Neural crest cell-specific <i>Brca2</i> deletion results in a craniofacial bone phenotype similar to that of the neural crest cell-specific <i>Brca1</i> deletion in mice.

<p>(A) Alcian blue- and alizarin red-stained skulls of control and <i>Brca2</i>:<i>Wnt1-Cre</i> mice at birth. Yellow broken lines indicate osteogenic fronts. Scale bar = 2mm. (B) Quantification of the area ratio of frontal foramen in the frontal bone area. White box represents the mean of each genotype. (C) Quantification of sagittal length. (D) Coronal sections of control and <i>Brca2</i>:<i>Wnt1-Cre</i> frontal bone primordia at E12.5 and E13.5 were double-labeled with RUNX2 (red) and BrdU (green). Broken line describes the osteogenic lineage cell population. Right charts show quantification of the ratio of BrdU-positive cells over RUNX2-positive cells. Scale bar = 50μm. (E) Immunostaining for RUNX2 and TUNEL assay of sections from control and <i>Brca2</i>:<i>Wnt1-Cre</i> embryos at E12.5. Broken line describes the osteogenic lineage cell population. White arrows indicate TUNEL-positive cells. Yellow color represents the non-specific signal from red blood cells. Scale bar = 100μm. as, alisphenoid; bo, basioccipital; bs, basisphenoid; eo, exoccipital; fb, frontal bone; ib, interparietal bone; jb, jugal bone; md, mandible; mx, maxilla; nb, nasal bone; p, palatine; pb, parietal bone; pmx, premaxilla; ppmx, palatal process of maxilla; ppp, palatal process of palatine; ptg, pterygoid; tr, tympanic ring.</p

Neural crest cell-specific <i>Brca1</i> deletion causes craniofacial bone abnormalities in mice.

<p>(A) Lateral views of control and <i>Brca1</i>:<i>Wnt1-Cre</i> mice at birth (NB). Scale bar = 5mm. (B) Alcian blue- and alizarin red-stained skulls of control and <i>Brca1</i>:<i>Wnt1-Cre</i> mice at birth. Yellow broken lines indicate osteogenic fronts. Note that the frontal bones of <i>Brca1</i>:<i>Wnt1-Cre</i> mice are separated by a large open space. Scale bar = 2mm. as, alisphenoid; bo, basioccipital; bs, basisphenoid; eo, exoccipital; fb, frontal bone; ib, interparietal bone; jb, jugal bone; md, mandible; mx, maxilla; nb, nasal bone; p, palatine; pb, parietal bone; pmx, premaxilla; ppmx, palatal process of maxilla; ppp, palatal process of palatine; ptg, pterygoid; tr, tympanic ring.</p

The <i>Brca1-p53</i> interaction is critical for craniofacial bone morphogenesis.

<p>(A) Immunoblot analysis of facial region tissue from E13.5 embryos. Each sample is from individual embryos. Right chart shows quantification of p53 relative production level. (B) Alcian blue- and alizarin red-stained skulls of control, <i>Brca1</i>^<i>-/-</i>:<i>Wnt1-Cre</i>, <i>Brca1</i>^<i>-/-</i>:<i>p53</i>^<i>+/-</i>:<i>Wnt1-Cre</i> and <i>Brca1</i>^<i>-/-</i>:<i>p53</i>^<i>-/-</i>:<i>Wnt1-Cre</i> mice at birth. Yellow broken lines indicate osteogenic fronts. Scale bar = 2mm. (C) Quantification of the area ratio of frontal foramen in the frontal bone area. White box represents the mean of each genotype. (D) Quantification of sagittal length. (E) Quantification of skull width. (F) Measurement schema of C, D and E. The shaded area surrounded by the red line is the measured frontal area in C. Sagittal length was divided into anterior and posterior part at the estimated anterior fontanelle (yellow). Data in A, D and E are represented as mean ±SD, n = 3 in A and n = 10 in C, D and E in each group. *P<0.05. N.S., not significant.</p

Photon Upconverted Emission Based on Dye-Sensitized Triplet–Triplet Annihilation in Silica Sol–Gel System

Photon upconverted emission based on dye-sensitized triplet–triplet annihilation was observed in silica gel systems containing Pt­(II) octaethylporphyrin (triplet sensitizer) and 9,10-diphenylanthracene (singlet emitter). The triplet sensitizer was encapsulated and highly dispersed in the silica gels prepared by the sol–gel method. The singlet emitter was adsorbed on the silica gel pores accessible to the outside. Phosphorescence of the triplet sensitizer was partially quenched, and the singlet emission was enhanced with an increase in the concentration of the singlet emitter. The emission intensity increased in proportion to the approximate square of the irradiation power. The triplet energy transfer from some of the encapsulated triplet sensitizer molecules to the adsorbed singlet emitter molecules was observed in the silica gels followed by the triplet–triplet annihilation and upconverted singlet emission. The phosphorescence quenching and upconverted singlet emission were more significantly observed in the gel dried at a lower temperature (wetter gel). The wetter gel contained higher amounts of solvent and water molecules, in which the triplet sensitizer and singlet emitter should collide and then the sensitized emitters should collide between themselves during their excited-state lifetime. The photon upconversion process required the triplet sensitizer and singlet emitter molecules to be in an environment similar to the solvents

Deletion of <i>p53</i> partially rescues the skull defects by preventing cell death.

