text:
Editor: Rustam Aminov
Dynamics of development and dispersal in sessile microbial communities: examples from Pseudomonas aeruginosa and Pseudomonas putida model biofilms
Version of Record online: 16 MAY 2006
DOI: 10.1111/j.1574-6968.2006.00280.x
FEMS Microbiology Letters
Volume 261, Issue 1, pages 1–11, August 2006
Additional Information
How to Cite
Klausen, M., Gjermansen, M., Kreft, J.-U. and Tolker-Nielsen, T. (2006), Dynamics of development and dispersal in sessile microbial communities: examples from Pseudomonas aeruginosa and Pseudomonas putida model biofilms. FEMS Microbiology Letters, 261: 1–11. doi: 10.1111/j.1574-6968.2006.00280.x
Publication History
- Issue online: 16 MAY 2006
- Version of Record online: 16 MAY 2006
- Received 1 March 2006, revised 10 April 2006; accepted 10 April 2006.First published online 16 May 2006.
- Abstract
- Article
- References
- Cited By
- biofilm;
- development;
- dynamics;
- evolution;
- Pseudomonas aeruginosa;
- Pseudomonas putida
Surface-associated microbial communities in many cases display dynamic developmental patterns. Model biofilms formed by Pseudomonas aeruginosa and Pseudomonas putida in laboratory flow-chamber setups represent examples of such behaviour. Dependent on the experimental conditions the bacteria in these model biofilms develop characteristic multicellular structures through a series of distinct steps where cellular migration plays an important role. Despite the appearance of these characteristic developmental patterns in the model biofilms the available evidence suggest that the biofilm forming organisms do not possess comprehensive genetic programs for biofilm development. Instead the bacteria appear to have evolved a number of different mechanisms to optimize surface colonization, of which they express a subset in response to the prevailing environmental conditions. These mechanisms include the ability to regulate cellular adhesiveness and migration in response to micro-environmental signals including those secreted by the bacteria themselves.
Bacteria in nature often exist in surface-associated sessile communities referred to as biofilms (Costerton et al., 1995). Microbial biofilms have been subject to intense study during the last decade mainly for two reasons. First, it is of basic scientific interest to understand how bacteria form and live in multicellular communities. Second, because bacteria in biofilms in many cases are difficult to eliminate via biocide or antibiotic treatment or host immune responses, biofilm formation causes considerable problems in industrial and medical settings (Costerton et al., 1999), and knowledge about the environmental cues, genetic elements, and molecular mechanism which are involved when bacteria form biofilms is the basis for rational design of strategies to control biofilm development.
A substantial part of the studies of microbial biofilms carried out during the last decade has involved laboratory setups such as microtitre trays and flow chambers. The model biofilms grown in these setups most likely differ a lot from the biofilms which are formed in industrial, clinical, and environmental settings. Nevertheless experiments with model biofilms have identified and provided information about a number of factors which are important during biofilm development and should be relevant beyond the model systems. These factors include bacterial attachment mechanisms, surface-associated spreading mechanisms, cell-to-cell interconnecting components, dispersion mechanisms, and the environmental cues and underlying genetic elements which are involved. In the present review we present examples of work carried out on these topics with model biofilms formed by Pseudomonas putida and Pseudomonas aeruginosa. These two pseudomonads within the controlled conditions of laboratory setups represent different pathways of biofilm development. Our current knowledge is presented in accordance with the biofilm developmental cycle, i.e. attachment, microcolony formation, maturation, and dispersion. Finally, we discuss whether the presented types of biofilm development should be regarded as a form of microbial development, and we touch upon issues of sociality and cooperation in biofilms.
Development and dynamics in P. aeruginosa biofilms
Studies of P. aeruginosa biofilm development and dynamics have provided illustrative examples of the diversity of ways surface-associated microbial multicellular structures can develop. Pseudomonas aeruginosa is capable of flagellum-driven swimming motility and type IV pili-mediated surface-associated twitching motility. Both swimming and twitching motility can contribute to the dynamic structural changes that occur during P. aeruginosa biofilm development.
Transport of P. aeruginosa cells to a surface before attachment is a dynamic equilibrium between diffusive, convective and active flagellum-driven transport (van Loosdrecht et al., 1990). Mutations in genes encoding flagella (O'Toole & Kolter, 1998; Sauer et al., 2002), type IV pili (Deziel et al., 2001; Chiang & Burrows, 2003) and cup fimbria (Vallet et al., 2001) were shown to affect the ability of P. aeruginosa to attach to surfaces in microtiter tray and flow-chamber setups. However, growth conditions under which lack of flagella and/or type IV pili did not affect surface attachment in a flow chamber have been reported as well (Klausen et al., 2003b). After P. aeruginosa has attached to a surface it either detaches, stay attached at the position of attachment, or move along the surface by twitching motility (Singh et al., 2002; Klausen et al., 2003a, b). The gene sadB (PA5346) has been shown to be involved in regulation of the attachment–detachment frequency in a flow-chamber setup (Caiazza & O'Toole, 2004).
Studies of microcolony formation of twitching and nontwitching cells by time-lapse confocal laser scanning microscopy (CLSM) have shown that cells attach to a surface and form microcolonies by keeping a balance between cell division, surface migration and detachment (Singh et al., 2002; Klausen et al., 2003a, b). Experiments with mixtures of cyan fluorescent protein-tagged and yellow fluorescent protein-tagged cells provided evidence that microcolonies developed by clonal growth in a flow-chamber system (Fig. 1) (Klausen et al., 2003a). On the contrary, microcolony formation by motility driven cell-to-cell aggregation has been observed in a microtitre tray setup (O'Toole & Kolter, 1998). Conditions that promote extensive twitching motility of P. aeruginosa have been shown to prevent microcolony formation in a flow-chamber setup. Interestingly, this could be a mechanism which prevents biofilm formation in humans as lactoferrin, a component of the innate immune system, induce extensive twitching motility in P. aeruginosa by chelating iron (Singh et al., 2002).
Extracellular polymeric substances (EPS), which consist of polysaccharide, extracellular DNA, and protein, have important cell-to-cell interconnecting functions in biofilms. Five gene clusters in P. aeruginosa potentially encode functions for exopolysaccharide biosynthesis. The alg biosynthesis operon (PA3540–PA3551) appears not to be expressed in P. aeruginosa flow-chamber biofilms (Wozniak et al., 2003), but evidently play a role in biofilm formation by P. aeruginosa in particular in the lungs of cystic fibrosis patients (Govan & Deretic, 1996). The psl cluster (PA2231–PA2245) is involved in the production of a mannose-rich exopolysaccharide, which was shown to play a role in P. aeruginosa biofilm formation (Friedman & Kolter, 2004b; Jackson et al., 2004; Matsukawa & Greenberg, 2004). Bacteria with a mutation in pslA displayed delayed microcolony formation in a flow chamber setup (Matsukawa & Greenberg, 2004). The psl locus was also found to be important for early biofilm formation in a microtitre tray assay (Friedman & Kolter, 2004b). The pel (PA3058–PA3064) cluster encodes production of a glucose rich matrix component which was shown to facilitate biofilm formation of some P. aeruginosa strains at the liquid air interface (Friedman & Kolter, 2004a). Mutations in the pel locus also made P. aeruginosa biofilms susceptible to shear force in a microtitre tray setup (Friedman & Kolter, 2004a). In addition, two other gene clusters (PA1381–PA1393 and PA3552–3558) potentially code for synthesis of exopolysaccharides which may have roles in biofilm development under some conditions (Matsukawa & Greenberg, 2004). Addition of DNase to young (24-h-old) flow chamber grown P. aeruginosa biofilms was shown to remove the biofilm cells, indicating an importance of extracellular DNA as a biofilm matrix component under some conditions (Whitchurch et al., 2002). Evidence has been presented that the extracellular DNA in P. aeruginosa biofilms may be generated via at least two different pathways. A basal level of extracellular DNA appears to be generated via a pathway which is not linked to quorum sensing, while a larger amount of extracellular DNA appears to be generated via a quorum-sensing regulated pathway (Allesen-Holm et al., 2006). Evidence has been presented that the extracellular DNA in P. aeruginosa biofilms originates from lysis of a small subpopulation of the cells (Allesen-Holm et al., 2006). The mutants lasIrhlI (PA1432–PA3476), pqsA (PA0996) and fliMpilA (PA1443–PA4525) formed biofilms with low extracellular DNA levels and showed increased susceptibility to treatment with SDS, indicating a stabilizing effect of the extracellular DNA in biofilms (Allesen-Holm et al., 2006). In addition, mature wild-type biofilms that were treated with DNase for a short time before SDS treatment showed elevated susceptibility to the SDS treatment (Allesen-Holm et al., 2006). In agreement with a role of quorum sensing in biofilm development Davies et al. (1998) found that, unlike the wild-type, a P. aeruginosa lasI mutant was unable to structurally mature into mushroom shaped SDS-resistant microcolonies, but developed a flat SDS-susceptible biofilm in a flow-chamber setup. Evidence is accruing that cup fimbria in addition to their role in initial biofilm formation also play a role as cell-to-cell interconnecting compounds in mature biofilms. The sadARS genes (PA3946–3948) codes for a putative sensor histidine kinase and two response regulators, and mutations in any of these genes were shown to result in biofilms with an altered mature structure (Kuchma et al., 2005). In another study, the sadARS genes (PA3946-3948, termed rocARS) were shown to regulate biosynthesis of cup fimbria (Kulasekara et al., 2005).
After formation of the initial microcolonies structural biofilm development by P. aeruginosa appears to be conditional. For example, a flat biofilm was formed in flow chambers irrigated with citrate minimal medium (Klausen et al., 2003b), while a heterogeneous biofilm with mushroom-shaped multicellular structures was formed in flow chambers irrigated with glucose minimal medium (Klausen et al., 2003a). Pseudomonas aeruginosa biofilm development in flow chambers with citrate as carbon source was shown to occur via formation of initial microcolonies by clonal growth of sessile cells at the substratum, followed by expansive migration of the bacteria out on the substratum, and the eventual formation of a flat biofilm (Fig. 1) (Klausen et al., 2003b). As biofilm formation by a P. aeruginosa pilA mutant (deficient in biogenesis of type IV pili) occurred without the expansive phase, and resulted in discrete protruding microcolonies, it was suggested that the expansive migration of the bacteria out on the substratum was type IV pili-driven. CLSM time-lapse microscopy indicated that the shift from nonmotile to migrating cells occurred when the initial microcolonies reached a certain size (Fig. 1), suggesting that the shift may by induced by some sort of limitation arising in the initial microcolonies. The formation of mushroom-shaped structures in glucose-grown P. aeruginosa biofilms was shown to occur in a sequential process involving a nonmotile bacterial subpopulation which formed the initial microcolonies by growth in certain foci of the biofilm, and a migrating bacterial subpopulation which initially formed a monolayer on the substratum, and subsequently formed the mushroom caps by aggregating on top of the microcolonies (mushroom stalks) via a type IV pili-dependent process (Klausen et al., 2003a). In colour-coded biofilms this type of dynamic development gave rise to mushroom-shaped structures with stalks composed of clonal subpopulations of one colour, and caps composed of aggregated subpopulations of mixed colour (Fig. 2a). Growth of the initial microcolonies in the glucose-grown biofilms continued past the point where spreading by twitching motility prevented further microcolony formation in the citrate-grown biofilms. In pilA/wild-type mixed biofilms formation of mushroom-shaped structures could occur also with citrate as the carbon source (Fig. 2b). In these citrate-grown mixed biofilms the pilA bacteria formed the mushroom stalks, and the wild-type bacteria accumulated on top of these stalks and formed the mushroom caps. The fact that a twitching deficient hyperpiliated P. aeruginosa pilT mutant formed biofilms with large mushroom-shaped structures (Chiang & Burrows, 2003), and our unpublished experiments with mixtures of pilA mutants and twitching deficient but piliated mutants, indicate that type IV piliation of the cells is required for mushroom cap formation, while type IV pili-driven motility, although not absolutely required, can facilitate the process substantially. The extracellular DNA in P. aeruginosa biofilms was shown to be present in high concentrations on the microcolonies in young biofilms and between the stalk-forming and the cap-forming subpopulations in mature biofilms (Fig. 3) (Allesen-Holm et al., 2006). Because type IV pili bind to DNA (Aas et al., 2002; Van Schaik et al., 2005), it is tempting to speculate that the high concentration of extracellular DNA on the mushroom stalks might cause accumulation of migrating (either twitching or swimming) piliated bacteria resulting in the formation of the mushroom caps.
