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Figure 2.

Longitudinal series of isolates. Each series (collected from patients P01 to P16) is depicted as a row of dots to represent sampling time points. Colors indicate different RAPD genotypes, and relative dot size indicates patient lung function (%FEV) at that time point. Gray circles indicate no associated %FEV (percentage of predicted forced expiratory volume in 1 sec) measurement (also see Supplemental Fig. S1 ).

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Table 1.

Summary of 16 longitudinal series

The majority of patients were colonized by subgroup A isolates (202 of 215 isolates) primarily from one of the major B. cenocepacia epidemic lineages. Except for the single patient P09 infected with subgroup B, all isolates from the same patient belonged to the same RAPD type. By using these 215 isolates, we conducted a comprehensive survey of clinically relevant phenotypes and carried out whole-genome sequence analysis of all isolates.

To understand how bacterial phenotypes change over time during chronic infections, we systematically cataloged phenotypic variation for all isolates. Specifically, we measured: growth in two liquid media types (lysogeny broth [LB]; and synthetic cystic fibrosis medium [SCFM]), swimming motility, biofilm formation, acute virulence in an insect model, and mucoidy morphotype ( Stuart Weitzman Woman Chicflat Suede Ballet Flats Mushroom Size 95 Stuart Weitzman Free Shipping Visit Sale Pay With Paypal Buy Cheap Outlet Store Buy Cheap Manchester Sale For Nice zeIWKM
). The results reveal extensive phenotypic variation among B. cenocepacia isolates both within and among RAPD types and within patient series. Detailed descriptions and results for each phenotype are in Supplemental Text S1 , Supplemental Table S1 , and Supplemental Figures S2 and S3 .

By considering these phenotypes on a global level, we found that RAPD genotype was a significant predictor of phenotypic variation, explaining between ∼20% and 50% of the variation ( Cheap Sale Visit Charlotte Olympia Woman Leathertrimmed Lurex Sandals Silver Size 35 Charlotte Olympia Sale Excellent View Enjoy Shopping 07leE
; Supplemental Fig. S4 ). However, considerable variation was also seen within patient series and within RAPD type. We also identified progressive phenotypic changes in many longitudinal series ( Crus Woven Leather Sandals Yuketen 2018 New Cheap Sale Get Authentic Really Cheap Shoes Online I1sEcNR
). Six of the 16 series showed progressive decreases in motility, five showed temporal changes in biofilm formation (two increasing and three decreasing), two showed decreases in acute virulence as measured in an insect model, none showed changes in mucoidy, and, finally, seven showed changes in one or more growth parameters.

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Table 2.

Correlation between RAPD type and clinically relevant phenotypes

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Table 3.

Percentage change per year in clinically relevant phenotypes across longitudinal series

Given that each individual phenotype could have a complex genetic basis, we examined pairwise phenotypic correlations across all isolates and for each RAPD type individually ( Mens Stamped Leather Venetian Drivers Harris Explore Cheap Price Sale Cheapest Outlet Locations Cheapest For Sale jI0vinVNs
). In general, most phenotypes we measured were positively correlated; with the exception that biofilm formation was negatively correlated with acute insect virulence and growth in LB ( Low Rise Skin Skinny Jean Blue blue Cheap Monday Explore For Sale Get VkseS
A). This is not surprising; biofilm has been associated with persistent infection and not with acute virulence ( Bowembellished satin sandals Loewe Clearance Recommend Buy Cheap Factory Outlet hRYHVO4Q
; Wu et al. 2015 ). Overall, motility was positively correlated with biofilm formation and mucoidy (Spearman correlation, ρ = 0.48 and 0.40, respectively, adjusted P -value <0.0001) ( Fig. 3 A). A strong positive correlation was found between motility and biofilm formation for most RAPD genotypes, except for a negative correlation was seen for subgroup B RAPD44 isolates (ρ = −0.85, adjusted P -value = 0.001) ( Fig. 3 B–G).

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Where a local authority feels that an applicant is refusing to cooperate and that the refusal is 'deliberate and unreasonable', it can follow a specific procedure for notifying the applicant.

The information on this page applies only to homelessness applications made on or after 3 April 2018 .

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In order for the applicant to be penalised, the refusal must have been deliberate and unreasonable.

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It may be found that certain steps in the PHP are not reasonable, or have become unreasonable. For example, even if requiring an applicant to search for accommodation in a particular area at first seemed reasonable, it may be s/he has tried all of the private landlords in an area and none of them are willing to offer accommodation. In these circumstances, it is unlikely to be reasonable to require her/him to keep looking. If steps are not reasonable, or are no longer reasonable, then the applicant should be notified and should not face consequences for failing to take them. Clearance Factory Outlet Free Shipping Many Kinds Of Mens Soft 8 LowTop Sneakers Ecco JN7uJGTNRh

It may emerge during the authority's investigations (see below) that the applicant did not understand one of the steps in the plan (perhaps because of a language difficulty) and if so, s/he should not be found to have deliberately and unreasonably refused to cooperate. Buy Online Authentic Sale Collections Beya loafer White Nicholas Kirkwood Popular Buy Cheap Explore FVfPS

The Code requires that local authorities take into account any particular difficulties the applicant has in managing communication. It suggests, for example, that cooperation may be particularly difficult where an applicant is street homeless or moving between temporary places to stay. [7] The problems may be around, for example, an applicant having a learning disability or more practical issues such as being required to contact landlords and not having telephone access. In circumstances such as these, it is less likely that failing to cooperate would be counted as deliberate or unreasonable.

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