal Area Precision Scheduling and Spacing (TAPSS) enhancement to the Traffic Management Advisor (TMA) arrival scheduling tool to coordinate high volume arrival traffic to intersecting runways. In the simulation, four en route sector controllers and four terminal radar approach control (TRACON) controllers managed traffic inbound to Newark International Airport's primary runway, 22L, and its intersecting overflow runway, 11. TAPSS was used to generate independent arrival schedules for each runway and a traffic management coordinator participant adjusted the arrival schedule for each runway 11 aircraft to follow one of the 22L aircraft. TAPSS also provided controller-managed spacing tools (slot markers with speed advisories and timelines) to assist the TRACON controllers in managing the arrivals that were descending on OPDs. Results showed that the tools significantly decreased the occurrence of runway violations (potential go-arounds) when compared with a Baseline condition with no tools. Further, the combined use of the tools with the new OPD produced a peak arrival rate of over 65 aircraft per hour using instrument flight rules (IFR), exceeding the current maximum arrival rate at Newark Liberty International Airport (EWR) of 52 per hour under visual flight rules (VFR). Although the participants rated the workload as relatively low and acceptable both with and without the tools, they rated the tools as reducing their workload further. Safety and coordination were rated by most participants as acceptable in both

  4. The position of mefloquine as a 21st century malaria chemoprophylaxis.

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    Schlagenhauf, Patricia; Adamcova, Miriam; Regep, Loredana; Schaerer, Martin T; Rhein, Hans-Georg

    2010-12-09

    Malaria chemoprophylaxis prevents the occurrence of the symptoms of malaria. Travellers to high-risk Plasmodium falciparum endemic areas need an effective chemoprophylaxis. A literature search to update the status of mefloquine as a malaria chemoprophylaxis. Except for clearly defined regions with multi-drug resistance, mefloquine is effective against the blood stages of all human malaria species, including the recently recognized fifth species, Plasmodium knowlesi. New data were found in the literature on the tolerability of mefloquine and the use of this medication by groups at high risk of malaria. Use of mefloquine for pregnant women in the second and third trimester is sanctioned by the WHO and some authorities (CDC) allow the use of mefloquine even in the first trimester. Inadvertent pregnancy while using mefloquine is not considered grounds for pregnancy termination. Mefloquine chemoprophylaxis is allowed during breast-feeding. Studies show that mefloquine is a good option for other high-risk groups, such as long-term travellers, VFR travellers and families with small children. Despite a negative media perception, large pharmaco-epidemiological studies have shown that serious adverse events are rare. A recent US evaluation of serious events (hospitalization data) found no association between mefloquine prescriptions and serious adverse events across a wide range of outcomes including mental disorders and diseases of the nervous system. As part of an in-depth analysis of mefloquine tolerability, a potential trend for increased propensity for neuropsychiatric adverse events in women was identified in a number of published clinical studies. This trend is corroborated by several cohort studies that identified female sex and low body weight as risk factors. The choice of anti-malarial drug should be an evidence-based decision that considers the profile of the individual traveller and the risk of malaria. Mefloquine is an important, first-line anti-malarial drug

  5. The position of mefloquine as a 21st century malaria chemoprophylaxis

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    Regep Loredana

    2010-12-01

    Full Text Available Abstract Background Malaria chemoprophylaxis prevents the occurrence of the symptoms of malaria. Travellers to high-risk Plasmodium falciparum endemic areas need an effective chemoprophylaxis. Methods A literature search to update the status of mefloquine as a malaria chemoprophylaxis. Results Except for clearly defined regions with multi-drug resistance, mefloquine is effective against the blood stages of all human malaria species, including the recently recognized fifth species, Plasmodium knowlesi. New data were found in the literature on the tolerability of mefloquine and the use of this medication by groups at high risk of malaria. Discussion Use of mefloquine for pregnant women in the second and third trimester is sanctioned by the WHO and some authorities (CDC allow the use of mefloquine even in the first trimester. Inadvertent pregnancy while using mefloquine is not considered grounds for pregnancy termination. Mefloquine chemoprophylaxis is allowed during breast-feeding. Studies show that mefloquine is a good option for other high-risk groups, such as long-term travellers, VFR travellers and families with small children. Despite a negative media perception, large pharmaco-epidemiological studies have shown that serious adverse events are rare. A recent US evaluation of serious events (hospitalization data found no association between mefloquine prescriptions and serious adverse events across a wide range of outcomes including mental disorders and diseases of the nervous system. As part of an in-depth analysis of mefloquine tolerability, a potential trend for increased propensity for neuropsychiatric adverse events in women was identified in a number of published clinical studies. This trend is corroborated by several cohort studies that identified female sex and low body weight as risk factors. Conclusion The choice of anti-malarial drug should be an evidence-based decision that considers the profile of the individual traveller and the

