Sophorolipid Butyl Ester Diacetate Does Not Affect Macrophage Polarization but Enhances Astrocytic Glial Fibrillary Acidic Protein Expression at Micromolar Concentrations in Vitro

Peritoneal macrophages (PMACs) and spinal cord astrocytes were exposed to varying concentrations of soluble sophorolipid butyl ester diacetate (SLBEDA) in vitro. Macrophages and astrocytes demonstrated no decrease in viability in response to SLBEDA. Studying pro- and anti-inflammatory genes, PMACs did not show a shift toward a pro-inflammatory phenotype. However, at higher concentrations (3 and 30 μM), astrocytes showed an increase in their expression of glial acidic fibrillary protein. This novel category of compounds poses low risk to PMAC and astrocyte viability; however, the effect on PMAC polarization and astrocyte reactivity requires more elucidation

Sensitivity and Specificity of PW-Doppler parameters t_s/t_T and v_max as predictors of plaque formation.

<p>Sensitivity and Specificity of PW-Doppler parameters t_s/t_T and v_max as predictors of plaque formation.</p

Changes in Lumen and IEL area upon carotid constriction.

<p>Data was analyzed by One Way ANOVA.</p><p>“*”indicates p<0.05 as compared to presurgery.</p>“†”<p>indicates p<0.05 as compared to 2 weeks post surgery.</p

Plaque length is accurately measured by ultrasound.

<p>(a) Representative sagittal B-mode images of RCCA from one animal prior to surgery (0) and 3, 6 and 9 weeks after cast placement. Plaque length is indicated with arrows. (b) Whole mount of the carotid artery in (a) at study termination. (c) Linear regression of ultrasound vs microcaliper measurements of plaque length (n = 6 cuffs, 10 casts). Dotted lines indicate 95% confidence intervals. Correlation coefficient = 0.92, n = 16. (d) Bland Altman analysis of plaque length as measured by microcaliper and UBM. Dotted lines represent 95% limits of agreement. Bias, indicated by solid line, was calculated as −0.27 mm+/−0.22 SD.</p

Percent stenosis calculated from UBM parameters correlates with histological measurements.

<p>(a) Transverse B-mode image of RCCA 2 and 9 weeks after surgery. Upper Images, Adventitia is marked “A”, plaque “P”, “Medial layer M”. Lower, images are identical, but the inner elastic lamina is outlined with dotted white line, lumen with a red dotted line. Trichrome-stained transverse sections of the same 9 wk artery; internal elastic lamina (IEL) and lumen are outlined in white and red, respectively. (b) IMT measurements of casted and cuffed animals after 2 and 9 weeks of surgery. n.s. Not significant. (c) Linear regression analysis of percent stenosis estimated by UBM and histology. See Materials and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0029944#s2" target="_blank">Methods</a> for calculations of percent stenosis. Dotted lines indicate 95% confidence intervals; correlation coefficient = 0.75, n = 10 casts. (d) Comparison of UBM and histology measurements using Bland-Altman analysis. Dotted lines indicate 95% limits of agreement. Bias is indicated by solid line, was calculated as −4.76 mm±13.3 SD.</p

Plaques Progress More Rapidly in Casted vs Cuffed Animals.

<p>The rate of plaque growth was determined by performing individual linear regression analysis of plaque length or stenosis over time for each animal. The slopes (rates) were compared by unpaired t-test. (a) Rate of plaque growth in length (b) Rate of plaque growth in terms of stenosis. Data are mean±SEM, n = 7 cuffs, 10 casts “*” indicates p<0.05. (See <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0029944#pone.0029944.s002" target="_blank">Fig. S2</a> for source data.)</p

Study time-line.

<p>Study time-line.</p

Pre-plaque PW-Doppler parameters correlate with plaque length at 9 weeks.

<p>(a) Representative PW-Doppler tracing of a proximal RCCA before and after surgery. Note the dramatic decrease in velocity. Measurements of v_max, time in systole (t_s) and t_T (total time) are indicated. (b) v_max and t_s/t_T were calculated from pre-surgery tracings and 2 and 9 weeks after device placement. Data (mean±SEM, n = 7 cuffs, 10 casts), were analyzed by 2 way ANOVA. There was a highly significant effect of device on v_max and t_s/t_T ratio (p<0.005), but no significant difference between cast and cuffs on either parameter. “*” p<0.05 compared to pre-surgery. (c) Using 39 animals fitted with casts, 2 week t_s/t_T and v_max values were calculated and plotted against plaque length at 9 weeks and the correlation determined by linear regression. t_s/t_T positively correlated with plaque length (p<0.0005, r = 0.66) between, v_max and plaque length were negatively correlated (p<0.0005, r = 0.59). Dotted lines indicate 95% confidence interval.</p