DataSheet_1_A self-compatible pear mutant derived from γ-irradiated pollen carries an 11-Mb duplication in chromosome 17.zip

Self-compatibility is a highly desirable trait for pear breeding programs. Our breeding program previously developed a novel self-compatible pollen-part Japanese pear mutant (Pyrus pyrifolia Nakai), ‘415-1’, by using γ-irradiated pollen. ‘415-1’ carries the S-genotype S4dS5S5, with “d” indicating a duplication of S5 responsible for breakdown of self-incompatibility. Until now, the size and inheritance of the duplicated segment was undetermined, and a reliable detection method was lacking. Here, we examined genome duplications and their inheritance in 140 F1 seedlings resulting from a cross between ‘515-20’ (S1S3) and ‘415-1’. Amplicon sequencing of S-RNase and SFBB18 clearly detected S-haplotype duplications in the seedlings. Intriguingly, 30 partially triploid seedlings including genotypes S1S4dS5, S3S4dS5, S1S5dS5, S3S5dS5, and S3S4dS4 were detected among the 140 seedlings. Depth-of-coverage analysis using ddRAD-seq showed that the duplications in those individuals were limited to chromosome 17. Further analysis through resequencing confirmed an 11-Mb chromosome duplication spanning the middle to the end of chromosome 17. The duplicated segment remained consistent in size across generations. The presence of an S3S4dS4 seedling provided evidence for recombination between the duplicated S5 segment and the original S4haplotype, suggesting that the duplicated segment can pair with other parts of chromosome 17. This research provides valuable insights for improving pear breeding programs using partially triploid individuals.</p

Human and mouse dermal microvascular endothelial cells have similar angiogenic potential <i>in vitro</i> and <i>in vivo</i>.

<p>(<b>A-D</b>) Representative photomicrographs of human dermal microvascular endothelial cells (HDMEC) and mouse dermal microvascular endothelial cells (MDMEC) stably transduced with GFP under light microscopy (<b>A</b> and <b>B</b>) and fluorescence microscopy (<b>C</b> and <b>D</b>). (<b>E</b> and <b>F</b>) Representative images of capillary sprouts formed by HDMEC (<b>E</b>) and MDMEC (<b>F</b>) on 3-D type I collagen matrices. (<b>G</b> and <b>H</b>) Photomicrographs of representative fields of GFP-immunostaining (red color) used to localize the blood vessels formed by HDMEC (<b>G</b>) and MDMEC (<b>G</b>) 14 days after implantation in immunodefficient mice. (<b>I</b>) The number of microvessels in implants populated with HDMEC or MDMEC. Microvessels were counted in 10 random microscopic fields/scaffold (200×) in 6 scaffolds per group. Results are representative of 3 independent experiments, n = 6.</p

<i>In vitro</i> response of human and mouse endothelial cells to a chemotherapeutic and an anti-angiogenesis drug.

<p>(<b>A</b> and <b>B</b>) The 48-hour cytotoxicity of cisplatin (<b>A</b>) and sunitinib (<b>B</b>) was evaluated by the SRB assay in HDMEC and MDMEC. Results are normalized against vehicle control and initial plating density. (<b>C</b> and <b>D</b>) Fold-change difference in the percentage of apoptotic cells upon treatment with cisplatin (<b>C</b>) or sunitinib (<b>D</b>) for 48 hours. Apoptosis was determined as the percentage of cells in Sub-G₀/G₁ by propidium iodide staining followed by flow cytometry. (<b>E</b> and <b>F</b>) Effect of cisplatin (<b>E</b>) or sunitinib (<b>F</b>) on the cell cycle of HDMEC and MDMEC. Results are representative of 3 independent experiments.</p

Tumor xenografts vascularized with human endothelial cells have similar microvessel density by grow faster than xenografts vascularized with mouse endothelial cells.