  6. Antibacterial Mechanism of 405-Nanometer Light-Emitting Diode against Salmonella at Refrigeration Temperature.

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    Kim, Min-Jeong; Yuk, Hyun-Gyun

    2017-03-01

    The aim of this study was to elucidate the antibacterial mechanism of 405 ± 5-nm light-emitting diode (LED) illumination against Salmonella at 4°C in phosphate-buffered saline (PBS) by determining endogenous coproporphyrin content, DNA oxidation, damage to membrane function, and morphological change. Gene expression levels, including of oxyR , recA , rpoS , sodA , and soxR , were also examined to understand the response of Salmonella to LED illumination. The results showed that Salmonella strains responded differently to LED illumination, revealing that S. enterica serovar Enteritidis (ATCC 13076) and S. enterica subsp. enterica serovar Saintpaul (ATCC 9712) were more susceptible and resistant, respectively, than the 16 other strains tested. There was no difference in the amounts of endogenous coproporphyrin in the two strains. Compared with that in nonilluminated cells, the DNA oxidation levels in illuminated cells increased. In illuminated cells, we observed a loss of efflux pump activity, damage to the glucose uptake system, and changes in membrane potential and integrity. Transmission electron microscopy revealed a disorganization of chromosomes and ribosomes due to LED illumination. The levels of the five genes measured in the nonilluminated and illuminated S Saintpaul cells were upregulated in PBS at a set temperature of 4°C, indicating that increased gene expression levels might be due to a temperature shift and nutrient deficiency rather than to LED illumination. In contrast, only oxyR in S Enteritidis cells was upregulated. Thus, different sensitivities of the two strains to LED illumination were attributed to differences in gene regulation. IMPORTANCE Bacterial inactivation using visible light has recently received attention as a safe and environmentally friendly technology, in contrast with UV light, which has detrimental effects on human health and the environment. This study was designed to understand how 405 ± 5-nm light-emitting diode (LED

  7. Conditional Function of Autoaggregative Protein Cah and Common cah Mutations in Shiga Toxin-Producing Escherichia coli.

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    Carter, Michelle Qiu; Brandl, Maria T; Kudva, Indira T; Katani, Robab; Moreau, Matthew R; Kapur, Vivek

    2017-10-20

    DH5α cells with a strong autoaggregative phenotype that is inversely correlated with its ability to form biofilms, and plays a strain-specific role in plant and animal colonization by STEC. Although cah is widespread in the STEC population, we detected a mutation rate of 31.3% in cah, which is similar to that reported in rpoS and in fimH. Formation of cell aggregates due to increased bacteria-to-bacteria interactions may be disadvantageous to bacterial populations under conditions that favor a planktonic state in STEC. Therefore, loss-of-function mutation in cah is likely a selective trait in STEC when autoaggregative properties become detrimental to bacterial cells, and may contribute to the adaptability of STEC to fluctuating environments. This is a work of the U.S. Government and is not subject to copyright protection in the United States. Foreign copyrights may apply.

  8. Cinnamon Oil Inhibits Shiga Toxin Type 2 Phage Induction and Shiga Toxin Type 2 Production in Escherichia coli O157:H7

    Science.gov (United States)

