Ultrahigh-Frequency, Wireless MEMS QCM Biosensor for Direct, Label-Free Detection of Biomarkers in a Large Amount of Contaminants.

Label-free biosensors, including conventional quartz-crystal-microbalance (QCM) biosensors, are seriously affected by nonspecific adsorption of contaminants involved in analyte solution, and it is exceptionally difficult to extract the sensor responses caused only by the targets. In this study, we reveal that this difficulty can be overcome with an ultrahigh-frequency, wireless QCM biosensor. The sensitivity of a QCM biosensor dramatically improves when the quartz resonator is thinned, which also makes the resonance frequency higher, causing high-speed surface movement. Contaminants weakly (nonspecifically) interact with the quartz surface, but they fail to follow the fast surface movement and cannot be detected as the loaded mass. The targets are, however, tightly captured by the receptor proteins immobilized on the surface, and they can move with the surface, contributing to the loaded mass and decreasing the resonant frequency. We have developed a MEMS QCM biosensor in which an AT-cut quartz resonator, 26 µm thick, is packaged without fixing, and we demonstrate this phenomenon by comparing the frequency changes of the fundamental (∼64 MHz) and ninth (∼576 MHz) modes. At ultrahigh-frequency operation with the ninth mode, the sensor response is independent of the amount of impurity proteins, and the binding affinity is unchanged. We then applied this method to the label-free and sandwich-free, direct detection of C-reactive protein (CRP) in serum and confirmed its applicability.

Novel hybrid organic thermoelectric materials:three-component hybrid films consisting of a nanoparticle polymer complex, carbon nanotubes, and vinyl polymer.

A novel class of hybrid organic thermoelectric materials is demonstrated for the first time for constructing flexible thermoelectric devices on polyimide substrates with high output power by using nanotechnology instead of conducting polymers such as poly(3,4-ethylenedioxythiophene). The hybrid organic thermoelectric materials are composed of nanoparticles of a polymer complex, carbon nanotubes, and poly(vinyl chloride), and show high performance (dimensionless thermoelectric figure-of-merit, ZT ≈ 0.3, based on the thermal conductivity through the film).

Cloning and expression of the turtle (Trachemys scripta) immunoglobulin joining (J)-chain cDNA.

The J-chain protein is a M(r) 15000 polypeptide associated with polymeric IgA and IgM. The complete cDNA sequences of human, mouse, cow, brushtail possum, chicken and frog J chains have been previously reported, but nothing is known about the cDNA and amino acid sequences of reptilian J chain. Here, we determined a turtle J-chain cDNA sequence by RT-PCR and RACE, and examined J-chain mRNA and protein expression by Northern blotting and immunohistochemistry. This turtle J-chain cDNA was 1934 bp and had an open reading frame of 477 nucleotides, encoding 159 amino acids. The mature J-chain protein is composed of 137 amino acids, M(r) approximately 15000. The deduced amino acid sequence of the turtle J chain was highly homologous to that of human (60%), mouse (61%), cow (60%), rabbit (60%), chicken (69%), brushtail possum (65%), Rana catesbeiana (47%) and Xenopus laevis (58%). Eight cysteine residues were located at the same positions as in these other species, with the exception of X. laevis. PROSITE database analysis indicated the presence of two N-glycosylation sites in turtle, one of which was novel. Northern blot analysis revealed that turtle J-chain mRNA was expressed in lung, stomach, spleen and intestine. In addition, immunohistochemistry showed J-chain-positive plasma cells in the intestine and spleen. These results suggest the presence of a mucosal immune system mainly composed of J-chain-containing Ig in the turtle.

Ring-opening reactions of cyclic acetals and 1,3-oxazolidines with halosilane equivalents.

Reactions of acetal and 1,3-oxazolidine rings were examined using two kinds of iodosilane equivalent reagents, a 1:2 mixture of Me3SiNEt2 and MeI (reagent 1a) and a 1:1 mixture of Et3SiH and MeI containing a catalytic amount of PdCl2 (reagent 1b). In the reactions of alkanone ethylene acetals with reagent 1a, a C-O bond in the acetal ring readily cleaved to give 2-(trimethylsiloxy)ethyl enol ethers. Similarly, the C-O bond of 1,3-oxazolidine rings cleaved to give ring-opened imine or enamine derivatives. The reactions of aromatic ketone ethylene acetals and cyclohexanone trimethylene acetal led to deprotection of the acetal unit to liberate free ketones. With reagent 1b, cycloalkanone ethylene acetal afforded a dimeric product with 2-iodoethyl alkenoate moieties, while aromatic ketone ethylene or trimethylene acetals produced deprotected ketones.

Ring-opening iodo- and bromosilation of lactones for the formation of silyl haloalkanoates.

Ring-opening halosilation of lactones with two types of reagents, Et(3)SiH/MeI(PdCl(2)) (1a) and Et(3)SiH/AllylBr(PdCl(2)) (1b), was studied. Cyclic esters such as gamma-butyrolactones, delta-valerolactone, and 6-hexanolide reacted with 1 equiv of 1a,b to give triethylsilyl omega-iodo- and omega-bromoalkanoates in good yields. Reaction of an acyclic ester, methyl benzoate, with 1a afforded triethylsilyl benzoate. O-Silyl-protected amino acids could be obtained by amination of the halosilation products, triethylsilyl omega-bromoalkanoates.

Cloning of the chicken immunoglobulin joining (J)-chain gene and characterization of its promoter region.

Three overlapping genomic clones of the chicken immunoglobulin joining (J) chain were isolated and then characterized using restriction enzyme analysis, Southern blot analysis with cDNA probes, and DNA sequencing. The gene consisted of four exons separated by a 2.6-kb intron 1, a 0.9-kb intron 2, and a 0.5-kb intron 3. A transcriptional initiation site was identified by a primer extension method using mRNA and cDNA, indicating that exon 1 was 86 bp encoding 20 amino acid residues. A TATA box was positioned at 29 approximately 25 bp upstream of exon 1. Exons, 2, and 3 consisted of 133 bp and 81 bp, encoding 43 and 26 amino acid residues of the mature protein, respectively. Exon 4 consisted of 202 bp encoding 66 amino acid residues and 1.2 kb of untranslated sequence. Deletion mutants of a 4.1-kb genomic fragment containing exon 1 showed high levels of promoter activities when examined in luciferase reporter assays following transfection into the DT-40 chicken B-cell line. These results suggest that the chicken J-chain gene consists of four exons and three introns and that the transcriptional regulatory elements may be present within 3.8 kb upstream of exon 1.