Geneplots of the probe sets influencing the MAPK pathway.

<p>Geneplots are shown for the 50 most influential probe sets from the KEGG MAPK pathway (A) and for the GO-term MAPKKK cascade (B) after stimulation with RA that contribute to the differential pathway expression in SoS and control. Probe sets are scaled to unit standard deviations and the height of the bars are the number of standard deviations above the cut-off level of 0.7. Higher bars indicate higher influence on the pathway. Probe sets with the highest influence on the pathway (i.e. FGF13 in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0049229#pone-0049229-g001" target="_blank">Figure 1A</a> and TNIK in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0049229#pone-0049229-g001" target="_blank">Figure 1B</a>) are depicted on the left. Corresponding gene names are written below each bar.</p

Ultrahigh-Throughput Production of Monodisperse and Multifunctional Janus Microparticles Using in-Air Microfluidics

Compartmentalized Janus microparticles advance many applications ranging from chemical synthesis to consumer electronics. Although these particles can be accurately manufactured using microfluidic droplet generators, the per-nozzle throughputs are relatively low (∼μL/min). Here, we use “in-air microfluidics” to combine liquid microjets in midair, thereby enabling orders of magnitude faster production of Janus microparticles (∼mL/min) as compared to chip-based microfluidics. Monodisperse Janus microparticles with diameters between 50 and 500 μm, tunable compartment sizes, and functional cargo are controllably produced. Furthermore, these microparticles are designed as magnetically steerable microreactors, which represents a novel tool to perform enzymatic cascade reactions within continuous fluid flow

Ultrahigh-Throughput Production of Monodisperse and Multifunctional Janus Microparticles Using in-Air Microfluidics

Compartmentalized Janus microparticles advance many applications ranging from chemical synthesis to consumer electronics. Although these particles can be accurately manufactured using microfluidic droplet generators, the per-nozzle throughputs are relatively low (∼μL/min). Here, we use “in-air microfluidics” to combine liquid microjets in midair, thereby enabling orders of magnitude faster production of Janus microparticles (∼mL/min) as compared to chip-based microfluidics. Monodisperse Janus microparticles with diameters between 50 and 500 μm, tunable compartment sizes, and functional cargo are controllably produced. Furthermore, these microparticles are designed as magnetically steerable microreactors, which represents a novel tool to perform enzymatic cascade reactions within continuous fluid flow

Sotos Syndrome Is Associated with Deregulation of the MAPK/ERK-Signaling Pathway

<div><p>Sotos syndrome (SoS) is characterized by tall stature, characteristic craniofacial features and mental retardation. It is caused by haploinsufficiency of the <em>NSD1</em> gene. In this study, our objective was to identify downstream effectors of NSD1 and to map these effectors in signaling pathways associated with growth. Genome-wide expression studies were performed on dermal fibroblasts from SoS patients with a confirmed <em>NSD1</em> abnormality. To substantiate those results, phosphorylation, siRNA and transfection experiments were performed. A significant association was demonstrated with the Mitogen-Activated Protein Kinase (MAPK) pathway. Members of the fibroblast growth factor family such as <em>FGF4</em> and <em>FGF13</em> contributed strongly to the differential expression in this pathway. In addition, a diminished activity state of the MAPK/ERK pathway was demonstrated in SoS. The Ras Interacting Protein 1 (RASIP1) was identified to exhibit upregulated expression in SoS. It was shown that RASIP1 dose-dependently potentiated bFGF induced expression of the MAPK responsive SBE reporter providing further support for a link between NSD1 and the MAPK/ERK signaling pathway. Additionally, we demonstrated <em>NSD1</em> expression in the terminally differentiated hypertrophic chondrocytes of normal human epiphyseal growth plates. In short stature syndromes such as hypochondroplasia and Noonan syndrome, the activation level of the FGF-MAPK/ERK-pathway in epiphyseal growth plates is a determining factor for statural growth. In analogy, we propose that deregulation of the MAPK/ERK pathway in SoS results in altered hypertrophic differentiation of <em>NSD1</em> expressing chondrocytes and may be a determining factor in statural overgrowth and accelerated skeletal maturation in SoS.</p> </div

Differentially expressed probe sets in SoS after stimulation with RA.

<p>Differentially expressed probe sets in SoS after stimulation with RA.</p

Protein phosphorylation.

<p>The results are shown for the phosphorylation levels for the MEK1 (A), ERK1/ERK2 (B), ERK2 (C), p38MAPK (D), cJUN (E), ATF2 (F), JNK (G), HSP27 (H) and p90RSK kinases (I). Bar heights depict the mean fluorescence intensity levels measured (MFI) and the p-values for the difference between SoS and control (after correction for total protein levels) are shown above the bars.</p

Differentially expressed probe sets in SoS in basal situation.

<p>Differentially expressed probe sets in SoS in basal situation.</p

Signaling pathways analysis results.

<p>Signaling pathways analysis results.</p

bFGF induced SBE reporter activation is potentiated by RASIP1.

<p>Values are expressed as fold induction compared to control. Cells were stimulated with bFGF for 24 hours. Control was not stimulated with bFGF and no RASIP1 was co-transfected. Co-tranfection of RASIP1 did not affect basal reporter activity. bFGF (10 ng/ml) significantly stimulated SBE reporter activity (indicated with #; p<0.05). RASIP1 enhanced bFGF induced reporter dose-dependently (indicated with *; p<0.05).</p

On-Chip Electrophoresis in Supported Lipid Bilayer Membranes Achieved Using Low Potentials

A micro supported lipid bilayer (SLB) electrophoresis method was developed, which functions at low potentials and appreciable operating times. To this end, (hydroxymethyl)-ferrocene (FcCH₂OH) was employed to provide an electrochemical reaction at the anode and cathode at low applied potential to avoid electrolysis of water. The addition of FcCH₂OH did not alter the SLB characteristics or affect biomolecule function, and pH and temperature variations and bubble formation were eliminated. Applying potentials of 0.25–1.2 V during flow gave homogeneous electrical fields and a fast, reversible, and strong build-up of a charged dye-modified lipid in the direction of the oppositely charged electrode. Moreover, streptavidin mobility could be modulated. This method paves the way for further development of analytical devices