The roof plate formation in the <i>mib</i> mutant.

<p>(A), Confocal images, lateral view of the spinal cord of <i>mib</i> mutant, 72 hpf. Dashed rectangular shows the magnified in (A′) region of the spinal cord with the absence of GFP-positive RP cells. (B, C), the orthogonal optical sections of confocal images of the spinal cord of <i>mib</i> mutant illustrate lack of the roof plate extension between 36 and 72 hpf. Immunofluorescent staining of the transverse sections of spinal cord using anti-GFP (green) and anti-GFAP (red) antibodies; wild-type embryo (D–F) and <i>mib</i> mutant (G–I).</p

Re-orientation of the RP cells.

<p>Confocal images of the spinal cord of SqET33 line at different developmental stages (A–C, dorsal view, J–L, R, lateral view, M–P, orthogonal optical sections) and outline of the dorsal cells (D–F) superimposed onto the GFP intensity chart. Note that RB cells undergo the lateral-medial displacement. (G–I), Whole-mount immunohistochemistry detecting Islet1 in RB cells (red, nuclei), dorsal view of the spinal cord. (Q), Transverse section of the spinal cord at high magnification showing a fine structure of the central canal. RP process is stained with anti-GFP (green) and nuclei are counterstained with DAPI (grey). (S), Confocal image of the spinal cord of SqET33-10 line expressing GFP in the roof and floor plates, lateral view. Abbreviations: cc, central canal; di, dorsal interneurons; fp, floor plate; rb, Rohon-Beard cells; rp, roof plate cells.</p

Critical events during stretching morphogenesis of the RP in zebrafish.

<p>The scheme illustrates the stretching morphogenesis of the RP, which takes place during late neurulation in correlation with reduction of the primitive lumen into the central canal. In zebrafish this process depends on activity of Zic6 and Rock.</p

Effect of Rock inhibition on the RP morphogenesis.

<p>Confocal images of 72 hpf larva, untreated (A) and after Y-27632 injection into hindbrain ventricle at 30 hpf (B). The arrows show an approximate position of the transverse sections. (C–E′), transverse sections of the spinal cord of larva, treated with Y-27632. Immunofluorescent staining with anti-GFP (green) and phalloidin (red). Whole mount <i>in situ</i> hybridization with <i>glyt1</i> antisense RNA probe, control (F) and after treatment with Y-27632 (G).</p

Conversion of primitive lumen into central canal.

<p>(A–L), Contraction of opposed apical surfaces reflects recession of the primitive lumen into a central canal between 24 and 72 hpf. Immunofluorescent staining by a combination of anti-GFP with anti-β-catenin antibody (A–D), anti-ZO-1 antibody (E–H), and phalloidin that detects F-actin (I–L). Increase in cell number and build-up of axonal tracts (*), plastic transverse sections of the spinal cord (M–P). (Q–T), Schematics corresponding to (M–P) showing cell nuclei.</p

Growth of the RP processes correlates with a shift of <i>glyt1</i> expression along D–V axis.

<p>(A–D), Double whole mount <i>in situ</i> hybridization (<i>glyt1</i>, dark purple) and immunostaining (GFP, green). <i>glyt1</i> is expressed in the midline glial cells except RP; GFP is expressed in the RP cells. (E–J), Whole mount <i>in situ</i> hybridization (<i>glyt1</i>, dark purple) and immunostaining (β-catenin, red), the transverse sections of the spinal cord. The arrow shows an approximate position of attachment of the RP process to the apical surface of central canal. Confocal images of the spinal cord of <i>pard6γb</i> heterozygote (K) and <i>pard6γb</i> mutant (L) transgenic fish. (M), double whole mount <i>in situ</i> hybridization (<i>glyt1</i>, dark purple) and immunostaining (GFP, green) of <i>pard6γb</i> mutant. Abbreviation: fp, floor plate.</p

Characterization of SqET33 transgenic line.