    Sheng, Lina; Rasco, Barbara

    2016-01-01

    ABSTRACT This study evaluated the inhibitory effect of cinnamon oil against Escherichia coli O157:H7 Shiga toxin (Stx) production and further explored the underlying mechanisms. The MIC and minimum bactericidal concentration (MBC) of cinnamon oil against E. coli O157:H7 were 0.025% and 0.05% (vol/vol), respectively. Cinnamon oil significantly reduced Stx2 production and the stx2 mRNA expression that is associated with diminished Vero cell cytotoxicity. Consistently, induction of the Stx-converting phage where the stx2 gene is located, along with the total number of phages, decreased proportionally to cinnamon oil concentration. In line with decreased Stx2 phage induction, cinnamon oil at 0.75× and 1.0× MIC eliminated RecA, a key mediator of SOS response, polynucleotide phosphorylase (PNPase), and poly(A) polymerase (PAP I), which positively regulate Stx-converting phages, contributing to reduced Stx-converting phage induction and Stx production. Furthermore, cinnamon oil at 0.75× and 1.0× MIC strongly inhibited the qseBC and luxS expression associated with decreased AI-2 production, a universal quorum sensing signaling molecule. However, the expression of oxidative stress response genes oxyR, soxR, and rpoS was increased in response to cinnamon oil at 0.25× or 0.5× MIC, which may contribute to stunted bacterial growth and reduced Stx2 phage induction and Stx2 production due to the inhibitory effect of OxyR on prophage activation. Collectively, cinnamon oil inhibits Stx2 production and Stx2 phage induction in E. coli O157:H7 in multiple ways. IMPORTANCE This study reports the inhibitory effect of cinnamon oil on Shiga toxin 2 phage induction and Shiga toxin 2 production. Subinhibitory concentrations (concentrations below the MIC) of cinnamon oil reduced Stx2 production, stx2 mRNA expression, and cytotoxicity on Vero cells. Subinhibitory concentrations of cinnamon oil also dramatically reduced both the Stx2 phage and total phage induction in E. coli O157:H7

  9. Circuitry linking the global Csr and σE-dependent cell envelope stress response systems.

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    Yakhnin, Helen; Aichele, Robert; Ades, Sarah E; Romeo, Tony; Babitzke, Paul

    2017-09-18

    CsrA of Escherichia coli is an RNA-binding protein that globally regulates a wide variety of cellular processes and behaviors including carbon metabolism, motility, biofilm formation, and the stringent response. CsrB and CsrC are sRNAs that sequester CsrA, thereby preventing CsrA-mRNA interaction. RpoE (σE) is the extracytoplasmic stress response sigma factor of E. coli Previous RNA-seq studies identified rpoE mRNA as a CsrA target. Here we explored the regulation of rpoE by CsrA and found that CsrA represses rpoE translation. Gel mobility shift, footprint and toeprint studies identified three CsrA binding sites in the rpoE leader transcript, one of which overlaps the rpoE Shine-Dalgarno (SD) sequence, while another overlaps the rpoE translation initiation codon. Coupled in vitro transcription-translation experiments showed that CsrA represses rpoE translation by binding to these sites. We further demonstrate that σE indirectly activates transcription of csrB and csrC, leading to increased sequestration of CsrA such that repression of rpoE by CsrA is reduced. We propose that the Csr system fine-tunes the σE-dependent cell envelope stress response. We also identified a 51 amino acid coding sequence whose stop codon overlaps the rpoE start codon, and demonstrate that rpoE is translationally coupled with this upstream open reading frame (ORF51). Loss of coupling reduces rpoE translation by more than 50%. Identification of a translationally coupled ORF upstream of rpoE suggests that this previously unannotated protein may participate in the cell envelope stress response. In keeping with existing nomenclature, we name ORF51 as rseD, resulting in an operon arrangement of rseD-rpoE-rseA-rseB-rseCIMPORTANCE CsrA posttranscriptionally represses genes required for bacterial stress responses, including the stringent response, catabolite repression, and the RpoS (σS)-mediated general stress response. We show that CsrA represses translation of rpoE, encoding the