Dispersion of biofilms in response to perturbation in the carbon availability has been reported for P. aeruginosa biofilms. Sauer et al. (2004) reported that P. aeruginosa biofilms grown in flow chambers on glutamate medium responded to an abrupt upshift in carbon availability by initiating a dispersion process that lead to the majority of the biomass being released from the biofilm. The extent of dispersion was dependent on the carbon source used and was associated with increased expression of flagella and down regulation of twitching motility. The production of rhamnolipid biosurfactant has also been associated with dispersal of cells from P. aeruginosa biofilms (Boles et al., 2005). During biofilm development of P. aeruginosa in rich medium local dispersion was observed as a hollowing out of some microcolonies (Purevdorj-Gage et al., 2005). Through careful microscopic inspection it was observed that two subpopulations existed in the microcolonies. On the outside of the microcolonies a wall forming subpopulation of nonmotile cells was present, and on the inside of the microcolonies a motile rapidly moving subpopulation was present. The motile subpopulation eventually found its way out of the microcolony which resulted in microcolonies with a central void. This phenomenon has been termed 'seeding dispersal' and was shown to be dependant on the microcolonies reaching a critical size.
Development and dynamics in P. putida biofilms
Initial attachment and colonization by P. putida on the surface of plant roots and fungal hyphae have been proposed to involve flagella (Yang et al., 1994; Turnbull et al., 2001). Flagella, however, do not seem to be required for efficient attachment and biofilm formation by P. putida under all conditions as a nonflagellated fliM (PP4358) mutant formed biofilms in a flow-chamber system within the same timeframe as the wild-type (Gjermansen et al., 2005). In addition, Choy et al. (2004) reported that for a P. putida morA (similar to PP0672) mutant, which was shown to be hyper-flagellated and hyper-motile, initial biofilm formation was impaired. In a transposon screen for P. putida mutants defective in adhesion to seeds Espinosa-Urgel et al. (2000) identified a mutant, mus-24 (PP0168), which had defects in both seed adhesion and adhesion to abiotic materials like glass and plastic. The mus-24 locus of P. putida is highly homologues to the lapA gene of Pseudomonas fluorescens (Pfl_0133 in strain PfO-1), and a detailed study of a P. fluorescens lapA mutant documented that LapA was necessary for nonreversible binding to surfaces (Hinsa et al., 2003). LapA belongs to a group of large surface proteins which contain highly repetitive sequences and share functional features such as adhesion to surfaces (recently reviewed by Lasa & Penades, 2006). Our unpublished results suggest that LapA also functions as cell-to-surface adhesin in flow-chamber-grown P. putida biofilms, and that it in addition is important for later stages in biofilm development as a part of the cell-to-cell interconnecting matrix.
Cellulose is an important constituent of the biofilm matrix material of many bacteria and has also been suggested to have a role in P. putida biofilms (Camesano & Abu-Lail, 2002; Chang & Halverson, 2003). In P. fluorescens an acetylated form of cellulose plays an important role in maintaining the structural rigidity of air–liquid interface biofilms (Spiers et al., 2003; Spiers & Rainey, 2005), and the production of this cellulose-like polymer confers an ecological advantage during the colonization of plant surfaces (Gal et al., 2003). Pseudomonas putida KT2440 has a complete cellulose biosynthetic operon (PP2634–2638), and the production of calcofluor white-stainable material in flow-chamber-grown P. putida biofilms suggests that cellulose is expressed in this system (Nelson et al., 2002; our unpublished results). In addition to cellulose and LapA protein P. putida has been reported to produce substantial amounts of extracellular DNA in the sessile mode of growth (Steinberger & Holden, 2005), which may function as biofilm stabilizing matrix material.
Structural biofilm development by P. putida in flow chambers irrigated with citrate minimal medium has been investigated using dual fluorescent labelling and CLSM (Tolker-Nielsen et al., 2000). Biofilm development occurs initially by the formation of compact microcolonies via clonal growth of nonmotile bacteria (Fig. 4a). When the compact microcolonies reach a critical size, the bacteria inside the microcolonies are no longer firmly interconnected and they move rapidly by means of flagellum-driven motility. The combination of local dissolution of the cell-to-cell interconnecting bonds and flagellum-driven motility, results in release of cells from inside of the microcolonies, and leads to a shift in the biofilm architecture from compact microcolonies to loose protruding structures (Fig. 5). The loose protruding structures in mature P. putida biofilms contain a mixture of bacteria from separate microcolonies (Fig. 4b) indicating that a substantial dynamic reorganization occurs during development (Tolker-Nielsen et al., 2000). Time-lapse microscopy revealed that the structural rearrangement was preceded by rapid movement of bacteria inside the microcolonies, and the lack of this movement in microcolonies formed by a fliM mutant suggested that it was flagellum-driven. Consistent with the described developmental steps in P. putida biofilms, Sauer and colleagues found that the expression of flagella in P. putida populations was down regulated quickly after surface attachment, but upregulated again at later stages of biofilm development (Sauer & Camper, 2001). Evidence has been provided that the local biofilm dissolution and release of cells from the interior of the microcolonies is induced by local carbon starvation (Gjermansen et al., 2005). It was also found that global carbon starvation led to rapid dispersal of the entire biofilm within few minutes after the onset of starvation (Fig. 6). Microscopic analysis indicated that the cells were swimming rapidly during the global dispersion process, but experiments with a nonmotile fliM mutant showed that dispersion did not occur because of the motility per se, but through modulation of the cell-to-cell interconnecting compounds (Gjermansen et al., 2005). Experiments with a biofilm dispersion deficient mutant indicated that the local and global dispersion process utilizes the same molecular mechanisms.
Genetic analysis of the dispersion process in P. putida OUS82 lead to the identification of an operon, consisting of the two genes PP0164 and PP0165, which regulates the adhesiveness of the cells (Gjermansen et al., 2005). Biofilms formed by a PP0164 mutant did not undergo the transition from compact microcolonies to loose protruding structures but instead the microcolonies continued to expand reaching almost complete substratum coverage. In addition, biofilms formed by the PP0164 mutant did not undergo dispersion in response to global carbon starvation. The PP0164 protein contains a putative signal protein for transport to the periplasm and a domain of unknown function, DUF920, which based on a bioinformatics approach has been proposed to function as a papain-like cysteine proteinase (Ginalski et al., 2004). Contrary to the PP0164 mutants, bacteria with mutations in the PP0165 gene were unable to initiate biofilm formation. The PP0165 gene encodes a putative inner membrane protein which contains an N-terminal periplasmic domain and C-terminal GGDEF and EAL domains. The GGDEF and EAL domains have been implicated in the turnover of the bacterial second messenger c-di-GMP. The GGDEF domain contains the diguanylate cyclase activity for synthesis of c-di-GMP, and the EAL domain contains a phosphodiesterase activity for degradation of c-di-GMP (Paul et al., 2004; Schmidt et al., 2005). Evidence that GGDEF and EAL domain proteins are involved in the regulation of cellular adhesiveness in diverse bacteria through modulation of the intracellular c-di-GMP levels is growing rapidly (D'Argenio & Miller, 2004; Römling et al., 2005). Comparative sequence analysis shows that PP0164/PP0165-homologues in a number of environmental species including P. putida, P. fluorescens and Shewanella oneidensis cluster together with genes encoding synthesis and export of proteins homologous to LapA. Our unpublished data suggest that PP0164 and PP0165 in conjunction regulate the expression or surface localization of the LapA protein and that this modulation play an important role in the formation/dispersal processes observed. Our experiments also suggest that LapA is not only a surface adhesin but also act as an important component of the matrix that interconnects the cells in the biofilm at later stages. One possible mechanism for the dispersion process could be dependant on the release of LapA from the bacterial surface in response to starvation by the action of the putative protease activity of the PP0164 protein. This hypothesis would be consistent with the very fast initiation of the dispersion process in response to starvation.
Is biofilm structure the outcome of a developmental program or the result of ecological adaptation?
Although P. putida and P. aeruginosa biofilm development can result in elaborate multicellular structures and appears to involve complex regulation, there is at present no evidence suggesting that these organisms possess a comprehensive genetic biofilm developmental program comparable to, for example, the fruiting body developmental program of myxobacteria (see Kaiser (2003) for a review about myxobacterial development). The elaborate biofilm structure may be nothing more than a spatial pattern emerging because of the actions of the individuals constituting the biofilm. Comparative analysis of the transcriptome or proteome of P. aeruginosa cells from biofilms and planktonic cultures (e.g. Whiteley et al., 2001; Sauer et al., 2002; Hentzer et al., 2004) has not identified a comprehensive genetic program for biofilm development. The fact that biofilm development is conditionally dependent on, among other things, the type and amount of carbon source, implies that the organisms are able to respond to environmental factors while they are forming the biofilm. It is therefore more likely that the bacteria are not committed to a special developmental program during biofilm formation, but that they have evolved mechanisms to optimize surface colonization in response to the environmental conditions. These mechanisms include the ability to regulate cellular adhesiveness and motility in response to micro-environmental signals including those secreted by the bacteria themselves.
From this perspective of ecological adaptation of individual cells driving the evolution of attachment, biofilm formation, and dispersal mechanisms, the degree of cooperative behaviour of the members of the multicellular biofilm is called into question. Perhaps the evolution of some of the apparently cooperative traits can be explained more simply by the fitness advantages of this behaviour for the individual bacterium? In the following, we will examine the evidence for and against the cooperative nature of the above reviewed behaviour of P. aeruginosa and P. putida in biofilms.
To what degree does biofilm formation involve multicellular cooperation?
Most aspects of biofilm formation reviewed in this article can be viewed either as social behaviour or as single-cell behaviour (for examples of such a dual interpretation, see Redfield, 2002). Social behaviour may be positive (cooperation and altruism) or negative (spite and cannibalism). The key problem for the evolution of cooperation is the emergence of cheaters. They benefit from the cooperating individuals' investment into public goods without investing into cooperation themselves. This creates a conflict of interest between the fitness of the individual and the fitness of the group (Velicer, 2003; Kreft, 2004b; Travisano & Velicer, 2004). The evolution of cooperation in microorganisms can be explained either by kin selection theory (Hamilton, 1964; Griffin et al., 2004) or by multilevel selection theory (Sober & Wilson, 1998). These two explanations complement rather than contradict each other (Queller, 2004). The predominantly clonal structure of the microcolonies, combined with refounding of them from single cells, acts as a purification mechanism to get rid of cheaters, and promotes the evolution of cooperation in biofilms (Kreft, 2004a; Kreft & Bonhoeffer, 2005). However, the mushroom caps in P. aeruginosa biofilms, and other multicellular structures which are formed via pathways that involve aggregation, are not clonal.
The production of the cell-to-cell interconnecting components in biofilms may be a cost each bacterium pays in order to contribute to the social activity of creating a protective biofilm domicile, or it may simply increase the adhesiveness of single cells allowing them to persist in an environment. Production of matrix components appears to be cooperative because cheaters who benefit from the existence of the matrix, but do not invest into its construction, may exist. Further, the matrix as a public good could be exploited by selfish mutants using it as a food source but damaging its integrity in turn.
The local biofilm dissolution and structural changes occurring during P. putida biofilm development may be a social process which creates optimal nutritional conditions to the biofilm population, or it may simply be a response of the individual cells to the local nutritional conditions. The latter view is supported by the experiments showing that almost all cells leave the biofilm if the flow, and therefore substrate supply, is switched off. The formation of regular mushroom-shaped structures in P. aeruginosa biofilms via a pathway that involves type IV pili-dependent cellular migration may be a coordinated social process that creates biofilms with architectures which allow optimal circulation and nutrient supply to the population, or it may, because nutrient gradients develop in biofilms, be the result of chemotaxis of individuals moving to a favourable nutrient-containing microenvironment.
