Fast and sensitive DNA analysis using changes in the FRET signals of molecular beacons in a PDMS microfluidic channel

A new DNA hybridization analytical method using a microfluidic channel and a molecular beacon-based probe (MB-probe) is described. A stem-loop DNA oligonucleotide labeled with two fluorophores at the 5′ and 3′ termini (a donor dye, TET, and an acceptor dye, TAMRA, respectively) was used to carry out a fast and sensitive DNA analysis. The MB-probe utilized the specificity and selectivity of the DNA hairpin-type probe DNA to detect a specific target DNA of interest. The quenching of the fluorescence resonance energy transfer (FRET) signal between the two fluorophores, caused by the sequence-specific hybridization of the MB-probe and the target DNA, was used to detect a DNA hybridization reaction in a poly(dimethylsiloxane) (PDMS) microfluidic channel. The azoospermia gene, DYS 209, was used as the target DNA to demonstrate the applicability of the method. A simple syringe pumping system was used for quick and accurate analysis. The laminar flow along the channel could be easily controlled by the 3-D channel structure and flow speed. By injecting the MB-probe and target DNA solutions into a zigzag-shaped PDMS microfluidic channel, it was possible to detect their sequence-specific hybridization. Surface-enhanced Raman spectroscopy (SERS) was also used to provide complementary evidence of the DNA hybridization. Our data show that this technique is a promising real-time detection method for label-free DNA targets in the solution phase. Figure FRET-based DNA hybridization detection using a molecular beacon in a zigzag-shaped PDMS microfluidic channel     

A new catalyst layer based on in situ pore generation of sulfonated poly(ether ether ketone) for PEMFC

A new concept of in situ pore generation to reduce water flooding in cathode catalyst layer (CCL) of polymer electrolyte membrane fuel cell (PEMFC) is proposed with the introduction of water soluble poly(ethylene glycol) (PEG) as a porogen to CCL based on sulfonated poly(ether ether ketone) (sPEEK). In this new type of CCL, PEG is directly removed by water produced during the cathode reaction. The new CCL exhibited much higher cell performance especially in mass transport limitation region compared to the pristine sPEEK-CCL. In addition, the presence of PEG in the new CCL lowered the glass transition temperature of the sPEEK binder, and it could improve the transference of catalyst layer onto the polymer electrolyte membrane.     

Rate constants for the quenching of metastable O2(1Σ) molecules

Singlet molecular oxygen, O₂(¹Σ), is one of the important intermediate species in the atmospheres of Earth, Mars, and Venus. To elucidate the chemistry of this excited molecular oxygen, a series of kinetic measurements have been undertaken using the flow-discharge/optical-emission technique. By monitoring the characteristic emission (762 nm for ¹Σ), the quenching rates for several important molecules have been obtained at room temperature. The following table summarizes measurements.

Yields of O2(b1Σ) from reactions of HO2

A discharge-flow apparatus with resonance fluorescence and chemiluminescence detection has been used to monitor O₂(b ¹σ) production from several reactions of the HO₂ radical at 300 K and 1-torr total pressure. O₂(b), HO₂, and OH were observed when F atoms were added to H₂O₂ in the gas phase. Signal strengths of O₂(b) were proportional to initial concentrations of H₂O₂ and HO₂. These observations were analyzed by using a simple three step mechanism and a more complete computer simulation with 22 reaction steps. The results indicate that the F + HO₂ reaction yields O₂(b) with an efficiency of (3.6 ± 1.4) × 10^?3. By monitoring [O₂(b)] and [HO₂] upon addition of an excess second reactant to HO₂, O₂(b) yields from the reactions of HO₂ with O, Cl, D, H, and OH were found to be <1 × 10^?2, <5 × 10^?4, <2 × 10^?3, <8 × 10^?3, and <1 × 10^?3, respectively. Yields of O₂(b) from the HO₂ ± HO₂ reaction were found to be less than 3 × 10^?2.     

Photoinitiator-Free Photopolymerization of Silicon-Containing Maleimides Based on Electron-Donor-Acceptor Systems

Summary: Photocured coatings promise enhanced performance, higher productivity and an improved environmental impact on the favorable reduction or near elimination of volatile organic compounds. Increasing interest in the photocuring industry has been imposed on the “photoinitiator-free photopolymerization [PIFP]” system based on the electron-donor-acceptor [EDA] concept, which commonly employs a mixture of a vinyl ether monomer (donor) and an N-substituted maleimide or bismaleimide monomer (acceptor). The PIFP systems employing silicon-containing maleimides together with vinyl ether monomers were firstly investigated and their photoreactions resulted in quite fast curing comparable to the conventional acrylate- based photocurable systems.     

Impact of Milling on Rice Constituents (Oryza Sativa L.): A Metabolomic Approach

Milling is an influential factor that affects the nutritional components in rice. However, the alteration of rice constituents by milling has not been thoroughly examined. In this study, rice with various degrees of milling was analyzed by gas chromatography–mass spectrometry and high-performance liquid chromatography–mass spectrometry. Principal component analysis and partial least square-discriminant analysis were performed to characterize the nutritional components that have significant changes during milling. The concentrations of sugars and sugar alcohols decreased while the phospholipids increased in accordance with the milling degree. These results provide a contrast to the common idea that brown rice is nutritionally superior to white rice. In conclusion, the knowledge of nutritional alterations related to milling may benefit rice production and consumption.     

Synthesis,Characterization, and Directional Binding of Anisotropic Biohybrid Microparticles for Multiplexed Biosensing

Anisotropic microarchitectures with different physicochemical properties have been developed as advanced materials for challenging industrial and biomedical applications including switchable displays, multiplexed biosensors and bioassays, spatially‐controlled drug delivery systems, and tissue engineering scaffolds. In this study, anisotropic biohybrid microparticles (MPs) spatio‐selectively conjugated with two different antibodies (Abs) are first developed for fluorescence‐based, multiplexed sensing of biological molecules. Poly(acrylamide‐co‐acrylic acid) is chemically modified with maleimide‐ or acetylene groups to introduce different targeting biological moieties into each compartment of anisotropic MPs. Modified polymer solutions containing two different fluorescent dyes are separately used for electrohydrodynamic co‐jetting with side‐by‐side needle geometry. The anisotropic MPs are chemically stabilized by thermal imidization, followed by bioconjugation of two different sets of polyclonal Abs with two individual compartments via maleimide‐thiol coupling reaction and Huisgen 1,3‐dipolar cycloaddition. Finally, two compartments of the anisotropic biohybrid MPs are spatio‐selectively associated with the respective monoclonal Ab‐immobilized substrate in the presence of the antigen by sandwich‐type immunocomplex formation, resulting in their ordered orientation due to the spatio‐specific molecular interaction, as confirmed by confocal laser scanning microscopy. In conclusion, anisotropic biohybrid MPs capable of directional binding have great potential as a new fluorescence‐based multiplexing biosensing system.

     

Bioorthogonal Cleavage and Exchange of Major Histocompatibility Complex Ligands by Employing Azobenzene‐Containing Peptides

Bioorthogonal cleavable linkers are attractive building blocks for compounds that can be manipulated to study biological and cellular processes. Sodium dithionite sensitive azobenzene‐containing (Abc) peptides were applied for the temporary stabilization of recombinant MHC complexes, which can then be employed to generate libraries of MHC tetramers after exchange with a novel epitope. This technology represents an important tool for high‐throughput studies of disease‐specific T cell responses.