Nonfouling Hydrophilic Poly(ethylene glycol) Engraftment Strategy for PDMS/SU‑8 Heterogeneous Microfluidic Devices

We report a novel nonfouling passivation method using poly­(ethylene glycol) (PEG) engraftment on the surfaces of poly­(dimethylsiloxane) (PDMS) microfluidic devices sealed with SU-8. To achieve bonding between the PDMS and SU-8 surfaces, the PDMS surface was first functionalized with amines by treatment with 3-aminopropyltrimethoxysilane (APTMS) for subsequent reaction with epoxide functional groups on SU-8 surfaces. To modify the heterogeneous surfaces of the resulting PDMS/SU-8 microfluidic device further, the remaining SU-8 surfaces were amino functionalized using ethylene diamine (EDA), followed by treating both amino-functionalized PDMS and SU-8 surfaces with mPEG-NHS (<i>N</i>-hydroxysuccinimide) through an amine-NHS reaction for facile PEG immobilizations, thus simultaneously modifying both PDMS and SU-8 surfaces in one reaction. Detailed surface analyses such as the water contact angle, X-ray photoelectron spectroscopy (XPS), and atomic force microscopy (AFM) were conducted to confirm the chemical reactions and characterize the resulting surface properties. To test the efficacy of this surface-modification strategy, we conducted nonspecific protein and particle binding tests using microfluidic devices with and without modifications. The PEG-modified PDMS/SU-8 device surfaces showed a 64.5% reduction in nonspecific bovine serum albumin (BSA) adsorption in comparison to that of the unmodified surfaces and 92.0 and 95.8% reductions in microbead adhesion under both stagnant and flowing conditions, respectively

MOESM1 of Subunits of human condensins are potential therapeutic targets for cancers

Additional file 1: Table S1. Subunits of human condensin I and condensin II that are involved in cancers

IspC is the antigen recognized by each MAb.

<p>Protein identification was performed on the protein immunoprecipitated by M2799 with MS. MASCOT software was used to match the observed MS/MS spectra against protein sequences in the NCBInr database. This analysis indicated that IspC is the most likely protein match, with a MASCOT score of 2538. Peptides which were identified by their MS/MS spectrum as matching IspC sequence are shown in red (A). Amino acid sequences shown in black were not detected during MS analysis, but are shown to illustrate the proportion of the IspC protein which was identified by MS. N-terminal sequencing of the immunoprecipitated protein yielded ten residues that aligned perfectly with residues 45–55 of the IspC protein (B). Fifteen previously generated MAbs <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0055098#pone.0055098-Lin1" target="_blank">[10]</a> reacted strongly on a western blot with recombinant IspC (C).</p

Epitopes for each MAb are localized to the C-terminal CBD of IspC.

<p>The left panel (A) provides a representation of each of the recombinant truncated IspC proteins that were produced in <i>E. coli</i>. The right panel (A) shows a summary of the ability of the corresponding truncated proteins to react with the MAbs. Group 1 consists of M2773, M2788, M2792, M2795 and M2800. Group 2 is composed of M2775 and M2797. Group 3 contains M2777 and M2778. Group 4 is composed of M2774 and M2779. The MAbs M2780, M2781, M2790 and M2799 are each in their own group since they were the only MAbs with their particular reaction profile. An illustration of the approximate location of the epitope for each MAb on the IspC protein is shown in (B).</p

Cross-reactions of anti-IspC MAbs with other <i>L. monocytogenes</i> serotypes.

a<p>Isolate details can be found in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0055098#pone.0055098.s001" target="_blank">Table S1</a>.</p>b<p>The interaction between each antibody-isolate pair was examined using ELISA in three independent experiments. Positive reactions were recorded if the average of the three OD₄₁₄ measurements was >25% of the OD₄₁₄ recorded for the same antibody when reacting with <i>L. monocytogenes</i> serotype 4b strain LI0521.</p>c<p>Negative reactions are reported if the average OD₄₁₄ of the three independent experiments was <25% of the OD₄₁₄ recorded for the same antibody when reacting with <i>L. monocytogenes</i> serotype 4b strain LI0521. In most cases a negative reaction could also be defined by OD₄₁₄ of all three experiments being below the 25% threshold. However, M2777, M2781, M2792, M2795, M2797 and M2799 each frequently had one or two of three the measurements above the 25% threshold, even though the average remained below 25%. The variability of each of these MAbs makes them poor candidates for diagnostics.</p

Sequential GW-modules in the CBD of IspC have areas of significant sequence homology.

<p>Matching residues are shaded.</p

Kinetic analysis of IspC and Fab interactions.