  10. Catabolic regulation analysis of Escherichia coli and its crp, mlc, mgsA, pgi and ptsG mutants

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    Yao Ruilian

    2011-08-01

    Full Text Available Abstract Background Most bacteria can use various compounds as carbon sources. These carbon sources can be either co-metabolized or sequentially metabolized, where the latter phenomenon typically occurs as catabolite repression. From the practical application point of view of utilizing lignocellulose for the production of biofuels etc., it is strongly desirable to ferment all sugars obtained by hydrolysis from lignocellulosic materials, where simultaneous consumption of sugars would benefit the formation of bioproducts. However, most organisms consume glucose prior to consumption of other carbon sources, and exhibit diauxic growth. It has been shown by fermentation experiments that simultaneous consumption of sugars can be attained by ptsG, mgsA mutants etc., but its mechanism has not been well understood. It is strongly desirable to understand the mechanism of metabolic regulation for catabolite regulation to improve the performance of fermentation. Results In order to make clear the catabolic regulation mechanism, several continuous cultures were conducted at different dilution rates of 0.2, 0.4, 0.6 and 0.7 h-1 using wild type Escherichia coli. The result indicates that the transcript levels of global regulators such as crp, cra, mlc and rpoS decreased, while those of fadR, iclR, soxR/S increased as the dilution rate increased. These affected the metabolic pathway genes, which in turn affected fermentation result where the specific glucose uptake rate, the specific acetate formation rate, and the specific CO2 evolution rate (CER were increased as the dilution rate was increased. This was confirmed by the 13C-flux analysis. In order to make clear the catabolite regulation, the effect of crp gene knockout (Δcrp and crp enhancement (crp+ as well as mlc, mgsA, pgi and ptsG gene knockout on the metabolism was then investigated by the continuous culture at the dilution rate of 0.2 h-1 and by some batch cultures. In the case of Δcrp (and also

  11. The importance of pretreatment tailoring on the performance of ultrafiltration membranes to treat two-phase olive mill wastewater

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    Ochando Pulido, J. M.

    2015-03-01

    Full Text Available In this work, the performance of an ultrafiltration (UF membrane in the treatment of the effluents by-produced by olive mills is addressed by applying different pretreatments on the raw effluents. By conducting a photo-catalytic process (UV/TiO2 PC after pH-temperature flocculation (pH-T F higher threshold flux values were observed for all feed stocks than by applying solely the pH-T F process, with an 18.8–34.2% increment. In addition, the performance of the UF membrane was also improved in terms of rejection efficiency, such that higher rejection values were yielded by the membrane for the organic pollutants (RCOD by 48.5 vs. 39.9% and 53.4 vs. 42.0%. The UF membrane performance was also improved in terms of the volume feed recovery factor (VFR, achieving up to 88.2 vs. 87.2% and 90.7 vs. 89.3%. Results in the same line were also observed when the highly polluted olives oil washing wastewater raw stream was previously mixed with the effluent stream coming from the washing of the olives. This permits the UF to permeate, achieving the standard limits to reuse the purified effluent for irrigation purposes (COD values below 1000 mg·L−1, which makes the treatment process cost-effective and results in making the olive oil production process environmentally friendly.En este estudio se aborda el rendimiento de una membrana de ultrafiltración (UF para el tratamiento de los efluentes generados por la industria oleícola, mediante la aplicación de distintos pretratamientos. Tras aplicar un proceso fotocatalítico (UV/TiO2 PC después de una floculación pH-temperatura (pH-T F se observaron flujos límite para todos los efluentes mayores que tras la aplicación únicamente del proceso pH-T F, con incrementos del 18.8–34.2 %. Además, el rendimiento de la membrana de UF mejoró en términos de eficiencia de rechazo, con mayores valores de rechazo respecto de los contaminantes orgánicos (RCOD, 48.5 vs. 39.9 % y 53.4 vs. 42.0 %. El rendimiento de

  12. Evaluating Alerting and Guidance Performance of a UAS Detect-And-Avoid System

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    Lee, Seung Man; Park, Chunki; Thipphavong, David P.; Isaacson, Douglas R.; Santiago, Confesor

    2016-01-01

    A key challenge to the routine, safe operation of unmanned aircraft systems (UAS) is the development of detect-and-avoid (DAA) systems to aid the UAS pilot in remaining "well clear" of nearby aircraft. The goal of this study is to investigate the effect of alerting criteria and pilot response delay on the safety and performance of UAS DAA systems in the context of routine civil UAS operations in the National Airspace System (NAS). A NAS-wide fast-time simulation study was conducted to assess UAS DAA system performance with a large number of encounters and a broad set of DAA alerting and guidance system parameters. Three attributes of the DAA system were controlled as independent variables in the study to conduct trade-off analyses: UAS trajectory prediction method (dead-reckoning vs. intent-based), alerting time threshold (related to predicted time to LoWC), and alerting distance threshold (related to predicted Horizontal Miss Distance, or HMD). A set of metrics, such as the percentage of true positive, false positive, and missed alerts, based on signal detection theory and analysis methods utilizing the Receiver Operating Characteristic (ROC) curves were proposed to evaluate the safety and performance of DAA alerting and guidance systems and aid development of DAA system performance standards. The effect of pilot response delay on the performance of DAA systems was evaluated using a DAA alerting and guidance model and a pilot model developed to support this study. A total of 18 fast-time simulations were conducted with nine different DAA alerting threshold settings and two different trajectory prediction methods, using recorded radar traffic from current Visual Flight Rules (VFR) operations, and supplemented with DAA-equipped UAS traffic based on mission profiles modeling future UAS operations. Results indicate DAA alerting distance threshold has a greater effect on DAA system performance than DAA alerting time threshold or ownship trajectory prediction method

  13. Using Natural Products to Treat Resistant and Persistent Bacterial Infections

    Science.gov (United States)

    Deering, Robert W.

