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    <p>(A) Rat muscle extracts were separated by SDS PAGE and probed by immunoblotting as described in the Methods. The analysis was limited to 15 animals (n = 8 biome enriched; lanes E1 through E8, and n = 7 biome depleted; lanes D1 through D7) due to size constraints of the gel. A control strip with no serum is labeled “C”, and indicates reactivity of the anti-IgM conjugate with muscle-derived antigens. (B) The number of bands recognized by natural IgM in individual sera (p = 0.0089) and the total reactivity of natural IgM from each serum sample (p = 0.0093) are shown, with the bars indicating the mean and standard error. (C) The distribution of bands as a function of band size is shown. For this analysis, the average number of bands in biome depleted and biome enriched rats (Y-axis) was plotted on linear and log scales (main figure and figure inset, respectively) versus different band sizes (X-axis).</p

    Relative concentration of DNP-specific antibody in the serum of biome depleted (n = 20) and biome enriched (n = 15) rats.

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    <p>The relative concentration of antibody was determined by ELISA as described in the Methods. Relative levels of (A) IgM, (B) IgG, and (C) subclasses of IgG are shown. The means and standard errors are shown. The <i>p</i>-values associated with comparing data from biome depleted and biome enriched animals using a t-test are shown. (NS = not significant)</p

    Natural anti-human serum albumin antibody levels in the serum of biome depleted (n = 20) and biome enriched (n = 15) rats.

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    <p>The relative concentration of antibody was determined by ELISA as described in the Methods. Relative levels of (A) IgM and (B) IgG are shown. Binding to human serum albumin (HSA) was used as a measure of reactivity toward a xenogeneic antigen for which the animals lacked previous exposure. The means, standard errors, and the <i>p</i>-values associated with comparing data from biome depleted and biome enriched animals using a t-test are shown. (NS = not significant)</p

    Schematic of the density-based tests to identify SCD.

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    <p>Both versions of the SCD-AMPS are designed to separate dense red blood cells present in SCD from whole blood. Blood passes through the phases—top (T) and bottom (B) for SCD-AMPS-2 and top (T), middle (M), and bottom (B) for SCD-AMPS-3—upon centrifugation. If sickled cells are present, they collect at the interface between the bottom phase and the seal (<i>B/S</i>), and provide a visual readout for the presence of SCD. In SCD-AMPS-3, the additional phase allows the discrimination of Hb SS from Hb SC by evaluating the distribution of red cells at the upper interfaces (between the top and middle phases (<i>T/M</i>) and the middle and bottom phases (<i>M/B</i>).</p

    Tabulation of Results of SCD-AMPS Tests Compared to Reference Test Results by Hemoglobin Electrophoresis.

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    <p>*Samples found to have>50% Hb S but non-zero levels of Hb A, potentially a result of Hb S with β-thalassemia or a transfused Hb SS subject.</p><p>Tabulation of Results of SCD-AMPS Tests Compared to Reference Test Results by Hemoglobin Electrophoresis.</p

    The sensitivity and specificity of SCD-AMPS as a function of the amount of time between collecting samples and running tests.

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    <p>The specificity shows a decline over each 24 hour increment, with a significant decline over 48 hours (p-value <0.0005). The sensitivity increased between the first and second time interval, but then decreased between the second and third interval (p-values <0.01). The sample size used for each time interval is provided below each bar.</p
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