Operation time and postoperative thrombosis in Microstent anastomosis.

<p>(<b>A</b>) Operation time of microvascular anastomoses in rats. A single surgeon performed anastomosis of the infrarenal aorta of the rat by 10–0 nylon suture and needle (<i>n</i> = 43) (blue) or by Microstent (<i>n</i> = 45) (orange). The lines demonstrate the changes in the ischemic time of the infrarenal aorta during anastomosis with the number of operations. The operation time for the Microstent anastomosis includes time for gluing by cyanoacrylate as well. The average ischemic time of the hand-sewn group was 40.9 min, while the average time of the Microstent group was 29.9 min. In some cases, the first operation to implant the Microstent failed, and a second or third one was required. In such cases, the ischemic time was prolonged (asterisks). There was a significant difference between the two groups (<i>p</i> < 0.00001). <i>p</i> values were determined by Student’s <i>t</i>-test. (<b>B</b>) The bar chart shows evidence of thrombosis a week after the operations (orange). Some of the rats died due to undefined reasons without anastomotic thrombosis (grey). The thrombosis rate was 2.3% in the hand-sewn group. No significant difference in the thrombosis rate was found between the hand-sewn group and the Microstent group (<i>p = 0</i>.<i>104)</i>. <i>p</i> values were determined by Fisher’s exact test. (<b>C</b>) Ultrasonography of an implanted Microstent (dashed square). The Microstent is visualized as a high-echoic region inside the aortic lumen (see also <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0181520#pone.0181520.s003" target="_blank">S2 Movie</a>). (<b>D</b>) Doppler studies of the peripheral aorta to the anastomotic site demonstrate patent blood flow a week after operation in the Microstent group.</p

Stent-assisted sutureless microvascular anastomoses.

<p>(<b>A</b>) Showing the traditional microvascular anastomosis using a needle and thread. Multiple interrupted sutures, usually 6–10, are required for anastomosis. The key to success is to maximize the vessel lumen (asterisk) by coaptation of both ends of the intima (arrowhead) without a gap and placing sutures at even intervals. (<b>B</b>) The Microstent system to perform sutureless microvascular anastomosis. The Microstent compressed by a 7–0 nylon thread squeezed in the holder is inserted into each side of the vessel ends (top). After both ends are coapted (center), the 7–0 nylon thread is cut to loosen the Microstent, and the holder is removed. The Microstent automatically expands to maintain the lumen without a gap (bottom). Finally, cyanoacrylate glue (green) is applied around the junction of the vessels (See also <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0181520#pone.0181520.s002" target="_blank">S1 Movie</a>). (<b>C</b>) The approximate diameter (<i>R</i>) of the compressed Microstent is determined by the number of branches (<i>n</i>), tube thickness (<i>d</i>), and branch width (<b><i>w</i></b>). When the Microstent is compressed to the maximum extent, the branches touch each other. (<b>D</b>) NiTi stent (Microstent) fully expanded (2 x 4 mm) (left) and a stent compressed by a 7–0 nylon thread (right). Note the continuously arranged branches in a Z-shape at the center in order to be compressed uniformly end to end. Bar: 1 mm. (<b>E</b>) Scanning electron microscope image of the Microstent before and after electrolytic polishing (ELP), Bars: 100 μm. (<b>F</b>) Intraoperative view of anastomosis of rat aorta using Microstent. The anastomotic site was sealed by cyanoacrylate (arrow). Bar: 1 mm.</p

Ex vivo analyses of wall compliance and mechanical strength of Microstent anastomosed vessels.

<p>(<b>A</b>) The anastomosed vessels are connected to an ex vivo cardiac pump system. The lines demonstrate the average change in blood pressure with the blood flow change of the vessel specimen at 2 weeks, 7 weeks, and 26 weeks in each group (n = 6). (<b>B</b>) The representative data of the pressure changes of the Microstent-anastomosed vessel under cardiac pump pulsation. Both groups could withstand the highest arterial pressure of over 400 mmHg (see also <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0181520#pone.0181520.s004" target="_blank">S3</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0181520#pone.0181520.s005" target="_blank">S4</a> Movies). (<b>C</b>) Photograph of Microstent-anastomosed vessels under different arterial pressures: 150, 200, 300, and 400 mmHg. (<b>D</b>) The representative tensile curve of the anastomosed vessel. The vessels are pulled under physiological pulsation and blood pressure (see also <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0181520#pone.0181520.s006" target="_blank">S5</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0181520#pone.0181520.s007" target="_blank">S6</a> Movies). (<b>E</b>) The average tensile strength of hand-sewn and Microstent-anastomosed vessels at 2, 7, and 26 weeks. There is a significant difference between the two groups at 26 weeks. <i>p</i> values were determined by Student’s <i>t</i>-test. *<i>p < 0</i>.<i>05</i>.</p

Histological examination of anastomosed vessels.