<p>(A), Confocal image of 3 dpf larva of SqET33 line, lateral view. Dashed lines depict the position of transverse sections shown in B–D. (B–D), transverse sections, immunofluorescent staining with anti-GFP antibodies. Arrow indicates the elongated central process of the roof plate cell. (E–J), whole-mount immunofluorescent staining with anti-GFP (midline RP cells and lateral dorsal interneurons) and anti-HuC/HuD (lateral dorsal neurons) antibodies, dorsal view of spinal cord. (K–M), immunofluorescent staining with anti-GFP (green) and anti-GFAP (red) antibodies, transverse section of the spinal cord. Abbreviations: cc, central canal; dt, dorsal thalamus; ha, habenula; m, meninx; nc, notochord; p, pallium; po, preoptic area; rp, roof plate.</p

Description of pH-sensitive MBMs used.

<p>(a) General scheme of preparation of MBMs using the LbL method: co-precipitation of SNARF-1-D (purple) into porous CaCO₃ cores (yellow); LbL assembly of microcapsule shell around the cores (only three layers of negatively charged polymer, three layers of positively charged polymer and final biocompatible layer are depicted); and dissolution of cores. (b) SEM image of porous CaCO₃ cores with incorporated SNARF-1-D. (c) Prepared MBMs under confocal laser scanning microscope LSM 700. (d) Examples of fluorescence spectra of microencapsulated SNARF-1-D with varying pH. (e) Calibration curve of MBMs containing SNARF-1-D with varying pH was built based on median values (emphasized by larger dark blue points); original values are also depicted (smaller light blue points).</p

Acid-Responsive Polymeric Doxorubicin Prodrug Nanoparticles Encapsulating a Near-Infrared Dye for Combined Photothermal-Chemotherapy

Combination therapy with high spatial and temporal resolution is highly promising for efficient medical treatment of cancer. In this study, doxorubicin (DOX) conjugated amphiphilic block copolymer with a terminal folic acid moiety was prepared, which could self-assemble into nanoparticles by encapsulating organic near-infrared (NIR) absorbing dye IR825 for combined photothermal-chemotherapy. The resulting PDOX/IR825 nanoparticles showed excellent colloidal stability and monodispersity in aqueous solution. Specifically, the conjugated DOX could be released quickly in weak acidic environment for chemotherapy due to the cleavage of acid-labile hydrazone bond. Meanwhile, the encapsulated dye could convert the NIR light energy into heat with high efficiency, which makes the self-assembled nanoparticles an effective platform for photothermal therapy. Confocal microscopy observations and flow cytometry analysis confirmed that the PDOX/IR825 nanoparticles could be efficiently endocytosed by HeLa cells and deliver DOX into the nuclei of cancer cells. The in vitro cell viability assays indicated that both DOX-sensitive HeLa cells and DOX-resistant A2780/DOX^R cells were completely killed by the treatment of PDOX/IR825 under NIR light irradiation. Significant tumor regression was also observed in the zebrafish liver hyperplasia model upon combinational therapy provided from the PDOX/IR825 nanoparticles. Hence, the PDOX/IR825 nanoparticles exhibited a great potential in site-specific combined photothermal-chemotherapy of tumor

Instant Room-Temperature Gelation of Crude Oil by Chiral Organogelators

Large-scale treatment of oily water arising from frequent marine oil spills presents a major challenge to scientists and engineers. Although the recently emerged phase-selective organogelators (PSOG) do offer very best promises for oil spill treatment, there still exists a number of technical barriers to overcome collectively, including gelators’ high solubility, high gelling ability, general applicability toward crude oil of various types, rapid gelation with room temperature operation, low toxicity, and low cost. Here, a denovo-designed unusually robust molecular gelling scaffold is used for facile construction of a PSOG library and for rapid identification of PSOGs with the most sought-after practical traits. The identified gelators concurrently overcome the existing technical hurdles, and for the first time enable instant room-temperature gelation of crude oil of various types in the presence of seawater. Remarkably, these excellent gelations were achieved with the use of only 0.058–0.18 L of environmentally benign carrier solvents and 7–35 g of gelator per liter of crude oil. Significantly, 2 out of 20 gelators could further congeal crude oil in the powder form at room temperature, highlighting another excellent potential of the developed modularly tunable system in searching for more powerful powder-based gelators for oil spill treatment