    -lactams, tetracyclines, aminoglycosides, and chloramphenicol. The efflux pump inhibitory mechanism was further proved through an accumulation assay with the Hoechst dye 33342. In chapter three, we report the discovery of a 1,2-benzisoxazole with new antibacterial activity against MDR A. baumannii, a pathogen with a critical need of new treatments. This compound was produced by bacterial fermentation and synthetic preparation and shows minimum inhibitory concentrations as low as 6.25 ?g/mL against a panel of four clinically relevant A. baumannii strains. Key structure activity relationships were demonstrated using synthetic analogs of the lead 1,2-benzisoxazole. We advocate for further studies to advance the development of this compound. The third study, describes an in vitro quiescent state of uropathogenic E. coli (UPEC) and bacteria-produced signals that can prevent this state. Quiescence was seen in the classic UPEC strain CFT073 only when grown on glucose M9 minimal medium agar plates seeded with ≤10 6 CFU. Interestingly, this quiescent state is seen in 80% of E. coli phylogenetic group B2 multilocus sequence type 73 strains, as well as 22.5% of randomly selected UPEC strains isolated from community acquired urinary tract infections in Denmark. Furthermore, it was determined that CFT073 forms a high persister cell fraction under these growth conditions. Both the persistent and quiescent states were inhibited significantly by a cocktail of lysine, tyrosine, and methionine at concentrations relevant to those in human urine. The use of CFT073 mini-Tn5 metabolic mutants ( gnd, gdhA, pykF, sdhA, and zwf) showed that both quiescence and persistence require a complete TCA cycle, but that the dormant states differ in that persistence requires a non-functional rpoS gene and quiescence does not. These results suggest that interference with these central metabolic pathways may be able to mitigate UPEC infections. In the fifth chapter, cranberry oligosaccharides and related compounds were

  14. Malaria Surveillance - United States, 2014.

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    Mace, Kimberly E; Arguin, Paul M

    2017-05-26

    . Less than 1.0% of patients were infected with two species. The infecting species was unreported or undetermined in 11.7% of cases. CDC provided diagnostic assistance for 14.2% of confirmed cases and tested 12.0% of P. falciparum specimens for antimalarial resistance markers. Of patients who reported purpose of travel, 57.5% were visiting friends and relatives (VFR). Among U.S. residents for whom information on chemoprophylaxis use and travel region was known, 7.8% reported that they initiated and adhered to a chemoprophylaxis drug regimen recommended by CDC for the regions to which they had traveled. Thirty-two cases were among pregnant women, none of whom had adhered to chemoprophylaxis. Among all reported cases, 17.0% were classified as severe illness, and five persons with malaria died. CDC received 137 P. falciparum-positive samples for the detection of antimalarial resistance markers (although some loci for chloroquine and mefloquine were untestable for up to nine samples). Of the 137 samples tested, 131 (95.6%) had genetic polymorphisms associated with pyrimethamine drug resistance, 96 (70.0%) with sulfadoxine resistance, 77 (57.5%) with chloroquine resistance, three (2.3%) with mefloquine drug resistance, one (https://www.cdc.gov/malaria/travelers/drugs.html). Malaria infections can be fatal if not diagnosed and treated promptly with antimalarial medications appropriate for the patient's age and medical history, likely country of malaria acquisition, and previous use of antimalarial chemoprophylaxis. Recent molecular laboratory advances have enabled CDC to identify and conduct molecular surveillance of antimalarial drug resistance markers (https://www.cdc.gov/malaria/features/ars.html) and improve the ability of CDC to track, guide treatment, and manage drug resistance in malaria parasites both domestically and globally. For this effort to be successful, specimens should be submitted for all cases diagnosed in the United States. Clinicians should consult CDC