Despite the increase of biofilm surface area owing to the formation of the loose protruding structures or the mushroom-shaped structures, the effective mass transfer area may actually decrease because only the top of the structures receives most of the substrate, unless convection in the valleys is dominant (Picioreanu et al., 2000). If we do assume that the formation of the mushroom-shaped structures or loose protruding structures increase substrate flux to the biofilms as a whole, which would imply a group level benefit of the structural changes, why does the stalk-forming subpopulation not evolve to become a cap-forming subpopulation or the wall-forming subpopulation not evolve to be part of the loose protruding structures, thereby increasing their substrate supply? The fitness of the subpopulations would differ under most circumstances, and natural selection would be expected to shift the balance in favour of the fitter subpopulation. The split into the phenotypically different subpopulations appears to be epigenetic, however, because isolated cap subpopulations are able to form biofilms with mushroom-shaped structures (our unpublished data), and evolution, therefore, cannot select the fitter subpopulation. Circumventing natural selection between the subpopulations subjects the combination of the subpopulations to natural selection. Therefore, a population may increase its fitness by evolving a mechanism for epigenetic formation of subpopulations whose interactions give rise to synergistic benefits. The relative frequency of the subpopulations might have evolved to depend on the environmental conditions, which could be underlying causes resulting in the structural differences seen in citrate-grown and glucose-grown P. aeruginosa biofilms.
While the observed diversification of the biofilm populations into subpopulations may result in population-level fitness benefits because of substrate transfer optimization, it could also be an insurance mechanism (Boles et al., 2004). For example, the bacteria in the caps or loose protruding structures could be able to benefit from better growth in good times, while the slower-growing or dormant bacteria in the stalks or walls could have better chances of survival in times of adversity (Wolf et al., 2005).
The release of DNA via lysis of a small subpopulation of P. aeruginosa cells may serve the purpose of biofilm stabilization, procurement of food, elimination of damaged cells, or provide a common gene pool for diversification of the population. Whatever the purpose, cell lysis may be altruism if it is autolysis (Lewis, 2000) or cannibalism/fratricide if the lysed cells are killed by their kin/siblings (Gonzalez-Pastor et al., 2003; Gilmore & Haas, 2005; Havarstein et al., 2006). Both altruism and cannibalism can give rise to group-level benefits such as diversification. The observed involvement of quorum sensing in DNA release from the lysing subpopulation, resulting in biofilm stabilization, is difficult not to interpret as social behaviour because the lysing cells that provide the DNA obviously do not benefit themselves directly.
Model biofilms formed by Pseudomonas aeruginosa and Pseudomonas putida in laboratory flow-chamber setups represent examples of surface-associated microbial communities which display dynamic developmental patterns. Dependent on the experimental conditions these model biofilms develop flat, mushroom-shaped, or loose protruding structures through a series of distinct steps where regulation of cellular migration and adhesiveness play important roles. Despite the appearance of these characteristic developmental patterns in the model biofilms, the available evidence suggests that these organisms do not possess a comprehensive genetic program for biofilm development. Instead, the bacteria appear to have evolved a number of different mechanisms to optimize surface attachment, microcolony formation, and dispersal, of which they express a subset in response to the prevailing environmental conditions. It appears that the increased adhesiveness that distinguishes biofilm cells from planktonic cells can be achieved by the expression of many different genes, and that the environmental conditions determine the actual cell-to-cell interconnecting components which are being expressed. While certain characteristics of biofilm formation, such as diversification of the population and DNA-release by cell lysis, presumably constitute social behaviour, the group benefits of the apparent biofilm structural engineering may not only lie in improved substrate supply but also in the establishment of subpopulations with different survival strategies.
This work was supported by grants from the Lundbeck foundation and the Danish Technical Research Council (grant no. 26-03-02) to T.T.N.
- 2002) Competence for natural transformation in Neisseria gonorrhoeae: components of DNA binding and uptake linked to type IV pilus expression. Mol Microbiol 46: 749–760. , , , , & (
- 2006) A characterization of DNA release in Pseudomonas aeruginosa cultures and biofilms. Mol Microbiol 59: 1114–1128. , , , , , , , & (
- 2004) Self-generated diversity produces 'insurance effects' in biofilm communities. Proc Natl Acad Sci USA 101: 16630–16635. , & (
- 2005) Rhamnolipids mediate detachment of Pseudomonas aeruginosa from biofilms. Mol Microbiol 57: 1210–1223. , & (
- 2004) SadB is required for the transition from reversible to irreversible attachment during biofilm formation by Pseudomonas aeruginosa PA14. J Bacteriol 186: 4476–4485. & (
- 2002) Heterogeneity in bacterial surface polysaccharides, probed on a single-molecule basis. Biomacromolecules 3: 661–667. & (
- 2003) Reduced water availability influences the dynamics, development, and ultrastructural properties of Pseudomonas putida biofilms. J Bacteriol 185: 6199–6204. & (
- 2003) Biofilm formation by hyperpilated mutants of Pseudomonas aeruginosa. J Bacteriol 185: 2374–2378. & (
- 2004) MorA defines a new class of regulators affecting flagellar development and biofilm formation in diverse Pseudomonas species. J Bacteriol 186: 7221–7228. , , , & (
- 1995) Microbial biofilms. Annu Rev Microbiol 49: 711–745. , , , & (
- 1999) Bacterial biofilms: a common cause of persistent infections. Science 284: 1318–1322. , & (
- 2004) Cyclic di-GMP as a bacterial second messenger. Microbiology 150: 2497–2502. & (
- 1998) The involvement of cell-to-cell signals in the development of a bacterial biofilm. Science 280: 295–298. , , , , & (
- 2001) Initiation of biofilm formation by Pseudomonas aeruginosa 57RP correlates with emergence of hyperpiliated and highly adherent phenotypic variants deficient in swimming, swarming, and twitching motilities. J Bacteriol 183: 1195–1204. , & (
- 2000) Genetic analysis of functions involved in adhesion of Pseudomonas putida to seeds. J Bacteriol 182: 2363–2369. , & (
- 2004a) Genes involved in matrix formation in Pseudomonas aeruginosa PA14 biofilms. Mol Microbiol 51: 675–690. & (
- 2004b) Two genetic loci produce distinct carbohydrate-rich structural components of the Pseudomonas aeruginosa biofilm matrix. J Bacteriol 186: 4457–4465. & (
- 2003) Genes encoding a cellulosic polymer contribute toward the ecological success of Pseudomonas fluorescens SBW25 on plant surfaces. Mol Ecol 12: 3109–3121. , , , & (
- 2005) The selective advantage of microbial fratricide. Proc Natl Acad Sci USA 102: 8401–8402. & (
- 2004) BTLCP proteins: a novel family of bacterial transglutaminase-like cysteine proteinases. Trends Biochem Sci 29: 392–395. , , & (
- 2005) Characterization of starvation-induced dispersion in Pseudomonas putida biofilms. Environ Microbiol 7: 894–906. , , , & (
- 2003) Cannibalism by sporulating bacteria. Science 301: 510–513. , & (
- 1996) Microbial pathogenesis in cystic fibrosis: mucoid Pseudomonas aeruginosa and Burkholderia cepacia. Microbiol Rev 60: 539–574. & (
- 2004) Cooperation and competition in pathogenic bacteria. Nature 430: 1024–1027. , & (
- 1964) The genetical evolution of social behaviour II. J Theor Biol 7: 17–52. (
- 2006) New insights into the pneumococcal fratricide: relationship to clumping and identification of a novel immunity factor. Mol Microbiol 59: 1297–1307. , , , & (
- 2004) Quorum sensing in Biofilms: gossip in the slime world? Microbial Biofilms (GhannoumM & O'TooleG, eds), pp. 118–140. ASM Press, Washington, DC. , & (
- 2003) Transition from reversible to irreversible attachment during biofilm formation by Pseudomonas fluorescens WCS365 requires an ABC transporter and a large secreted protein. Mol Microbiol 49: 905–918. , , & (
- 2004) Identification of psl, a locus encoding a potential exopolysaccharide that is essential for Pseudomonas aeruginosa PAO1 biofilm formation. J Bacteriol 186: 4466–4475. , , , & (
- 2003) Coupling cell movement to multicellular development in myxobacteria. Nat Rev Microbiol 1: 45–54. (
- 2003a) Involvement of bacterial migration in the development of complex multicellular structures in Pseudomonas aeruginosa biofilms. Mol Microbiol 50: 61–68. , , & (
- 2003b) Biofilm formation by Pseudomonas aeruginosa wild type, flagella and type IV pili mutants. Mol Microbiol 48: 1511–1524. , , , , , & (
- 2004a) Biofilms promote altruism. Microbiology 150: 2751–2760. (
- 2004b) Conflicts of interest in biofilms. Biofilms 1: 265–276. (
- 2005) The evolution of groups of cooperating bacteria and the growth rate versus yield trade-off. Microbiology 151: 637–641. & (
- 2005) A three-component regulatory system regulates biofilm maturation and type III secretion in Pseudomonas aeruginosa. J Bacteriol 187: 1441–1454. , & (
- 2005) A novel two-component system controls the expression of Pseudomonas aeruginosa fimbrial cup genes. Mol Microbiol 55: 368–380. , , , , & (
- 2006) Bap: a family of surface proteins involved in biofilm formation. Res Microbiol 157: 99–107. & (
- 2000) Programmed death in bacteria. Microbiol Mol Biol Rev 64: 503–514. (
- 2004) Putative exopolysaccharide synthesis genes influence Pseudomonas aeruginosa biofilm development. J Bacteriol 186: 4449–4456. & (
- 2002) Complete genome sequence and comparative analysis of the metabolically versatile Pseudomonas putida KT2440. Environ Microbiol 4: 799–808. , , , et al. (
- 1998) Flagellar and twitching motility are necessary for Pseudomonas aeruginosa biofilm development. Mol Microbiol 30: 295–304. & (
- 2004) Cell cycle-dependent dynamic localization of a bacterial response regulator with a novel di-guanylate cyclase output domain. Genes Dev 18: 715–727. , , , , , & (
- 2000) A theoretical study on the effect of surface roughness on mass transport and transformation in biofilms. Biotech Bioeng 68: 355–369. , & (
- 2005) Phenotypic differentiation and seeding dispersal in non-mucoid and mucoid Pseudomonas aeruginosa biofilms. Microbiology 151: 1569–1576. , & (
- 2004) Social evolution: kinship is relative. Nature 430: 975–976. (
- 2002) Is quorum sensing a side effect of diffusion sensing? Trends Microbiol 10: 365–370. (
- 2005) C-di-GMP: the dawning of a novel bacterial signalling system. Mol Microbiol 57: 629–639. , & (
- 2001) Characterization of phenotypic changes in Pseudomonas putida in response to surface-associated growth. J Bacteriol 183: 6579–6589. & (
- 2002) Pseudomonas aeruginosa displays multiple phenotypes during development as a biofilm. J Bacteriol 184: 1140–1154. , , , & (
- 2004) Characterization of nutrient-induced dispersion in Pseudomonas aeruginosa PAO1 biofilm. J Bacteriol 186: 7312–7326. , , , , & (
- 2005) The ubiquitous protein domain EAL is a cyclic diguanylate-specific phosphodiesterase: enzymatically active and inactive EAL domains. J Bacteriol 187: 4774–4781. , & (
- 2002) A component of innate immunity prevents bacterial biofilm development. Nature 417: 552–555. , , & (
- 1998) Unto Others: The Evolution and Psychology of Unselfish Behaviour. Harvard University Press, Cambridge, MA. & (
- 2003) Biofilm formation at the air–liquid interface by the Pseudomonas fluorescens SBW25 wrinkly spreader requires an acetylated form of cellulose. Mol Microbiol 50: 15–27. , , & (
- 2005) The Pseudomonas fluorescens SBW25 wrinkly spreader biofilm requires attachment factor, cellulose fibre and LPS interactions to maintain strength and integrity. Microbiology 151: 2829–2839. & (
- 2005) Extracellular DNA in single- and multiple-species unsaturated biofilms. Appl Environ Microbiol 71: 5404–5410. & (
- 2000) Development and dynamics of Pseudomonas sp. biofilms. J Bacteriol 182: 6482–6489. , , , , & (
- 2004) Strategies of microbial cheater control. Trends Microbiol 12: 72–78. & (
- 2001) The role of motility in the in vitro attachment of Pseudomonas putida PaW8 to wheat roots. FEMS Microbiol Ecol 35: 57–65. , , & (
- 2001) The chaperone/usher pathways of Pseudomonas aeruginosa: identification of fimbrial gene clusters (cup) and their involvement in biofilm formation. Proc Natl Acad Sci USA 98: 6911–6916. , , , & (
- 1990) Influence of interfaces on microbial activity. Microbiol Rev 54: 75–87. , , & (
- 2005) DNA binding: a novel function of Pseudomonas aeruginosa type IV pili. J Bacteriol 187: 1455–1464. , , , , , , & (
- 2003) Social strife in the microbial world. Trends Microbiol 11: 330–337. (
- 2002) Extracellular DNA required for bacterial biofilm formation. Science 295: 1487. , , & (
- 2001) Gene expression in Pseudomonas aeruginosa biofilms. Nature 413: 860–864. , , , , , & (
- 2005) Diversity in times of adversity: probabilistic strategies in microbial survival games. J Theor Biol 234: 227–253. , & (
- 2003) Alginate is not a significant component of the extracellular polysaccharide matrix of PA14 and PAO1 Pseudomonas aeruginosa biofilms. Proc Natl Acad Sci 100: 7907–7912. , , , , , & (
- 1994) Mutations affecting hyphal colonization and pyoverdine production in pseudomonads antagonistic toward Phytophthora parasitica. Appl Environ Microbiol 60: 473–481. , & (


