Monitoring molecular beacon DNA probe hybridization at the single-molecule level

We have monitored the reaction dynamics of the DNA hybridization process on a liquid/solid interface at the single-molecule level by using a hairpin-type molecular beacon DNA probe. Fluorescence images of single DNA probes were recorded by using total internal reflection fluorescence microscopy. The fluorescence signal of single DNA probes during the hybridization to individual complementary DNA probes was monitored over time. Among 400 molecular beacon DNA probes that we tracked, 349 molecular beacons (87.5 %) were hybridized quickly and showed an abrupt fluorescence increase, while 51 probes (12.5 %) reacted slowly, resulting in a gradual fluorescence increase. This ratio stayed about the same when varying the concentrations of cDNA in MB hybridization on the liquid/surface interface. Statistical data of the 51 single-molecule hybridization images showed that there was a multistep hybridization process. Our results also showed that photostability for the dye molecules associated with the double-stranded hybrids was better than that for those with the single-stranded molecular beacon DNA probes. Our results demonstrate the ability to obtain a better understanding of DNA hybridization processes using single-molecule techniques, which will improve biosensor and biochip development where surface-immobilized molecular beacon DNA probes provide unique advantages in signal transduction.     

Porphyrin hetero-dimer as charge separating system for photocurrent generation

Introducing a porphyrin bearing electron acceptor onto a self-assembled monolayer (SAM) using a supramolecular method to form a hetero-dimer increased the photocurrent value compared with using porphyrin without an electron acceptor.     

Polymer-supported bisBINOL ligands for the immobilization of multicomponent asymmetric catalysts

[reaction: see text] Polymer-supported bisBINOL ligands were successfully utilized for the immobilization of multicomponent asymmetric catalysts. The polymer-supported Al-Li-bis(binaphthoxide) (ALB) catalyst was more effective than the dendrimer-supported ALB in the Michael reaction of 2-cyclohexen-1-one with dibenzyl malonate affording the adduct in 91% yield with 96% ee. The polymer was also effective for the immobilization of a mu-oxodititanium complex that promoted carbonyl-ene reaction of ethyl glyoxalate with alpha-methyl styrene to provide the adduct with up to 98% ee.     

Evaluation of Superharmonic Functions Using Limits Along Lines

If u is a superharmonic function on R², then [formula] for all (x, y) R². This follows from the fact that a line segmentin R² is non-thin at each of its constituent points. (See Doob[1, 1.XI] or Helms [7, Chapter 10] for an account of thin setsand the fine topology.) The situation is different in higherdimensions. For example, if u is the Newtonian potential onR³ defined by [formula] then [formula] Corollary 2 below will show that, nevertheless, for nearly everyvertical line L, the value of a superharmonic function at anypoint X of L is determined by its lower limit along L at X. Throughout this paper, we let n 3. A typical point of Rⁿ willbe denoted by X or (X', x), where X'R^n–1 and xR. Givenany function f:Rⁿ [–,+] and any point X, we define thevertical cluster set of f at X by [formula] and the fine cluster set of f at X by [formula] 1991 Mathematics Subject Classification 31B05.     

Integrability of superharmonic functions in a John domain

The integrability of positive superharmonic functions on a bounded fat John domain is established. No exterior conditions are assumed. For a general bounded John domain the -integrability is proved with the estimate of in terms of the John constant.

     

Simple Analog Complementary Metal Oxide Semiconductor Circuit for Generating Motion Signal

We proposed in this study a novel analog complementary metal oxide semiconductor (CMOS) circuit for generating a motion signal when an object moves, which is a simple structure. The proposed unit circuit was constructed using a previously proposed edge detection circuit and a novel proposed circuit for generating a motion signal which accepts an edge signal. The part for generating the motion signal was constructed using six metal oxide semiconductor (MOS) transistors and one capacitor. Results obtained by the simulation program with integrated circuit emphasis (SPICE) and the measured results of a test circuit constructed with discrete MOS transistors and the test circuit fabricated with a 1.2 μm CMOS process showed that the proposed unit circuit can output pulsed current (motion signal) when an object moves on the circuit. It was clarified from the SPICE results that the two-dimensional network constructed with proposed unit circuits can output motion signals. The size of the novel unit circuit is expected to be about 110 × 110μm² obtained by the 1.2 μm CMOS process. It is possible to arrange 90 × 90 unit circuits on a chip which has an area of 1 × 1cm². The aperture ratio is expected to be about 21%, which is twice as large as that of the previously proposed circuit. An integrated circuit for image processing in real time can thus be realized by applying the two-dimensional network constructed with the proposed circuits.     

Isomeric states in 214Th and 213Th

Isomeric states in ²¹⁴Th and ²¹³Th were identified by means of γ -rays measured in delayed coincidence with the implanted evaporation residues. These were produced in irradiations of ¹⁶⁴Dy with ⁵⁴Cr projectiles and separated in-flight by the velocity filter SHIP. An isomeric state of I ^π = 8⁺ with a half-life of (1.24±0.12) μs was identified in ²¹⁴Th . The configuration π[1h _9/2 ⊗ 2f _7/2] was assigned to this state. An isomeric state with a half-life of (1.4±0.4) μs was observed in ²¹³Th . Tentatively it was assigned to an I ^π = 13/2⁺ state.     

High-Quality Protein Crystal Growth of Mouse Lipocalin-Type Prostaglandin D Synthase in Microgravity

Lipocalin-type prostaglandin (PG) D synthase (L-PGDS) catalyzes the isomerization of PGH(2) to PGD(2) and is involved in the regulation of pain and of nonrapid eye movement sleep and the differentiation of male genital organs and adipocytes, etc. L-PGDS is secreted into various body fluids and binds various lipophilic compounds with high affinities, acting also as an extracellular transporter. Mouse L-PGDS with a C65A mutation was previously crystallized with citrate or malonate as a precipitant, and the X-ray crystallographic structure was determined at 2.0 ? resolution. To obtain high-quality crystals, we tried, unsuccessfully, to crystallize the C65A mutant in microgravity under the same conditions used in the previous study. After further purifying the protein and changing the precipitant to polyethylene glycol (PEG) 8000, high-quality crystals were grown in microgravity. The precipitant solution was 40% (w/v) PEG 8000, 100 mM sodium chloride, and 100 mM HEPES-NaOH (pH 7.0). Crystals grew on board the International Space Station for 11 weeks in 2007, yielding single crystals of the wild-type L-PGDS and the C65A mutant, both of which diffracted at around 1.0 ? resolution. The crystal quality was markedly improved through the use of a high-viscosity precipitant solution in microgravity, in combination with the use of a highly purified protein.