<p>(<b>A</b>) A cross-section of a hand-sewn rat infrarenal aorta at 26 weeks. Low-magnification view (H&E stain). High-magnification view (elastic Van Gieson stain) shows hyperplasia of the tunica media. Black bar, 1 mm; yellow bar, 250 μm. (<b>B</b>) A cross-section of a Microstent-anastomosed rat infrarenal aorta at 26 weeks. High-magnification view shows neointimal proliferation limited to the region around the stent branches. (<b>C</b>) A SEM view of the lumen of a hand-sewn vessel at 26 weeks showing thread exposure into the lumen. Bar, 200 μm. (<b>D</b>) A SEM view of the lumen of a Microstent-anastomosed vessel at 26 weeks. The vessel is half-cut. All branches of the Microstent are covered with endothelium. Left bar, 500 μm; right bars, 100 μm.</p

Quantitative Trait Loci (QTL) Associated with Resistance to a Monogenean Parasite (<i>Benedenia seriolae</i>) in Yellowtail (<i>Seriola quinqueradiata</i>) through Genome Wide Analysis

<div><p>Benedenia infections caused by the monogenean fluke ectoparasite <i>Benedenia seriolae</i> seriously impact marine finfish aquaculture. Genetic variation has been inferred to play a significant role in determining the susceptibility to this parasitic disease. To evaluate the genetic basis of Benedenia disease resistance in yellowtail (<i>Seriola quinqueradiata</i>), a genome-wide and chromosome-wide linkage analyses were initiated using F₁ yellowtail families (n = 90 per family) based on a high-density linkage map with 860 microsatellite and 142 single nucleotide polymorphism (SNP) markers. Two major quantitative trait loci (QTL) regions on linkage groups Squ2 (<i>BDR-1</i>) and Squ20 (<i>BDR-2</i>) were identified. These QTL regions explained 32.9–35.5% of the phenotypic variance. On the other hand, we investigated the relationship between QTL for susceptibility to <i>B. seriolae</i> and QTL for fish body size. The QTL related to growth was found on another linkage group (Squ7). As a result, this is the first genetic evidence that contributes to detailing phenotypic resistance to Benedenia disease, and the results will help resolve the mechanism of resistance to this important parasitic infection of yellowtail.</p></div

Localization of significant markers for Benedenia disease resistance in linkage group Squ2F and Squ20F with family A.

<p>Squ(linkage group)F; marker distance in female map. (A) Squ2F, (B) Squ20F. Map positions and LOD scores are based on a simple interval mapping QTL analysis using the software MapQTL 5. Marker absolute map distances are given in (cM). 95% confidence probability LOD support interval was indicated as Gray bold line. Horizontal lines across each plot indicate LOD siginificance threshold, <i>P_g</i>; genome-wide significance threshold.</p

Significant markers for Benedenia disease resistance multiple-QTL model mapping in linkage group Squ2F and Squ20F with family A.

<p>Squ(linkage group)F; marker distance in female map. (A) Squ2F, (B) Squ20F. This figure was described using by MapQTL 5.</p

Significant markers for Benedenia disease resistance using Kruskal–Wallis analysis with A and B families.

<p>Signif.; Significance levels:</p>*<p><0.1</p>**<p><0.05</p>***<p><0.01</p>****<p><0.005</p>*****<p><0.001</p>******<p><0.0005</p>*******<p><0.0001.</p><p>NS; not significant, -; not informative in this locus.</p><p>Squ(linkage group)F; F is dam allele in female linkage group. Squ(linkage group)M; M is sire allele in male linkage group.</p

Multiple QTL model mapping results of the significant markers for Benedenia disease resistance in linkage group 2 and 20 in family A.

<p>Locus; marker name, LOD; Lod scores, % Var; percent of variance explained, Effect; estimated effect. Squ(linkage group)F; F is dam allele in female linkage group. Values in bold are LOD max in peak of each QTL marker position and each value.</p

Between fish size and number of parasites in pairwise Pearson correlations.

<p>Values in bold are different from a significance level <i>P</i> = 0.001.</p