<p>SPR sensorgrams showing FAb binding to immobilized IspC at concentrations of: 1, 2.5, 5, 10, 10, 25, 50 and 100 nM for M2773, 2.5, 5, 7.5, 10, 25, 50, 50 and 100 nM for M2774, 1, 2.5, 5, 10, 10, 25 and 50 nM for M2775, 0.5, 1, 2.5, 5, 10, 10 and 25 nM for M2777, 5, 10, 25, 50, 50, 100, 250, and 500 nM for M2779, 2.5, 5, 10, 25, 50, 50, 100 and 250 nM for M2780, 0.5, 1, 2.5, 5, 10, 10, 25 and 50 nM for M2781, 5, 10, 25, 50, 50, 100, 250, 500, and 1000 nM for M2788, 2.5, 5, 10, 25, 50, 50, 100, 250, 500 and 1000 nM for M2790, 1, 2.5, 5, 10, 10, 25, 50 and 100 nM for M2792, 1, 2.5, 5, 7.5, 10, 10, 25, 50 and 100 nM forM2795, 2.5, 5, 10, 10, 25, 50 and 100 nM for M2797, 1, 2.5, 5, 7.5, 10, 10, 25, 50 and 100 nM for M2800 are shown in (A). Black lines represent raw data measurements and red lines represent fitted curves. Rate and affinity constants are given in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0055098#pone-0055098-t001" target="_blank">Table 1</a>. A rate plane plot with iso-affinity diagonals is shown in (B). This kinetic map summarizes the respective affinities of each MAb for IspC as determined by SPR. Blue is used to denote high affinity MAbs, while red shows moderately high affinity MAbs and black is used to label lower affinity MAbs.</p

Additional file 1: Figure S1. of Characterization of early transcriptional responses to cadmium in the root and leaf of Cd-resistant Salix matsudana Koidz

Plant screening and growth. (a) Seeds cultured in 100 μM Cd medium for 1 month. (b) Plant growth when exposed to 50 μM Cd for 1 month. Figure S2. Measurement of chlorophyll and cadmium. (a) Chlorophyll a, b, and a + b content in the root, stem, and leaf of S. matsudana Koidz. (b) Cd concentration in the root, stem, and leaf treated with 50 μM Cd for 1 month. Figure S3. Characteristics of the similarity search of unigenes against Nr databases. (a) E-value distribution of BlastX hits for each unigene with an E-value threshold of 10E −5. (b) Similarity distribution of the top BLAST hits for each unigene. (c) Species distribution is shown as a percentage of total homologous sequences with an E-value of at least 1.0E −5. (d) Clusters of orthologous group functional classification of all unigenes. (e) GO classifications of assembled unigenes. Figure S4. Metabolism pathways of sulfur metabolism. Figure S5. Metabolism pathways of flavonoid biosynthesis. (PPT 732 kb

Additional file 2: Table S1. of Characterization of early transcriptional responses to cadmium in the root and leaf of Cd-resistant Salix matsudana Koidz

Unigene primers used for QRT-PCR analysis. Table S2. Sequencing output statistics. Table S3. Statistics of assembly quality. Table S4. Unigene classification by clusters of orthologous groups function. Table S5. Unigenes assembled by Gene Ontology classification. Table S6. Gene analysis by MapMan in the leaf. Table S7. Gene analysis by MapMan in the root. Table S8. Up-regulated genes in response to cadmium in leaf. Table S9. Up-regulated genes in response to cadmium in root. Table S10. Genes used for cluster analysis. Table S11. Down-regulated genes in response to cadmium in leaf. Table S12. Down-regulated genes in response to cadmium in root. (DOC 751 kb

Synthesis and Biological Evaluation of Novel σ₁ Receptor Ligands for Treating Neuropathic Pain: 6‑Hydroxypyridazinones

By use of the 6-hydroxypyridazinone framework, a new series of potent σ₁ receptor ligands associated with pharmacological antineuropathic pain activity was synthesized and is described in this article. In vitro receptor binding studies revealed high σ₁ receptor affinity (<i>K</i>_i σ₁ = 1.4 nM) and excellent selectivity over not only σ₂ receptor (1366-fold) but also other CNS targets (adrenergic, μ-opioid, sertonerigic receptors, etc.) for 2-(3,4-dichlorophenyl)-6-(3-(piperidin-1-yl)­propoxy)­pyridazin-3­(2<i>H</i>)-one (compound <b>54</b>). Compound <b>54</b> exhibited dose-dependent antiallodynic properties in mouse formalin model and rats chronic constriction injury (CCI) model of neuropathic pain. In addition, functional activity of compound <b>54</b> was evaluated using phenytoin and indicated that the compound was a σ₁ receptor antagonist. Moreover, no motor impairments were found in rotarod tests at antiallodynic doses and no sedative side effect was evident in locomotor activity tests. Last but not least, good safety and favorable pharmacokinetic properties were also noted. These profiles suggest that compound <b>54</b> may be a member of a novel class of candidate drugs for treatment of neuropathic pain