<p>(A) Coronal sections of control, <i>Brca1</i>^<i>-/-</i>:<i>Wnt1-Cre</i> and <i>Brca1</i>^<i>-/-</i>:<i>p53</i>^<i>-/-</i>:<i>Wnt1-Cre</i> frontal bone primordia at E12.5 were double-labeled with RUNX2 (red) and BrdU (green). Broken line describes the osteogenic lineage cell population. Right charts show quantification of the ratio of BrdU-positive cells over RUNX2-positive cells. Scale bar = 50μm. (B) Immunostaining for RUNX2 and TUNEL assay of sections from control, <i>Brca1</i>^<i>-/-</i>:<i>Wnt1-Cre</i> and <i>Brca1</i>^<i>-/-</i>:<i>p53</i>^<i>-/-</i>:<i>Wnt1-Cre</i> embryos at E12.5. Broken line describes the osteogenic lineage cell population. White arrows indicate TUNEL-positive cells. Yellow color represents the non-specific signal from red blood cells. Right chart shows quantification of the percentage of TUNEL-positive cells in the frontal bone primordium. (C) Immunostaining for γ-H2AX and Cleaved Caspase-3 and/or p-Chk2 of sections from control, <i>Brca1</i>^<i>-/-</i>:<i>Wnt1-Cre</i> and <i>Brca1</i>^<i>-/-</i>:<i>p53</i>^<i>-/-</i>:<i>Wnt1-Cre</i> embryos at E12.5. Broken line describes the osteogenic lineage cell population. White arrows indicate double-positive cells for γ-H2AX/Cleaved Caspase-3 and/or γ-H2AX/p-Chk2. Right chart shows quantification of the percentage of γ-H2AX, Cleaved Caspase-3 and p-Chk2 positive cells in the frontal bone primordium. Scale bar = 100μm. Data in A, B and C are represented as mean ±SD, n = 3 in each group. *P<0.05. N.S., not significant.</p

Primer sequences for Quantitative RT-PCR analysis.

<p>Primer sequences for Quantitative RT-PCR analysis.</p

<i>Brca1</i> is indispensable for osteoblast proliferation and survival at mid-gestation.

<p>(A) Coronal sections of control and <i>Brca1</i>:<i>Wnt1-Cre</i> frontal bone primordia at E12.5 and E13.5 were double-labeled with RUNX2 (red) and BrdU (green) to detect osteogenic cells and proliferative cells, respectively. Broken line describes the osteogenic lineage cell population. Right charts show quantification of the ratio of BrdU-positive cells over RUNX2-positive cells. Scale bar = 50μm. (B) Immunostaining for RUNX2 and TUNEL assay of sections from control and <i>Brca1</i>:<i>Wnt1-Cre</i> embryos at E12.5. Broken line describes the osteogenic lineage cell population. White arrows indicate TUNEL-positive cells. Yellow color represents the non-specific signal from red blood cells. Scale bar = 100μm. (C) Immunostaining for γ-H2AX and Cleaved Caspase-3 and/or p-Chk2 of sections from control and <i>Brca1</i>:<i>Wnt1-Cre</i> embryos at E12.5. Broken line describes the osteogenic lineage cell population. White arrows indicate γ-H2AX- and/or Cleaved Caspase-3-positive cells. Scale bar = 100μm. (D) Quantification of the percentage of γ-H2AX-, Cleaved Caspase3- and TUNEL-positive cells in the frontal bone primordium. Data in A and D are represented as mean ±SD, n = 3 in each group. *P<0.05.</p

Accuracy assessment of MRI and endoscopy.

<p>A, The graph shows percent accuracy of individual scans when assessed by each technique: visual endoscopy and MRI. B, The graph compares average accuracy of each technique as evaluated by the blinded study. MRI; magnetic resonance imaging.</p

Comparative analysis of a single animal with histology, endoscopy and MRI.

<p>A, Hemotoxylin and eosin staining (magnification×10) shows histological confirmation of EAC. B, Endoscopic evaluation of the same rat. Arrow designates suspected area of tumor identified by endoscopic study participants. C, Axial MRI assessment. The arrow shows an abnormal mass on esophageal wall selected by MRI study participants. D, Coronal MRI image. Arrow identifies corresponding cross section of suspected tumor in anastomotic area of interest. MRI; magnetic resonance imaging, EAC; esophageal adenocarcinoma.</p

The Levrat Model.

<p>Gross image of esophageal tumor (animal no. 13) induced by modified Levrat model.</p