Demographic characteristics.

<p>UPDRS = Unified Parkinson's Disease Rating Scale; LEU = L-dopa equivalent units; DA = dopamine agonists. All values are mean ± SD. Significant differences are labelled with “*”. P-values refer to columns indicated in brackets. Controls (column 1), PD+ICB (column 2), PD−ICB (column 3).</p

Behavioral results.

<p>All error bars are 1 s.e.m. A. Emotion bias for each group. B. Evidence vs. choice curve showing increased selection of happy faces when evidence supports angry face. C. Percent correct for each group.</p

Learning from positive vs. negative feedback.

<p>Error bars show 1 s.e.m. A. Parameter values from the Bayesian model in PD+ICBs. B. Parameter values in PDs.</p

Cytokine production in response to polyclonal stimuli.

<p>PBMCs were extracted from subjects with an MenC SBA titer <8 one year post vaccination and matched controls and stimulated for 96 hours with plate bound αCD3±αCD28 and/or the TI-II antigen mimic, α-δ-dex. Supernatants were collected and cytokine concentrations assessed by cytometric bead array. n = 10. One-way ANOVA+ Bonferroni selected pairs post-test (SBA low vs. SBA high).</p

T-cell CD3 and CD28 expression and B-cell memory pool distribution.

<p>PBMCs were extracted from subjects with a MenC SBA titer <8 and matched controls one year post vaccination and levels of T-cell CD3 and CD28 expression B-cell memory subsets were examined by flowcytometry. n = 11, One-way ANOVA+ Bonferroni selected pairs post-test (SBA low vs. SBA high).</p

Additional file 1: of Distinct clinical and neuropathological features of G51D SNCA mutation cases compared with SNCA duplication and H50Q mutation

A, Double immunofluorescence images of ubiquitin (green) with α-synuclein (red) in a representative G51D case (CA3), the duplication case (SN) and the H50Q case (EC). B, Representative immunofluorescence image of α-synuclein (red) and p62 (green) in G51D (CA1), the duplication case (substantia nigra) and the H50Q case (Temporal cortex). Scale bar represents 50 μm. (TIF 5,615 kb

STm COPS size characterization and the effect of pH on rhamnose and abequose O-acetylation in 1925wzzB-COPS.

<p>(A) SDS-PAGE and Pro-Q staining for LPS from CVD 1925 (lane 1) and CVD 1925 (pSEC10-<i>wzzB</i>) (lane 2). (B) Chromatogram of purified D65 COPS (dashed line) and 1925wzzB-COPS (solid line) assessed by HPLC-SEC with detection by refractive index. (C) ¹H NMR analysis of 1925wzzB-COPS after exposure to different pH levels. Peaks for rhamnose C2/C3 and abequose C2 O-acetyls are indicated. (D) Hestrin analysis of residual O-acetylation of 1925wzzB-COPS after incubation at different pH levels. (E) ELISA reactivity of 1925wzzB-COPS and dOAc-1925wzzB-COPS with monoclonal antibodies against O4 (α-STm O4) and O5 (α-STm-O5) or polyclonal sera recognizing O1,12 and core epitopes (α-SE). Error bars represent s.d. and were derived from technical replicates from one experiment.</p

Immunogenicity, protective efficacy and functional analyses of vaccine-induced antibody for mice immunized with conjugates of native or dOAc-1925wzzB-COPS with CRM₁₉₇.

<p>(A) Serum IgG titers for 1925wzzB-COPS (black), dOAc-1925wzzB-COPS (grey) or SE COPS (open) from mice (<i>n</i> = 40/group) immunized with PBS, STm-COPS^KDO:CRM₁₉₇ or dOAc-STm-COPS^KDO:CRM₁₉₇ as indicated. Each point represents an individual mouse. Red squares indicate mice that succumbed after challenge. Solid bars indicate the GMT; comparisons between groups were accomplished by either a two-tailed Mann-Whitney U test (PBS vs. conjugate, conjugate 1 vs. conjugate 2) or two-tailed Wilcoxon signed-rank test (paired serological analyses). (B) Kaplan-Meier survival curves of mice immunized with STm-COPS^KDO:CRM₁₉₇ (circle), dOAc-STm-COPS^KDO:CRM₁₉₇ (square) or PBS (diamond) after challenge (<i>n</i> = 20/group) with 1x10⁶ CFU (open symbol, dashed line) or 5x10⁶ CFU (closed symbol, solid line) of STm D65. Adjustments for multiple comparisons were not made. *<i>P</i> ≤ 0.0001 for indicated comparisons or for vaccinated mice relative to the respective PBS challenge group. ^#<i>P</i> ≤ 0.05, ^##<i>P</i> ≤ 0.005 for STm-COPS^KDO:CRM₁₉₇ vaccinated mice relative to the respective challenge dose group immunized with dOAc-STm-COPS^KDO:CRM₁₉₇. (C) Representative serum anti-COPS epitope profiles (identified by symbol) from mice immunized with either STm-COPS^KDO:CRM₁₉₇ or dOAc-STm-COPS^KDO:CRM₁₉₇. Selected sera were chosen to assess serum bactericidal antibodies (“SBA”, D) and opsonophagocytic antibodies (“OPA”, E) and were diluted such that each sample contained equivalent anti-1925wzzB-COPS IgG EU. Curves were fitted to each serial dilution of serum and were compared using nonlinear regression analysis. Dashed line indicates 0% killing. Results are representative of two independent assays, error bars represent s.d. and were derived from technical replicates; n.s., not significant.</p

Chemical structure of the OPS used for computational simulations and chemical models and sampled volumes of the most prevalent conformational cluster “11111111”.

<p>(A) Structure of the base (O:4,12) 3-repeat STm polysaccharide unit used for computational analyses. Individual monosaccharide units are indicated numerically, with the reducing end rhamnose (Rha) designated as (1, blue). The representative O-acetylated polysaccharide was constructed with the hydroxyl group substituted by an acetyl at the C2 position of both α-D-Abequ<i>p</i> and α-L-Rha<i>p</i> (-OH, red). (B,C) Structural models of GL cluster “11111111” displaying 90° of rotation around the carbohydrate stem for the base (B) and O-acetylated base (C) polysaccharides. The acetyl group is shown in brown, glucose in blue, mannose in green, galactose in yellow, abequose in purple, and rhamnose in cyan. For clarity, all hydrogen atoms are omitted except those on the acetyl group. (D,E,F,G) 3D spatial distributions (wire frame) displaying 90° of rotation around the carbohydrate stem for the base (D,E) and O-acetylated base (F,G) polysaccharides for all non-hydrogen atoms. The reducing end Rha (1, blue), indicated at the base, is approximated as linked to the core polysaccharide or preceding OPS repeat. Conformational flexibility increases progressively relative to the reducing end anchor point. The base polysaccharide backbone is identified by a transparent grey surface overlaid with the individual O-acetyl groups in color for monosaccharides (1) [blue], (4) [red], (5) [orange], (8) [tan], (9) [green] and (12) [purple]. Contour levels are set to 10⁻⁶ for the full polysaccharide and to 10⁻⁵ for the individual acetyl groups.</p

Biophysical and biochemical attributes of STm COPS conjugates used in this study.

<p>Biophysical and biochemical attributes of STm COPS conjugates used in this study.</p