Magnetic measurements and microstructure studies of highly oriented Bi2Sr2Ca2Cu3O8 bulks

A highly oriented structure for Bi₂Sr₂Ca₂Cu₃O₈ bulk superconductors was successfully made by hot-forging. The J_c values were closely related to the degree of orientation. Microstructure observations and magnetization measurements revealed that the properties of the highly oriented BSCCO reflected the two-dimensionality in this layered structure material.     

Microfluidic baker's transformation device for three-dimensional rapid mixing

We developed a new passive-type micromixer based on the baker's transformation and realized a fast mixing of a protein solution, which has lower diffusion constant. The baker's transformation is an ideal mixing method, but there is no report on the microfluidic baker's transformation (MBT), since it is required to fabricate the complicated three-dimensional (3D) structure to realize the MBT device. In this note, we successfully fabricate the MBT device by using precision diamond cutting of an oxygen-free copper substrate for the mould fabrication and PDMS replication. The MBT device with 10.4 mm mixing length enables us to achieve complete mixing of a FITC solution (D = 2.6 × 10(-10) m(2) s(-1)) within 51 ms and an IgG solution (D = 4.6 × 10(-11) m(2) s(-1)) within 306 ms. Its mixing speed is 70-fold higher for a FITC solution and 900-fold higher for an IgG solution than the mixing speed by the microchannel without MBT structures. The Péclet number to attain complete mixing in the MBT device is estimated to be 6.9 × 10(4).     

Rapid qualitative evaluation of DNA transcription factor NF-κB by microchip electrophoretic mobility shift assay in mammalian cells

We have developed a separation technique for DNA-protein complex based on electrophoretic mobility shift assay (EMSA) by microchip electrophoresis, which we call microchip electrophoretic mobility shift assay (μEMSA). To evaluate the μEMSA, we employed recombinant human nuclear factor-κB (rhNF-κB) and its consensus double-stranded oligonucleotide (dsOligo) fluorescently labeled with Cy5. We carried out the electrophoretic separation of the consensus dsOligo-rhNF-κB complex and the unbound dsOligo in methylcellulose solution and confirmed rapid (～200 s) and reliable identification and semi-quantitation of the specific interaction between dsOligo and rhNF-κB. The binding specificity of rhNF-κB was confirmed by introducing non-fluorescently labeled consensus oligonucleotide as a competitor. The progression of the binding reaction under various incubation times was monitored, and it was found that the dsOligo and rhNF-κB complex formation reached equilibrium (ca. 90% of the dsOligo was bound to rhNF-κB) after 5 min. Furthermore, without any purification process, even crude NF-κB in nuclear extracts from HeLa cells was specifically detected within 120 s by the μEMSA.     

A touch-and-go lipid wrapping technique in microfluidic channels for rapid fabrication of multifunctional envelope-type gene delivery nanodevices

Multifunctional envelope-type gene delivery nanodevices (MENDs) are promising non-viral vectors for gene therapy. Though MENDs remain strong in prolonged exposure to blood circulation, have low immunogenic response, and are suitable for gene targeting, their fabrication requires labor-intensive processes. In this work, a novel approach has been developed for rapid fabrication of MENDs by a touch-and-go lipid wrapping technique in a polydimethylsiloxane (PDMS)/glass microfluidic device. The MEND was fabricated on a glass substrate by introduction of a condensed plasmid DNA core into microfluidic channels that have multiple lipid bilayer films. The principle of the MEND fabrication in the microfluidic channels is based on electrostatic interaction between the condensed plasmid DNA cores and the coated lipid bilayer films. The constructed MEND was collected off-chip and characterized by dynamic light scattering. The MEND was constructed within 5 min with a narrow size distribution centered around 200 nm diameter particles. The size of the MEND showed strong dependence on flow velocity of the condensed plasmid DNA core in the microfluidic channels, and thus, could be controlled to provide the optimal size for medical applications. This approach was also proved possible for fabrication of a MEND in multiple channels at the same time. This on-chip fabrication of the MEND was very simple, rapid, convenient, and cost-effective compared with conventional methods. Our results strongly indicated that MENDs fabricated with our microfluidic device have a good potential for medical use. Moreover, MENDs fabricated by this microfluidic device have a great potential for clinical use because the devices are autoclavable and all the fabrication steps can be completed inside closed microfluidic channels without any external contamination.     

