Composition for personal growth : program design and evaluation.

<p>Two categories were used for cross-validation of the model, either type 1 or type 2. Clinical isolates were treated as an unknown class and cross-validated sensitivity, specificity, and class error were based on their classification prediction score with their respective reference strain control class. CV, cross-validated.</p><p>Cross-validated PLS-DA modeling statistics for the prediction performance for NA-SERS typing of individual type 1 and 2 <i>M</i>. <i>pneumoniae</i> clinical isolates.</p

Versatile Methodology for Glycosurfaces: Direct Ligation of Nonderivatized Reducing Saccharides to Poly(pentafluorophenyl acrylate) Grafted Surfaces via Hydrazide Conjugation

In this work, we report a convenient and versatile strategy for surface-grafted glycopolymer constructs with the goal of surface modification that controls the chemical presentation and grafting density of carbohydrate side chains. This approach employs a difunctional hydrazine linker, chemically modified to an active ester containing poly­(pentafluorophenyl acrylate) grafted scaffold, to conjugate a variety of saccharides through the reducing end. The successive conjugation steps are carried out under mild conditions and yield high surface densities of sugars, as high as 4.8 nmol·cm^–2, capable of multivalency, with an intact structure and retained bioactivity. We also demonstrate that this glycosylated surface can bind specific lectins according to the structure of its pendant carbohydrate. To demonstrate bioactivity, this surface platform is used to study the binding events of a human respiratory tract pathogen, Mycoplasma pneumoniae, on surfaces conjugated with sialylated sugars

PCR analysis of serial dilutions of <i>M. pneumoniae</i> strain II-3.

<p>Dilutions 10⁻¹⁰ to 10⁰ are indicated; std, DNA size standard (350 kbp); +, positive control (277 kbp). Starting concentration, 1.8×10⁹ CFU/ml.</p

Differentiation of <i>M. pneumoniae</i> strains.

<p>(<b>A</b>) Average spectra (n = 15) of <i>M. pneumoniae</i> strains with formalin background, baseline corrected and offset; and (<b>B</b>) first derivative spectra from panel A demonstrating strong similarities but also clear differences (boxes) among the strains.</p

PLS-DA of throat swabs spiked with serial dilutions of <i>M. pneumoniae</i>.

a<p>abbreviations same as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0013633#pone-0013633-t001" target="_blank">Table 1</a>.</p

PLS-DA of true clinical throat swab samples from different individuals.

<p>Gray symbols, samples previously shown to be <i>M. pneumoniae-</i>negative (Mp-negative) by culture and real-time PCR from five persons; open symbols, samples previously shown to be <i>M. pneumoniae-</i>positive (Mp-positive) by culture and real-time PCR from five persons. Five or six spectra were collected for each sample, in duplicate.</p

PLS-DA of NA-SERS specificity and sensitivity in discriminating three <i>M. pneumoniae</i> strains and two negative controls (formalin and water).

a<p>abbreviations the same as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0013633#pone-0013633-t001" target="_blank">Table 1</a>; single model generated using 12 latent variables accounting for 88% of the total variance for all serial dilutions of each strain.</p

Principal Component and Hierarchal Cluster Analyses.

<p>Chemometric analysis was conducted on the spectral data from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0013633#pone-0013633-g002" target="_blank">Figure 2</a>. (<b>A</b>) PC scores plot 1vs. 3 of <i>M. pneumoniae</i> strains FH, M129, and II-3, as indicated. 77% of the variance was captured in PC1 and 3% in PC3 to distinguish the strains. (<b>B</b>) HCA of pre-processed spectra of <i>M. pneumoniae</i> strains FH (dashed lines), M129 (solid lines), and II-3 (bold lines). Four spectra from strain II-3 were misclassified with FH (at left).</p

Reproducibility of spectra.

<p>Raw spectra (<b>A</b>) of <i>M. pneumoniae</i> strain FH collected from five random spots from three separate NA substrate wells, baseline corrected and offset. (<b>B</b>) First derivative spectra for strain FH from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0013633#pone-0013633-g001" target="_blank">Figure 1A</a>.</p

Differentiation of spiked and control throat swab samples.

<p>Representative difference spectra of spiked, pooled throat swab samples after subtraction of the spectrum of the un-spiked, pooled throat swab control. Bold and thin solid lines, spiked samples (8.2×10⁴ and 8.2×10³ CFU <i>M. pneumoniae</i> M129/µl, respectively); dashed line, representative spectrum of <i>M. pneumoniae</i> M129.</p