Additional file 1: Figure S1. of Detection of Mycobacterium ulcerans by real-time PCR with improved primers

Sequence alignment of the IS2404 elements from the genome of M. ulcerans strain Agy99. (PDF 1040 kb

Imaging plane for blood flow rate measurement in the bilateral internal carotid arteries and the basilar artery.

<p>The plane is set as close to perpendicular as possible to these three arteries using the lateral view of the maximum intensity projection image of a three-dimensional magnetic resonance angiograph. Thus, the blood flow rates were measured in the presellar or posterior vertical segment of the cavernous portion of the internal carotid artery and basilar artery. The arteries in these anatomical locations run in a fairly straight manner and have low turbulence.</p

Subjects' demographic data.

<p>Subjects' demographic data.</p

Flow rates of the stented internal carotid artery (solid line) and contralateral internal carotid artery (dotted line).

<p>Horizontal bars and boxes represent mean and mean +/- standard deviation, respectively. Compared to preoperative values, the flow rates of the stented internal carotid artery after carotid artery stenting (CAS) are significantly larger. Compared to preoperative values, the flow rate of the contralateral carotid artery at 3 months after CAS is significantly smaller.</p

Sum of flow rates of the bilateral internal carotid arteries (ICAs) and the basilar artery.

<p>The sum increases after CAS, then decreases over the next 3 months.</p

Flow rates of the middle cerebral artery ipsilateral to the stented internal carotid artery (solid line) and contralateral middle cerebral artery (dotted line).

<p>The flow rate of the stented side middle cerebral artery increases after CAS and its trend is similar to that of the stented internal carotid artery. The flow rate of the contralateral one does not show significant changes.</p

Flow rate of the basilar artery.

<p>Compared to the preoperative value, the flow rate at 3 months after CAS is significantly smaller.</p

Activated p53 with Histone Deacetylase Inhibitor Enhances L-Fucose-Mediated Drug Delivery through Induction of Fucosyltransferase 8 Expression in Hepatocellular Carcinoma Cells

<div><p>Background</p><p>The prognosis of advanced hepatocellular carcinoma (HCC) is dismal, underscoring the need for novel effective treatments. The α1,6-fucosyltransferase (fucosyltransferase 8, FUT8) has been reported to accelerate malignant potential in HCC. Our study aimed to investigate the regulation of FUT8 expression by p53 and develop a novel therapeutic strategy for targeting HCC cells using L-fucose-mediated drug delivery.</p><p>Methods</p><p>Binding sites for p53 were searched for within the <i>FUT8</i> promoter region. FUT8 expression was assessed by immunoblotting. Chromatin immunoprecipitation (ChIP) assays were performed to analyze p53 binding to the <i>FUT8</i> promoter. The delivery of Cy5.5-encapsulated L-fucose-liposomes (Fuc-Lip-Cy5.5) to a <i>Lens Culinaris</i> agglutinin-reactive fraction of α-fetoprotein (AFP-L3)-expressing HCC cells was analyzed by flow cytometry. The induction of FUT8 by histone deacetylase inhibitor (HDACi) -inducing acetylated -p53 was evaluated by immunoblotting. Flow cytometric analysis was performed to assess whether the activation of p53 by HDACi affected the uptake of Fuc-Lip-Cy5.5 by HCC cells. The cytotoxicity of an L-fucose-bound liposome carrying sorafenib (Fuc-Lip-sorafenib) with HDACi was assessed <i>in vivo</i> and <i>in vitro</i>.</p><p>Results</p><p>The knock down of p53 with siRNA led to decreased FUT8 expression. ChIP assays revealed p53 binds to the <i>FUT8</i> promoter region. Flow cytometric analyses demonstrated the specific uptake of Fuc-Lip-Cy5.5 into AFP-L3-expressing HCC cells in a p53- and FUT8-dependent manner. HDACi upregulated the uptake of Fuc-Lip-Cy5.5 by HCC cells by increasing FUT8 via acetylated -p53. The addition of a HDACi increased apoptosis induced by Fuc-Lip-sorafenib in HCC cells.</p><p>Conclusions</p><p>Our findings reveal that <i>FUT8</i> is a p53 target gene and suggest that p53 activated by HDACi induces Fuc-Lip-sorafenib uptake by HCC cells, highlighting this pathway as a promising therapeutic intervention for HCC.</p></div

Combination therapy with HDACi enhances the cytotoxicity of Fuc-Lip-sorafenib.

<p>(A) AFP-L3 expressing HepG2, and JHH7 cells, and AFP-L3 non-expressing JHH6 cells were treated for 2 h with F0-Lip-sorafenib or F50-Lip-sorafenib. The cells were then, washed, incubated for 48 h, and cell viability was measured by BrdU assay. The percentage of viable cells is shown compared with untreated cells. (B) HCC cells were treated with F0-Lip-sorafenib or F50-Lip-sorafenib (5 μM) for 2 h with or without SAHA (1 μM) and, then washed and incubated for 48 h. Cell viability was measured by BrdU assay. Experiments were carried out in triplicate and repeated three times. NT: no treatment. * <i>P</i> < 0.05.</p

p53 and FUT8 regulate the introduction of Fuc-Lip-Cy5.5 into HCC cells.

<p>(A) Expression levels of p53 and FUT8 in HCC cells were examined by western blot. (B) Quantification of AFP-L3 values. (C) Determination of the introduction of Fuc-Lip-Cy5.5 into HCC cells. After siRNA transfection, HepG2 and JHH7 cells were exposed to 1 μM SAHA for 6 h, then treated with F50-Lip-Cy5.5 for 30 min and analyzed by flow cytometry. One siRNA was used for p53 (si-p53) and two were used for FUT8 (si-FUT8-1 and si-FUT8-2). Experiments were carried out in triplicate and repeated at least three times.</p