text:
Drew Peterson: Untouchable
| ||
1:27 min|
Redbox Play Pass eligible
Plot Summary
When police Sgt. Drew Peterson meets Stacy Cales, he has it all: charm, money...and a wife, Kathleen. His old marriage falls by the wayside as Drew woos and weds Stacy, 30 years his junior. After his ex turns up dead in an apparent accident, Stacy is shocked as she sees a new side of Drew in his obsessive, controlling jealousy. Stacy vanishes, and her disappearance sets off a national media frenzy focused on Drew, who remains eerily certain that he's utterly untouchable.
Portions of this website Copyright ©2014 Baseline
This game can be purchased at:
Note: Do not reserve online. Simply go to the Box, select the game and buy it.
|Get Directions
Related Movies

subset: commoncrawl


text:
Revolving Door
July 30, 2012: Meredith Blake has been named New York-based entertainment writer at Los Angeles Times. She had been freelance Web producer at abcnews.com. (release)
August 2, 2011: Eleanor Hong has been named senior manager, content integration at ESPN3. She had been audience growth manager and SEO editor at ABCNews.com. (mb)
April 19, 2011: Brian Braiker has been named news editor at Adweek. He had been general assignment reporter at ABCNews.com. (mb)
October 29, 2010: Laiea Smith has been named managing editor at ABCNews.com. She had been senior producer at ABC News. (WebNewser)
October 29, 2010: Jon Dube has been named senior vice president and general manager at AOL News and Information. He had been vice president at ABCNews.com. (FBNY)
January 12, 2010: Eleanor Hong has been named lead editor, search engine optimization at ABCNews.com. She had been coordinating producer there. (mb)
November 16, 2009: Ed O'Keefe has been named managing editor at ABCNews.com. (WebNewser)
November 16, 2009: Deirdre Michalopoulos has been named senior managing editor, shows and show integration, at ABCNews.com. She had been managing editor there. (WebNewser)
November 6, 2009: Mark Schone has been named managing editor for investigative site Blotter at ABCNews.com. He had been executive news editor at Salon. (WebNewser)
September 23, 2009: Mark Forscher has been named creative director at ABCNews.com. He had been creative director at Newsweek.com. (mb)
August 19, 2008: Jonathan Dube has been named vice president of ABCNEWS.com. He had been director of digital media at CBC News. (release)
June 21, 2007: Ed O'Keefe has been named senior political editor at ABCNews.com. He had been a producer there for This Week. (TVNewser)
June 6, 2007: Chris Sheridan has been named executive producer at ABCNEWS.com. He had been executive producer at ABC News' Weekend News Content. (release)
December 13, 2006: Amanda Congdon is now at ABCNews.com, where she is making online video segments.(Guardian)
January 20, 2006: Steve Alperin has been named senior broadcast producer for ABCNEWS.com. He had been a writer and producer for ABC. (TVNewser)
August 30, 2005: Mark Cardwell has been named editorial director at ABCNEWS.com. He had been executive producer of the digital group at Associated Press since October 1, 1995. (mb)
May 6, 2005: Adam Glenn has been named a web consultant at NBC Universal. He was previously a senior producer at ABCNews.com. (mb)


