Atypical PKC and MAPK were involved in GSK-3 inactivation in EGF-promoted glioma invasion.

<p>(A) EGF-induced increase in pSer-GSK-3 levels was eliminated by inhibitors of aPKC and MAPK. <i>Left</i>: representative western blots of U251 lysates collected 3 hrs after wound recovery in the presence of EGF and indicated agents. <i>Right</i>: quantification of changes in pSer-GSK-3levels normalized to GSK-3 levels and then compared with the group without EGF treatment. (B) BothGSK-3β and PKCζ were present in the same complexes. Western blot analysis of the complexes precipitated by the indicated antibodies (U251 lysates collected at 2 hrs after EGF stimulation). (C) Inhibition of aPKC and MAPK suppressed U87 cell invasion in the transwell assay. #<i>P</i><0.05 versus DMSO; * <i>P</i><0.05, ** <i>P</i><0.01 versus DMSO+EGF. Wortmannin, 1μM; LY294002, 5μM; Gö6976, 0.1μM; Gö6983, 10μM; U0126, 10μM, EGF, 40ng/ml. Data were mean ± SD of three independent experiments in triplicate.</p

Inhibiting GSK-3 blocked migration and invasion of human glioma cells.

<p>(A) Wound recovery of U251 and U87 cells in the presence of indicated agents. ** <i>P</i><0.01 versus DMSO. (B and C) Blockade of cell migration and invasion in the presence of GSK-3 inhibitors. (B) Representative images of U251 cells in the transwell invasion assay in different treatments. <i>Original magnification, 100×</i>. (C) Quantifications of transwell migration in invasion assays of U251 and U87 cells. ** <i>P</i><0.01 versus DMSO. (D) Western blot analysis of U87 cell lysates transfected with indicated constructs. The values indicated the amount of indicated proteins normalized to that of GAPDH and compared with non-sense (<i>si-ctl</i>) or pcDNA3 (<i>ctl</i>) control group. (E) GSK-3 down-regulation by small interfering RNA inhibited U87 cell migration and invasion in the transwell assay. * <i>P</i><0.05 versus non-sense control group (<i>si-ctl</i>). (F) Uniform up- or down-regulation of GSK-3β activities inhibited U87 cell migration and invasion in the transwell assay. ** <i>P</i><0.01 versus pcDNA3 (<i>ctl</i>). si-3α, siRNA against GSK-3α; si-3β, siRNA against GSK-3β; WT, wild-type form of GSK-3β; S9A, constitutively active form of GSK-3β; GID, inhibitory peptide of GSK-3β; LP, mutant of GID without inhibitory effect. LiCl, 20mM, SB216763, 20μM, NaCl, 20mM. Data were mean ± SD of three independent experiments in triplicate. </p

Roles of GSK-3 in EGF-promoted glioma cell invasion.

<p>(A) EGF promoted U87 cell invasion in the transwell assay. ** <i>P</i><0.01 versus DMSO .(B) GSK-3 knock-down and uniform up-regulation of GSK-3β activities inhibited U87 cell invasion. * <i>P</i><0.05, ** <i>P</i><0.01 versus pcDNA3 (<i>ctl</i>) or non-sense control (<i>si-ctl</i>). (C) Up-regulation of p-Ser-GSK-3 levels during EGF-induced wound recovery of U251 cells. <i>Left</i>: representative western blots of U251 cell lysates collected at indicated time points after scratching. <i>Right</i>: statistics of pSer-GSK-3 levels normalized to GSK-3 levels and then compared with the value at 0hr. *<i>P</i><0.05, ** <i>P</i><0.01 versus the value of pGSK-3/GSK-3 at 0hr. (D) Location of increased pGSK-3 staining in EGF-induced wound recovery of U251 cells 3 hrs after scratching. <i>Green</i>: anti-pSer21-GSK-3αor anti-pSer9-GSK-3β; <i>Red</i>: rhodamine phalloidin. <i>Original magnification, 400×</i>. si-3α, siRNA against GSK-3α; si-3β, siRNA against GSK-3β; WT, wild-type form of GSK-3β; S9A, constitutively active form of GSK-3β. Data were mean ± SD of three independent experiments. EGF, 40ng/ml.</p

Inhibition of GSK-3 influenced directional persistence and locomotion speed of human glioma cell migration.

<p>(A) Scheme to define the speed and relative step angles as described in <i>Materials and Method</i>. (B) Morphological changes of the U87 cells treated with LiCl. <i>Red</i>: rhodamine phalloidin. <i>Original magnification, 400×</i>. (C) Decreased locomotion speed of the U87 cells upon LiCl treatment. Data were mean ± SD of ten cells in each group ** <i>P</i><0.01 versus DMSO. (D) The representative migration traces of the cells in the presence or absence of LiCl, with the starting point of migration superimposed at the origin of the X-Y plot. (E) The relative step angle of representative four migrating cells in the presence or absence of LiCl. Relative step angle and direction change were defined as described in <i>Materials and Methods</i>. Negative or positive values in <i>y-axis</i> indicated angles in clockwise or counter-clockwise turn, respectively. Red line at 60°or -60° outlined time points when a direction change occurred (beyond the field of -60°<⊿α<60°). LiCl, 20mM.</p

Localized inactivation of GSK-3β was associated with the formation of cell polarity.