<p>(A) Representative photographs of HeLa tumors vascularized with either HDMEC or MDMEC. (B-D) Graphs depicting tumor growth of xenografts of HN12 (B), HeLa (C), UM-SCC-17B (D) vascularized with HDMEC or MDMEC. (E) Graph depicting tumor volume at the end of the experimental period. Asterisk depicts p<0.05. (F and G) Representative photomicrographs of tumors generated with UM-SCC-17B and HDMEC or MDMEC. Immunohistochemistry for GFP was performed to identify GFP-tagged endothelial cells. (H) Microvessel density of xenograft tumors generated with UM-SCC-17B and HDMEC or MDMEC. Results are representative of 3 independent experiments, n = 8.</p

Table_1_A self-compatible pear mutant derived from γ-irradiated pollen carries an 11-Mb duplication in chromosome 17.xlsx

Self-compatibility is a highly desirable trait for pear breeding programs. Our breeding program previously developed a novel self-compatible pollen-part Japanese pear mutant (Pyrus pyrifolia Nakai), ‘415-1’, by using γ-irradiated pollen. ‘415-1’ carries the S-genotype S4dS5S5, with “d” indicating a duplication of S5 responsible for breakdown of self-incompatibility. Until now, the size and inheritance of the duplicated segment was undetermined, and a reliable detection method was lacking. Here, we examined genome duplications and their inheritance in 140 F1 seedlings resulting from a cross between ‘515-20’ (S1S3) and ‘415-1’. Amplicon sequencing of S-RNase and SFBB18 clearly detected S-haplotype duplications in the seedlings. Intriguingly, 30 partially triploid seedlings including genotypes S1S4dS5, S3S4dS5, S1S5dS5, S3S5dS5, and S3S4dS4 were detected among the 140 seedlings. Depth-of-coverage analysis using ddRAD-seq showed that the duplications in those individuals were limited to chromosome 17. Further analysis through resequencing confirmed an 11-Mb chromosome duplication spanning the middle to the end of chromosome 17. The duplicated segment remained consistent in size across generations. The presence of an S3S4dS4 seedling provided evidence for recombination between the duplicated S5 segment and the original S4haplotype, suggesting that the duplicated segment can pair with other parts of chromosome 17. This research provides valuable insights for improving pear breeding programs using partially triploid individuals.</p

Human tumor xenografts vascularized with mouse endothelial cells is more responsive to a chemotherapeutic and an anti-angiogenesis drug.

<p>Xenograft tumors were engineered in immunodefficient mice by the co-transplantation of human tumor cells and human endothelial cells or human tumor cells and mouse endothelial cells seeded in biodegradable scaffolds measuring 6×6×1 mm. (A) Graph depicting the growth of human xenografts tumors (UM-SCC-17B cells) vascularized with HDMEC or MDMEC. As soon as we observed growth of the tumors beyond the size of the scaffold (<i>i.e.</i> when average tumor volume was 180 mm³), mice began to receive either 4 mg/kg cisplatin (i.p.) every 5 days, or 40 mg/kg sunitinib (o.r.) daily. (B) Fold change difference between the pre-treatment volume of the tumors and the volume of the same tumors after 30 days of treatment with cisplatin or sunitinib. (C) Tumor volume at the end of treatment with cisplatin or sunitinib. (D) Graph depicting the number of microvessels in tumor xenografts vascularized with HDMEC or MDMEC after treatment with cisplatin or sunitinib. Asterisk depicts P<0.05, as compared with controls. Results are representative of 3 independent experiments, n = 8.</p

Aberrant Methylation Inactivates Somatostatin and Somatostatin Receptor Type 1 in Head and Neck Squamous Cell Carcinoma