text:
Jobs in fashion
Graduate - Manchester area, North West (UK)
Our client is asuccessful childnreswear supplierwho are lookng to emply a graduate within their merchandising team.
You will have a keen interested in fashion, preferably with a placeemnt year working within a supplier or retailer.
Please send cvs to firstname.lastname@example.com
Salary: £12000 - £14000
Job Added by In Focus S&S
Date Added: 7th July 2010
Closing Date: 1st September 2010
Upcoming Events
TFN Linkcubator
Sat 31st December 2016


text:
Select Local Merchants
Perfecta Salon & SpaRead MoreNot everyone can brush the tangles from their own hair, let alone style it. That's why Perfecta Salon & Spa employs a team of stylists who are experts in the intricacies of hair––everything from thickness and texture to color and astrological sign. In the comfort of the cozy studio, clients can request face-flattering layers, or rewire their head's circuit boards with bold highlights.3800 West 144th AvenueBroomfield, CO US
Salon TaoRead MoreBook your next day spa appointment at Salon Tao in Broomfield and enjoy a day filled with pampering beauty treatments. Whether you prefer natural or bold nails, this spa offers nail treatments for everyone. The staff at this spa is always familiar with the latest hair trends, making them some of the most creative and innovative stylists in the area. For your hair removal needs, head to Salon Tao for quick and painless services. Just pick a time that works for you and come into the spa. Walks ins are valued here. If taking care of your mind and body is high on your list of priorities, swing by for some relaxation and renewal at a price you can afford. Salon Tao is more than willing to accept your major credit card as a form of payment for your beauty services. The parking lot near Salon Tao will have you in and out in a jiffy. Book an appointment at Salon Tao and put your nails in the best care possible.13654 Xavier LaneBroomfield, CO US
Bella Pelle LaserRead MoreLooking for a great place to get your Zen on? Check out the selection of day spa services at Bella Pelle Laser in Broomfield. Soften, brighten, and nourish your skin with a rejuvenating facial from this spa. This establishment offers professional hair removal services for both men and women. Don't let your stress get you down! Head to this spa for the ultimate massage treatment. This spa offers hair care for men, women and children of all ages. Feel nourished and refreshed this weekend with a visit to Bella Pelle Laser.13383 Briarwood DriveBroomfield, CO US
The Right Touch Massage Therapy LLCRead MoreCobalt blue and redolent with the fragrance of brand-name essential oils, the treatment room of The Right Touch Massage Therapy LLC lavishes visitors in tranquil luxury. The massage therapist calls upon his extensive experience when conducting hot stone and relaxation massages, slicking his skilled hands with Young Living oils before kneading stress-addled muscles. In durations of 60, 90, and 120 minutes, each session seeks to restore balance and thoroughly relax its recipient.12995 Sheridan Blvd #101Broomfield, CO US


text:
Loading hotel availability
Hotel Description:Experience the charm of Boston's storied Seaport District at the acclaimed Renaissance Boston Waterfront Hotel. Our luxury hotel offers guests upscale, modern accommodations with breathtaking views of the Boston Harbor and the city. Our nautically-inspired guest rooms feature deluxe amenities like marble bathrooms, designer bedding, and flat-screen TVs. While nearby activities abound, travelers can easily spend a day enjoying our state-of-the-art fitness center or relaxing in our indoor pool and sauna. Enjoy casual dining at our Italian restaurant, Sap re Ristorante & Bar, or savor a local craft brew at our Capiz Bar. While we offer guests convenient access to conventions at Boston's Convention and Exhibition Center, our hotel's 20,000 square feet of dazzling venue space is a fantastic option for corporate events and weddings alike. Whether for business or pleasure, come experience the luxury of waterfront accommodations here at the Renaissance Boston Waterfront Hotel.
Check In: 4:00 PM
Check Out: 12:00 PM
Native currency: American Dollars
Number of floors: 21
Number of rooms: 471
Number of suites: 21
Check Out: 12:00 PM
Native currency: American Dollars
Number of floors: 21
Number of rooms: 471
Number of suites: 21
Restaurants:Sap re Ristorante & Bar: Sap re Ristorante & Bar invites you to enjoy modern interpretations of the classic rustic flavors of Italy with a focus on indigenous and seasonal ingredients of New England.
Capiz Bar: The unique spirit of Boston's Seaport is captured at Capiz Bar, our neighborhood pub serving craft cocktails and local brews in a nautical atmosphere.
Starbucks : Serving hot and cold coffee beverages, to-go snacks, and light menu items, Starbucks is located conveniently in our hotel in the heart of Boston's Seaport District.
Bowling alley
Hiking trail
Horseback riding
Mountain biking trail
Putt putt golf
Rock climbing
Squash court
Indoor pool
Children's program
Fitness center on-site
Health club
Room Info:The amenities listed in this description represent a sample of amenities available in some or all guest rooms in this hotel: High speed internet connection, Internet access, Wireless internet connection, High speed wireless, Non-smoking, Coffee/Tea maker, Instant hot water, Alarm clock, Air conditioning, Air conditioning individually controlled in room, Bottled water, Safe, Shower, Bathtub/shower combination, Separate tub and shower, Marble bathroom, Private bathroom, Weight scale, Bathrobe, Hairdryer, Upgraded bathroom amenities, Double vanity, Separate closet, Walk-in closet, Closets in room, Iron, Bathroom amenities, Down/feather pillows, Foam pillows, Pillow top mattress, Duvet, Feather bed, Sofa bed, Cribs, Rollaway bed, Electrical adaptors available, Local calls, Free toll free calls, Turn down service, Pantry, Tables and chairs, Mini-refrigerator, Color television, Cable television, Plug & play panel, Premium movie channels, CNN availaible, HBO available, ESPN available, Pay per view movies on TV, AM/FM radio, Remote control television, Large desk, Chair provided with desk, Desk with electrical outlet, Spare electrical outlet available at desk, Telephone, Voice mail, Multi-line phone, Speaker phone, Cordless phone, Connecting rooms
Meeting Facilities:Pacific Grand Ballroom:
Pacific A-D or E-H:
Pacific A-C or D or E or F-H:
Pacific A:
Pacific B:
Pacific C:
Pacific D:
Pacific E:
Pacific F:
Pacific G:
Pacific H:
Atlantic Junior Ballroom:
Atlantic 1:
Atlantic 2:
Atlantic 3:
Aegean Room:
Bering Board Room:
Caspian Room:
Mediterranean Room:
Thompson Room:
Spectacle Room:
Georges Room:
Brewster Room:
Hotel Amenities:(additional fees may apply to certain amenities)
- 24-hour front desk
- ATM/Cash machine
- Accessible facilities
- Barber shop
- Beauty shop/salon
- Boarding pass print-out available
- Bookstore
- Buffet breakfast
- Business center
- Business services
- Car rental desk
- Check cashing policy
- Cocktail lounge
- Cocktail lounge with entertainment
- Cocktail lounge with light fare
- Coffee shop
- Complimentary in-room coffee or tea
- Complimentary newspaper delivered to room
- Complimentary newspaper in lobby
- Concierge desk
- Continental breakfast
- Currency exchange
- Drugstore/pharmacy
- Dry cleaning
- Elevators
- Executive floor
- Exercise gym
- Express check-in
- Express check-out
- Florist
- Food and beverage outlets
- Front desk
- Full american breakfast
- Full service housekeeping
- Gift/News stand
- Hair dryer
- Health club
- High speed internet access
- High speed wireless
- Housekeeping - daily
- Ice machine
- Individual Efolio available
- Indoor pool
- Interior corridors
- Ironing board
- Laundry/Valet service
- Lounges/bars
- Meeting rooms
- Motorcycle parking
- Non-smoking rooms (generic)
- Off-Site parking
- Pool
- Public area air conditioned
- Restaurant
- Room service
- Safe deposit box
- Sauna
- Security
- Spa
- Steam bath
- Turndown service
- Valet cleaning
- Valet parking
- Vending machines
- Video checkout
- Video review billing
- Wakeup service
- Wedding services
- Whirlpool
- Wireless internet connection in public areas
Photo Gallery:
Boston MA AttractionsHotels near Boston Opera HouseHotels near Massachusetts General HospitalHotels near TD Banknorth Garden - Celtics and BruinsHotels near Tufts New England Medical CenterHotels near World Trade Center BostonHotels near Massachusetts Convention Center AuthorityHotels near Hynes Convention CenterHotels near Berklee College of MusicHotels near Northeastern UniversityHotels near Fenway Park - Boston Red SoxHotels near Wentworth Institute of TechnologyHotels near Children's Hospital BostonHotels near General Edward Lawrence Logan International Airport (BOS)Hotels near Boston UniversityHotels near Brigham and Women's Hospital