<p>(A) Up-regulation of pSer-GSK-3 during the wound recovery of U251 cells. <i>Left</i>: representative western blot results of U251 cell lysates collected at indicated time points after scratching. <i>Right</i>: statistics of p-Ser-GSK-3 levels normalized to GSK-3 levels and compared with the value at 0hr. *<i>P</i><0.05, ** <i>P</i><0.01 versus the value of pGSK-3/GSK-3 at 0hr. (B) Location of increased pGSK-3 staining in the wound recovery of U251 cells at 0 hr or 3 hrs after scratching. Green: anti-pSer21-GSK-3α or anti-pSer9-GSK-3β; Red: rhodamine phalloidin. <i>Original magnification, 400×</i>.(C) MTOC mis-orientation in U251 caused by GSK-3 inhibitors. <i>Left</i>: representative images of MTOC in different treatments. Green: anti-α-tubulin; Red: anti-pericentrin; Blue: Hoechst 33258. <i>Original magnification, 400×</i>. <i>Right</i>: quantification of centrosome polarization as described in <i>Materials and Methods</i> in U251 with or without GSK-3β inhibitors. ** <i>P</i><0.01 versus DMSO. LiCl 20mM, SB216763, 20μM, NaCl, 20 mM. Data were mean ± SD of three independent experiments.</p

Stiffer but More Healable Exponential Layered Assemblies with Boron Nitride Nanoplatelets

Self-healing ability and the elastic modulus of polymeric materials may seem conflicting because of their opposite dependence on chain mobility. Here, we show that boron nitride (BN) nanoplatelets can simultaneously enhance these seemingly contradictory properties in exponentially layer-by-layer-assembled nanocomposites as both surface coatings and free-standing films. On one hand, embedding hard BN nanoplatelets into a soft hydrogen bonding network can enhance the elastic modulus and ultimate strength through effective load transfer strengthened by the incorporation of interfacial covalent bonding; on the other hand, during a water-enabled self-healing process, these two-dimensional flakes induce an anisotropic diffusion, maintain the overall diffusion ability of polymers at low loadings, and can be “sealing” agents to retard the out-of-plane diffusion, thereby hampering polymer release into the solution. A detailed mechanism study supported by a theoretical model reveals the critical parameters for achieving a complete self-healing process. The insights gained from this work may be used for the design of high-performance smart materials based on other two-dimensional fillers

Strong and Stiff Aramid Nanofiber/Carbon Nanotube Nanocomposites

Small but strong carbon nanotubes (CNTs) are fillers of choice for composite reinforcement owing to their extraordinary modulus and strength. However, the mechanical properties of the nanocomposites are still much below those for mechanical parameters of individual nanotubes. The gap between the expectation and experimental results arises not only from imperfect dispersion and poor load transfer but also from the unavailability of strong polymers that can be effectively utilized within the composites of nanotubes. Aramid nanofibers (ANFs) with analogous morphological features to nanotubes represent a potential choice to complement nanotubes given their intrinsic high mechanical performance and the dispersible nature, which enables solvent-based processing methods. In this work, we showed that composite films made from ANFs and multiwalled CNTs (MWCNTs) by vacuum-assisted flocculation and vacuum-assisted layer-by-layer assembly exhibited high ultimate strength of up to 383 MPa and Young’s modulus (stiffness) of up to 35 GPa, which represent the highest values among all the reported random CNT nanocomposites. Detailed studies using different imaging and spectroscopic characterizations suggested that the multiple interfacial interactions between nanotubes and ANFs including hydrogen bonding and π–π stacking are likely the key parameters responsible for the observed mechanical improvement. Importantly, our studies further revealed the attractive thermomechanical characteristics of these nanocomposites with high thermal stability (up to 520 °C) and ultralow coefficients of thermal expansion (2–6 ppm·K^–1). Our results indicated that ANFs are promising nanoscale building blocks for functional ultrastrong and stiff materials potentially extendable to nanocomposites based on other nanoscale fillers

Four-phasic Doppler flow spectra of the MHV in the pulmonary hypertension (PH) group.

<p>Doppler spectra of MHV obtained from a patient with ventricular septal defect with the MPAP of 80 mmHg. The four-phasic waveform includes S wave, D wave, A wave and ventricular reversal (V wave) waves. Compared with the control group, the spectra had a higher value of the peak flow velocity and the velocity time integral (VTI) of A wave, and a lower value of peak flow velocity and VTI of S wave and D wave.</p

Hybrid Au/ZnO Hexagonal Pyramid Nanostructures: Preferred Growth on the Apexes of the Basal Plane than on the Tip

Nanoscale materials having size- and shape-dependent interactions with light provide flexible opportunities for harvesting solar energy. Photocatalysts based on semiconductor nanoparticles (NPs) have been the most effective materials for the conversion of light into chemical energy, the efficiency of which can be further enhanced by the incorporation of metallic NPs forming hybrid nanostructures. The structural parameters of not only constituent components but also the resultant hybrid nanostructures are critical for the optimization of photocatalytic performance of composite catalysts. Here we demonstrated the successful size control over ZnO hexagonal pyramids (HPs) for the first time. The smallest HPs showing the best photocatalytic properties were used for further Au attachment. Interestingly, we found that most of the Au NPs preferred to grow on the apexes of the basal plane. Very occasionally, Au NPs at the tip of ZnO HPs can be observed. The role of light in promoting the reduction of gold salt by sodium citrate was also revealed. Quantum mechanical calculations were used to explain the site-specific growth of Au on the surface of ZnO HPs. Enhanced degradation rates over organic dyes were found for Au/ZnO hybrids under both UV and visible light irradiation

Bi-phasic Doppler flow spectra of the MHV in the PH group.

<p>Doppler spectra of MHV obtained from a patient with patent ductus arteriosus with the MPAP of 96 mmHg. The bi-phasic waveform of MHV includes S wave and A wave.</p