<div><p>Purpose</p><p>The aim of this study was to define somatostatin (<i>SST</i>) and somatostatin receptor type 1 (<i>SSTR1</i>) methylation profiles for head and neck squamous cell carcinoma (HNSCC) tumors at diagnosis and follow up and to evaluate their prognostic significance and value as a biomarker.</p><p>Methods</p><p>Gene expression was measured by quantitative RT-PCR. Promoter methylation status was determined by quantitative methylation-specific PCR (Q-MSP) in HNSCC.</p><p>Results</p><p>Methylation was associated with transcription inhibition. <i>SST</i> methylation in 81% of HNSCC tumor specimens significantly correlated with tumor size (<i>P</i> = 0.043), stage (<i>P</i> = 0.008), galanin receptor type 2 (<i>GALR2</i>) methylation (<i>P</i> = 0.041), and tachykinin-1 (<i>TAC1</i>) (<i>P</i> = 0.040). <i>SSTR1</i> hypermethylation in 64% of cases was correlated with tumor size (<i>P</i> = 0.037), stage (<i>P</i> = 0.037), <i>SST</i> methylation (<i>P</i> < 0.001), and expression of <i>galanin</i> (<i>P</i> = 0.03), <i>GALR2</i> (<i>P</i> = 0.014), <i>TAC1</i> (<i>P</i> = 0.023), and tachykinin receptor type 1 (<i>TACR1</i>) (<i>P</i> = 0.003). <i>SST</i> and <i>SSTR1</i> promoter hypermethylation showed highly discriminating receiver operator characteristic curve profiles, which clearly distinguished HNSCC from adjacent normal mucosal tissues. Concurrent hypermethylation of <i>galanin</i> and <i>SSTR1</i> promoters correlated with reduced disease-free survival (log-rank test, <i>P</i> = 0.0001). Among patients with oral cavity and oropharynx cancer, methylation of both <i>SST</i> and <i>SSTR1</i> promoters correlated with reduced disease-free survival (log-rank test, P = 0.028). In multivariate logistic-regression analysis, concomitant methylation of <i>galanin</i> and <i>SSTR1</i> was associated with an odds ratio for recurrence of 12.53 (95% CI, 2.62 to 59.8; <i>P</i> = 0.002).</p><p>Conclusions</p><p>CpG hypermethylation is a likely mechanism of <i>SST</i> and <i>SSTR1</i> gene inactivation, supporting the hypothesis that <i>SST</i> and <i>SSTR1</i> play a role in the tumorigenesis of HNSCC and that this hypermethylation may serve as an important biomarker.</p></div

Retrato del duque de San Carlos

Postal que reproduce una pintura del año 1815 de Francisco de Goya y Lucientes. Representa a José Miguel de Carvajal Vargas y Manrique de Lara, duque de San Carlos. Se muestra de cuerpo entero, ataviado con traje militar de color negro entorchado, medias blancas, un vistoso fajín rojo a la cintura y numerosas condecoraciones pendiendo de la casaca: el Toisón de Oro, la banda y la insignia de la orden de Carlos III y otras medallas. Con su brazo derecho sostiene el sombrero y una carta en la mano, mientras que la izquierda, más separada del cuerpo, se apoya sobre un bastón de mando, que otorga a la pose del duque un porte distinguido. Es el rostro la parte mejor ejecutada de la obra, realizado a partir de un estudio del natural http://www.museodezaragoza.es/wp-content/uploads/2015/09/Duque-de-San-Carlos_fondogris.pdfEn la postal aparece el siguiente texto: Museo de Bellas Artes. Zaragoz

Additional file 3: of The neuropeptide genes SST, TAC1, HCRT, NPY, and GAL are powerful epigenetic biomarkers in head and neck cancer: a site-specific analysis

Figure S2. ROC curves for the methylation markers in head and neck carcinomas versus adjacent normal mucosal tissue. On the basis of the ROC curve analysis, the sensitivity, specificity, and cutoff level were determined to be 80.6%, 94.4%, and 0.046 for SST (a); 72.2%, 97.2%, and 0.08 for TAC1 (b); 67.6%, 97.2%, and 0.099 for HCRT (c); 50.0%, 97.2%, and 0.041 for NPY (d); and 25.0%, 86.1%, and 0.100 for GAL (e). (EPS 1057 kb

Additional file 5: of The neuropeptide genes SST, TAC1, HCRT, NPY, and GAL are powerful epigenetic biomarkers in head and neck cancer: a site-specific analysis

Figure S3. Kaplan-Meier survival curves for the 58 patients with hypopharyngeal cancer, according to the methylation status of the five target genes. Disease-free survival for (a) SST, (b) TAC1, (c) HCRT, (d) NPY, and (e) GAL in the case of methylated (red lines) and unmethylated (blue lines) genes. (f) Joint analysis of the 5 genes. Blue line: patients with 0–2 methylated genes; red line: patients with 3–5 methylated genes. A probability of < 0.05 (*P < 0.05) was considered to represent a statistically significant difference. (EPS 1460 kb