text: Here Are the Republicans Who Voted 'No' on Hurricane Sandy Relief FundsRobert Kessler1/04/13 5:25pmFiled to: PolidicksHurricane SandyCongressPoliticsPaul RyanJohn BoehnerGettypic288EditInvite manuallyPromoteDismissUndismissHideShare to KinjaToggle Conversation toolsGo to permalink Friday, Congress finally approved a $9.7 billion package to pay flood insurance claims from Hurricane Sandy. The measure was supposed to come to a vote earlier in the week, but was tabled by House Speaker John Boehner, drawing much criticism from both Democrats and his fellow Republicans alike.The measure passed unanimously through the Senate, but 67 members of the House of Representatives voted "no" to assisting people who were left, at best, powerless or homeless by a hurricane in November. All 67 are Republicans: Justin Amash (R-MI)Andy Barr (R-KY)Dan Benishek (R-MI)Kerry Bentivolio (R-MI)Marsha Blackburn (R-TN)Jim Bridenstine (R-OK)Mo Brooks (R-AL)Paul Broun (R-GA)Steve Chabot (R-OH)Doug Collins (R-GA)Mike Conaway (R-TX)Tom Cotton (R-AR)Steve Daines (R-MT)Ron DeSantis (R-FL)Scott DesJarlais (R-TN)Sean Duffy (R-WI)Jeff Duncan (R-SC)Jimmy Duncan (R-TN)Stephen Fincher (R-TN)John Fleming (R-LA)Bill Flores (R-TX)Virginia Foxx (R-NC)Trent Franks (R-AZ)Louie Gohmert (R-TX)Bob Goodlatte (R-VA)Paul Gosar (R-AZ)Trey Gowdy (R-SC)Tom Graves (R-GA)Sam Graves (R-MO)Andrew Harris (R-MD)George Holding (R-NC)Richard Hudson (R-NC)Tim Huelskamp (R-KS)Randy Hultgren (R-IL)Lynn Jenkins (R-KS)Jim Jordan (R-OH)Doug Lamborn (R-CO)Kenny Marchant (R-TX)Thomas Massie (R-KY)Tom McClintock (R-CA)Mark Meadows (R-NC)Markwayne Mullin (R-OK)Mick Mulvaney (R-SC)Randy Neugebauer (R-TX)Steven Palazzo (R-MS)Steve Pearce (R-NM)Scott Perry (R-PA)Tom Petri (R-WI)Mike Pompeo (R-KS)Tom Price (R-GA)Phil Roe (R-TN)Todd Rokita (R-IN)Keith Rothfus (R-PA)Ed Royce (R-CA)Paul Ryan (R-WI)Matt Salmon (R-AZ)David Schweikert (R-AZ)Jim Sensenbrenner (R-WI)Marlin Stutzman (R-IN)Mac Thornberry (R-TX)Randy Weber (R-TX)Brad Wenstrup (R-OH)Roger Williams (R-TX)Joe Wilson (R-SC)Rob Woodall (R-GA)Kevin Yoder (R-KS)Ted Yoho (R-FL) The most high-profile Congressman on the list is Paul Ryan, failed vice presidential nominee. It also includes Mo Brooks, Ted Yoho, Ron DeSantis, Steven Palazzo and John Fleming from Alabama, Florida, Louisiana and Mississippi, all of which are states that received much-needed federal aid following Hurricane Katrina, the only hurricane in American history that was more costly than Sandy. Speaker Boehner did not vote. This list also includes the entire Republican representation from Arizona, Kansas, Maryland, Montana and New Mexico (Maryland, Montana and New Mexico are only represented by one Republican each). While it took Congress more than two months to approve any federal aid for Sandy victims, it took just 10 days after Hurricane Katrina in 2005. Sponsored This aid package is only, however, the tip of the iceberg, as Congress will take another vote on an additional $51 billion aid package on January 15. In a joint statement, New York and New Jersey governors Andrew Cuomo and Chris Christie called the $9.7 billion a good start: "Today's action by the House was a necessary and critical first step towards delivering aid to the people of New Jersey and New York. While we are pleased with this progress, today was just a down payment and it is now time to go even further and pass the final and more complete, clean disaster aid bill. We are trusting Congress to act accordingly on January 15th and pass the final $51 billion instrumental for long-term rebuilding in order for New Jersey, New York and our people to recover after the severe devastation of Hurricane Sandy." [Image via Getty]Reply288 repliesLeave a reply


text:
BICESTER'S youth council has been forced to close after "losing its way," councillors said at a meeting on Monday.
Due to a lack of interest from young people, the council has been forced to disband after old members left the organisation.
The youth council was not run by the town council, but instead was designed to work alongside it to ensure young people's views got across to town leaders.
However, it was thought that the youth council was becoming more and more focussed on organising events, rather than representing young people in local politics.
Its numbers had also dwindled as those involved moved on in their lives.
Member of Bicester Youth Council and former chairman Jason Slaymaker said he had enjoyed his time at the organisation.
The 22-year-old Bicester resident said: "I don't think it is a case of decline, I think it had been going from strength to strength.
"But the people involved from day one – they have gone through it and moved on to university of colleges in other places.
"I enjoyed my time there and it gave me lots of opportunities to work with other local organisations, like OYAP, Grassroots Bicester and the town council."
But he added: "I think it has come to a natural end. Now it will give young people in Bicester the chance to decide how they want their voices to be heard."
But he said he was not sure how any new incarnation of the youth council would work.
Bicester town councillor Melanie Magee, who has worked with the youth council, said: "I am incredibly disappointed, but it had lost its way.
"The whole group underpinning it had moved on and been replaced by those who only wanted to do events.
"There is only so much we can do, it had lost its sense of purpose."
Elections for new members could be held later this year, she said.
Councillor Jolanta Lis said there had been no choice but to disband the group.
She said: "They have no members left to continue to run the council."
She added the money would be given back to the town council to look after until new elections had taken place.
The youth council was made up of volunteers, aged between 11 and 19, living in Bicester and the surrounding villages.
The idea of the group was to give young people a voice and a chance to make a positive impact in the community.
This year they helped organise the town's Big Lunch event and carnival and met to discuss issues with town councillors.
Councillor James Porter said: "This is a very sad state of affairs. When it was put forward it was the right voice, in the right place at the right time.
"But now teenagers have a plethora of ways to put their views across."
He said that schools had been reluctant to get involved in the venture because they had their own councils, which worked to improve the schools, but not the town.
For more information or to get involved visit bicesteryouthcouncil.co.uk
Do you want alerts delivered straight to your phone via our WhatsApp service? Text NEWS or SPORT or NEWS AND SPORT, depending on which services you want, and your full name to 07767 417704. Save our number into your phone's contacts as Oxford Mail WhatsApp and ensure you have WhatsApp installed.
- Send your Letter to the Editor: What do you think? We welcome letters from our readers on a wide variety of subjects and you can send us a letter for publication here
Our top stories
- Five things you need to know in Oxfordshire today
- Management consultants Newton Europe named Business of the Year at 2016 Oxfordshire Business Awards
- Eating Weetabix and brown rice could help prevent an early death say experts
- VIDEO: Kerry Reeves' mother bravely faces her 'beautiful' daughter's killers in emotional tribute
- Prisons' shake-up will be at heart of today's Queen's Speech
- Five things you need to know in Oxfordshire today


text:
I received the auto-update notification for the latest version of TOP and updated. The update said it was successful and when I went to return to the dashboard it would only load a blank page for my site.
I tried loading my site on multiple machines in multiple browsers after clearing caches, etc. I disabled my caching plugin (W3TC) via FTP and it still wouldn't load. Finally, after I renamed the Tweet Old Post plugin folder was I able to get back in. I really like this plugin and welcome suggestions to get it working again so that it doesn't bring my site down.


