sj-pdf-2-jcb-10.1177_0271678X231202594 - Supplemental material for Randomized placebo-controlled trial of CL2020, an allogenic muse cell–based product, in subacute ischemic stroke

Supplemental material, sj-pdf-2-jcb-10.1177_0271678X231202594 for Randomized placebo-controlled trial of CL2020, an allogenic muse cell–based product, in subacute ischemic stroke by Kuniyasu Niizuma, Shin-Ichiro Osawa, Hidenori Endo, Shin-Ichi Izumi, Kota Ataka, Akihiro Hirakawa, Masao Iwano and Teiji Tominaga in Journal of Cerebral Blood Flow & Metabolism</p

sj-pdf-1-jcb-10.1177_0271678X231202594 - Supplemental material for Randomized placebo-controlled trial of CL2020, an allogenic muse cell–based product, in subacute ischemic stroke

Supplemental material, sj-pdf-1-jcb-10.1177_0271678X231202594 for Randomized placebo-controlled trial of CL2020, an allogenic muse cell–based product, in subacute ischemic stroke by Kuniyasu Niizuma, Shin-Ichiro Osawa, Hidenori Endo, Shin-Ichi Izumi, Kota Ataka, Akihiro Hirakawa, Masao Iwano and Teiji Tominaga in Journal of Cerebral Blood Flow & Metabolism</p

sj-pdf-3-jcb-10.1177_0271678X231202594 - Supplemental material for Randomized placebo-controlled trial of CL2020, an allogenic muse cell–based product, in subacute ischemic stroke

Supplemental material, sj-pdf-3-jcb-10.1177_0271678X231202594 for Randomized placebo-controlled trial of CL2020, an allogenic muse cell–based product, in subacute ischemic stroke by Kuniyasu Niizuma, Shin-Ichiro Osawa, Hidenori Endo, Shin-Ichi Izumi, Kota Ataka, Akihiro Hirakawa, Masao Iwano and Teiji Tominaga in Journal of Cerebral Blood Flow & Metabolism</p

Specificity and sensitivity of immuno-northern blotting.

<p>(A) The sequences of synthesized 15-mer ssRNA containing either m⁶A or A at the center position. (B) SYBR staining and INB with anti-m⁶A antibody. Serially diluted m⁶A- and A-containing 15-mer ssRNA (10, 4, 1.6, 0.64, and 0.25 pmol) were analyzed. (C) Relative band intensities by INB of m⁶A-containing ssRNA. Densitometric analysis was performed to quantitate the band intensities. Data were expressed relative to the band intensity of each 10 pmol sample, which was taken as 1.0 arbitrary units (AU), N = 3. Values are shown as the mean ± SD.</p

Immuno-northern blotting protocol.

<p>In this method, RNAs were separated by denaturing acrylamide or agarose gel electrophoresis, transferred onto a positively charged nylon membrane followed by UV cross-linking, and then incubated with the primary antibodies against the specific modified nucleoside as well as the secondary antibody. The specific bands were visualized by chemiluminescence.</p

Immuno-northern blot analysis using the anti-m⁵C antibody with acrylamide and agarose gel separation.

<p>(A) Total RNA (0.6 μg) isolated from mouse liver was analyzed by SYBR staining and INB with anti-m⁵C antibody after electrophoresis in the 12% polyacrylamide or 1% agarose denaturing gel. The INB image using acrylamide gel is a long-exposure image. Arrowheads denote the positive signals, which appear to be integrated in the INB image using the acrylamide gel. (B) Total RNA isolated from mouse liver (1.2 μg) and <i>E</i>. <i>coli</i> HST04 (0.6 μg) was analyzed by SYBR staining and INB with anti-m⁵C antibody or isotype IgG. As a negative control for INB, isotype IgG was used instead of anti-m⁵C antibody at the same concentration. (C, D) The effect of DNase I treatment on the anti-m⁵C positive signal. Total RNAs isolated from mouse liver (1.2 μg, C), <i>E</i>. <i>coli</i> HST04 (0.6 μg, D), and yeast BY4742 (1.0 μg, D) were treated with or without DNase I, and then it was analyzed by SYBR staining and INB with anti-m⁵C antibody using the indicated gel. Arrowheads denote the positive signals.</p

Immuno-northern blotting using antibodies against modified nucleosides.

<p>(A) Structural formulas of 1-methyladenosine (m¹A), <i>N</i>6-methyladenosine (m⁶A), pseudouridine (Ψ), and 5-methylcytidine (m⁵C). (B) Total RNA (0.6 μg) isolated from the indicated samples was separated in the 12% polyacrylamide-8 M urea denaturing gel and analyzed by SYBR staining and immuno-northern blotting (INB) using the anti-m¹A, anti-m⁶A, anti-Ψ and anti-m⁵C antibodies. The total RNA samples (0.6 μg) were also analyzed by the dot blot assay using each antibody, as indicated in the bottom panels.</p

Real-world variability in the prediction of intracranial aneurysm wall shear stress: The 2015 International Aneurysm CFD Challenge

Purpose—Image-based computational fluid dynamics (CFD) is widely used to predict intracranial aneurysm wall shear stress (WSS), particularly with the goal of improving rupture risk assessment. Nevertheless, concern has been expressed over the variability of predicted WSS and inconsistent associations with rupture. Previous challenges, and studies from individual groups, have focused on individual aspects of the image-based CFD pipeline. The aim of this Challenge was to quantify the total variability of the whole pipeline. Methods—3D rotational angiography image volumes of five middle cerebral artery aneurysms were provided to participants, who were free to choose their segmentation methods, boundary conditions, and CFD solver and settings. Participants were asked to fill out a questionnaire about their solution strategies and experience with aneurysm CFD, and provide surface distributions of WSS magnitude, from which we objectively derived a variety of hemodynamic parameters. Results—A total of 28 datasets were submitted, from 26 teams with varying levels of self-assessed experience. Wide variability of segmentations, CFD model extents, and inflow rates resulted in interquartile ranges of sac average WSS up to 56%, which reduced to < 30% after normalizing by parent artery WSS. Sac-maximum WSS and low shear area were more variable, while rank-ordering of cases by low or high shear showed only modest consensus among teams. Experience was not a significant predictor of variability. Conclusions—Wide variability exists in the prediction of intracranial aneurysm WSS. While segmentation and CFD solver techniques may be difficult to standardize across groups, our findings suggest that some of the variability in image-based CFD could be reduced by establishing guidelines for model extents, inflow rates, and blood properties, and by encouraging the reporting of normalized hemodynamic parameters