Observation of B decays to two kaons

Using 449x10(6) BB[over ] pairs collected with the Belle detector at the KEKB asymmetric-energy e(+)e(-) collider, we observe clear signals for B(+)-->K[over ](0)K(+) and B(0)-->K[over ](0)K(0) decays with 5.3sigma and 6.0sigma significance, respectively. We measure the branching fractions B(B(+)-->K[over ](0)K(+))=(1.22(-0.28-0.16)(+0.32+0.13))x10(-6) and B(B(0)-->K[over ](0)K(0))=(0.87(-0.20)(+0.25)+/-0.09)x10(-6), and partial-rate asymmetries A(CP)(B(+)-->K[over ](0)K(+))=0.13(-0.24)(+0.23)+/-0.02 and A(CP)(B(0)-->K[over ](0)K(0))=-0.58(-0.66)(+0.73)+/-0.04. From a simultaneous fit, we also obtain B(B(+)-->K(0)pi(+))=(22.8(-0.7)(+0.8)+/-1.3)x10(-6) and A(CP)(B(+)-->K(0)pi(+))=0.03+/-0.03+/-0.01. The first and second error in the branching fractions and the partial-rate asymmetries are statistical and systematic, respectively. No signal is observed for B(0)-->K(+)K(-) decays, and for this branching fraction, we set an upper limit of 4.1x10(-7) at the 90% confidence level.     

Evidence for CP violation in B0-->D+D- decays

We report measurements of the branching fraction and CP violation parameters in B(0)-->D+ D- decays. The results are based on a data sample that contains 535 x 10(6) BB pairs collected at the Upsilon(4S) resonance, with the Belle detector at the KEKB asymmetric-energy e+e- collider. We obtain [1.97+/-0.20(stat) +/- 0.20(syst)] x 10(-4) for the branching fraction of B0-->D+D-. The measured values of the CP violation parameters are S=-1.13+/-0.37+/-0.09, A=0.91+/-0.23+/-0.06, where the first error is statistical and the second is systematic. We find evidence of CP violation in B0-->D+D- at the 4.1sigma confidence level. While the value of S is consistent with expectations from other measurements, the value of the parameter A favors large direct CP violation at the 3.2sigma confidence level, in contradiction to standard model expectations.     

Measurement of D0-D0 mixing parameters in D0 --> Ks pi+ pi- decays

We report a measurement of D0-D(0) mixing parameters in D(0) --> K(s)(0) pi(+) pi(-) decays using a time-dependent Dalitz-plot analysis. We first assume CP conservation and subsequently allow for CP violation. The results are based on 540 fb(-1) of data accumulated with the Belle detector at the KEKB e(+)e(-) collider. Assuming negligible CP violation, we measure the mixing parameters x = (0.80 +/- 0.29(-0.07-0.14)(+0.09+0.10))% and y = (0.33+/-0.24(-0.12-0.08)(+0.08+0.06))%, where the errors are statistical, experimental systematic, and systematic due to the Dalitz decay model, respectively. Allowing for CP violation, we obtain the CP-violating parameters |q / p| = 0.86(-0.29-0.03)(+0.30+0.06) +/- 0.08 and arg(q/p) = (-14(-18-3-4)(+16+5+2)) degrees .     

Quinolone analogues 9. Synthesis of 7‐methylsulfanyl‐ and 7‐methanesulfonylpyridazino[3,4‐b]quinoxalin‐4(1H)‐ones

The reaction of 7‐chloro‐1‐methylpyridazino[3,4‐b]quinoxalin‐4(1H)‐ones 3a‐5a with sodium methylthiolate gave 1‐methyl‐7‐methylsulfanylpyridazino[3,4‐b]quinoxalin‐4(1H)‐ones 8a‐c , whose reaction with m‐chloroperbenzoic acid afforded the 7‐methanesulfonyl‐1‐methylpyridazino[3,4‐b]‐quinoxalin‐4(1H)‐ones 9a‐c , respectively. The above substituent change at the 7‐position resulted in the activity alteration to microorganisms.     

Determination of human blood glucose levels using microchip electrophoresis

A high-performance monitoring system for human blood glucose levels was developed using microchip electrophoresis with a plastic chip. The combination of reductive amination as glucose labeling with fluorescent 2-aminoacridone (AMAC) and glucose-borate complex formation realized the highly selective detection of glucose even in a complex matrix such as a blood sample. The migration time of a single peak, observed on an electropherogram of AMAC-labeled plasma, closely resembled that of glucose standard solution. The treatment of plasma with hexokinase or glucokinase for glucose phosphorylation resulted in a peak shift from approximately 145 to 70 s, corresponding to glucose and glucose-6-phosphate, respectively. A double-logarithm plot revealed a linear relationship between glucose concentration and fluorescence intensity in the range of 1-300 microM of glucose (r(2) = 0.9963; p <0.01), and the detection limit was 0.92 microM. Furthermore, blood glucose concentrations estimated from the standard curves of three subjects were compared with results obtained by conventional colorimetric analysis using glucose dehydrogenase. Good correlation was observed between methods according to simple linear regression analysis (p <0.05). The reproducibility of the assay was about 6.3-9.1% (RSD) and the within-days and between-days reproducibility were 1.6-8.4 and 5.2-7.2%, respectively. This system enables us to determine blood glucose with high sensitivity and accuracy, and will be applicable to clinical diagnosis.