text:
Similar items in stock
- Rollback
- Rollback
- Rollback$39.88 Was $49.88Save $10.00
- Rollback$29.99 Was $79.88Save $49.89
- Rollback$19.88 Was $29.99Save $10.11
- Rollback$29.99 Was $49.88Save $19.89
- Rollback$55.00 Was $89.99Save $34.99
- $25.99
- Rollback$9.88 Was $49.88Save $40.00
- Clearance$3.99
- $249.99 List price $270.00Save $20.01
Straight Talk Samsung Galaxy Centura S738C
Consider these popular products
- Rollback$149.99 Was $299.00Save $149.01
- Rollback$19.99 Was $59.99Save $40.00
- Rollback$199.99 Was $247.92Save $47.93
- $11.96 Was $47.83Save $35.87
- Rollback$49.88 Was $79.92Save $30.04
- Rollback$9.99 Was $39.99Save $30.00
- Rollback$129.00 Was $149.00Save $20.00
- Rollback$39.88
- Rollback$14.88 Was $29.92Save $15.04
- Rollback$19.88 Was $29.88Save $10.00
- Rollback
- Rollback$69.82 Was $79.99Save $10.17
- Rollback$299.00 Was $399.00Save $100.00
- $14.99 List price $19.99Save $5.00
- Rollback$69.99 Was $99.99Save $30.00
- Rollback$9.88 Was $49.88Save $40.00
- Rollback
- Rollback$25.99 Was $39.88Save $13.89
- $126.99 List price $149.99Save $23.00
- Rollback$9.74
About this item
About this item
Important Made in USA Origin Disclaimer: For certain items sold by Walmart on Walmart.com, the displayed country of origin information may not be accurate or consistent with manufacturer information. For updated, accurate country of origin data, it is recommended that you rely on product packaging or manufacturer information.
Improve your mobile life on the go with the Straight Talk Samsung S738C Galaxy Centura. It comes pre-loaded with Google Mobile Services, so wherever you go, your entertainment comes along for the ride. Android 4.0, Ice Cream Sandwich lets you customize your screen for faster access to the widgets you use most. And, with the vibrant clarity of the brilliant 3.5" touchscreen, youll never look at your Straight Talk Samsung S738C Galaxy Centura the same way again.
Straight Talk Samsung Galaxy Centura S738C:
- The Straight Talk Samsung Galaxy Centura has Android 4.0 Ice Cream Sandwich
- 3.5" Touch Screen
- APP Capable
- Mobile Web, MP3 Player
- Bluetooth Wireless Technology
- 3.0 Megapixel Camera & Video Recorder
- MMS Picture Messaging
- Multitasking
- Instant Messaging
- GPS Capabilities
- 3G/WI-FI Connectivity
- MP3 Player (cable not included)
- FM Radio
- Supports microSD Card up to 32GB (not included)
- Phonebook entries up to storage limit
- Hearing Aid Compatible (HAC) Rating: M3/T3
- Battery Talk Time up to 7.22 hours
- Standby Time up to 10.33 days
- Straight Talk Samsung Galaxy Centura S738C comes with Battery, Charger, Activation Card, Service Guide and User Manual
- Note: The Straight Talk Samsung Galaxy Phone will ONLY work with the Straight Talk $45/30-Day Unlimited Service Plan or the $60/30-Day Unlimited International Service Plan.
|Cell Phone Service Provider:
|Has Bluetooth:
|Manufacturer Part Number:
|Contained Battery Type:
|Lithium Ion
|Straight Talk
|Assembled Product Dimensions (L x W x H):
|6.00 x 1.75 x 9.20 IN
Customer Reviews | 315 reviews | 4.1 out of 5
Customer Reviews
Customer review by JohnnyOfFlorida
I bought this phone as a Black Friday deal with the intent on replacing my LG slider phone. The LG was running on an older Gingerbread version of Android and had FREQUENT memory problems due to its low amount of internal memory. I did like the QWERTY slider keyboard on the LG but thought I'd be able to adapt to only using the display keyboard on the new Samsung given some time. Boy was I wrong - the on-display keyboard is easy to use and I had no problem adjusting to it. Even better is the option to use Google voice-recognition to compose text messages and WOW is it accurate!! So much quicker than typing! The Samsung Galaxy Centura uses Android Jellybean and is so much faster than my old phone. It also has more internal memory (sorry, can't recall off the top of my head how much) but I've installed a few apps and have dozens of text message on the phone and still haven't encountered the dreaded low memory message! The display is clear and large, I give this phone 5 *'s!!!!
Customer review by PaulyEddyFreddy
About 6 months ago, my wife's Straight Talk (ST) LG Optimus Logic went bust on a trip, so I got her the ST Centura at a Walmart. At the time I was using the ST Samsung Proclaim. Her Centura worked very well, especially since it had Android 4.0 instead of 2.1. When we got home, the Thanksgiving Black Friday ad came out and when I noticed that the phone was only $29 instead of the $99 that I paid for my wife's phone, I could not stop myself from buying one for me :-). Because her LG phone went bust on a trip, I realized that we needed to carry a spare phone with us---That now is my ST Proclaim. The phone works very well. It does have one problem I had read about and noticed myself. Sometimes the clock shows a different time—right out of the blue. I have not looked that hard for a fix yet, but just reboot the phone and its OK. The Android 4.0 means that you can download more apps than with a 2.1 phone. I haven't tried all the features, but it seems to have more features than my Proclaim. Really nice ST phone.
Read all 315 reviews
Customer Q&A
Ask a question
If you would like to share feedback with us about pricing, delivery or other customer service issues, please contact customer service directly.
Preview Question
Thank you.
Questions will be checked against our question guidelines and posted within five to seven business days.
- I owned the $99 phone and shattered the screen bought the $29 phone and I noticed no differenceby An anonymous customer12/16/2013Was this answer helpful? (0) (0)Was this answer helpful?
- Nothing other than price. Black Friday and Cyber Monday had specials on these phones.by debbiek31112/15/2013Was this answer helpful? (0) (0)Was this answer helpful?
- Nothing this was just a special price for black Friday and cyber weekby kkk201212/15/2013Was this answer helpful? (0) (0)Was this answer helpful?
- The sale price.by putnut12/15/2013Was this answer helpful? (0) (0)Was this answer helpful?
- The page showing $29 is a sale price. If out of stock, check back frequently. That's what I did.by towercaller12/15/2013Was this answer helpful? (0) (0)Was this answer helpful?
Q. Is this Samsung Galaxy Centura S738C listed for $29.00 the same as one listed for $99.99?
- Hi Kel. It is confusing as they have listed both. Many in my family have this model of phone. a few at the $99 price and few at the $29 price from this week. This phone was sold in the stores Thanksgiving night for a 2 hour deal for $29. With Cyber Monday (looks like week now) it is $29. We have seen no difference whatsoever to make us believe it is a different phone. As far as the networks. There is no Straight talk network. All of these phones are made by the same company that makes the Trac fone. Straight Talk can offer the cheap service because it does not own any towers. Different phones operate on different networks. An example on this one on the front of the box in the lower left corner I believe it says CDMA-V. CDMA is the type of network and V is for Verizon towers it uses. It does not use a SIM card. I use an LG-L40G. Mine said GSM-A. GSM is the network and it uses AT&T towers. GSM phones do use a SIM card. Best of luck. Verizon does not suit my personal needs anymore. If it does yours this is an AWESOME deal. Do not let this get away. This phone is worth way more than $29by Purpledemon12/10/2013Was this answer helpful? (6) (0)Was this answer helpful?
- I BOUGHT 3 FROM THE STORE AND THEY WERE ALL SMART PHONES AND HAD SOME GEAT APPS PRE INSTALLED AND I HAVE DOWN LOADED SEVERAL MORE SO I ORDERED ANOTHER ONE ON LINE WHILE I COULDby Randy12/10/2013Was this answer helpful? (2) (1)Was this answer helpful?
- I called Corperate and they told me that theyre not really sure what the difference is but they did tell me that it might be that one is a smartphone and one is not (which doesnt really help considering that they seem alike.) I guess the best way to find out is to wait until it comes in and see whats in the box. There is a shred of light however they told me that if its not what Im looking for then all I would need to do is just take it to my local walmart store and they would refund me the money as long as I had proof of purchase...so I guess its just a waiting game. Good Luck to all who purchase this phone!by Alsome199212/3/2013Was this answer helpful? (2) (0)Was this answer helpful?
- Kel, you've done your research! I've been wondering about this for a few days now wondering if there is some type of misprint or if they will take this page down because it is a mistake. I hadn't done as much nitpicking as you have so I kept thinking that they are the exact same phone but now that you've brought to light that there are some subtle differences, I'm not so sure. I answered a few inquiries below that this is the exact same phone being sold for $99.88 as the general model numbers are actually the same and they seem to have the exact same features. I'm now leaning toward this page being a mistake. One of the reasons I am thinking it's a mistake because Walmart has the phone advertised in their Sunday ad in the area I'm in as "IN STORE ONLY ...Valid Dec 1 - Dec 7, $99.88" This leads me to ask "why would they be selling the exact same phone online for $29 then?" Let's see if anybody in authority answers ...lolby richardarr12/10/2013Was this answer helpful? (6) (1)Was this answer helpful?
- Noby Edenblessing11/30/2013Was this answer helpful? (0) (0)Was this answer helpful?
- noby WYCase11/29/2013Was this answer helpful? (0) (0)Was this answer helpful?
- No this phone is Not a slider. It is a touchscreen. Hope this helps you.by 2013Mustang11/29/2013Was this answer helpful? (0) (0)Was this answer helpful?
- No, it is not.by mariansc11/29/2013Was this answer helpful? (0) (0)Was this answer helpful?


text:
Hotels in Gallipolis
Search Gallipolis Hotels
Trending now in Gallipolis
Hotels in Gallipolis
- Most bookedHampton Inn Gallipolis
22 people looked at this hotel in the last hour
Most recent review
"Front desk staff gets an A+ and the breakfast was excellent. The condition of the hotel does not measure up. I have never stayed in a hotel before with squeaky floors overhead. It was unreal! Thankfully, the folks on the second floor retired about the same time I did. The bathroom was lovely exce..."
A Traveller, Dec 2015, US
18 people looked at this hotel in the last hour
Most recent review
"Room was very comfortable and clean. Staff was very friendly. The breakfast was good. "
A Traveller, May 2016, US
- Knights Inn GallipolisNo Guest Reviews yet
7 people looked at this hotel in the last hour
- Super 8 Gallipolis Point PleasantNo Guest Reviews yet
7 people looked at this hotel in the last hour
Hotels within 25 miles of Gallipolis
20 people looked at this hotel in the last hour
Most recent review
"For medical reasons, Asked for ground floor. Close to door. Put me in a Smoking Room which smelled like it. My clothes stunk after that!!"
A Traveller, Jun 2016, US
- Jackson-Days InnNo Guest Reviews yet
3 people looked at this hotel in the last hour
Best Gallipolis Hotels
Our 5 most booked Gallipolis hotels from the last hour are:
1. Hampton Inn Gallipolis
2. Quality Inn
3. Knights Inn Gallipolis
4. Super 8 Gallipolis Point Pleasant
5. Red Roof Inn Jackson, OH
But, low prices and a great selection are not the only thing Hotels.com offers. We also have 192 genuine Hotels.com guest reviews for Gallipolis.
Why use Hotels.com to book your Gallipolis accommodation?
We can help you to find the perfect Gallipolis hotel room for almost any occasion. With our fast and secure hotel search you may compare different Gallipolis hotels by:
- - Price
- - Availability
- - Gallipolis neighbourhoods
- - Nearby Gallipolis attractions and landmarks
- - 192 genuine Hotels.com guest reviews for Gallipolis
- - Star rating
- - Amenities
- - Hotel name & chain
- - Theme
Alternatively, you can use our detailed Gallipolis maps showing hotel locations in Gallipolis, using all the filters listed above.
Cheap Gallipolis hotels - getting the best deal
On Hotels.com you can always be sure you're getting the best deal.
We make it easy to save money on every booking. We offer our Best Price Guarantee on most bookings. Plus we offer 3 more ways to save:
1. Hotels.com Rewards - Collect 10 nights, get 1 free*
2. Hotels.com Deals - Search over 10,000 deals!
3. Hotels.com voucher codes - All our official promo and discount codes in one place.


text:
In 1975, Children's National Health System opened one of the first intensive care units dedicated for children in the United States. While advances in technology have changed the look of our Pediatric Intensive Care Unit (PICU), an enduring hallmark of our care is an emphasis on comfort for patients and compassion for the entire family.
Children's National serves as the primary referral center in the Washington, DC, region for pediatric emergency, trauma, cardiac, and critical care. Annually, the PICU treats more than 1,400 patients with a wide variety of conditions from across the United States and abroad, including:
The PICU is a state-of-the-art facility designed with the family in mind. In addition to creating a child-friendly environment, there are special waiting and resting areas for families so you can stay close by while your child is with us.
Our team offers the most current therapies for cardiac, pulmonary, and neurologic injury. Children's was one of the first pediatric hospitals to use the CereTom Portable CT Scanner, which delivers high-resolution images that help diagnose head and neck injuries. We can even use the scanner at the patient's bedside without having to move the child. This limits the risk to the child because we do not have to move him/her for diagnostic testing.
The ICU's Baier Media Room has a large-format screen that permits seamless, simultaneous display of real time and stored images from multiple sources - improving care and increasing our telemedicine capabilities.
The PICU's multidisciplinary team works side-by-side with families and providers. Our comprehensive approach to care has helped Children's achieve patient outcomes among the best in the nation. The team includes:
Your child's care team is also your care team. We recognize that when a child is ill, it affects the entire family, and we do everything possible to meet your needs as well.
Among the support resources for parents and families at Children's National, we created Our Stories as a way for parents to learn from others' past experiences and arm themselves with the knowledge to make it through this challenging process. It can be extremely difficult to face a health issue with your child, but parents who have been there can provide compassion, guide, and comfort parents in need.
All three of our Intensive Care Units (ICUs) received the Beacon Award from the American Association of Critical-Care Nurses (AACN). The PICU and CICU are designated gold-level and honored with a 3-year designation. The NICU is designated silver-level and has a 2-year designation. The Beacon Award signifies exceptional care through improved outcomes and greater overall satisfaction. For our critical care nurses and clinical teams, the Beacon Award means a positive and supportive work environment with greater collaboration between colleagues and leaders, higher morale and lower turnover. Congratulations to all of our caregivers and leadership teams across our Intensive Care Units for working together to meet and exceed the high standards set forth by the Beacon Award for Excellence.
How ECMO helped Emilee not only survive, but thrive.
Read More of Emilee's Story
Did you know we have 4 locations in Northern Virginia?
Find a Children's Location Near You
Learn more about the causes, symptoms and treatments for lysosomal storage disease, a genetic condition.
Genetics expert Marshall Summar, MD, interviewed Jess Thoene, one of the founders and pioneers in the field of inborn errors in metabolism in rare diseases.
Dr. Marshall Summar interviews Hemoshear CEO James C. Powers about technological advances in the rare disease field.


