Fig 1: A comparison of S100A12 concentrations in plasma within risk groups. (A) Comparison of plasma concentrations of S100A12 in UCB patients divided into groups according to their risk score. The standard error of the mean is represented by error bars. (B) A ROC analysis of plasma S100A12 concentrations in high risk UCB patients vs. healthy group. (C) A ROC analysis of plasma S100A12 concentrations of intermediate risk UCB patients against study healthy. ns, not significant, ***p ≤ 0.0001 and *p ≤ 0.05.
Fig 2: (A) Heat map of S100A12 as most significantly regulated gene in risk groups. The gene and samples are clustered using Centroid linkage and Euclidean distance metric by ArrayStar software. The diagram row represents the S100A12 gene and the risk groups are presented in the columns. Color saturation reflects the differences in gene expression between tumor samples; red is higher than control group expression (blue). The intensity of color indicates the degree of gene expression from red (high expression fold) to yellow (low fold expression). qPCR validation of expression was done for selected genes. Normalized ratio ± ratio error determined for cDNA pools of study risk (B) and prognostic groups (C) by calculation of the ratio of the samples (target/reference) divided by the ratio of run calibrator (target/reference) (healthy samples). Quantitative PCR was performed for S100A12 (black), S100A8 (gray), NAMPT (light gray), JUP (red), KLRF1 (blue), and PTGDR (green). New positive, New UCB cases; recurrent, Cases with recurrence of the disease within 5 years after treatment; previously positive, Cases with UCB history but negative at the time the sample. ***p ≤ 0.0001 and **p ≤ 0.01.
Fig 3: Representative immunohistochemical analysis for tumors of T1, T2, and T3. IHC staining patterns for S100A12 in cancer tissue array tumors stained withanti-S100A12.
Fig 4: Comparison of S100A12 levels in UCB patient prognostics groups. (A) S100A12 concentrations in plasma from UCB patients sorted according to their prognosis. The standard error of the mean is represented by error bars. (B) The ROC analysis of plasma S100A12 concentrations of recurrent group against new positive group of patients. The AUC is 0.793 with standard error 0.09, p = 0.019 and the 95% confidence interval appeared to be 0.6114–0.9759. The best cut-off in this comparison was found to be 512.5 ng/mL, giving 71.4% sensitivity and 66.67% specificity. (C) A ROC analysis of S100A12 concentration of previously positive group (recurrent-negative at sample) compared to new positive group. The AUC is 0.725 with standard error 0.095, p = 0.058 and 95% confidence interval between 0.5387 and 0.911. Using the cut-off 462.5 ng/mL the sensitivity is 68.4% and specificity will be 55.56%. (D) ROC analysis of S100A12 of new positive UCB patients against all healthy. ns, not significant, **p ≤ 0.01 and *p ≤ 0.05.
Fig 5: Quantitation of S100A12 in UCB plasma samples by BLI (A). The concentration of S100A12 was measured from plasma samples of each group (UCB patients and Healthy) using the biolayer interferometry (BLI). For each group, the mean value for of S100A12 is shown and standard error of the mean is represented by error bars. (B) A ROC analysis of plasma S100A12 concentrations in all UCB patients vs. all healthy in study cohort ***p ≤ 0.0001.
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