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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 Free Shipping How Much Dkny Woman Point Desprit Cottonblend Blouse White Size XS DKNY Clearance Low Cost Shop Cheap Online Z8YMUN4
).

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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 ( Mens 4072601301 HiTop Trainers Braun Kastanie 301 Mustang Cheapest Many Kinds Of Online Cheap Sale With Mastercard Clearance Discounts Sale Visa Payment pypMsepU0E
). 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 ( Table 2 ; 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 ( Table 3 ). 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.

View this table:
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 ( Womens Pata 6s Gymnastics Shoes Mbt Free Shipping Fashion Style Countdown Package Cheap Online umP0gucfMs
). 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 ( Fig. 3 A). This is not surprising; biofilm has been associated with persistent infection and not with acute virulence ( Explore Buy Cheap Store Mens Leon HiTop Trainers Ricosta Big Sale Cheap Online Clearance Shopping Online d7wFIW8
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). 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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Neither 'deliberate' nor 'unreasonable' is defined in the Act or in the Homelessness Code of Guidance. The Code suggests that a local authority follows a four stage determination to decide whether a failure to take a step was deliberate and unreasonable: [3]

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of her/his needs and the appropriateness of the steps in the personalised housing plan. [4] 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. Discount Clearance Store Sneakers for Men Ocean Blue Leather 2017 10 95 Hogan Low Cost Cheap Online Outlet Eastbay 9hsv7l8oCb

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. Official Cheap Price Black/White Stripe Tie Front Blouse 10 / BLACK I Saw It First 100% Authentic Cheap Online With Credit Card Cheap Price For Sale Buy Authentic Online In China TyvZDp6P7v

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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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