text:
Sign up today and activate a
Moving day is full of excitement and also a lot of hard work. The weeks prior to the big day can be hectic, but the process will be easier if you follow a checklist to make sure every task gets completed in a timely manner. As you get ready to settle into a new home, check off your list to keep yourself sane and on track.
Research moving companies. Get estimates and decide if you want the movers to pack your belongings or just transport them. Find out what they will and won't move and what their policies are for reimbursing you for damage. After you've decided on a company, get an estimate in writing.
Sort your belongings. Clean out your house, garage, and other storage areas and get rid of things you don't want to take with you. Donate stuff in good condition to charity or list it on an online site for classified ads. It's a good idea to begin this process even before you have an exact moving date, since you'll be very busy after you set the date.
Make a list of your valuables. Be sure to include an estimated replacement cost for each item and have estimates done for anything you're unsure of. Note serial numbers and other identification marks on expensive items. You need this list for reference if anything gets lost or damaged in the move.
If you're a renter, inform your landlord of your move-out date. Many landlords require at least one month's notice, so check in with yours as soon as you have a date. This is also the time to reread the lease and confirm the requirements for getting a refund on your security deposit.
Confirm your moving date with the moving company. Set up the times the packers will arrive and get the estimated time the truck will leave the old house and arrive at your new home.
Order packing supplies if necessary. If you plan to do your own packing, you'll need a lot of supplies, such as boxes, packing paper, bubble wrap, and tape.
Gather important family documents. If you're moving a long distance, you may need birth certificates and other records readily available for school registration and to get new driver's licenses. You'll also want medical and dental records, tax documents, financial records, and veterinarian records. Pack these documents yourself in a waterproof box.
Research your kids' new schools. Arrange to transfer school records, and find out what documentation your new schools require, such as birth certificates and immunization records.
Make a change-of-address list. In addition to having the post office forward mail to your new address, you'll need to make some contacts directly by email, by phone, and in writing.
Schedule changes for your utilities. You'll need to set disconnection dates for your old home and connection dates for your new home. There will most likely be some overlap because you'll want to have everything turned on as soon as you get to your new place.
Use up perishable items. Sort the food in your freezer, refrigerator, and pantry. Discard items that are out of date. Use the rest for family meals, buying as little extra as possible until you've moved.
Cancel services you receive at your old home. These include newspaper subscriptions, lawn care, snow removal, and housecleaning services.
Have cars checked by your mechanic. If your move involves a long road trip, your car needs to be road-ready. Have the tires checked and have a tune-up if it's been a while since the car was last serviced.
Clean and defrost the refrigerator. If you plan to take the refrigerator to your new home, check the owner's manual for instructions on prepping it for the move.
Prep any appliances you're taking with you. The owner's manuals and manufacturers' websites have information on preparing large appliances for transport.
Get your payments ready. Even if you can pay for most expenses with a credit card, you may need cash for some transactions, like tipping the furniture movers.
Refill prescriptions. Make a list of your family's medications and make sure you have enough for a few weeks, so you can avoid a frantic hunt for a new pharmacy. This is also a good time to refresh and update your family's first-aid kit.
Get rid of flammable materials. Drain lawnmowers and other small machines of gasoline. Find a community resource for discarding the flammables, including paint and oil.
Pack things you'll take with you in the car or on the plane. Prepare as you would for a vacation, packing a suitcase for each family member with clothes, toiletries, medications, and snacks. Gather any documents and valuables, such as jewelry, that you want to keep with you, too.
Pack and label an "open first" box. This can be a box you keep with you or one you load on the moving truck. It contains the items you'll need to feel comfortable right away in your new home. Some people pack cleaning supplies so they can clean the kitchen and bathrooms immediately. Others pack one set of bedding for each bed.
Have instructions ready for the moving staff. If the company will pack your belongings, tell the packers how you want the boxes labeled; many people like to have the destination room marked on each box. Double check the new address and the estimated arrival time with the van driver.
Make a final inspection. Walk through the house, garage, and yard after the movers have finished, making sure nothing is getting left behind and no tasks are being left undone.
Lock up the house. Close the blinds and drapes, and unplug any appliances you're leaving in the house.
Shop Overstock™ and find the best online deals on everything for your home and your family. We work every day to bring you discounts on new products across our entire store. Whether you're looking for memorable gifts or everyday essentials, you can buy them here for less. Not just anyone's mobile outlet, your mobile outlet.O.com ® Ethical Pricepoints ™


text:
BAML bumps up rates with Reynolds
Craig Reynolds will join Bank of America Merrill Lynch (BAML) as managing director and head of North America rates trading in August. Reynolds will be based in New York and report to David Gu, head of global rates and currencies (GRC) at the US bank. The appointment comes along with internal moves that have seen Peter Antico become head of global G10 currency trading, while Chris Hodson will relocate to London and become head of rates trading for Europe, the Middle East and Africa. Reynolds joins...
More on Structured Products
7 days in 60 seconds
Smart contracts, Brexit and sovereign risk
The week on Risk.net, July 22–28, 2016Receive this by email
Back to Top


text: Humax USB Wi-Fi dongle allows you to connect your Humax HDR-2000Tor HDR-1000S to your home network wirelessly. Use the dongle to access catch-up TV and the internet via your Smart TV. Speeds of up to 150 Mbps and fully secure using WEP, WPA and WPA2 encryption. Compatible with 802.11n/g/b standards.


text: Celebrity News July 23, 2012 'Extra' Raw! 'Call Me Maybe' Teen Choice Awards Mash-Up "Extra's' Terri Seymour got a few of the 2012 Teen Choice Award stars to sing a mash-up of the hit Carly Rae Jepsen song, "Call Me Maybe." Check it out! Comments Around the Web More In Celebrity News Getty Images Celebrity News Lindsay Lohan Fights with Fiancé, Tweets, 'I Am Pregnant' Instagram @frenchmontana Celebrity News Kim Kardashian & Calvin Harris Light Up J.Lo's Pre-Birthday Party Instagram @kimkardashian Celebrity News Kim Kardashian Pours Gasoline on Taylor Swift Feud, Raps Along to Kanye's… Instagram @taylorswift Celebrity News Taylor Swift Returns to Instagram, Hangs with Tom Hiddleston in L.A. AMC Celebrity News New 'Walking Dead' Season 7 Trailer, Revelations, and More from Comic-Con Celebrity News Family Sources Say Lamar Odom Intervention Was Unsuccessful Getty Celebrity News Rumor Bust! Khloé Kardashian Is NOT Pregnant with Lamar Odom's Baby Load More


text:
What are the best brands for salsa dance shoes?
I saw this symbols beside some dance tracks. Any idea what they stand for
Is Swing and Jive the same type of music ? Can I use the music for one for the other ?
What brands do you go for standard and latin shoes ?
Hi I just started learning dancing. I was just wondering is there any point for me to go social dancing because I find that when I dance at...
I am looking for an mp3 player that can slow down the temp of the music without changing the pitch with an external microphone. Budget is about...
Do your dance studio usually allows you to videotape their lessons ?
Does anyone have Corky Ballas instructional DVDs ? I have the whole set to gold and I don't understand why I can't find the lock step for Cha Cha...
I am a male and I a query. After how many lessons then is it appropriate for me to ask my female dance partner from my dance class that she...
Are there any timing CDs for ballroom/latin music ? I know there is one for salsa
Is there a recommended sequence of learning the ballroom and latin dances ?
Does anyone have any good ballroom dance videos on tehcnique and styling to suggest ?
I am learning the IDTA syllabus and in Cha Cha there is something called the lock steps. I tried looking it up in Corky Ballas instructional DVDS...
What are the best DVDs for learning the 3 major types of salsa
I am ordering some dance DVDs from dance vision in US that costs about 750 US dollars in total and am deciding whether to get it shipped either to...
I would like to find out what are the major dance certifications around world wide. I am in UK and the main one I am familiar with is IDTA....
Is it possible to learn both the Bronze and Silver Medals at the same time for both latin and ballroom dancing ?
What's the best way to learn to hear the beat in music
I have been searching for good DVDs for learning and these are the few that I come up with
http://www.dancesport.uk.com/video/fb-ddc1698.htm -...
Separate names with a comma.


text:
Win Ueli Steck with the BMC! Last Chance!by Jack Geldard - UKC Chief Editor Dec/2012
This news story has been read 4,315 times
Go to the page, hit 'Like' - then email firstname.lastname@example.com with the answer to this question:
Q.�What is the total weight (in pounds) of Mountain Hardwear's Ueli Steck system?
Find it out here: Mountain Hardware
Email your answer here: email@example.com
There's loads of other free gifts - have a look at the BMC's Website for full details about this competition.
Our Friday Night Video this week is a beautifully filmed piece by actor, singer-songwriter and director Jared Leto. With stunning... Read more
Shawn Raboutou, 18, has repeated Monkey wedding and Spray of light, both ~8C, in Rocklands. Rumours have been... Read more
Adam Ondra is now back in Flatanger where he intends to work some of his remaining projects. On his way there he made a couple of... Read more
Today the British Mountaineering Council has announced a change of name to 'Climb Britain', while its... Read more
This week's Friday Night Video gives us a glimpse into some of the adventures and fun that the climbers at the Women's... Read more
This weekend the 2016 British Bouldering Championships will be held at Devonshire Green in Sheffield. For those of you who... Read more


text:
Translation of "respectful" - English-Portuguese dictionary
(Translation of "respectful" from the Cambridge English–Portuguese Dictionary © Cambridge University Press)
Get a quick, free translation!
Create and share your own word lists and quizzes for free!
Word of the Day
a type of heavy cloth that has a pattern woven into it that is the same colour as the cloth